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Sample records for dsb ic3 train

  1. Characterization of DsbD in Neisseria meningitidis

    PubMed Central

    Kumar, Pradeep; Sannigrahi, Soma; Scoullar, Jessica; Kahler, Charlene M.; Tzeng, Yih-Ling

    2011-01-01

    Proper periplasmic disulfide bond formation is important for folding and stability of many secreted and membrane proteins, and is catalyzed by three DsbA oxidoreductases in Neisseria meningitidis. DsbD provides reducing power to DsbC that shuffles incorrect disulfide bond in misfolded proteins as well as to the periplasmic enzymes that reduce apo-cytochrome c (CcsX) or repair oxidative protein damages (MrsAB). The expression of dsbD, but not other dsb genes, is positively regulated by the MisR/S two-component system. qRT-PCR analyses showed significantly reduced dsbD expression in all misR/S mutants, which was rescued by genetic complementation. The direct and specific interaction of MisR with the upstream region of the dsbD promoter was demonstrated by EMSA, and the MisR-binding sequences were mapped. Further, the expression of dsbD was found to be induced by dithiothrietol (DTT), through the MisR/S regulatory system. Surprisingly, we revealed that inactivation of dsbD can only be achieved in a strain carrying an ectopically located dsbD, in the dsbA1A2 double mutant or in the dsbA1A2A3 triple mutant, thus DsbD is indispensable for DsbA-catalyzed oxidative protein folding in N. meningitidis. The defects of the meningococcal dsbA1A2 mutant in transformation and resistance to oxidative stress were more severe in the absence of dsbD. PMID:21219471

  2. Comparative sequence, structure and redox analyses of Klebsiella pneumoniae DsbA show that anti-virulence target DsbA enzymes fall into distinct classes.

    PubMed

    Kurth, Fabian; Rimmer, Kieran; Premkumar, Lakshmanane; Mohanty, Biswaranjan; Duprez, Wilko; Halili, Maria A; Shouldice, Stephen R; Heras, Begoña; Fairlie, David P; Scanlon, Martin J; Martin, Jennifer L

    2013-01-01

    Bacterial DsbA enzymes catalyze oxidative folding of virulence factors, and have been identified as targets for antivirulence drugs. However, DsbA enzymes characterized to date exhibit a wide spectrum of redox properties and divergent structural features compared to the prototypical DsbA enzyme of Escherichia coli DsbA (EcDsbA). Nonetheless, sequence analysis shows that DsbAs are more highly conserved than their known substrate virulence factors, highlighting the potential to inhibit virulence across a range of organisms by targeting DsbA. For example, Salmonella enterica typhimurium (SeDsbA, 86 % sequence identity to EcDsbA) shares almost identical structural, surface and redox properties. Using comparative sequence and structure analysis we predicted that five other bacterial DsbAs would share these properties. To confirm this, we characterized Klebsiella pneumoniae DsbA (KpDsbA, 81 % identity to EcDsbA). As expected, the redox properties, structure and surface features (from crystal and NMR data) of KpDsbA were almost identical to those of EcDsbA and SeDsbA. Moreover, KpDsbA and EcDsbA bind peptides derived from their respective DsbBs with almost equal affinity, supporting the notion that compounds designed to inhibit EcDsbA will also inhibit KpDsbA. Taken together, our data show that DsbAs fall into different classes; that DsbAs within a class may be predicted by sequence analysis of binding loops; that DsbAs within a class are able to complement one another in vivo and that compounds designed to inhibit EcDsbA are likely to inhibit DsbAs within the same class. PMID:24244651

  3. Comparative Sequence, Structure and Redox Analyses of Klebsiella pneumoniae DsbA Show That Anti-Virulence Target DsbA Enzymes Fall into Distinct Classes

    PubMed Central

    Kurth, Fabian; Rimmer, Kieran; Premkumar, Lakshmanane; Mohanty, Biswaranjan; Duprez, Wilko; Halili, Maria A.; Shouldice, Stephen R.; Heras, Begoña; Fairlie, David P.; Scanlon, Martin J.; Martin, Jennifer L.

    2013-01-01

    Bacterial DsbA enzymes catalyze oxidative folding of virulence factors, and have been identified as targets for antivirulence drugs. However, DsbA enzymes characterized to date exhibit a wide spectrum of redox properties and divergent structural features compared to the prototypical DsbA enzyme of Escherichia coli DsbA (EcDsbA). Nonetheless, sequence analysis shows that DsbAs are more highly conserved than their known substrate virulence factors, highlighting the potential to inhibit virulence across a range of organisms by targeting DsbA. For example, Salmonella enterica typhimurium (SeDsbA, 86 % sequence identity to EcDsbA) shares almost identical structural, surface and redox properties. Using comparative sequence and structure analysis we predicted that five other bacterial DsbAs would share these properties. To confirm this, we characterized Klebsiella pneumoniae DsbA (KpDsbA, 81 % identity to EcDsbA). As expected, the redox properties, structure and surface features (from crystal and NMR data) of KpDsbA were almost identical to those of EcDsbA and SeDsbA. Moreover, KpDsbA and EcDsbA bind peptides derived from their respective DsbBs with almost equal affinity, supporting the notion that compounds designed to inhibit EcDsbA will also inhibit KpDsbA. Taken together, our data show that DsbAs fall into different classes; that DsbAs within a class may be predicted by sequence analysis of binding loops; that DsbAs within a class are able to complement one another in vivo and that compounds designed to inhibit EcDsbA are likely to inhibit DsbAs within the same class. PMID:24244651

  4. Becoming a crossover-competent DSB.

    PubMed

    Lake, Cathleen M; Hawley, R Scott

    2016-06-01

    The proper execution of meiotic recombination (or crossing over) is essential for chromosome segregation during the first meiotic division, and thus this process is regulated by multiple, and often elaborate, mechanisms. Meiotic recombination begins with the programmed induction of DNA double-strand breaks (DSBs), of which only a subset are selected to be repaired into crossovers. This crossover selection process is carried out by a number of pro-crossover proteins that regulate the fashion in which DSBs are repaired. Here, we highlight recent studies regarding the process of DSB fate selection by a family of pro-crossover proteins known as the Zip-3 homologs. PMID:26806636

  5. Structure of a DsbF homologue from Corynebacterium diphtheriae

    PubMed Central

    Um, Si-Hyeon; Kim, Jin-Sik; Lee, Kangseok; Ha, Nam-Chul

    2014-01-01

    Disulfide-bond formation, mediated by the Dsb family of proteins, is important in the correct folding of secreted or extracellular proteins in bacteria. In Gram-negative bacteria, disulfide bonds are introduced into the folding proteins in the periplasm by DsbA. DsbE from Escherichia coli has been implicated in the reduction of disulfide bonds in the maturation of cytochrome c. The Gram-positive bacterium Mycobacterium tuberculosis encodes DsbE and its homologue DsbF, the structures of which have been determined. However, the two mycobacterial proteins are able to oxidatively fold a protein in vitro, unlike DsbE from E. coli. In this study, the crystal structure of a DsbE or DsbF homologue protein from Corynebacterium diphtheriae has been determined, which revealed a thioredoxin-like domain with a typical CXXC active site. Structural comparison with M. tuberculosis DsbF would help in understanding the function of the C. diphtheriae protein. PMID:25195886

  6. Diversity of the Epsilonproteobacteria Dsb (disulfide bond) systems

    PubMed Central

    Bocian-Ostrzycka, Katarzyna M.; Grzeszczuk, Magdalena J.; Dziewit, Lukasz; Jagusztyn-Krynicka, Elżbieta K.

    2015-01-01

    The bacterial proteins of the Dsb family—important components of the post-translational protein modification system—catalyze the formation of disulfide bridges, a process that is crucial for protein structure stabilization and activity. Dsb systems play an essential role in the assembly of many virulence factors. Recent rapid advances in global analysis of bacteria have thrown light on the enormous diversity among bacterial Dsb systems. While the Escherichia coli disulfide bond-forming system is quite well understood, the mechanisms of action of Dsb systems in other bacteria, including members of class Epsilonproteobacteria that contain pathogenic and non-pathogenic bacteria colonizing extremely diverse ecological niches, are poorly characterized. Here we present a review of current knowledge on Epsilonproteobacteria Dsb systems. We have focused on the Dsb systems of Campylobacter spp. and Helicobacter spp. because our knowledge about Dsb proteins of Wolinella and Arcobacter spp. is still scarce and comes mainly from bioinformatic studies. Helicobacter pylori is a common human pathogen that colonizes the gastric epithelium of humans with severe consequences. Campylobacter spp. is a leading cause of zoonotic enteric bacterial infections in most developed and developing nations. We focus on various aspects of the diversity of the Dsb systems and their influence on pathogenicity, particularly because Dsb proteins are considered as potential targets for a new class of anti-virulence drugs to treat human infections by Campylobacter or Helicobacter spp. PMID:26106374

  7. Small molecule inhibitors of disulfide bond formation by the bacterial DsbA-DsbB dual enzyme system.

    PubMed

    Halili, Maria A; Bachu, Prabhakar; Lindahl, Fredrik; Bechara, Chérine; Mohanty, Biswaranjan; Reid, Robert C; Scanlon, Martin J; Robinson, Carol V; Fairlie, David P; Martin, Jennifer L

    2015-04-17

    The DsbA:DsbB redox machinery catalyzes disulfide bond formation in secreted proteins and is required for bacterial virulence factor assembly. Both enzymes have been identified as targets for antivirulence drugs. Here, we report synthetic analogues of ubiquinone (dimedone derivatives) that inhibit disulfide bond formation (IC50∼1 μM) catalyzed by E. coli DsbA:DsbB. The mechanism involves covalent modification of a single free cysteine leaving other cysteines unmodified. A vinylogous anhydride in each inhibitor is cleaved by the thiol, which becomes covalently modified to a thioester by a propionyl substituent. Cysteines and lysines on DsbA and DsbB and a nonredox enzyme were modified in a manner that implies some specificity. Moreover, human thioredoxin was not inhibited under the same conditions that inhibited EcDsbA. This proof of concept work uses small molecules that target specific cysteines to validate the DsbA and DsbB dual enzyme system as a viable and potentially druggable antivirulence target. PMID:25603425

  8. Evidence that the pathway of disulfide bond formation in Escherichia coli involves interactions between the cysteines of DsbB and DsbA.

    PubMed Central

    Guilhot, C; Jander, G; Martin, N L; Beckwith, J

    1995-01-01

    Disulfide bond formation is catalyzed in the periplasm of Escherichia coli. This process involves at least two proteins: DsbA and DsbB. Recent evidence suggests that DsbA, a soluble periplasmic protein directly catalyzes disulfide bond formation in proteins, whereas DsbB, an inner membrane protein, is involved in the reoxidation of DsbA. Here we present direct evidence of an interaction between DsbA and DsbB. (Kishigami et al. [Kishigami, S., Kanaya, E., Kikuchi, M. & Ito, K. (1995) J. Biol. Chem. 270, 17072-17074] have described similar findings.) We isolated a dominant negative mutant of dsbA, dsbAd, where Cys-33 of the DsbA active site is changed to tyrosine. Both DsbAd and DsbA are able to form a mixed disulfide with DsbB, which may be an intermediate in the reoxidation of DsbA. This complex is more stable with DsbAd. The dominance can be suppressed by increasing the production of DsbB. By using mutants of DsbB in which one or two cysteines have been changed to alanine, we show that only Cys-104 is important for complex formation. Therefore, we suggest that in vivo, reduced DsbA forms a complex with DsbB in which Cys-30 of DsbA is disulfide-bonded to Cys-104 of DsbB. Cys-104 is rapidly replaced by Cys-33 of DsbA to generate the oxidized form of this protein. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:7568240

  9. Preferential binding of an unfolded protein to DsbA.

    PubMed Central

    Frech, C; Wunderlich, M; Glockshuber, R; Schmid, F X

    1996-01-01

    The oxidoreductase DsbA from the periplasm of escherichia coli introduces disulfide bonds into proteins at an extremely high rate. During oxidation, a mixed disulfide is formed between DsbA and the folding protein chain, and this covalent intermediate reacts very rapidly either to form the oxidized protein or to revert back to oxidized DsbA. To investigate its properties, a stable form of the intermediate was produced by reacting the C33A variant of DsbA with a variant of RNase T1. We find that in this stable mixed disulfide the conformational stability of the substrate protein is decreased by 5 kJ/mol, whereas the conformational stability of DsbA is increased by 5 kJ/mol. This reciprocal effect suggests strongly that DsbA interacts with the unfolded substrate protein not only by the covalent disulfide bond, but also by preferential non-covalent interactions. The existence of a polypeptide binding site explains why DsbA oxidizes protein substrates much more rapidly than small thiol compounds. Such a very fast reaction is probably important for protein folding in the periplasm, because the accessibility of the thiol groups for DsbA can decrease rapidly when newly exported polypeptide chains begin to fold. PMID:8617214

  10. Expression and crystallization of DsbA from Staphylococcus aureus

    SciTech Connect

    Heras, B. Kurz, M.; Jarrott, R.; Byriel, K. A.; Jones, A.; Thöny-Meyer, L.; Martin, J. L.

    2007-11-01

    Free-interface diffusion crystallization chips were used to identify crystallization conditions for S. aureus DsbA, representing the first Gram-positive DsbA to be crystallized. Native and selenomethionine-derivative crystals diffracted to 2.1 and 2.4 Å resolution, respectively. Bacterial Dsb proteins catalyse the in vivo formation of disulfide bonds, a critical step in the stability and activity of many proteins. Most studies on Dsb proteins have focused on Gram-negative bacteria and thus the process of oxidative folding in Gram-positive bacteria is poorly understood. To help elucidate this process in Gram-positive bacteria, DsbA from Staphylococcus aureus (SaDsbA) has been focused on. Here, the expression, purification, crystallization and preliminary diffraction analysis of SaDsbA are reported. SaDsbA crystals diffract to a resolution limit of 2.1 Å and belong to the hexagonal space group P6{sub 5} or P6{sub 1}, with unit-cell parameters a = b = 72.1, c = 92.1 Å and one molecule in the asymmetric unit (64% solvent content)

  11. The gene ICS3 from the yeast Saccharomyces cerevisiae is involved in copper homeostasis dependent on extracellular pH.

    PubMed

    Alesso, C A; Discola, K F; Monteiro, G

    2015-09-01

    In the yeast Saccharomyces cerevisiae, many genes are involved in the uptake, transport, storage and detoxification of copper. Large scale studies have noted that deletion of the gene ICS3 increases sensitivity to copper, Sortin 2 and acid exposure. Here, we report a study on the Δics3 strain, in which ICS3 is related to copper homeostasis, affecting the intracellular accumulation of this metal. This strain is sensitive to hydrogen peroxide and copper exposure, but not to other tested transition metals. At pH 6.0, the Δics3 strain accumulates a larger amount of intracellular copper than the wild-type strain, explaining the sensitivity to oxidants in this condition. Unexpectedly, sensitivity to copper exposure only occurs in acidic conditions. This can be explained by the fact that the exposure of Δics3 cells to high copper concentrations at pH 4.0 results in over-accumulation of copper and iron. Moreover, the expression of ICS3 increases in acidic pH, and this is correlated with CCC2 gene expression, since both genes are regulated by Rim101 from the pH regulon. CCC2 is also upregulated in Δics3 in acidic pH. Together, these data indicate that ICS3 is involved in copper homeostasis and is dependent on extracellular pH. PMID:26127016

  12. Structural and Biochemical Characterization of the Oxidoreductase NmDsbA3 from Neisseria meningitidis

    SciTech Connect

    Vivian, Julian P.; Scoullar, Jessica; Robertson, Amy L.; Bottomley, Stephen P.; Horne, James; Chin, Yanni; Wielens, Jerome; Thompson, Philip E.; Velkov, Tony; Piek, Susannah; Byres, Emma; Beddoe, Travis; Wilce, Matthew C.J.; Kahler, Charlene M.; Rossjohn, Jamie; Scanlon, Martin J.

    2009-09-02

    DsbA is an enzyme found in the periplasm of Gram-negative bacteria that catalyzes the formation of disulfide bonds in a diverse array of protein substrates, many of which are involved in bacterial pathogenesis. Although most bacteria possess only a single essential DsbA, Neisseria meningitidis is unusual in that it possesses three DsbAs, although the reason for this additional redundancy is unclear. Two of these N. meningitidis enzymes (NmDsbA1 and NmDsbA2) play an important role in meningococcal attachment to human epithelial cells, whereas NmDsbA3 is considered to have a narrow substrate repertoire. To begin to address the role of DsbAs in the pathogenesis of N. meningitidis, we have determined the structure of NmDsbA3 to 2.3-{angstrom} resolution. Although the sequence identity between NmDsbA3 and other DsbAs is low, the NmDsbA3 structure adopted a DsbA-like fold. Consistent with this finding, we demonstrated that NmDsbA3 acts as a thiol-disulfide oxidoreductase in vitro and is reoxidized by Escherichia coli DsbB (EcDsbB). However, pronounced differences in the structures between DsbA3 and EcDsbA, which are clustered around the active site of the enzyme, suggested a structural basis for the unusual substrate specificity that is observed for NmDsbA3.

  13. Structural and biochemical characterization of the oxidoreductase NmDsbA3 from Neisseria meningitidis.

    PubMed

    Vivian, Julian P; Scoullar, Jessica; Robertson, Amy L; Bottomley, Stephen P; Horne, James; Chin, Yanni; Wielens, Jerome; Thompson, Philip E; Velkov, Tony; Piek, Susannah; Byres, Emma; Beddoe, Travis; Wilce, Matthew C J; Kahler, Charlene M; Rossjohn, Jamie; Scanlon, Martin J

    2008-11-21

    DsbA is an enzyme found in the periplasm of Gram-negative bacteria that catalyzes the formation of disulfide bonds in a diverse array of protein substrates, many of which are involved in bacterial pathogenesis. Although most bacteria possess only a single essential DsbA, Neisseria meningitidis is unusual in that it possesses three DsbAs, although the reason for this additional redundancy is unclear. Two of these N. meningitidis enzymes (NmDsbA1 and NmDsbA2) play an important role in meningococcal attachment to human epithelial cells, whereas NmDsbA3 is considered to have a narrow substrate repertoire. To begin to address the role of DsbAs in the pathogenesis of N. meningitidis, we have determined the structure of NmDsbA3 to 2.3-A resolution. Although the sequence identity between NmDsbA3 and other DsbAs is low, the NmDsbA3 structure adopted a DsbA-like fold. Consistent with this finding, we demonstrated that NmDsbA3 acts as a thiol-disulfide oxidoreductase in vitro and is reoxidized by Escherichia coli DsbB (EcDsbB). However, pronounced differences in the structures between DsbA3 and EcDsbA, which are clustered around the active site of the enzyme, suggested a structural basis for the unusual substrate specificity that is observed for NmDsbA3. PMID:18715864

  14. PREFACE: 8th International Conference on 3D Radiation Dosimetry (IC3DDose)

    NASA Astrophysics Data System (ADS)

    Olsson, Lars E.; Bäck, S.; Ceberg, Sofie

    2015-01-01

    IC3DDose 2014, the 8th International Conference on 3D Radiation Dosimetry was held in Ystad, Sweden, from 4-7 September 2014. This grew out of the DosGel series, which began as DosGel99, the 1st International Workshop on Radiation Therapy Gel Dosimetry in Lexington, Kentucky. Since 1999 subsequent DoSGel conferences were held in Brisbane, Australia (2001), Ghent, Belgium (2004), Sherbrooke, Canada (2006) and Crete, Greece (2008). In 2010 the conference was held on Hilton Head Island, South Carolina and underwent a name-change to IC3DDose. The 7th and last meeting was held in Sydney, Australia from 4-8 November 2012. It is worth remembering that the conference series started at the very beginning of the intensity modulated radiotherapy era and that the dosimeters being developed then were, to some extent, ahead of the clinical need of radiotherapy. However, since then the technical developments in radiation therapy have been dramatic, with dynamic treatments, including tracking, gating and volumetric modulated arc therapy, widely introduced in the clinic with the need for 3D dosimetry thus endless. This was also reflected by the contributions at the meeting in Ystad. Accordingly the scope of the meeting has also broadened to IC3DDOSE - I See Three-Dimensional Dose. A multitude of dosimetry techniques and radiation detectors are now represented, all with the common denominator: three-dimensional or 3D. Additionally, quality assurance (QA) procedures and other aspects of clinical dosimetry are represented. The implementation of new dosimetric techniques in radiotherapy is a process that needs every kind of caution, carefulness and thorough validation. Therefore, the clinical needs, reformulated as the aims for IC3DDOSE - I See Three-Dimensional Dose, are: • Enhance the quality and accuracy of radiation therapy treatments through improved clinical dosimetry. • Investigate and understand the dosimetric challenges of modern radiation treatment techniques. • Provide

  15. Disulfide Bond Oxidoreductase DsbA2 of Legionella pneumophila Exhibits Protein Disulfide Isomerase Activity

    PubMed Central

    Kpadeh, Zegbeh Z.; Jameson-Lee, Max; Yeh, Anthony J.; Chertihin, Olga; Shumilin, Igor A.; Dey, Rafik; Day, Shandra R.

    2013-01-01

    The extracytoplasmic assembly of the Dot/Icm type IVb secretion system (T4SS) of Legionella pneumophila is dependent on correct disulfide bond (DSB) formation catalyzed by a novel and essential disulfide bond oxidoreductase DsbA2 and not by DsbA1, a second nonessential DSB oxidoreductase. DsbA2, which is widely distributed in the microbial world, is phylogenetically distinct from the canonical DsbA oxidase and the DsbC protein disulfide isomerase (PDI)/reductase of Escherichia coli. Here we show that the extended N-terminal amino acid sequence of DsbA2 (relative to DsbA proteins) contains a highly conserved 27-amino-acid dimerization domain enabling the protein to form a homodimer. Complementation tests with E. coli mutants established that L. pneumophila dsbA1, but not the dsbA2 strain, restored motility to a dsbA mutant. In a protein-folding PDI detector assay, the dsbA2 strain, but not the dsbA1 strain, complemented a dsbC mutant of E. coli. Deletion of the dimerization domain sequences from DsbA2 produced the monomer (DsbA2N), which no longer exhibited PDI activity but complemented the E. coli dsbA mutant. PDI activity was demonstrated in vitro for DsbA2 but not DsbA1 in a nitrocefin-based mutant TEM β-lactamase folding assay. In an insulin reduction assay, DsbA2N activity was intermediate between those of DsbA2 and DsbA1. In L. pneumophila, DsbA2 was maintained as a mixture of thiol and disulfide forms, while in E. coli, DsbA2 was present as the reduced thiol. Our studies suggest that DsbA2 is a naturally occurring bifunctional disulfide bond oxidoreductase that may be uniquely suited to the majority of intracellular bacterial pathogens expressing T4SSs as well as in many slow-growing soil and aquatic bacteria. PMID:23435972

  16. PREFACE: 7th International Conference on 3D Radiation Dosimetry (IC3DDose)

    NASA Astrophysics Data System (ADS)

    Thwaites, David; Baldock, Clive

    2013-06-01

    IC3DDose 2013, the 7th International Conference on 3D Radiation Dosimetry held in Sydney, Australia from 4-8 November 2012, grew out of the DosGel series, which began as DosGel99, the 1st International Workshop on Radiation Therapy Gel Dosimetry in Lexington, Kentucky. Since 1999 subsequent DoSGel conferences were held in Brisbane, Australia (2001), Ghent, Belgium (2004), Sherbrooke, Canada (2006) and Crete, Greece (2008). In 2010 the conference was held on Hilton Head Island, South Carolina and underwent a name-change to IC3DDose. The aim of the first workshop was to bring together individuals, both researchers and users, with an interest in 3D radiation dosimetry techniques, with a mix of presentations from basic science to clinical applications, which has remained an objective for all of the meetings. One rationale of DosGel99 was stated as supporting the increasing clinical implementation of gel dosimetry, as the technique appeared, at that time, to be leaving the laboratories of gel dosimetry enthusiasts and entering clinical practice. Clearly by labelling the first workshop as the 1st, there was a vision of a continuing series, which has been fulfilled. On the other hand, the expectation of widespread clinical use of gel dosimetry has perhaps not been what was hoped for and anticipated. Nevertheless the rapidly increasing demand for advanced high-precision 3D radiotherapy technology and techniques has continued apace. The need for practical and accurate 3D dosimetry methods for development and quality assurance has only increased. By the 6th meeting, held in South Carolina in 2010, the Conference Scientific Committee recognised the wider developments in 3D systems and methods and decided to widen the scope, whilst keeping the same span from basic science to applications. This was signalled by a change of name from 'Dosgel' to 'IC3DDose', a name that has continued to this latest conference. The conference objectives were: to enhance the quality and accuracy of

  17. Structural and functional characterization of the oxidoreductase alpha-DsbA1 from Wolbachia pipientis.

    PubMed

    Kurz, Mareike; Iturbe-Ormaetxe, Iñaki; Jarrott, Russell; Shouldice, Stephen R; Wouters, Merridee A; Frei, Patrick; Glockshuber, Rudi; O'Neill, Scott L; Heras, Begoña; Martin, Jennifer L

    2009-07-01

    The alpha-proteobacterium Wolbachia pipientis is a highly successful intracellular endosymbiont of invertebrates that manipulates its host's reproductive biology to facilitate its own maternal transmission. The fastidious nature of Wolbachia and the lack of genetic transformation have hampered analysis of the molecular basis of these manipulations. Structure determination of key Wolbachia proteins will enable the development of inhibitors for chemical genetics studies. Wolbachia encodes a homologue (alpha-DsbA1) of the Escherichia coli dithiol oxidase enzyme EcDsbA, essential for the oxidative folding of many exported proteins. We found that the active-site cysteine pair of Wolbachia alpha-DsbA1 has the most reducing redox potential of any characterized DsbA. In addition, Wolbachia alpha-DsbA1 possesses a second disulfide that is highly conserved in alpha-proteobacterial DsbAs but not in other DsbAs. The alpha-DsbA1 structure lacks the characteristic hydrophobic features of EcDsbA, and the protein neither complements EcDsbA deletion mutants in E. coli nor interacts with EcDsbB, the redox partner of EcDsbA. The surface characteristics and redox profile of alpha-DsbA1 indicate that it probably plays a specialized oxidative folding role with a narrow substrate specificity. This first report of a Wolbachia protein structure provides the basis for future chemical genetics studies. PMID:19265485

  18. Glycosylation of DsbA in Francisella tularensis subsp. tularensis.

    PubMed

    Thomas, Rebecca M; Twine, Susan M; Fulton, Kelly M; Tessier, Luc; Kilmury, Sara L N; Ding, Wen; Harmer, Nicholas; Michell, Stephen L; Oyston, Petra C F; Titball, Richard W; Prior, Joann L

    2011-10-01

    In Francisella tularensis subsp. tularensis, DsbA has been shown to be an essential virulence factor and has been observed to migrate to multiple protein spots on two-dimensional electrophoresis gels. In this work, we show that the protein is modified with a 1,156-Da glycan moiety in O-linkage. The results of mass spectrometry studies suggest that the glycan is a hexasaccharide, comprised of N-acetylhexosamines, hexoses, and an unknown monosaccharide. Disruption of two genes within the FTT0789-FTT0800 putative polysaccharide locus, including a galE homologue (FTT0791) and a putative glycosyltransferase (FTT0798), resulted in loss of glycan modification of DsbA. The F. tularensis subsp. tularensis ΔFTT0798 and ΔFTT0791::Cm mutants remained virulent in the murine model of subcutaneous tularemia. This indicates that glycosylation of DsbA does not play a major role in virulence under these conditions. This is the first report of the detailed characterization of the DsbA glycan and putative role of the FTT0789-FTT0800 gene cluster in glycan biosynthesis. PMID:21803994

  19. A new light on the meiotic DSB catalytic complex.

    PubMed

    Robert, Thomas; Vrielynck, Nathalie; Mézard, Christine; de Massy, Bernard; Grelon, Mathilde

    2016-06-01

    Meiotic recombination is initiated by the formation of programmed DNA double-strand breaks (DSBs). More than 15 years ago, Spo11 was identified as the protein responsible for meiotic DSB formation, notably because of its striking similarities with the A subunit of topoisomerase VI (TopoVI). TopoVI are enzymes that modify DNA topology by generating transient DSBs and are active as heterotetramers, composed of two A and two B subunits. A2 dimers catalyse the DNA cleavage reaction, whereas the B subunits regulate A2 conformation, DNA capture, cleavage and re-ligation. The recent identification in plants and mammals of a B-like TopoVI subunit that interacts with SPO11 and is required for meiotic DSB formation makes us to reconsider our understanding of the meiotic DSB catalytic complex. We provide here an overview of the knowledge on TopoVI structure and mode of action and we compare them with their meiotic counterparts. This allows us to discuss the nature, structure and functions of the meiotic TopoVI-like complex during meiotic DSB formation. PMID:26995551

  20. Homeostatic regulation of meiotic DSB formation by ATM/ATR

    SciTech Connect

    Cooper, Tim J.; Wardell, Kayleigh; Garcia, Valerie; Neale, Matthew J.

    2014-11-15

    Ataxia–telangiectasia mutated (ATM) and RAD3-related (ATR) are widely known as being central players in the mitotic DNA damage response (DDR), mounting responses to DNA double-strand breaks (DSBs) and single-stranded DNA (ssDNA) respectively. The DDR signalling cascade couples cell cycle control to damage-sensing and repair processes in order to prevent untimely cell cycle progression while damage still persists [1]. Both ATM/ATR are, however, also emerging as essential factors in the process of meiosis; a specialised cell cycle programme responsible for the formation of haploid gametes via two sequential nuclear divisions. Central to achieving accurate meiotic chromosome segregation is the introduction of numerous DSBs spread across the genome by the evolutionarily conserved enzyme, Spo11. This review seeks to explore and address how cells utilise ATM/ATR pathways to regulate Spo11-DSB formation, establish DSB homeostasis and ensure meiosis is completed unperturbed.

  1. Cancer TARGETases: DSB repair as a pharmacological target.

    PubMed

    Samadder, Pounami; Aithal, Rakesh; Belan, Ondrej; Krejci, Lumir

    2016-05-01

    Cancer is a disease attributed to the accumulation of DNA damages due to incapacitation of DNA repair pathways resulting in genomic instability and a mutator phenotype. Among the DNA lesions, double stranded breaks (DSBs) are the most toxic forms of DNA damage which may arise as a result of extrinsic DNA damaging agents or intrinsic replication stress in fast proliferating cancer cells. Accurate repair of DSBs is therefore paramount to the cell survival, and several classes of proteins such as kinases, nucleases, helicases or core recombinational proteins have pre-defined jobs in precise execution of DSB repair pathways. On one hand, the proper functioning of these proteins ensures maintenance of genomic stability in normal cells, and on the other hand results in resistance to various drugs employed in cancer therapy and therefore presents a suitable opportunity for therapeutic targeting. Higher relapse and resistance in cancer patients due to non-specific, cytotoxic therapies is an alarming situation and it is becoming more evident to employ personalized treatment based on the genetic landscape of the cancer cells. For the success of personalized treatment, it is of immense importance to identify more suitable targetable proteins in DSB repair pathways and also to explore new synthetic lethal interactions with these pathways. Here we review the various alternative approaches to target the various protein classes termed as cancer TARGETases in DSB repair pathway to obtain more beneficial and selective therapy. PMID:26899499

  2. Tolerance is dependent on complement C3 fragment iC3b binding to antigen-presenting cells

    PubMed Central

    Sohn, Jeong-Hyeon; Bora, Puran S.; Suk, Hye-Jung; Molina, Hector; Kaplan, Henry J.; Bora, Nalini S.

    2007-01-01

    Systemic tolerance can be induced by the introduction of antigen into an immune-privileged site. Here we investigated the role of complement in the induction of tolerance after intraocular injection. We found that the development of antigen-specific tolerance is dependent on a complement activation product. The ligation of the complement C3 activation product iC3b to complement receptor type 3 (the iC3b receptor) on antigen-presenting cells resulted in the sequential production of transforming growth factor-β2 and interleukin-10, which is essential for the induction of tolerance. These observations may extend to the development of both neonatal tolerance and other forms of acquired tolerance. PMID:12514742

  3. DNA DSB repair pathway choice: an orchestrated handover mechanism

    PubMed Central

    Kakarougkas, A

    2014-01-01

    DNA double strand breaks (DSBs) are potential lethal lesions but can also lead to chromosome rearrangements, a step promoting carcinogenesis. DNA non-homologous end-joining (NHEJ) is the major DSB rejoining process and occurs in all cell cycle stages. Homologous recombination (HR) can additionally function to repair irradiation-induced two-ended DSBs in G2 phase. In mammalian cells, HR predominantly uses a sister chromatid as a template for DSB repair; thus HR functions only in late S/G2 phase. Here, we review current insight into the interplay between HR and NHEJ in G2 phase. We argue that NHEJ represents the first choice pathway, repairing approximately 80% of X-ray-induced DSBs with rapid kinetics. However, a subset of DSBs undergoes end resection and repair by HR. 53BP1 restricts resection, thereby promoting NHEJ. During the switch from NHEJ to HR, 53BP1 is repositioned to the periphery of enlarged irradiation-induced foci (IRIF) via a BRCA1-dependent process. K63-linked ubiquitin chains, which also form at IRIF, are also repositioned as well as receptor-associated protein 80 (RAP80), a ubiquitin binding protein. RAP80 repositioning requires POH1, a proteasome component. Thus, the interfacing barriers to HR, 53BP1 and RAP80 are relieved by POH1 and BRCA1, respectively. Removal of RAP80 from the IRIF core is required for loss of the ubiquitin chains and 53BP1, and for efficient replication protein A foci formation. We propose that NHEJ is used preferentially to HR because it is a compact process that does not necessitate extensive chromatin changes in the DSB vicinity. PMID:24363387

  4. Evolutionary domain fusion expanded the substrate specificity of the transmembrane electron transporter DsbD

    PubMed Central

    Katzen, Federico; Deshmukh, Meenal; Daldal, Fevzi; Beckwith, Jon

    2002-01-01

    Modular organization of proteins has been postulated as a widely used strategy for protein evolution. The multidomain transmembrane protein DsbD catalyzes the transfer of electrons from the cytoplasm to the periplasm of Escherichia coli. Most bacterial species do not have DsbD, but instead their genomes encode a much smaller protein, CcdA, which resembles the central hydrophobic domain of DsbD. We used reciprocal heterologous complementation assays between E.coli and Rhodobacter capsulatus to show that, despite their differences in size and structure, DsbD and CcdA are functional homologs. While DsbD transfers reducing potential to periplasmic protein disulfide bond isomerases and to the cytochrome c thioreduction pathway, CcdA appears to be involved only in cytochrome c biogenesis. Our findings strongly suggest that, by the acquisition of additional thiol-redox active domains, DsbD expanded its substrate specificity. PMID:12145197

  5. Dehydrohalogenation and Dehydration Reactions of i-C3H7Br and i-C3H7OH by Sodium Ions Studied by Guided Ion Beam Techniques and Quantum Chemical Methods.

    PubMed

    López, E; Lucas, J M; de Andrés, J; Albertí, M; Bofill, J M; Aguilar, A

    2016-07-14

    Dehydrohalogenation and dehydration reactions of gas-phase i-C3H7Br and i-C3H7OH molecules induced by collision with Na(+), all participants being in their electronic ground state, were studied experimentally in our laboratory using a radiofrequency-guided ion beam apparatus and covering the 0.10-10.00 eV center of mass (CM) energy range. In Na(+) + i-C3H7Br collisions the formation of [C3H6-Na](+) and [HBr-Na](+) by dehydrohalogenation was observed and quantified, as well as that of the ion-molecule adduct [Na-i-C3H7Br](+) together with its decomposition products C3H7(+) and NaBr. In Na(+) + i-C3H7OH collisions the dehydration product [H2O-Na](+) was also found, while [C3H6-Na](+) was hardly detected. Moreover, the [Na-i-C3H7OH](+) adduct formation as well as its decomposition into C3H7(+) and NaOH were also quantified. For all these processes, absolute reaction cross sections were measured as a function of the CM collision energy. From measured excitation functions, rate constants for the formation of [C3H6-Na](+), [HBr-Na](+), and [H2O-Na](+) at 303 K were obtained. Complementing the experiments, exhaustive ab initio structure calculations at the MP2 level of theory were performed, giving information on the most relevant features of the potential energy surfaces (PESs) where the dehydrohalogenation, dehydration, and decomposition reactions take place adiabatically for both collision systems. On these PESs different stationary points associated with potential energy minima and transition state barriers were characterized, and their connectivity was ensured using the intrinsic-reaction-coordinate method. The main topology features of the ab initio calculated PESs allowed a qualitative interpretation of the experimental data also exposing the role of the sodium ion as a catalyst in elimination reactions. PMID:26811987

  6. The Structure of the Bacterial Oxidoreductase Enzyme DsbA in Complex with a Peptide Reveals a Basis for Substrate Specificity in the Catalytic Cycle of DsbA Enzymes

    SciTech Connect

    Paxman, Jason J.; Borg, Natalie A.; Horne, James; Thompson, Philip E.; Chin, Yanni; Sharma, Pooja; Simpson, Jamie S.; Wielens, Jerome; Piek, Susannah; Kahler, Charlene M.; Sakellaris, Harry; Pearce, Mary; Bottomley, Stephen P.; Rossjohn, Jamie; Scanlon, Martin J.

    2010-09-07

    Oxidative protein folding in Gram-negative bacteria results in the formation of disulfide bonds between pairs of cysteine residues. This is a multistep process in which the dithiol-disulfide oxidoreductase enzyme, DsbA, plays a central role. The structure of DsbA comprises an all helical domain of unknown function and a thioredoxin domain, where active site cysteines shuttle between an oxidized, substrate-bound, reduced form and a DsbB-bound form, where DsbB is a membrane protein that reoxidizes DsbA. Most DsbA enzymes interact with a wide variety of reduced substrates and show little specificity. However, a number of DsbA enzymes have now been identified that have narrow substrate repertoires and appear to interact specifically with a smaller number of substrates. The transient nature of the DsbA-substrate complex has hampered our understanding of the factors that govern the interaction of DsbA enzymes with their substrates. Here we report the crystal structure of a complex between Escherichia coli DsbA and a peptide with a sequence derived from a substrate. The binding site identified in the DsbA-peptide complex was distinct from that observed for DsbB in the DsbA-DsbB complex. The structure revealed details of the DsbA-peptide interaction and suggested a mechanism by which DsbA can simultaneously show broad specificity for substrates yet exhibit specificity for DsbB. This mode of binding was supported by solution nuclear magnetic resonance data as well as functional data, which demonstrated that the substrate specificity of DsbA could be modified via changes at the binding interface identified in the structure of the complex.

  7. Real-time Monitoring of Intermediates Reveals the Reaction Pathway in the Thiol-Disulfide Exchange between Disulfide Bond Formation Protein A (DsbA) and B (DsbB) on a Membrane-immobilized Quartz Crystal Microbalance (QCM) System*

    PubMed Central

    Yazawa, Kenjiro; Furusawa, Hiroyuki; Okahata, Yoshio

    2013-01-01

    Disulfide bond formation protein B (DsbBS-S,S-S) is an inner membrane protein in Escherichia coli that has two disulfide bonds (S-S, S-S) that play a role in oxidization of a pair of cysteine residues (SH, SH) in disulfide bond formation protein A (DsbASH,SH). The oxidized DsbAS-S, with one disulfide bond (S-S), can oxidize proteins with SH groups for maturation of a folding preprotein. Here, we have described the transient kinetics of the oxidation reaction between DsbASH,SH and DsbBS-S,S-S. We immobilized DsbBS-S,S-S embedded in lipid bilayers on the surface of a 27-MHz quartz crystal microbalance (QCM) device to detect both formation and degradation of the reaction intermediate (DsbA-DsbB), formed via intermolecular disulfide bonds, as a mass change in real time. The obtained kinetic parameters (intermediate formation, reverse, and oxidation rate constants (kf, kr, and kcat, respectively) indicated that the two pairs of cysteine residues in DsbBS-S,S-S were more important for the stability of the DsbA-DsbB intermediate than ubiquinone, an electron acceptor for DsbBS-S,S-S. Our data suggested that the reaction pathway of almost all DsbASH,SH oxidation processes would proceed through this stable intermediate, avoiding the requirement for ubiquinone. PMID:24145032

  8. Real-time monitoring of intermediates reveals the reaction pathway in the thiol-disulfide exchange between disulfide bond formation protein A (DsbA) and B (DsbB) on a membrane-immobilized quartz crystal microbalance (QCM) system.

    PubMed

    Yazawa, Kenjiro; Furusawa, Hiroyuki; Okahata, Yoshio

    2013-12-13

    Disulfide bond formation protein B (DsbBS-S,S-S) is an inner membrane protein in Escherichia coli that has two disulfide bonds (S-S, S-S) that play a role in oxidization of a pair of cysteine residues (SH, SH) in disulfide bond formation protein A (DsbASH,SH). The oxidized DsbAS-S, with one disulfide bond (S-S), can oxidize proteins with SH groups for maturation of a folding preprotein. Here, we have described the transient kinetics of the oxidation reaction between DsbASH,SH and DsbBS-S,S-S. We immobilized DsbBS-S,S-S embedded in lipid bilayers on the surface of a 27-MHz quartz crystal microbalance (QCM) device to detect both formation and degradation of the reaction intermediate (DsbA-DsbB), formed via intermolecular disulfide bonds, as a mass change in real time. The obtained kinetic parameters (intermediate formation, reverse, and oxidation rate constants (kf, kr, and kcat, respectively) indicated that the two pairs of cysteine residues in DsbBS-S,S-S were more important for the stability of the DsbA-DsbB intermediate than ubiquinone, an electron acceptor for DsbBS-S,S-S. Our data suggested that the reaction pathway of almost all DsbASH,SH oxidation processes would proceed through this stable intermediate, avoiding the requirement for ubiquinone. PMID:24145032

  9. Negative Regulation of DsbA-L Gene Expression by the Transcription Factor Sp1

    PubMed Central

    Fang, Qichen; Yang, Wenjing; Li, Huating; Hu, Wenxiu; Chen, Lihui; Jiang, Shan; Dong, Kun; Song, Qianqian; Wang, Chen; Chen, Shuo; Liu, Feng

    2014-01-01

    Disulfide-bond A oxidoreductase-like protein (DsbA-L) possesses beneficial effects such as promoting adiponectin multimerization and stability, increasing insulin sensitivity, and enhancing energy metabolism. The expression level of DsbA-L is negatively correlated with obesity in mice and humans, but the underlying mechanisms remain unknown. To address this question, we generated reporter gene constructs containing the promoter sequence of the mouse DsbA-L gene. Deletion analysis showed that the proximal promoter of mouse DsbA-L is located between −186 and −34 bp relative to the transcription start site. In silico analysis identified a putative Sp1 transcription factor binding site in the first intron of the DsbA-L gene. Electrophoretic mobility shift assay and chromatin immunoprecipitation analysis indicated that Sp1 bound to this intron region in vitro and in intact cells. Overexpression of Sp1 or suppressing Sp1 expression by siRNA reduced or increased DsbA-L promoter activity, respectively. The binding activity of Sp1 was gradually decreased during 3T3-L1 cell differentiation and was significantly increased in adipose tissues of obese mice. Our results identify Sp1 as an inhibitor of DsbA-L gene transcription, and the Sp1-mediated inhibition of DsbA-L gene expression may provide a mechanism underlying obesity-induced adiponectin downregulation and insulin resistance. PMID:25024375

  10. Cleavage of Complement C3b to iC3b on the Surface of Staphylococcus aureus Is Mediated by Serum Complement Factor I

    PubMed Central

    Cunnion, K. M.; Hair, P. S.; Buescher, E. S.

    2004-01-01

    Complement-mediated opsonization of Staphylococcus aureus bearing the dominant capsule serotypes, serotypes 5 and 8, remains incompletely understood. We have previously shown that complement plays a vital role in the efficient phagocytosis of a serotype 5 S. aureus strain and that the opsonic fragments of the central complement protein C3, C3b and iC3b, were present on the bacterial surface after incubation in human serum. In the present studies, C3b and iC3b were found on several serotype 5 and 8 S. aureus strains after incubation in human serum. Using purified classical activation pathway complement proteins and the Western blot assay, we showed that when C3b was generated on the S. aureus surface no iC3b fragments were found, suggesting that other serum proteins may be required for cleaving C3b to iC3b. When C3b-coated S. aureus was incubated with serum factor I, a complement regulatory protein, iC3b was generated. Purified factor H, a serum protein cofactor for factor I, did not enhance factor I-mediated cleavage of C3b. These findings suggest that C3b cleavage to iC3b on S. aureus is mediated by serum factor I and does not require factor H. PMID:15102797

  11. Biochemical and structural study of the homologues of the thiol-disulfide oxidoreductase DsbA in Neisseria meningitidis.

    PubMed

    Lafaye, Céline; Iwema, Thomas; Carpentier, Philippe; Jullian-Binard, Céline; Kroll, J Simon; Collet, Jean-François; Serre, Laurence

    2009-10-01

    Bacterial virulence depends on the correct folding of surface-exposed proteins, a process catalyzed by the thiol-disulfide oxidoreductase DsbA, which facilitates the synthesis of disulfide bonds in Gram-negative bacteria. The Neisseria meningitidis genome possesses three genes encoding active DsbAs: DsbA1, DsbA2 and DsbA3. DsbA1 and DsbA2 have been characterized as lipoproteins involved in natural competence and in host interactive biology, while the function of DsbA3 remains unknown. This work reports the biochemical characterization of the three neisserial enzymes and the crystal structures of DsbA1 and DsbA3. As predicted by sequence homology, both enzymes adopt the classic Escherichia coli DsbA fold. The most striking feature shared by all three proteins is their exceptional oxidizing power. With a redox potential of -80 mV, the neisserial DsbAs are the most oxidizing thioredoxin-like enzymes known to date. Consistent with these findings, thermal studies indicate that their reduced form is also extremely stable. For each of these enzymes, this study shows that a threonine residue found within the active-site region plays a key role in dictating this extraordinary oxidizing power. This result highlights how residues located outside the CXXC motif may influence the redox potential of members of the thioredoxin family. PMID:19631659

  12. Compounds targeting disulfide bond forming enzyme DsbB of Gram-negative bacteria

    PubMed Central

    Landeta, Cristina; Blazyk, Jessica L.; Hatahet, Feras; Meehan, Brian M.; Eser, Markus; Myrick, Alissa; Bronstain, Ludmila; Minami, Shoko; Arnold, Holly; Ke, Na; Rubin, Eric J.; Furie, Barbara C.; Furie, Bruce; Beckwith, Jon; Dutton, Rachel; Boyd, Dana

    2015-01-01

    In bacteria, disulfide bonds confer stability on many proteins exported to the cell envelope or beyond. These proteins include numerous bacterial virulence factors. Thus, bacterial enzymes that promote disulfide bond formation represent targets for compounds inhibiting bacterial virulence. Here, we describe a novel target- and cell-based screening methodology for identifying compounds that inhibit the disulfide bond-forming enzymes E. coli DsbB (EcDsbB) or M. tuberculosis VKOR (MtbVKOR). MtbVKOR can replace EcDsbB although the two are not homologues. Initial screening of 51,487 compounds yielded six specifically inhibiting EcDsbB. These compounds share a structural motif and do not inhibit MtbVKOR. A medicinal chemistry approach led us to select related compounds some of which are much more effective DsbB inhibitors than those found in the screen. These compounds inhibit purified DsbB and prevent anaerobic E. coli growth. Furthermore, these compounds inhibit all but one of the DsbBs of nine other gram-negative pathogenic bacteria tested. PMID:25686372

  13. Functional and Bioinformatics Analysis of Two Campylobacter jejuni Homologs of the Thiol-Disulfide Oxidoreductase, DsbA

    PubMed Central

    Grabowska, Anna D.; Wywiał, Ewa; Dunin-Horkawicz, Stanislaw; Łasica, Anna M.; Wösten, Marc M. S. M.; Nagy-Staroń, Anna; Godlewska, Renata; Bocian-Ostrzycka, Katarzyna; Pieńkowska, Katarzyna; Łaniewski, Paweł; Bujnicki, Janusz M.; van Putten, Jos P. M.; Jagusztyn-Krynicka, E. Katarzyna

    2014-01-01

    Background Bacterial Dsb enzymes are involved in the oxidative folding of many proteins, through the formation of disulfide bonds between their cysteine residues. The Dsb protein network has been well characterized in cells of the model microorganism Escherichia coli. To gain insight into the functioning of the Dsb system in epsilon-Proteobacteria, where it plays an important role in the colonization process, we studied two homologs of the main Escherichia coli Dsb oxidase (EcDsbA) that are present in the cells of the enteric pathogen Campylobacter jejuni, the most frequently reported bacterial cause of human enteritis in the world. Methods and Results Phylogenetic analysis suggests the horizontal transfer of the epsilon-Proteobacterial DsbAs from a common ancestor to gamma-Proteobacteria, which then gave rise to the DsbL lineage. Phenotype and enzymatic assays suggest that the two C. jejuni DsbAs play different roles in bacterial cells and have divergent substrate spectra. CjDsbA1 is essential for the motility and autoagglutination phenotypes, while CjDsbA2 has no impact on those processes. CjDsbA1 plays a critical role in the oxidative folding that ensures the activity of alkaline phosphatase CjPhoX, whereas CjDsbA2 is crucial for the activity of arylsulfotransferase CjAstA, encoded within the dsbA2-dsbB-astA operon. Conclusions Our results show that CjDsbA1 is the primary thiol-oxidoreductase affecting life processes associated with bacterial spread and host colonization, as well as ensuring the oxidative folding of particular protein substrates. In contrast, CjDsbA2 activity does not affect the same processes and so far its oxidative folding activity has been demonstrated for one substrate, arylsulfotransferase CjAstA. The results suggest the cooperation between CjDsbA2 and CjDsbB. In the case of the CjDsbA1, this cooperation is not exclusive and there is probably another protein to be identified in C. jejuni cells that acts to re-oxidize CjDsbA1. Altogether

  14. Virtual Screening of Peptide and Peptidomimetic Fragments Targeted to Inhibit Bacterial Dithiol Oxidase DsbA.

    PubMed

    Duprez, Wilko; Bachu, Prabhakar; Stoermer, Martin J; Tay, Stephanie; McMahon, Róisín M; Fairlie, David P; Martin, Jennifer L

    2015-01-01

    Antibacterial drugs with novel scaffolds and new mechanisms of action are desperately needed to address the growing problem of antibiotic resistance. The periplasmic oxidative folding system in Gram-negative bacteria represents a possible target for anti-virulence antibacterials. By targeting virulence rather than viability, development of resistance and side effects (through killing host native microbiota) might be minimized. Here, we undertook the design of peptidomimetic inhibitors targeting the interaction between the two key enzymes of oxidative folding, DsbA and DsbB, with the ultimate goal of preventing virulence factor assembly. Structures of DsbB--or peptides--complexed with DsbA revealed key interactions with the DsbA active site cysteine, and with a hydrophobic groove adjacent to the active site. The present work aimed to discover peptidomimetics that target the hydrophobic groove to generate non-covalent DsbA inhibitors. The previously reported structure of a Proteus mirabilis DsbA active site cysteine mutant, in a non-covalent complex with the heptapeptide PWATCDS, was used as an in silico template for virtual screening of a peptidomimetic fragment library. The highest scoring fragment compound and nine derivatives were synthesized and evaluated for DsbA binding and inhibition. These experiments discovered peptidomimetic fragments with inhibitory activity at millimolar concentrations. Although only weakly potent relative to larger covalent peptide inhibitors that interact through the active site cysteine, these fragments offer new opportunities as templates to build non-covalent inhibitors. The results suggest that non-covalent peptidomimetics may need to interact with sites beyond the hydrophobic groove in order to produce potent DsbA inhibitors. PMID:26225423

  15. Kinetics of DSB rejoining and formation of simple chromosome exchange aberrations

    NASA Technical Reports Server (NTRS)

    Cucinotta, F. A.; Nikjoo, H.; O'Neill, P.; Goodhead, D. T.

    2000-01-01

    PURPOSE: To investigate the role of kinetics in the processing of DNA double strand breaks (DSB), and the formation of simple chromosome exchange aberrations following X-ray exposures to mammalian cells based on an enzymatic approach. METHODS: Using computer simulations based on a biochemical approach, rate-equations that describe the processing of DSB through the formation of a DNA-enzyme complex were formulated. A second model that allows for competition between two processing pathways was also formulated. The formation of simple exchange aberrations was modelled as misrepair during the recombination of single DSB with undamaged DNA. Non-linear coupled differential equations corresponding to biochemical pathways were solved numerically by fitting to experimental data. RESULTS: When mediated by a DSB repair enzyme complex, the processing of single DSB showed a complex behaviour that gives the appearance of fast and slow components of rejoining. This is due to the time-delay caused by the action time of enzymes in biomolecular reactions. It is shown that the kinetic- and dose-responses of simple chromosome exchange aberrations are well described by a recombination model of DSB interacting with undamaged DNA when aberration formation increases with linear dose-dependence. Competition between two or more recombination processes is shown to lead to the formation of simple exchange aberrations with a dose-dependence similar to that of a linear quadratic model. CONCLUSIONS: Using a minimal number of assumptions, the kinetics and dose response observed experimentally for DSB rejoining and the formation of simple chromosome exchange aberrations are shown to be consistent with kinetic models based on enzymatic reaction approaches. A non-linear dose response for simple exchange aberrations is possible in a model of recombination of DNA containing a DSB with undamaged DNA when two or more pathways compete for DSB repair.

  16. Virtual Screening of Peptide and Peptidomimetic Fragments Targeted to Inhibit Bacterial Dithiol Oxidase DsbA

    PubMed Central

    Stoermer, Martin J.; Tay, Stephanie; McMahon, Róisín M.; Fairlie, David P.; Martin, Jennifer L.

    2015-01-01

    Antibacterial drugs with novel scaffolds and new mechanisms of action are desperately needed to address the growing problem of antibiotic resistance. The periplasmic oxidative folding system in Gram-negative bacteria represents a possible target for anti-virulence antibacterials. By targeting virulence rather than viability, development of resistance and side effects (through killing host native microbiota) might be minimized. Here, we undertook the design of peptidomimetic inhibitors targeting the interaction between the two key enzymes of oxidative folding, DsbA and DsbB, with the ultimate goal of preventing virulence factor assembly. Structures of DsbB - or peptides - complexed with DsbA revealed key interactions with the DsbA active site cysteine, and with a hydrophobic groove adjacent to the active site. The present work aimed to discover peptidomimetics that target the hydrophobic groove to generate non-covalent DsbA inhibitors. The previously reported structure of a Proteus mirabilis DsbA active site cysteine mutant, in a non-covalent complex with the heptapeptide PWATCDS, was used as an in silico template for virtual screening of a peptidomimetic fragment library. The highest scoring fragment compound and nine derivatives were synthesized and evaluated for DsbA binding and inhibition. These experiments discovered peptidomimetic fragments with inhibitory activity at millimolar concentrations. Although only weakly potent relative to larger covalent peptide inhibitors that interact through the active site cysteine, these fragments offer new opportunities as templates to build non-covalent inhibitors. The results suggest that non-covalent peptidomimetics may need to interact with sites beyond the hydrophobic groove in order to produce potent DsbA inhibitors. PMID:26225423

  17. Disarming Burkholderia pseudomallei: Structural and Functional Characterization of a Disulfide Oxidoreductase (DsbA) Required for Virulence In Vivo

    PubMed Central

    McMahon, Róisín M.; Marshall, Laura E.; Halili, Maria; Furlong, Emily; Tay, Stephanie; Sarkar-Tyson, Mitali

    2014-01-01

    Abstract Aims: The intracellular pathogen Burkholderia pseudomallei causes the disease melioidosis, a major source of morbidity and mortality in southeast Asia and northern Australia. The need to develop novel antimicrobials is compounded by the absence of a licensed vaccine and the bacterium's resistance to multiple antibiotics. In a number of clinically relevant Gram-negative pathogens, DsbA is the primary disulfide oxidoreductase responsible for catalyzing the formation of disulfide bonds in secreted and membrane-associated proteins. In this study, a putative B. pseudomallei dsbA gene was evaluated functionally and structurally and its contribution to infection assessed. Results: Biochemical studies confirmed the dsbA gene encodes a protein disulfide oxidoreductase. A dsbA deletion strain of B. pseudomallei was attenuated in both macrophages and a BALB/c mouse model of infection and displayed pleiotropic phenotypes that included defects in both secretion and motility. The 1.9 Å resolution crystal structure of BpsDsbA revealed differences from the classic member of this family Escherichia coli DsbA, in particular within the region surrounding the active site disulfide where EcDsbA engages with its partner protein E. coli DsbB, indicating that the interaction of BpsDsbA with its proposed partner BpsDsbB may be distinct from that of EcDsbA-EcDsbB. Innovation: This study has characterized BpsDsbA biochemically and structurally and determined that it is required for virulence of B. pseudomallei. Conclusion: These data establish a critical role for BpsDsbA in B. pseudomallei infection, which in combination with our structural characterization of BpsDsbA will facilitate the future development of rationally designed inhibitors against this drug-resistant organism. Antioxid. Redox Signal. 20, 606–617. PMID:23901809

  18. Preliminary crystallographic data of the three homologues of the thiol–disulfide oxidoreductase DsbA in Neisseria meningitidis

    PubMed Central

    Lafaye, Céline; Iwena, Thomas; Ferrer, Jean-Luc; Kroll, J. Simon; Griat, Mickael; Serre, Laurence

    2008-01-01

    Bacterial virulence depends on the correct folding of surface-exposed proteins, a process that is catalyzed by the thiol-disulfide oxidoreductase DsbA, which facilitates the synthesis of disulfide bonds in Gram-negative bacteria. Uniquely among bacteria, the Neisseria meningitidis genome possesses three genes encoding active DsbAs: DsbA1, DsbA2 and DsbA3. DsbA1 and DsbA2 have been characterized as lipoproteins involved in natural competence and in host-interactive biology, while the function of DsbA3 remains unknown. In an attempt to shed light on the reason for this multiplicity of dsbA genes, the three enzymes from N. meningitidis have been purified and crystallized in the presence of high concentrations of ammonium sulfate. The best crystals were obtained using DsbA1 and DsbA3; they belong to the orthorhombic and tetragonal systems and diffract to 1.5 and 2.7 Å resolution, respectively. PMID:18259062

  19. Preliminary crystallographic data of the three homologues of the thiol-disulfide oxidoreductase DsbA in Neisseria meningitidis.

    PubMed

    Lafaye, Céline; Iwema, Thomas; Iwena, Thomas; Ferrer, Jean-Luc; Kroll, J Simon; Griat, Mickael; Serre, Laurence

    2008-02-01

    Bacterial virulence depends on the correct folding of surface-exposed proteins, a process that is catalyzed by the thiol-disulfide oxidoreductase DsbA, which facilitates the synthesis of disulfide bonds in Gram-negative bacteria. Uniquely among bacteria, the Neisseria meningitidis genome possesses three genes encoding active DsbAs: DsbA1, DsbA2 and DsbA3. DsbA1 and DsbA2 have been characterized as lipoproteins involved in natural competence and in host-interactive biology, while the function of DsbA3 remains unknown. In an attempt to shed light on the reason for this multiplicity of dsbA genes, the three enzymes from N. meningitidis have been purified and crystallized in the presence of high concentrations of ammonium sulfate. The best crystals were obtained using DsbA1 and DsbA3; they belong to the orthorhombic and tetragonal systems and diffract to 1.5 and 2.7 A resolution, respectively. PMID:18259062

  20. A new role for Escherichia coli DsbC protein in protection against oxidative stress.

    PubMed

    Denoncin, Katleen; Vertommen, Didier; Arts, Isabelle S; Goemans, Camille V; Rahuel-Clermont, Sophie; Messens, Joris; Collet, Jean-François

    2014-05-01

    We report a new function for Escherichia coli DsbC, a protein best known for disulfide bond isomerization in the periplasm. We found that DsbC regulates the redox state of the single cysteine of the L-arabinose-binding protein AraF. This cysteine, which can be oxidized to a sulfenic acid, mediates the formation of a disulfide-linked homodimer under oxidative stress conditions, preventing L-arabinose binding. DsbC, unlike the homologous protein DsbG, reduces the intermolecular disulfide, restoring AraF binding properties. Thus, our results reveal a new link between oxidative protein folding and the defense mechanisms against oxidative stress. PMID:24634211

  1. The dsbB gene product is required for protease production by Burkholderia cepacia.

    PubMed Central

    Abe, M; Nakazawa, T

    1996-01-01

    Burkholderia cepacia KF1, isolated from a pneumonia patient, produces a 37-kDa extracellular metalloprotease. A protease-deficient and lipase-proficient mutant, KFT1007, was complemented by a clone having an open reading frame coding for a 170-amino-acid polypeptide which showed significant homology to Escherichia coli DsbB. KFT1007, a presumed dsbB mutant, also failed to show motility, and both protease secretion and motility were restored by the introduction of the cloned dsbB gene of B. cepacia. The mutant KFT1007 excreted a 43-kDa polypeptide that is immunologically related to the 37-kDa mature protease. These results suggested that the dsbB mutant secretes a premature and catalytically inactive form of protease and that disulfide formation is required for the production of extracellular protease by B. cepacia. PMID:8926116

  2. Immune evasion of Enterococcus faecalis by an extracellular gelatinase that cleaves C3 and iC3b.

    PubMed

    Park, Shin Yong; Shin, Yong Pyo; Kim, Chong Han; Park, Ho Jin; Seong, Yeon Sun; Kim, Byung Sam; Seo, Sook Jae; Lee, In Hee

    2008-11-01

    Enterococcus faecalis (Ef) accounts for most cases of enterococcal bacteremia, which is one of the principal causes of nosocomial bloodstream infections (BSI). Among several virulence factors associated with the pathogenesis of Ef, an extracellular gelatinase (GelE) has been known to be the most common factor, although its virulence mechanisms, especially in association with human BSI, have yet to be demonstrated. In this study, we describe the complement resistance mechanism of Ef mediated by GelE. Using purified GelE, we determined that it cleaved the C3 occurring in human serum into a C3b-like molecule, which was inactivated rapidly via reaction with water. This C3 convertase-like activity of GelE was shown to result in a consumption of C3 and thus inhibited the activation of the complement system. Also, GelE was confirmed to degrade an iC3b that was deposited on the Ag surfaces without affecting the bound C3b. This proteolytic effect of GelE against the major complement opsonin resulted in a substantial reduction in Ef phagocytosis by human polymorphonuclear leukocytes. In addition, we verified that the action of GelE against C3, which is a central component of the complement cascade, was human specific. Taken together, it was suggested that GelE may represent a promising molecule for targeting human BSI associated with Ef. PMID:18941224

  3. DSB:Ce3+ scintillation glass for future

    NASA Astrophysics Data System (ADS)

    Auffray, E.; Akchurin, N.; Benaglia, A.; Borisevich, A.; Cowden, C.; Damgov, J.; Dormenev, V.; Dragoiu, C.; Dudero, P.; Korjik, M.; Kozlov, D.; Kunori, S.; Lecoq, P.; Lee, S. W.; Lucchini, M.; Mechinsky, V.; Pauwels, K.

    2015-02-01

    One of the main challenges for detectors at future high-energy collider experiments is the high precision measurement of hadron and jet energy and momentum. One possibility to achieve this is the dual-readout technique, which allows recording simultaneously scintillation and Cherenkov light in an active medium in order to extract the electromagnetic fraction of the total shower energy on an event- by-event basis. Making use of this approach in the high luminosity LHC, however, puts stringent requirements on the active materials in terms of radiation hardness. Consequently, the R&D carried out on suitable scintillating materials focuses on the detector performance as well as on radiation tolerance. Among the different scintillating materials under study, scintillating glasses can be a suitable solution due to their relatively simple and cost effective production. Recently a new type of inorganic scintillating glass: Cerium doped DSB has been developed by Radiation Instruments and New Components LLC in Minsk for oil logging industry. This material can be produced either in form of bulk or fiber shape with diameter 0.3-2mm and length up to 2000 mm. It is obtained by standard glass production technology at temperature 1400°C with successive thermal annealing treatment at relatively low temperature. The production of large quantities is relatively easy and the production costs are significantly lower compared to crystal fibers. Therefore, this material is considered as an alternative and complementary solution to crystal fibers in view of a production at industrial scale, as required for a large dual readout calorimeter. In this paper, the first results on optical, scintillation properties as well as the radiation damage behaviour obtained on different samples made with different raw materials and various cerium concentrations will be presented.

  4. Peptide inhibitors of the Escherichia coli DsbA oxidative machinery essential for bacterial virulence.

    PubMed

    Duprez, Wilko; Premkumar, Lakshmanane; Halili, Maria A; Lindahl, Fredrik; Reid, Robert C; Fairlie, David P; Martin, Jennifer L

    2015-01-22

    One approach to address antibiotic resistance is to develop drugs that interfere with bacterial virulence. A master regulator of virulence in Gram-negative bacteria is the oxidative folding machinery comprising DsbA and DsbB. A crystal structure at 2.5 Å resolution is reported here for Escherichia coli DsbA complexed with PFATCDS, a heptapeptide derived from the partner protein Escherichia coli DsbB. Details of the peptide binding mode and binding site provide valuable clues for inhibitor design. Structure-activity relationships for 30 analogues were used to produce short peptides with a cysteine that bind tightly to EcDsbA (Kd = 2.0 ± 0.3 μM) and inhibit its activity (IC50 = 5.1 ± 1.1 μM). The most potent inhibitor does not bind to or inhibit human thioredoxin that shares a similar active site. This finding suggests that small molecule inhibitors can be designed to exploit a key interaction of EcDsbA, as the basis for antivirulence agents with a novel mechanism of action. PMID:25470204

  5. Radio-over-fiber DSB-to-SSB conversion using semiconductor lasers at stable locking dynamics.

    PubMed

    Hsieh, Kun-Lin; Hung, Yu-Han; Hwang, Sheng-Kwang; Lin, Chien-Chung

    2016-05-01

    In radio-over-fiber systems, optical single-sideband (SSB) modulation signals are preferred to optical double-sideband (DSB) modulation signals for fiber distribution in order to mitigate the microwave power fading effect. However, typically adopted modulation schemes generate DSB signals, making DSB-to-SSB conversion necessary before or after fiber distribution. This study investigates a semiconductor laser at stable locking dynamics for such conversion. The conversion relies solely on the nonlinear dynamical interaction between an input DSB signal and the laser. Only a typical semiconductor laser is therefore required as the key conversion unit, and no pump or probe signal is necessary. The conversion can be achieved for a broad tunable range of microwave frequency up to at least 60 GHz. In addition, the conversion can be carried out even when the microwave frequency, the power of the input DSB signal, or the frequency of the input DSB signal fluctuates over a wide range, leading to high adaptability and stability of the conversion system. After conversion, while the microwave phase quality, such as linewidth and phase noise, is mainly preserved, a bit-error ratio down to 10-9 is achieved for a data rate up to at least 8 Gb/s with a detection sensitivity improvement of more than 1.5 dB. PMID:27137598

  6. Gradual implementation of the meiotic recombination program via checkpoint pathways controlled by global DSB levels.

    PubMed

    Joshi, Neeraj; Brown, M Scott; Bishop, Douglas K; Börner, G Valentin

    2015-03-01

    During meiosis, Spo11-induced double-strand breaks (DSBs) are processed into crossovers, ensuring segregation of homologous chromosomes (homologs). Meiotic DSB processing entails 5' end resection and preferred strand exchange with the homolog rather than the sister chromatid (homolog bias). In many organisms, DSBs appear gradually along the genome. Here we report unexpected effects of global DSB levels on local recombination events. Early-occurring, low-abundance "scout" DSBs lack homolog bias. Their resection and interhomolog processing are controlled by the conserved checkpoint proteins Tel1(ATM) kinase and Pch2(TRIP13) ATPase. Processing pathways controlled by Mec1(ATR) kinase take over these functions only above a distinct DSB threshold, resulting in progressive strengthening of the homolog bias. We conclude that Tel1(ATM)/Pch2 and Mec1(ATR) DNA damage response pathways are sequentially activated during wild-type meiosis because of their distinct sensitivities to global DSB levels. Moreover, relative DSB order controls the DSB repair pathway choice and, ultimately, recombination outcome. PMID:25661491

  7. Crystallization and preliminary diffraction analysis of a DsbA homologue from Wolbachia pipientis

    SciTech Connect

    Kurz, M.; Iturbe-Ormaetxe, I.; Jarrott, R.; O’Neill, S. L.; Byriel, K. A.; Martin, J. L. Heras, B.

    2008-02-01

    The first crystallization of a W. pipientis protein, α-DsbA1, was achieved using hanging-drop and sitting-drop vapour diffusion. α-DsbA1 is one of two DsbA homologues encoded by the Gram-negative α-proteobacterium Wolbachia pipientis, an endosymbiont that can behave as a reproductive parasite in insects and as a mutualist in medically important filarial nematodes. The α-DsbA1 protein is thought to be important for the folding and secretion of Wolbachia proteins involved in the induction of reproductive distortions. Crystals of native and SeMet α-DsbA1 were grown by vapour diffusion and belong to the monoclinic space group C2, with unit-cell parameters a = 71.4, b = 49.5, c = 69.3 Å, β = 107.0° and one molecule in the asymmetric unit (44% solvent content). X-ray data were recorded from native crystals to a resolution of 2.01 Å using a copper anode and data from SeMet α-DsbA1 crystals were recorded to 2.45 Å resolution using a chromium anode.

  8. Complement Receptor 3 Binds the Borrelia burgdorferi Outer Surface Proteins OspA and OspB in an iC3b-Independent Manner

    PubMed Central

    Garcia, Rodolfo C.; Murgia, Rossella; Cinco, Marina

    2005-01-01

    Persistence of borreliae within the vertebrate host depends on the fate of interactions between the spirochetes and target cells. The present work demonstrates the direct binding of the Borrelia burgdorferi outer surface proteins OspA and OspB to CR3 and that this binding is independent of iC3b. PMID:16113335

  9. Engineering of Helicobacter pylori Dimeric Oxidoreductase DsbK (HP0231)

    PubMed Central

    Bocian-Ostrzycka, Katarzyna M.; Grzeszczuk, Magdalena J.; Banaś, Anna M.; Jastrząb, Katarzyna; Pisarczyk, Karolina; Kolarzyk, Anna; Łasica, Anna M.; Collet, Jean-François; Jagusztyn-Krynicka, Elżbieta K.

    2016-01-01

    The formation of disulfide bonds that are catalyzed by proteins of the Dsb (disulfide bond) family is crucial for the correct folding of many extracytoplasmic proteins. Thus, this formation plays an essential, pivotal role in the assembly of many virulence factors. The Helicobacter pylori disulfide bond-forming system is uncomplicated compared to the best-characterized Escherichia coli Dsb pathways. It possesses only two extracytoplasmic Dsb proteins named HP0377 and HP0231. As previously shown, HP0377 is a reductase involved in the process of cytochrome c maturation. Additionally, it also possesses disulfide isomerase activity. HP0231 was the first periplasmic dimeric oxidoreductase involved in disulfide generation to be described. Although HP0231 function is critical for oxidative protein folding, its structure resembles that of dimeric EcDsbG, which does not confer this activity. However, the HP0231 catalytic motifs (CXXC and the so-called cis-Pro loop) are identical to that of monomeric EcDsbA. To understand the functioning of HP0231, we decided to study the relations between its sequence, structure and activity through an extensive analysis of various HP0231 point mutants, using in vivo and in vitro strategies. Our work shows the crucial role of the cis-Pro loop, as changing valine to threonine in this motif completely abolishes the protein function in vivo. Functioning of HP0231 is conditioned by the combination of CXXC and the cis-Pro loop, as replacing the HP0231 CXXC motif by the motif from EcDsbG or EcDsbC results in bifunctional protein, at least in E. coli. We also showed that the dimerization domain of HP0231 ensures contact with its substrates. Moreover, the activity of this oxidase is independent on the structure of the catalytic domain. Finally, we showed that HP0231 chaperone activity is independent of its redox function. PMID:27507968

  10. DNA DSB measurements and modelling approaches based on gamma-H2AX foci time evolution

    NASA Astrophysics Data System (ADS)

    Esposito, Giuseppe; Campa, Alessandro; Antonelli, Francesca; Mariotti, Luca; Belli, Mauro; Giardullo, Paola; Simone, Giustina; Antonella Tabocchini, Maria; Ottolenghi, Andrea

    DNA double strand breaks (DSBs) induced by ionising radiation are considered the main dam-age related to the deleterious consequences in the cells. Unrepaired or mis-repaired DSBs can cause mutations or loss of chromosome regions which can eventually lead to cell death or neo-plastic transformation. Quantification of the number and complexity of DSBs induced by low doses of radiation remains a complex problem. About ten years ago Rogakou et al. proposed an immunofluorescent technique able to detect even a single DSB per cell. This approach is based on the serine 139 phosphorylation of many molecules (up to 2000) of histone H2AX (γg-H2AX) following the induction of a DSB in the DNA. DSB can be visualized as foci by immunofluores-cence by using phospho-specific antibodies, so that enumeration of foci can be used to measure DSB induction and processing. It is still not completely clear how γ-H2AX dephosphorylation takes place; however it has been related with DSB repair, in particular with the efficiency of DSB repair. In this work we analyse the H2AX phosphorylation-dephosphorylation kinetics after irradiation of primary human fibroblasts (AG1522 cell line) with radiation of differing quality, that is γ-rays and α-particles (125 keV/µm), with the aim of comparing the time evolution of γ-H2AX foci. Our results show that, after a dose of 0.5 Gy, both γ-rays and α-particles induce the maximum number of γ-H2AX foci within 30 minutes from irradiation, that this number depends on the radiation type and is consistent with the number of track traversal in α-irradiated nuclei, that the dephosphorylation kinetics are very different, being the α-induced foci rate of disappearence slower than that of γ-induced foci. In this work a modellistic approach to estimate the number of DSB induced by γ-rays detectable by using the γ-H2AX assay is presented. The competing processes of appearance and disappearance of visible foci will be modeled taking into account the

  11. De novo design and evolution of artificial disulfide isomerase enzymes analogous to the bacterial DsbC.

    PubMed

    Arredondo, Silvia; Segatori, Laura; Gilbert, Hiram F; Georgiou, George

    2008-11-14

    The Escherichia coli disulfide isomerase, DsbC is a V-shaped homodimer with each monomer comprising a dimerization region that forms part of a putative peptide-binding pocket and a thioredoxin catalytic domain. Disulfide isomerases from prokaryotes and eukaryotes exhibit little sequence homology but display very similar structural organization with two thioredoxin domains facing each other on top of the dimerization/peptide-binding region. To aid the understanding of the mechanistic significance of thioredoxin domain dimerization and of the peptide-binding cleft of DsbC, we constructed a series of protein chimeras comprising unrelated protein dimerization domains fused to thioredoxin superfamily enzymes. Chimeras consisting of the dimerization domain and the alpha-helical linker of the bacterial proline cis/trans isomerase FkpA and the periplasmic oxidase DsbA gave rise to enzymes that catalyzed the folding of multidisulfide substrate proteins in vivo with comparable efficiency to E. coli DsbC. In addition, expression of FkpA-DsbAs conferred modest resistance to CuCl2, a phenotype that depends on disulfide bond isomerization. Selection for resistance to elevated CuCl2 concentrations led to the isolation of FkpA-DsbA mutants containing a single amino acid substitution that changed the active site of the DsbA domain from CPHC into CPYC, increasing the similarity to the DsbC active site (CGYC). Unlike DsbC, which is resistant to oxidation by DsbB-DsbA and does not normally catalyze disulfide bond formation under physiological conditions, the FkpA-DsbA chimeras functioned both as oxidases and isomerases. The engineering of these efficient artificial isomerases delineates the key features of catalysis of disulfide bond isomerization and enhances our understanding of its evolution. PMID:18782764

  12. De Novo Design and Evolution of Artificial Disulfide Isomerase Enzymes Analogous to the Bacterial DsbC*S⃞

    PubMed Central

    Arredondo, Silvia; Segatori, Laura; Gilbert, Hiram F.; Georgiou, George

    2008-01-01

    The Escherichia coli disulfide isomerase, DsbC is a V-shaped homodimer with each monomer comprising a dimerization region that forms part of a putative peptide-binding pocket and a thioredoxin catalytic domain. Disulfide isomerases from prokaryotes and eukaryotes exhibit little sequence homology but display very similar structural organization with two thioredoxin domains facing each other on top of the dimerization/peptide-binding region. To aid the understanding of the mechanistic significance of thioredoxin domain dimerization and of the peptide-binding cleft of DsbC, we constructed a series of protein chimeras comprising unrelated protein dimerization domains fused to thioredoxin superfamily enzymes. Chimeras consisting of the dimerization domain and the α-helical linker of the bacterial proline cis/trans isomerase FkpA and the periplasmic oxidase DsbA gave rise to enzymes that catalyzed the folding of multidisulfide substrate proteins in vivo with comparable efficiency to E. coli DsbC. In addition, expression of FkpA-DsbAs conferred modest resistance to CuCl2, a phenotype that depends on disulfide bond isomerization. Selection for resistance to elevated CuCl2 concentrations led to the isolation of FkpA-DsbA mutants containing a single amino acid substitution that changed the active site of the DsbA domain from CPHC into CPYC, increasing the similarity to the DsbC active site (CGYC). Unlike DsbC, which is resistant to oxidation by DsbB-DsbA and does not normally catalyze disulfide bond formation under physiological conditions, the FkpA-DsbA chimeras functioned both as oxidases and isomerases. The engineering of these efficient artificial isomerases delineates the key features of catalysis of disulfide bond isomerization and enhances our understanding of its evolution. PMID:18782764

  13. Application of fragment-based screening to the design of inhibitors of Escherichia coli DsbA.

    PubMed

    Adams, Luke A; Sharma, Pooja; Mohanty, Biswaranjan; Ilyichova, Olga V; Mulcair, Mark D; Williams, Martin L; Gleeson, Ellen C; Totsika, Makrina; Doak, Bradley C; Caria, Sofia; Rimmer, Kieran; Horne, James; Shouldice, Stephen R; Vazirani, Mansha; Headey, Stephen J; Plumb, Brent R; Martin, Jennifer L; Heras, Begoña; Simpson, Jamie S; Scanlon, Martin J

    2015-02-01

    The thiol-disulfide oxidoreductase enzyme DsbA catalyzes the formation of disulfide bonds in the periplasm of Gram-negative bacteria. DsbA substrates include proteins involved in bacterial virulence. In the absence of DsbA, many of these proteins do not fold correctly, which renders the bacteria avirulent. Thus DsbA is a critical mediator of virulence and inhibitors may act as antivirulence agents. Biophysical screening has been employed to identify fragments that bind to DsbA from Escherichia coli. Elaboration of one of these fragments produced compounds that inhibit DsbA activity in vitro. In cell-based assays, the compounds inhibit bacterial motility, but have no effect on growth in liquid culture, which is consistent with selective inhibition of DsbA. Crystal structures of inhibitors bound to DsbA indicate that they bind adjacent to the active site. Together, the data suggest that DsbA may be amenable to the development of novel antibacterial compounds that act by inhibiting bacterial virulence. PMID:25556635

  14. Heavy ion induced DNA-DSB in yeast and mammalian cells

    NASA Technical Reports Server (NTRS)

    Loebrich, M.; Ikpeme, S.; Kiefer, J.

    1994-01-01

    Molecular changes at the DNA are assumed to be the main cause for radiation effects in a number of organisms. During the course of the last decades techniques have been developed for measuring DNA double-strand breaks (dsb), generally assumed to be the most critical DNA lesions. The outcome of all those different approaches portrays a collection of data useful for a theoretical description of radiation action mechanisms. However, in the case of heavy ion induced DNA dsb the picture is not quite clear yet and further projects and strategies have to be developed. The biological systems studied in our group are yeast and mammalian cells. While in the case of yeast cells technical and methodical reasons highlight these organisms mammalian cells reach greater importance when dsb repair studies are performed. In both types of organisms the technique of pulsed-field gel electrophoresis (PFGE) is applied, although with different modifications and evaluation procedures mainly due to the different genome sizes.

  15. Rv2969c, essential for optimal growth in Mycobacterium tuberculosis, is a DsbA-like enzyme that interacts with VKOR-derived peptides and has atypical features of DsbA-like disulfide oxidases

    SciTech Connect

    Premkumar, Lakshmanane Heras, Begoña; Duprez, Wilko; Walden, Patricia; Halili, Maria; Kurth, Fabian; Fairlie, David P.; Martin, Jennifer L.

    2013-10-01

    The gene product of M. tuberculosis Rv2969c is shown to be a disulfide oxidase enzyme that has a canonical DsbA-like fold with novel structural and functional characteristics. The bacterial disulfide machinery is an attractive molecular target for developing new antibacterials because it is required for the production of multiple virulence factors. The archetypal disulfide oxidase proteins in Escherichia coli (Ec) are DsbA and DsbB, which together form a functional unit: DsbA introduces disulfides into folding proteins and DsbB reoxidizes DsbA to maintain it in the active form. In Mycobacterium tuberculosis (Mtb), no DsbB homologue is encoded but a functionally similar but structurally divergent protein, MtbVKOR, has been identified. Here, the Mtb protein Rv2969c is investigated and it is shown that it is the DsbA-like partner protein of MtbVKOR. It is found that it has the characteristic redox features of a DsbA-like protein: a highly acidic catalytic cysteine, a highly oxidizing potential and a destabilizing active-site disulfide bond. Rv2969c also has peptide-oxidizing activity and recognizes peptide segments derived from the periplasmic loops of MtbVKOR. Unlike the archetypal EcDsbA enzyme, Rv2969c has little or no activity in disulfide-reducing and disulfide-isomerase assays. The crystal structure of Rv2969c reveals a canonical DsbA fold comprising a thioredoxin domain with an embedded helical domain. However, Rv2969c diverges considerably from other DsbAs, including having an additional C-terminal helix (H8) that may restrain the mobility of the catalytic helix H1. The enzyme is also characterized by a very shallow hydrophobic binding surface and a negative electrostatic surface potential surrounding the catalytic cysteine. The structure of Rv2969c was also used to model the structure of a paralogous DsbA-like domain of the Ser/Thr protein kinase PknE. Together, these results show that Rv2969c is a DsbA-like protein with unique properties and a limited

  16. Structural and Biochemical Characterization of Xylella fastidiosa DsbA Family Members: New insightsinto the Enzyme-Substrate Interaction

    SciTech Connect

    Rinaldi, F.; Meza, A; Gulmarges, B

    2009-01-01

    Disulfide oxidoreductase DsbA catalyzes disulfide bond formation in proteins secreted to the periplasm and has been related to the folding process of virulence factors in many organisms. It is among the most oxidizing of the thioredoxin-like proteins, and DsbA redox power is understood in terms of the electrostatic interactions involving the active site motif CPHC. The plant pathogen Xylella fastidiosa has two chromosomal genes encoding two oxidoreductases belonging to the DsbA family, and in one of them, the canonical motif CPHC is replaced by CPAC. Biochemical assays showed that both X. fastidiosa homologues have similar redox properties and the determination of the crystal structure of XfDsbA revealed substitutions in the active site of X. fastidiosa enzymes, which are proposed to compensate for the lack of the conserved histidine in XfDsbA2. In addition, electron density maps showed a ligand bound to the XfDsbA active site, allowing the characterization of the enzyme interaction with an 8-mer peptide. Finally, surface analysis of XfDsbA and XfDsbA2 suggests that X. fastidiosa enzymes may have different substrate specificities.

  17. Targeting Bacterial Dsb Proteins for the Development of Anti-Virulence Agents.

    PubMed

    Smith, Roxanne P; Paxman, Jason J; Scanlon, Martin J; Heras, Begoña

    2016-01-01

    Recent years have witnessed a dramatic increase in bacterial antimicrobial resistance and a decline in the development of novel antibiotics. New therapeutic strategies are urgently needed to combat the growing threat posed by multidrug resistant bacterial infections. The Dsb disulfide bond forming pathways are potential targets for the development of antimicrobial agents because they play a central role in bacterial pathogenesis. In particular, the DsbA/DsbB system catalyses disulfide bond formation in a wide array of virulence factors, which are essential for many pathogens to establish infections and cause disease. These redox enzymes are well placed as antimicrobial targets because they are taxonomically widespread, share low sequence identity with human proteins, and many years of basic research have provided a deep molecular understanding of these systems in bacteria. In this review, we discuss disulfide bond catalytic pathways in bacteria and their significance in pathogenesis. We also review the use of different approaches to develop inhibitors against Dsb proteins as potential anti-virulence agents, including fragment-based drug discovery, high-throughput screening and other structure-based drug discovery methods. PMID:27438817

  18. Glycosylation of DsbA in Francisella tularensis subsp. tularensis▿†

    PubMed Central

    Thomas, Rebecca M.; Twine, Susan M.; Fulton, Kelly M.; Tessier, Luc; Kilmury, Sara L. N.; Ding, Wen; Harmer, Nicholas; Michell, Stephen L.; Oyston, Petra C. F.; Titball, Richard W.; Prior, Joann L.

    2011-01-01

    In Francisella tularensis subsp. tularensis, DsbA has been shown to be an essential virulence factor and has been observed to migrate to multiple protein spots on two-dimensional electrophoresis gels. In this work, we show that the protein is modified with a 1,156-Da glycan moiety in O-linkage. The results of mass spectrometry studies suggest that the glycan is a hexasaccharide, comprised of N-acetylhexosamines, hexoses, and an unknown monosaccharide. Disruption of two genes within the FTT0789-FTT0800 putative polysaccharide locus, including a galE homologue (FTT0791) and a putative glycosyltransferase (FTT0798), resulted in loss of glycan modification of DsbA. The F. tularensis subsp. tularensis ΔFTT0798 and ΔFTT0791::Cm mutants remained virulent in the murine model of subcutaneous tularemia. This indicates that glycosylation of DsbA does not play a major role in virulence under these conditions. This is the first report of the detailed characterization of the DsbA glycan and putative role of the FTT0789-FTT0800 gene cluster in glycan biosynthesis. PMID:21803994

  19. Structure of reduced DsbA from Escherichia coli in solution.

    PubMed

    Schirra, H J; Renner, C; Czisch, M; Huber-Wunderlich, M; Holak, T A; Glockshuber, R

    1998-05-01

    The three-dimensional structure of reduced DsbA from Escherichia coli in aqueous solution has been determined by nuclear magnetic resonance (NMR) spectroscopy and is compared with the crystal structure of oxidized DsbA [Guddat, L. W., Bardwell, J. C. A., Zander, T., and Martin, J. L. (1997) Protein Sci. 6, 1148-1156]. DsbA is a monomeric 21 kDa protein which consists of 189 residues and is required for disulfide bond formation in the periplasm of E. coli. On the basis of sequence-specific 1H NMR assignments, 1664 nuclear Overhauser enhancement distance constraints, 118 hydrogen bond distance constraints, and 293 dihedral angle constraints were obtained as the input for the structure calculations by simulated annealing with the program X-PLOR. The enzyme is made up of two domains. The catalytic domain has a thioredoxin-like fold with a five-stranded beta-sheet and three alpha-helices, and the second domain consists of four alpha-helices and is inserted into the thioredoxin motif. The active site between Cys30 and Cys33 is located at the N terminus of the first alpha-helix in the thioredoxin-like domain. The solution structure of reduced DsbA is rather similar to the crystal structure of the oxidized enzyme but exhibits a different relative orientation of both domains. In addition, the conformations of the active site and a loop between strand beta5 and helix alpha7 are slightly different. These structural differences may reflect important functional requirements in the reaction cycle of DsbA as they appear to facilitate the release of oxidized polypeptides from reduced DsbA. The extremely low pKa value of the nucleophilic active site thiol of Cys30 in reduced DsbA is most likely caused by its interactions with the dipole of the active site helix and the side chain of His32, as no other charged residues are located next to the sulfur atom of Cys30 in the solution structure. PMID:9572841

  20. Rv2969c, essential for optimal growth in Mycobacterium tuberculosis, is a DsbA-like enzyme that interacts with VKOR-derived peptides and has atypical features of DsbA-like disulfide oxidases

    PubMed Central

    Premkumar, Lakshmanane; Heras, Begoña; Duprez, Wilko; Walden, Patricia; Halili, Maria; Kurth, Fabian; Fairlie, David P.; Martin, Jennifer L.

    2013-01-01

    The bacterial disulfide machinery is an attractive molecular target for developing new antibacterials because it is required for the production of multiple virulence factors. The archetypal disulfide oxidase proteins in Escherichia coli (Ec) are DsbA and DsbB, which together form a functional unit: DsbA introduces disulfides into folding proteins and DsbB re­oxidizes DsbA to maintain it in the active form. In Mycobacterium tuberculosis (Mtb), no DsbB homologue is encoded but a functionally similar but structurally divergent protein, MtbVKOR, has been identified. Here, the Mtb protein Rv2969c is investigated and it is shown that it is the DsbA-like partner protein of MtbVKOR. It is found that it has the characteristic redox features of a DsbA-like protein: a highly acidic catalytic cysteine, a highly oxidizing potential and a destabilizing active-site disulfide bond. Rv2969c also has peptide-oxidizing activity and recognizes peptide segments derived from the periplasmic loops of MtbVKOR. Unlike the archetypal EcDsbA enzyme, Rv2969c has little or no activity in disulfide-reducing and disulfide-isomerase assays. The crystal structure of Rv2969c reveals a canonical DsbA fold comprising a thioredoxin domain with an embedded helical domain. However, Rv2969c diverges considerably from other DsbAs, including having an additional C-terminal helix (H8) that may restrain the mobility of the catalytic helix H1. The enzyme is also characterized by a very shallow hydrophobic binding surface and a negative electrostatic surface potential surrounding the catalytic cysteine. The structure of Rv2969c was also used to model the structure of a paralogous DsbA-like domain of the Ser/Thr protein kinase PknE. Together, these results show that Rv2969c is a DsbA-like protein with unique properties and a limited substrate-binding specificity. PMID:24100317

  1. Rv2969c, essential for optimal growth in Mycobacterium tuberculosis, is a DsbA-like enzyme that interacts with VKOR-derived peptides and has atypical features of DsbA-like disulfide oxidases.

    PubMed

    Premkumar, Lakshmanane; Heras, Begoña; Duprez, Wilko; Walden, Patricia; Halili, Maria; Kurth, Fabian; Fairlie, David P; Martin, Jennifer L

    2013-10-01

    The bacterial disulfide machinery is an attractive molecular target for developing new antibacterials because it is required for the production of multiple virulence factors. The archetypal disulfide oxidase proteins in Escherichia coli (Ec) are DsbA and DsbB, which together form a functional unit: DsbA introduces disulfides into folding proteins and DsbB reoxidizes DsbA to maintain it in the active form. In Mycobacterium tuberculosis (Mtb), no DsbB homologue is encoded but a functionally similar but structurally divergent protein, MtbVKOR, has been identified. Here, the Mtb protein Rv2969c is investigated and it is shown that it is the DsbA-like partner protein of MtbVKOR. It is found that it has the characteristic redox features of a DsbA-like protein: a highly acidic catalytic cysteine, a highly oxidizing potential and a destabilizing active-site disulfide bond. Rv2969c also has peptide-oxidizing activity and recognizes peptide segments derived from the periplasmic loops of MtbVKOR. Unlike the archetypal EcDsbA enzyme, Rv2969c has little or no activity in disulfide-reducing and disulfide-isomerase assays. The crystal structure of Rv2969c reveals a canonical DsbA fold comprising a thioredoxin domain with an embedded helical domain. However, Rv2969c diverges considerably from other DsbAs, including having an additional C-terminal helix (H8) that may restrain the mobility of the catalytic helix H1. The enzyme is also characterized by a very shallow hydrophobic binding surface and a negative electrostatic surface potential surrounding the catalytic cysteine. The structure of Rv2969c was also used to model the structure of a paralogous DsbA-like domain of the Ser/Thr protein kinase PknE. Together, these results show that Rv2969c is a DsbA-like protein with unique properties and a limited substrate-binding specificity. PMID:24100317

  2. Overexpression of the rhodanese PspE, a single cysteine-containing protein, restores disulfide bond formation to an Escherichia coli strain lacking DsbA

    PubMed Central

    Chng, Shu-Sin; Dutton, Rachel J.; Denoncin, Katleen; Vertommen, Didier; Collet, Jean-Francois; Kadokura, Hiroshi; Beckwith, Jonathan

    2012-01-01

    Summary Escherichia coli uses the DsbA/DsbB system for introducing disulfide bonds into proteins in the cell envelope. Deleting either dsbA or dsbB or both reduces disulfide bond formation but does not entirely eliminate it. Whether such background disulfide bond forming activity is enzyme-catalyzed is not known. To identify possible cellular factors that might contribute to the background activity, we studied the effects of overexpressing endogenous proteins on disulfide bond formation in the periplasm. We find that overexpressing PspE, a periplasmic rhodanese, partially restores substantial disulfide bond formation to a dsbA strain. This activity depends on DsbC, the bacterial disulfide bond isomerase, but not on DsbB. We show that overexpressed PspE is oxidized to the sulfenic acid form and reacts with substrate proteins to form mixed disulfide adducts. DsbC either prevents the formation of these mixed disulfides or resolves these adducts subsequently. In the process, DsbC itself gets oxidized and proceeds to catalyze disulfide bond formation. Although this PspE/DsbC system is not responsible for the background disulfide bond forming activity, we suggest that it might be utilized in other organisms lacking the DsbA/DsbB system. PMID:22809289

  3. Phosphorylation of Ku dictates DNA double-strand break (DSB) repair pathway choice in S phase

    PubMed Central

    Lee, Kyung-Jong; Saha, Janapriya; Sun, Jingxin; Fattah, Kazi R.; Wang, Shu-Chi; Jakob, Burkhard; Chi, Linfeng; Wang, Shih-Ya; Taucher-Scholz, Gisela; Davis, Anthony J.; Chen, David J.

    2016-01-01

    Multiple DNA double-strand break (DSB) repair pathways are active in S phase of the cell cycle; however, DSBs are primarily repaired by homologous recombination (HR) in this cell cycle phase. As the non-homologous end-joining (NHEJ) factor, Ku70/80 (Ku), is quickly recruited to DSBs in S phase, we hypothesized that an orchestrated mechanism modulates pathway choice between HR and NHEJ via displacement of the Ku heterodimer from DSBs to allow HR. Here, we provide evidence that phosphorylation at a cluster of sites in the junction of the pillar and bridge regions of Ku70 mediates the dissociation of Ku from DSBs. Mimicking phosphorylation at these sites reduces Ku's affinity for DSB ends, suggesting that phosphorylation of Ku70 induces a conformational change responsible for the dissociation of the Ku heterodimer from DNA ends. Ablating phosphorylation of Ku70 leads to the sustained retention of Ku at DSBs, resulting in a significant decrease in DNA end resection and HR, specifically in S phase. This decrease in HR is specific as these phosphorylation sites are not required for NHEJ. Our results demonstrate that the phosphorylation-mediated dissociation of Ku70/80 from DSBs frees DNA ends, allowing the initiation of HR in S phase and providing a mechanism of DSB repair pathway choice in mammalian cells. PMID:26712563

  4. Crystal Structure of DsbA from Corynebacterium diphtheriae and Its Functional Implications for CueP in Gram-Positive Bacteria

    PubMed Central

    Um, Si-Hyeon; Kim, Jin-Sik; Song, Saemee; Kim, Nam Ah; Jeong, Seong Hoon; Ha, Nam-Chul

    2015-01-01

    In Gram-negative bacteria in the periplasmic space, the dimeric thioredoxin-fold protein DsbC isomerizes and reduces incorrect disulfide bonds of unfolded proteins, while the monomeric thioredoxin-fold protein DsbA introduces disulfide bonds in folding proteins. In the Gram-negative bacteria Salmonella enterica serovar Typhimurium, the reduced form of CueP scavenges the production of hydroxyl radicals in the copper-mediated Fenton reaction, and DsbC is responsible for keeping CueP in the reduced, active form. Some DsbA proteins fulfill the functions of DsbCs, which are not present in Gram-positive bacteria. In this study, we identified a DsbA homologous protein (CdDsbA) in the Corynebacterium diphtheriae genome and determined its crystal structure in the reduced condition at 1.5 Å resolution. CdDsbA consists of a monomeric thioredoxin-like fold with an inserted helical domain and unique N-terminal extended region. We confirmed that CdDsbA has disulfide bond isomerase/reductase activity, and we present evidence that the N-terminal extended region is not required for this activity and folding of the core DsbA-like domain. Furthermore, we found that CdDsbA could reduce CueP from C. diphtheriae. PMID:26082031

  5. Crystal structure of the dithiol oxidase DsbA enzyme from proteus mirabilis bound non-covalently to an active site peptide ligand.

    PubMed

    Kurth, Fabian; Duprez, Wilko; Premkumar, Lakshmanane; Schembri, Mark A; Fairlie, David P; Martin, Jennifer L

    2014-07-11

    The disulfide bond forming DsbA enzymes and their DsbB interaction partners are attractive targets for development of antivirulence drugs because both are essential for virulence factor assembly in Gram-negative pathogens. Here we characterize PmDsbA from Proteus mirabilis, a bacterial pathogen increasingly associated with multidrug resistance. PmDsbA exhibits the characteristic properties of a DsbA, including an oxidizing potential, destabilizing disulfide, acidic active site cysteine, and dithiol oxidase catalytic activity. We evaluated a peptide, PWATCDS, derived from the partner protein DsbB and showed by thermal shift and isothermal titration calorimetry that it binds to PmDsbA. The crystal structures of PmDsbA, and the active site variant PmDsbAC30S were determined to high resolution. Analysis of these structures allows categorization of PmDsbA into the DsbA class exemplified by the archetypal Escherichia coli DsbA enzyme. We also present a crystal structure of PmDsbAC30S in complex with the peptide PWATCDS. The structure shows that the peptide binds non-covalently to the active site CXXC motif, the cis-Pro loop, and the hydrophobic groove adjacent to the active site of the enzyme. This high-resolution structural data provides a critical advance for future structure-based design of non-covalent peptidomimetic inhibitors. Such inhibitors would represent an entirely new antibacterial class that work by switching off the DSB virulence assembly machinery. PMID:24831013

  6. Crystal Structure of the Dithiol Oxidase DsbA Enzyme from Proteus Mirabilis Bound Non-covalently to an Active Site Peptide Ligand

    PubMed Central

    Kurth, Fabian; Duprez, Wilko; Premkumar, Lakshmanane; Schembri, Mark A.; Fairlie, David P.; Martin, Jennifer L.

    2014-01-01

    The disulfide bond forming DsbA enzymes and their DsbB interaction partners are attractive targets for development of antivirulence drugs because both are essential for virulence factor assembly in Gram-negative pathogens. Here we characterize PmDsbA from Proteus mirabilis, a bacterial pathogen increasingly associated with multidrug resistance. PmDsbA exhibits the characteristic properties of a DsbA, including an oxidizing potential, destabilizing disulfide, acidic active site cysteine, and dithiol oxidase catalytic activity. We evaluated a peptide, PWATCDS, derived from the partner protein DsbB and showed by thermal shift and isothermal titration calorimetry that it binds to PmDsbA. The crystal structures of PmDsbA, and the active site variant PmDsbAC30S were determined to high resolution. Analysis of these structures allows categorization of PmDsbA into the DsbA class exemplified by the archetypal Escherichia coli DsbA enzyme. We also present a crystal structure of PmDsbAC30S in complex with the peptide PWATCDS. The structure shows that the peptide binds non-covalently to the active site CXXC motif, the cis-Pro loop, and the hydrophobic groove adjacent to the active site of the enzyme. This high-resolution structural data provides a critical advance for future structure-based design of non-covalent peptidomimetic inhibitors. Such inhibitors would represent an entirely new antibacterial class that work by switching off the DSB virulence assembly machinery. PMID:24831013

  7. Kinetic analysis of the interactions of complement receptor 2 (CR2, CD21) with its ligands C3d, iC3b, and the EBV glycoprotein gp350/220.

    PubMed

    Sarrias, M R; Franchini, S; Canziani, G; Argyropoulos, E; Moore, W T; Sahu, A; Lambris, J D

    2001-08-01

    The molecular mechanisms involved in the interaction of complement receptor 2 (CR2) with its natural ligands iC3b and C3d are still not well understood. In addition, studies regarding the binding site(s) of the receptor on C3 as well as the affinities of the C3 fragments for CR2 have produced contradictory results. In the present study, we have used surface plasmon resonance technology to study the interaction of CR2 with its ligands C3d, iC3b, and the EBV surface glycoprotein gp350/220. We measured the kinetics of binding of the receptor to its ligands, examined the influence of ionic contacts on these interactions, and assessed whether immobilized and soluble iC3b bound with similar kinetics to CR2. Our results indicate that 1) gp350 binding to CR2 follows a simple 1:1 interaction, whereas that of the C3 fragments is more complex and involves more than one intramolecular component; 2) kinetic differences exist between the binding of C3d and iC3b to CR2, which may be due to an additional binding site found on the C3c region of iC3b; and 3) iC3b binds to CR2 with different kinetics, depending on whether the iC3b is in solution or immobilized on the surface. These findings suggest that binding of CR2 to iC3b and C3d is more complex than previously thought. PMID:11466369

  8. Intravenous and standard immune serum globulin preparations interfere with uptake of /sup 125/I-C3 onto sensitized erythrocytes and inhibit hemolytic complement activity

    SciTech Connect

    Berger, M.; Rosenkranz, P.; Brown, C.Y.

    1985-02-01

    Antibody-sensitized sheep erythrocytes were used as a model to determine the effects of therapeutic immune serum globulin (ISG) preparations on the ability of this particulate activator to fix C3 and initiate hemolysis. Both standard and intravenous forms of ISG inhibit uptake of /sup 125/I-C3, presumably by competing for the deposition of ''nascent'' C3b molecules onto the erythrocytes. Both forms of ISG also inhibit hemolytic activity of whole serum or purified complement components. The inhibition appears to be a specific property of IgG itself, since similar inhibition was not caused by equivalent concentrations of human serum albumin, and was not affected by the buffer in which the ISG was dissolved. Interference with C3 uptake onto antibody-sensitized platelets and/or inhibition of hemolytic complement activity could contribute to the efficacy of high dose intravenous ISG in idiopathic thrombocytopenic purpura.

  9. Lymphoblastoid cell supernatants increase expression of C3b receptors on human polymorphonuclear leucocytes: direct binding studies with 125I-C3b.

    PubMed Central

    Berger, M; Cross, A S

    1984-01-01

    Human PMN incubated in culture supernatants of the Raji long-term human lymphoblastoid cell line showed increased rosette formation with sheep erythrocytes coated with C3b (EIgM C4b3b) but no change in rosette formation with IgG-coated erythrocytes. This suggested a specific increase in cell surface C3b receptors, which was further investigated using 125I-C3b for direct binding studies. The results confirmed that specific binding of 125I-C3b to PMN incubated in culture supernatants increased up to three- to four-fold over binding to PMN incubated in control media alone. Scatchard analysis revealed that the apparent Ka for supernatant-treated cells, 3.36 +/- 0.89 X 10(7) L/M did not differ from the Ka for cells incubated in control media, 3.76 +/- 0.75 X 10(7) L/M, suggesting an increase in a single class of C3b receptors. Kinetic studies revealed that the active factor was present within 24 hr of culture of the Raji cells, and that neutrophils incubated in culture supernatants increased their C3b receptors continuously for up to 4 hr, the longest interval tested. The effect of the culture supernatant was lost with dilution beyond eight- to 10-fold. The results suggest that culture supernatants of this long-term lymphoblastoid cell line contain soluble factors that induce increased expression of C3b receptors on PMN and may thus serve as a model for study of important physiologic effects of lymphocyte products on PMN in vivo. PMID:6230308

  10. Modulation of Biofilm-Formation in Salmonella enterica Serovar Typhimurium by the Periplasmic DsbA/DsbB Oxidoreductase System Requires the GGDEF-EAL Domain Protein STM3615

    PubMed Central

    Römling, Ute; Rhen, Mikael

    2014-01-01

    In Salmonella enterica serovar Typhimurium (S. Typhimurium), biofilm-formation is controlled by the cytoplasmic intracellular small-molecular second messenger cyclic 3′, 5′-di- guanosine monophosphate (c-di-GMP) through the activities of GGDEF and EAL domain proteins. Here we describe that deleting either dsbA or dsbB, respectively encoding a periplasmic protein disulfide oxidase and a cytoplasmic membrane disulfide oxidoreductase, resulted in increased biofilm-formation on solid medium. This increased biofilm-formation, defined as a red, dry and rough (rdar) colony morphotype, paralleled with enhanced expression of the biofilm master regulator CsgD and the biofilm-associated fimbrial subunit CsgA. Deleting csgD in either dsb mutant abrogated the enhanced biofilm-formation. Likewise, overexpression of the c-di-GMP phosphodiesterase YhjH, or mutationally inactivating the CsgD activator EAL-domain protein YdiV, reduced biofilm-formation in either of the dsb mutants. Intriguingly, deleting the GGDEF-EAL domain protein gene STM3615 (yhjK), previously not connected to rdar morphotype development, also abrogated the escalated rdar morphotype formation in dsb mutant backgrounds. Enhanced biofilm-formation in dsb mutants was furthermore annulled by exposure to the protein disulfide catalyst copper chloride. When analyzed for the effect of exogenous reducing stress on biofilm-formation, both dsb mutants initially showed an escalated rdar morphotype development that later dissolved to reveal a smooth mucoid colony morphotype. From these results we conclude that biofilm-development in S. Typhimurium is affected by periplasmic protein disulphide bond status through CsgD, and discuss the involvement of selected GGDEF/EAL domain protein(s) as signaling mediators. PMID:25153529

  11. Protein folding in the periplasm in the absence of primary oxidant DsbA: modulation of redox potential in periplasmic space via OmpL porin

    PubMed Central

    Dartigalongue, Claire; Nikaido, Hiroshi; Raina, Satish

    2000-01-01

    Disulfide bond formation in Escherichia coli is a catalyzed reaction accomplished by DsbA. We found that null mutations in a new porin gene, ompL, allowed a total bypass of the DsbA requirement for protein oxidation. These mutations acted as extragenic null suppressors for dsbA, and restored normal folding of alkaline phosphatase and relieved sensitivity to dithiothreitol. ompL dsbA double mutants were completely like wild-type mutants in terms of motility and lack of mucoidy. This suppression was not dependent on DsbC and DsbG, since the oxidation status of these proteins was unaltered in ompL dsbA strains. Purified OmpL allowed diffusion of small solutes, including sugars, but the suppression was not dependent on the carbon sources used. Suppression by ompL null mutations required DsbB, leading us to propose a hypothesis that DsbB oxidizes yet unidentified, low-molecular-weight redox agents in the periplasm. These oxidized agents accumulate and substitute for DsbA if their leakage into the medium is prevented by the absence of OmpL, presumed to form a specific channel for their diffusion. PMID:11080145

  12. Biochemical Kinetics Model of DSB Repair and GammaH2AX FOCI by Non-homologous End Joining

    NASA Technical Reports Server (NTRS)

    Cucinotta, Francis, A.; Pluth, Janice M.; Anderson, Jennifer A.; Harper, Jane V.; O'Neill, Peter

    2007-01-01

    We developed a biochemical kinetics approach to describe the repair of double strand breaks (DSB) produced by low LET radiation by modeling molecular events associated with the mechanisms of non-homologous end-joining (NHEJ). A system of coupled non-linear ordinary differential equations describes the induction of DSB and activation pathways for major NHEJ components including Ku(sub 70/80), DNA-PK(sub cs), and the Ligase IV-XRCC4 hetero-dimer. The autophosphorylation of DNA-PK(sub cs and subsequent induction of gamma-H2AX foci observed after ionizing radiation exposure were modeled. A two-step model of DNA-PK(sub cs) regulation of repair was developed with the initial step allowing access of other NHEJ components to breaks, and a second step limiting access to Ligase IV-XRCC4. Our model assumes that the transition from the first to second-step depends on DSB complexity, with a much slower-rate for complex DSB. The model faithfully reproduced several experimental data sets, including DSB rejoining as measured by pulsed-field electrophoresis (PFGE), quantification of the induction of gamma-H2AX foci, and live cell imaging of the induction of Ku(sub 70/80). Predictions are made for the behaviors of NHEJ components at low doses and dose-rates, where a steady-state is found at dose-rates of 0.1 Gy/hr or lower.

  13. Critical energies for SSB and DSB induction in plasmid DNA: Studies using synchrotron radiation

    SciTech Connect

    Michael, B.D.; Prise, K.M.; Folkard, M.; Vojnovic, B.; Brocklehurst, B.; Munro, I.H.; Hopkirk, A.

    1995-12-31

    The most frequent energy depositions along the charged-particle tracks associated with high-energy ionizing radiations are on the order of 10`s of eV. There continues to be a need for experimental data that relate the magnitudes of energy-deposition events to the molecular damages that result from them. The energy range between a few and several hundred eV is of particular importance. In addition, the determination of the yields of various types of damage induced in DNA as functions of the energy deposited in it (action spectra) may provide useful insights into the initial pathways of lesion induction. With much interest currently focused on the role of lesion complexity in relation to biological effectiveness, it is important to know to what extent single energy deposition events can independently initiate damages involving more than one molecular site, for example a DNA dsb. This paper reports some of the experimental methods and initial data, also with dry plasmid DNA, obtained using VUV from a synchrotron radiation source at selected energies in the range 7 to 25 eV, determining action spectra for ssb and dsb induction.

  14. Recurrent gross mutations of the PTEN tumor suppressor gene in breast cancers with deficient DSB repair

    PubMed Central

    Saal, Lao H; Gruvberger-Saal, Sofia K; Persson, Camilla; Lövgren, Kristina; Jumppanen, Mervi; Staaf, Johan; Jönsson, Göran; Pires, Maira M; Maurer, Matthew; Holm, Karolina; Koujak, Susan; Subramaniyam, Shivakumar; Vallon-Christersson, Johan; Olsson, Haökan; Su, Tao; Memeo, Lorenzo; Ludwig, Thomas; Ethier, Stephen P; Krogh, Morten; Szabolcs, Matthias; Murty, Vundavalli VVS; Isola, Jorma; Hibshoosh, Hanina; Parsons, Ramon; Borg, Åke

    2010-01-01

    Basal-like breast cancer (BBC) is a subtype of breast cancer with poor prognosis1–3. Inherited mutations of BRCA1, a cancer susceptibility gene involved in double-strand DNA break (DSB) repair, lead to breast cancers that are nearly always of the BBC subtype3–5; however, the precise molecular lesions and oncogenic consequences of BRCA1 dysfunction are poorly understood. Here we show that heterozygous inactivation of the tumor suppressor gene Pten leads to the formation of basal-like mammary tumors in mice, and that loss of PTEN expression is significantly associated with the BBC subtype in human sporadic and BRCA1-associated hereditary breast cancers. In addition, we identify frequent gross PTEN mutations, involving intragenic chromosome breaks, inversions, deletions and micro copy number aberrations, specifically in BRCA1-deficient tumors. These data provide an example of a specific and recurrent oncogenic consequence of BRCA1-dependent dysfunction in DNA repair and provide insight into the pathogenesis of BBC with therapeutic implications. These findings also argue that obtaining an accurate census of genes mutated in cancer will require a systematic examination for gross gene rearrangements, particularly in tumors with deficient DSB repair. PMID:18066063

  15. Effects of indirect actions and oxygen on relative biological effectiveness: estimate of DSB induction and conversion induced by gamma rays and helium ions.

    PubMed

    Tsai, Ju-Ying; Chen, Fang-Hsin; Hsieh, Tsung-Yu; Hsiao, Ya-Yun

    2015-07-01

    Clustered DNA damage other than double-strand breaks (DSBs) can be detrimental to cells and can lead to mutagenesis or cell death. In addition to DSBs induced by ionizing radiation, misrepair of non-DSB clustered damage contributes extra DSBs converted from DNA misrepair via pathways for base excision repair and nucleotide excision repair. This study aimed to quantify the relative biological effectiveness (RBE) when DSB induction and conversion from non-DSB clustered damage misrepair were used as biological endpoints. The results showed that both linear energy transfer (LET) and indirect action had a strong impact on the yields for DSB induction and conversion. RBE values for DSB induction and maximum DSB conversion of helium ions (LET = 120 keV/μm) to (60)Co gamma rays were 3.0 and 3.2, respectively. These RBE values increased to 5.8 and 5.6 in the absence of interference of indirect action initiated by addition of 2-M dimethylsulfoxide. DSB conversion was ∼1-4% of the total non-DSB damage due to gamma rays, which was lower than the 10% estimate by experimental measurement. Five to twenty percent of total non-DSB damage due to helium ions was converted into DSBs. Hence, it may be possible to increase the yields of DSBs in cancerous cells through DNA repair pathways, ultimately enhancing cell killing. PMID:25902742

  16. Effects of indirect actions and oxygen on relative biological effectiveness: estimate of DSB induction and conversion induced by gamma rays and helium ions

    PubMed Central

    Tsai, Ju-Ying; Chen, Fang-Hsin; Hsieh, Tsung-Yu; Hsiao, Ya-Yun

    2015-01-01

    Clustered DNA damage other than double-strand breaks (DSBs) can be detrimental to cells and can lead to mutagenesis or cell death. In addition to DSBs induced by ionizing radiation, misrepair of non-DSB clustered damage contributes extra DSBs converted from DNA misrepair via pathways for base excision repair and nucleotide excision repair. This study aimed to quantify the relative biological effectiveness (RBE) when DSB induction and conversion from non-DSB clustered damage misrepair were used as biological endpoints. The results showed that both linear energy transfer (LET) and indirect action had a strong impact on the yields for DSB induction and conversion. RBE values for DSB induction and maximum DSB conversion of helium ions (LET = 120 keV/μm) to 60Co gamma rays were 3.0 and 3.2, respectively. These RBE values increased to 5.8 and 5.6 in the absence of interference of indirect action initiated by addition of 2-M dimethylsulfoxide. DSB conversion was ∼1–4% of the total non-DSB damage due to gamma rays, which was lower than the 10% estimate by experimental measurement. Five to twenty percent of total non-DSB damage due to helium ions was converted into DSBs. Hence, it may be possible to increase the yields of DSBs in cancerous cells through DNA repair pathways, ultimately enhancing cell killing. PMID:25902742

  17. Roles of a conserved arginine residue of DsbB in linking protein disulfide-bond-formation pathway to the respiratory chain of Escherichia coli

    PubMed Central

    Kadokura, Hiroshi; Bader, Martin; Tian, Hongping; Bardwell, James C. A.; Beckwith, Jon

    2000-01-01

    The active-site cysteines of DsbA, the periplasmic disulfide-bond-forming enzyme of Escherichia coli, are kept oxidized by the cytoplasmic membrane protein DsbB. DsbB, in turn, is oxidized by two kinds of quinones (ubiquinone for aerobic and menaquinone for anaerobic growth) in the electron-transport chain. We describe the isolation of dsbB missense mutations that change a highly conserved arginine residue at position 48 to histidine or cysteine. In these mutants, DsbB functions reasonably well aerobically but poorly anaerobically. Consistent with this conditional phenotype, purified R48H exhibits very low activity with menaquinone and an apparent Michaelis constant (Km) for ubiquinone seven times greater than that of the wild-type DsbB, while keeping an apparent Km for DsbA similar to that of wild-type enzyme. From these results, we propose that this highly conserved arginine residue of DsbB plays an important role in the catalysis of disulfide bond formation through its role in the interaction of DsbB with quinones. PMID:11005861

  18. Functional and evolutionary analyses of Helicobacter pylori HP0231 (DsbK) protein with strong oxidative and chaperone activity characterized by a highly diverged dimerization domain

    PubMed Central

    Bocian-Ostrzycka, Katarzyna M.; Łasica, Anna M.; Dunin-Horkawicz, Stanisław; Grzeszczuk, Magdalena J.; Drabik, Karolina; Dobosz, Aneta M.; Godlewska, Renata; Nowak, Elżbieta; Collet, Jean-Francois; Jagusztyn-Krynicka, Elżbieta K.

    2015-01-01

    Helicobacter pylori does not encode the classical DsbA/DsbB oxidoreductases that are crucial for oxidative folding of extracytoplasmic proteins. Instead, this microorganism encodes an untypical two proteins playing a role in disulfide bond formation – periplasmic HP0231, which structure resembles that of EcDsbC/DsbG, and its redox partner, a membrane protein HpDsbI (HP0595) with a β-propeller structure. The aim of presented work was to assess relations between HP0231 structure and function. We showed that HP0231 is most closely related evolutionarily to the catalytic domain of DsbG, even though it possesses a catalytic motif typical for canonical DsbA proteins. Similarly, the highly diverged N-terminal dimerization domain is homologous to the dimerization domain of DsbG. To better understand the functioning of this atypical oxidoreductase, we examined its activity using in vivo and in vitro experiments. We found that HP0231 exhibits oxidizing and chaperone activities but no isomerizing activity, even though H. pylori does not contain a classical DsbC. We also show that HP0231 is not involved in the introduction of disulfide bonds into HcpC (Helicobacter cysteine-rich protein C), a protein involved in the modulation of the H. pylori interaction with its host. Additionally, we also constructed a truncated version of HP0231 lacking the dimerization domain, denoted HP0231m, and showed that it acts in Escherichia coli cells in a DsbB-dependent manner. In contrast, HP0231m and classical monomeric EcDsbA (E. coli DsbA protein) were both unable to complement the lack of HP0231 in H. pylori cells, though they exist in oxidized forms. HP0231m is inactive in the insulin reduction assay and possesses high chaperone activity, in contrast to EcDsbA. In conclusion, HP0231 combines oxidative functions characteristic of DsbA proteins and chaperone activity characteristic of DsbC/DsbG, and it lacks isomerization activity. PMID:26500620

  19. Involvement of DNA-PK(sub cs) in DSB Repair Following Fe-56 Ion Irradiation

    NASA Technical Reports Server (NTRS)

    O'Neill, Peter; Harper, Jane; Anderson, Jennifer a.; Cucinnota, Francis A.

    2007-01-01

    When cells are exposed to radiation, cellular lesions are induced in the DNA including double strand breaks (DSBs), single strand breaks and clustered DNA damage, which if not repaired with high fidelity may lead to detrimental biological consequences. Complex DSBs are induced by ionizing radiation and characterized by the presence of base lesions close to the break termini. They are believed to be one of the major causes of the biological effects of IR. The complexity of DSBs increases with the ionization density of the radiation and these complex DSBs are distinct from the damage induced by sparsely ionizing gamma-radiation. It has been hypothesized that complex DSBs produced by heavy ions in space pose problems to the DNA repair machinery. We have used imm uno-cyto-chemical staining of phosphorylated histone H2AX (gamma-H2AX) foci, as a marker of DSBs. We have investigated the formation and loss of gamma-H2AX foci and RAD51 foci (a protein involved in the homologous recombination pathway) in mammalian cells induced by low fluences of low-LET gamma-radiation and high-LET Fe-56 ions (1GeV/n, 151 keV/micron LET). M059J and M059K cells, which are deficient and proficient in DNA-PK(sub cs) activity respectively, were used to examine the role of DNA-PK(sub cs), a key protein in the non-homologous end joining (NHEJ) pathway of DSB repair, along with HF19 human fibroblasts. Followi ng irradiation with Fe-56 ions the rate of repair was slower in M059J cells compared with that in M059K, indicating a role for DNA-PK(sub cs) in the repair of DSB induced by Fe-56 ions. However a small percentage of DSBs induced are rejoined within 5 h although many DSBs still persist up to 24 h. When RAD51 was examined in M059J/K cells, RAD51 foci are visible 24 hours after irradiation in approximately 40% of M059J cells compared with <5% of M059K cells indicating that persistent DSBs or those formed at stalled replication forks recruit RAD51 in DNA-PK(sub cs) deficient cells. Following 1 Gy

  20. Studies on human blood lymphocytes with iC3b (type 3) complement receptors: III. Abnormalities in patients with active systemic lupus erythematosus.

    PubMed Central

    Gray, J D; Lash, A; Bakke, A C; Kitridou, R C; Horwitz, D A

    1987-01-01

    Lymphocytes displaying iC3b (Type 3) complement receptors (CR3) were quantified by flow cytometry in patients with systemic lupus erythematosus. The percentages and absolute numbers were compared to age and sex matched controls. Total CR3+ lymphocytes identified by the monoclonal antibodies OKM1 or Leu 15 were significantly decreased in patients with symptomatic arthritis, serositis or vasculitis and those with lupus nephritis, whereas values for CR3+ lymphocytes in patients with inactive disease were similar to normal donors. The phenotype of CR3+ lymphocytes was markedly different in patients with active SLE. In normals granular lymphocytes bearing Fc receptors for IgG (L cells) comprised two-thirds of CR3+ lymphocytes. However, in SLE this subset was reduced to 20% and there was a corresponding increase in CR3+ lymphocytes co-expressing the T3 marker. Percentages of CR3 T4+ but not CR3+ T8+ lymphocytes were significantly increased in SLE. Although patients with active disease were lymphopenic, absolute numbers of CR3+ lymphocytes co-expressing T cell markers were similar to normal controls. Since L cells are non-specific suppressors of Ig production, the reduction of this subset along with the increase in CR3 T4+ cells could contribute to unregulated antibody production characteristic of SLE. PMID:2955974

  1. Structure of the Acinetobacter baumannii Dithiol Oxidase DsbA Bound to Elongation Factor EF-Tu Reveals a Novel Protein Interaction Site

    PubMed Central

    Premkumar, Lakshmanane; Kurth, Fabian; Duprez, Wilko; Grøftehauge, Morten K.; King, Gordon J.; Halili, Maria A.; Heras, Begoña; Martin, Jennifer L.

    2014-01-01

    The multidrug resistant bacterium Acinetobacter baumannii is a significant cause of nosocomial infection. Biofilm formation, that requires both disulfide bond forming and chaperone-usher pathways, is a major virulence trait in this bacterium. Our biochemical characterizations show that the periplasmic A. baumannii DsbA (AbDsbA) enzyme has an oxidizing redox potential and dithiol oxidase activity. We found an unexpected non-covalent interaction between AbDsbA and the highly conserved prokaryotic elongation factor, EF-Tu. EF-Tu is a cytoplasmic protein but has been localized extracellularly in many bacterial pathogens. The crystal structure of this complex revealed that the EF-Tu switch I region binds to the non-catalytic surface of AbDsbA. Although the physiological and pathological significance of a DsbA/EF-Tu association is unknown, peptides derived from the EF-Tu switch I region bound to AbDsbA with submicromolar affinity. We also identified a seven-residue DsbB-derived peptide that bound to AbDsbA with low micromolar affinity. Further characterization confirmed that the EF-Tu- and DsbB-derived peptides bind at two distinct sites. These data point to the possibility that the non-catalytic surface of DsbA is a potential substrate or regulatory protein interaction site. The two peptides identified in this work together with the newly characterized interaction site provide a novel starting point for inhibitor design targeting AbDsbA. PMID:24860094

  2. AdnAB: a new DSB-resecting motor-nuclease from mycobacteria.

    PubMed

    Sinha, Krishna Murari; Unciuleac, Mihaela-Carmen; Glickman, Michael S; Shuman, Stewart

    2009-06-15

    The resection of DNA double-strand breaks (DSBs) in bacteria is a motor-driven process performed by a multisubunit helicase-nuclease complex: either an Escherichia coli-type RecBCD enzyme or a Bacillus-type AddAB enzyme. Here we identify mycobacterial AdnAB as the founder of a new family of heterodimeric helicase-nucleases with distinctive properties. The AdnA and AdnB subunits are each composed of an N-terminal UvrD-like motor domain and a C-terminal nuclease module. The AdnAB ATPase is triggered by dsDNA with free ends and the energy of ATP hydrolysis is coupled to DSB end resection by the AdnAB nuclease. The mycobacterial nonhomologous end-joining (NHEJ) protein Ku protects DSBs from resection by AdnAB. We find that AdnAB incises ssDNA by measuring the distance from the free 5' end to dictate the sites of cleavage, which are predominantly 5 or 6 nucleotides (nt) from the 5' end. The "molecular ruler" of AdnAB is regulated by ATP, which elicits an increase in ssDNA cleavage rate and a distal displacement of the cleavage sites 16-17 nt from the 5' terminus. AdnAB is a dual nuclease with a clear division of labor between the subunits. Mutations in the nuclease active site of the AdnB subunit ablate the ATP-inducible cleavages; the corresponding changes in AdnA abolish ATP-independent cleavage. Complete suppression of DSB end resection requires simultaneous mutation of both subunit nucleases. The nuclease-null AdnAB is a helicase that unwinds linear plasmid DNA without degrading the displaced single strands. Mutations of the phosphohydrolase active site of the AdnB subunit ablate DNA-dependent ATPase activity, DSB end resection, and ATP-inducible ssDNA cleavage; the equivalent mutations of the AdnA subunit have comparatively little effect. AdnAB is a novel signature of the Actinomycetales taxon. Mycobacteria are exceptional in that they encode both AdnAB and RecBCD, suggesting the existence of alternative end-resecting motor-nuclease complexes. PMID:19470566

  3. DNA-PK: a dynamic enzyme in a versatile DSB repair pathway

    PubMed Central

    Davis, Anthony J.; Chen, Benjamin P.C.; Chen, David J.

    2014-01-01

    DNA double stranded breaks (DSBs) are the most cytoxic DNA lesion as the inability to properly repair them can lead to genomic instability and tumorigenesis. The prominent DSB repair pathway in humans is non-homologous end-joining (NHEJ). In the simplest sense, NHEJ mediates the direct re-ligation of the broken DNA molecule. However, NHEJ is a complex and versatile process that can repair DSBs with a variety of damages and ends via the utilization of a significant number of proteins. In this review we will describe the important factors and mechanisms modulating NHEJ with emphasis given to the versatility of this repair process and the DNA-PK complex. PMID:24680878

  4. Identification and characterization of the Escherichia coli gene dsbB, whose product is involved in the formation of disulfide bonds in vivo.

    PubMed Central

    Missiakas, D; Georgopoulos, C; Raina, S

    1993-01-01

    We have identified and characterized the Escherichia coli gene dsbB, whose product is required for disulfide bond formation of periplasmic proteins, by using two different approaches: (i) screening of a multicopy plasmid library for clones which protect E. coli from the lethal effects of dithiothreitol (DTT), and (ii) screening of insertion libraries of E. coli for DTT-sensitive mutants. Mapping and characterization of mutations conferring a DTT-sensitive phenotype also identified the dsbA, trxA, and trxB genes, whose products are involved in different oxidation-reduction pathways. Null mutations in dsbB conferred pleiotropic phenotypes such as sensitivity to benzylpenicillin and inability to support plaque formation of filamentous phages, and they were shown to severely affect disulfide bond oxidation of secreted proteins such as OmpA and beta-lactamase. These phenotypes resemble the phenotype of bacteria carrying either a null mutation in the dsbA gene or the double mutation dsbA dsbB. Sequencing and expression of the dsbB gene revealed that it encodes a 20-kDa protein predicted to possess an "exchangeable" disulfide bond in -Cys-Val-Leu-Cys-. The dsbB gene maps at 26.5 min on the genetic map of the E. coli chromosome, and its transcription is directed from two promoters, neither of which resembles the canonical E sigma 70-recognized promoter. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:7688471

  5. Cytogenetic Response to Ionizing Radiation Exposure in Human Fibroblasts with Suppressed Expression of Non-DSB Repair Genes

    NASA Technical Reports Server (NTRS)

    Zhang, Ye; Rohde, Larry H.; Emami, Kamal; Hammond, Dianne; Mehta, Satish K.; Jeevarajan, Antony S.; Pierson, Duane L.; Wu, Honglu

    2009-01-01

    Changes of gene expression profile are one of the most important biological responses in living cells after ionizing radiation (IR) exposure. Although some studies have shown that genes up-regulated by IR may play important roles in DNA damage repair, the relationship between the regulation of gene expression by IR, particularly genes not known for their roles in double-strand break (DSB) repair, and its impact on cytogenetic responses has not been well studied. The purpose of this study is to identify new roles of IR inducible genes in radiation-induced chromosome aberrations and micronuclei formation. In the study, the expression of 25 genes selected on the basis of their transcriptional changes in response to IR was individually knocked down by small interfering RNA in human fibroblast cells. Frequencies of micronuclei (MN) formation and chromosome aberrations were measured to determine the efficiency of cytogenetic repair, and the fraction of bi-nucleated cells in the MN analysis was used as a marker for cell cycle progression. In response to gamma radiation, the formation of MN was significantly increased by suppressed expression of five genes: Ku70 (DSB repair pathway), XPA (nucleotide excision repair pathway), RPA1 (mismatch repair pathway), RAD17 and RBBP8 (cell cycle control). Knocked-down expression of four genes (MRE11A, RAD51 in the DSB pathway, SESN1, and SUMO1) significantly inhibited cell cycle progression, possibly because of severe impairment of DNA damage repair. Moreover, decreased XPA, p21, or MLH1 expression resulted in both significantly enhanced cell cycle progression and increased yields of chromosome aberrations, indicating that these gene products modulate both cell cycle control and DNA damage repair. Nine of these eleven genes, whose knock-down expression affected cytogenetic repair, were up-regulated in cells exposed to gamma radiation, suggesting that genes transcriptionally modulated by IR were critical to regulate IR

  6. DNA Fragmentation and DSB correlation Induced in Human Fibroblasts by Accelerated 56Fe Ions of Differing Energies

    NASA Astrophysics Data System (ADS)

    Antonelli, F.; Belli, M.; Campa, A.; Dini, V.; Esposito, G.; Furusawa, Y.; Simone, G.; Sorrentino, E.; Tabocchini, M. A.

    HZE particles from space radiation raise an important protection concern during long-term astronauts travels Although these particles are less abundant than protons they are more effective in damaging biological systems It is thought that this is due to the frequent production of spatially correlated DNA damaged sites particularly double strand breaks DSB since this correlation can strongly affect the repair capability of the cells In this work we have studied the DNA fragmentation induced in human fibroblasts by accelerated 56 Fe ions of four different energies i e 115 MeV u 414 MeV u 1 GeV u and 5 GeV u and by gamma-rays used as reference radiation DNA fragmentation was studied in various size ranges varying from 1 to 5700 kbp using Pulsed or Constant Field Gel Electrophoresis The DSB yields have been derived from fragmentation in the overall range as well as in the two ranges 1-23 and 23-5700 kbp The overall DSB yield slightly increased with the ion energy maily due to the contribution of the 23-5700 kbp fragments while that of small fragments 1-23 kbp was almost constant Accordingly the relative biological effectiveness RBE for DSB induction increased with energy from about 1 3 at 115 MeV u to about 1 8 at about 5 GeV u i e less than the RBE for chromosome aberration and cell inactivation The degree of spatial correlation of DSB was evaluated through the departure from the randomness of the fragment distribution with a simple theoretical tool that we have recently introduced To this aim a parameter R was used

  7. Comparing Benign and Malignant Neoplasia and DSB Induction for Low-and High-LET Radiation

    NASA Astrophysics Data System (ADS)

    Burns, Fredric; (Eric) Tang, Moon-Shong; Wu, Feng

    One-and 2-stage models based on DNA double strand breaks (DSBs) have been developed to describe the dose and LET dependence of cancer induction in rat skin exposed to the Bragg plateau of several ion beams or electron radiation. Data are presented showing that carcinomas (malignant) and fibromas (benign) are induced differently by low and high LET radiation. DSBs are subject to complex repair processes, including homologous and non-homologous end joining, that slowly eliminate broken chromosome ends but at the expense of elevating genomic instability that increases the risk of neoplasia. In this formulation the initial molecular lesion in radiation carcinogenesis is assumed to be a DNA double strand break (DSB). The 2-event model assumes that pairs of DSBs join to create cellular genomic instability that eventually progresses to malignancy. The 1-event model assumes that joining is insignificant but that unrepaired DSBs remain and are sufficiently destabilizing to produce low-grade neoplasias. The respective expected relationships between neoplasia yield (Y), radiation dose (D) and LET (L) are: Y(D) = CLD + BD2 (A) for 2-events and Y(D) = CLD (B) for 1-event. Respective B and C values have been evaluated empirically for carcinomas, fibromas and DSBs, the latter via the -H2Ax technique in surrogate keratinocytes, for several types of radiations, including, 40Ar ions, 56Fe ions, 20Ne ions, protons, electrons and x-rays. Fibromas outnumber carcinomas by about 6:1 but are more sensitive than carcinomas to the cytolethal effect of the radiations. The 2-event model agrees well with carcinoma yields in rat skin but fails to model fibromas correctly. Instead the fibroma yields best fitted with the 1-event model for the high LET ion radiations, but at very low LET (electron radiation), an empirical D3 component becomes apparent which is not currently incorporated into the theoretical model. At higher LET values, the D3 component was not detected. The overall results are

  8. DNA-DSB in CHO-K1 cells induced by heavy-ions: Break rejoining and residual damage (GSI)

    NASA Technical Reports Server (NTRS)

    Taucher-Scholz, G.; Heilmann, J.; Becher, G.; Kraft, G.

    1994-01-01

    DNA double strand breaks (DSB's) are the critical lesions involved in cellular effects of ionizing radiation. Therefore, the evaluation of DSB induction in mammalian cells after heavy ion irradiation is an essential task for the assessment of high-LET radiation risk in space. Of particular interest has been the question of how the biological efficiency for the cellular inactivation endpoint relates to the initial lesions (DSBs) at varying LETs. For cell killing, an increased Relative Biological Efficiency (RBE) has been determined for highLET radiation around 100-200 keV/mu m. At higher LET, the RBE's decrease again to values below one for the very heavy particles. At GSI, DSB-induction was measured in CHO-K1 cells following irradiation with accelerated particles covering a wide LET range. The electrophoretic elution of fragmented DNA out of agarose plugs in a constant electrical field was applied for the detection of DSB's. The fraction of DNA retained was determined considering the relative intensities of ethidium bromide fluorescence in the well and in the gel lane. Dose-effect curves were established, from which the RBE for DSB induction was calculated at a fraction of 0.7 of DNA retained In summary, these rejoining studies are in line with an enhanced severity of the DNA DSB's at higher LET's, resulting in a decreased repairability of the induced lesions. However, no information concerning the fidelity of strand breaks rejoining is provided in these studies. To assess correct rejoining of DNA fragments an experimental system involving individual DNA hybridization bands has been set up. In preliminary experiments Sal I generated DNA fragments of 0.9 Mbp were irradiated with xrays and incubated for repair However, restitution of the original signals was not observed, probably due to the high radiation dose necessary for breakage of a fragment of this size. A banding pattern with NotI hybridization signals in a higher MW range (3Mbp) has been obtained by varying

  9. Identification of Novel Radiosensitizers in a High-Throughput, Cell-Based Screen for DSB Repair Inhibitors

    PubMed Central

    Goglia, Alexander G.; Delsite, Robert; Luz, Antonio N.; Shahbazian, David; Salem, Ahmed F.; Sundaram, Ranjini K.; Chiaravalli, Jeanne; Hendrikx, Petrus J.; Wilshire, Jennifer A.; Jasin, Maria; Kluger, Harriet; Glickman, J. Fraser; Powell, Simon N.; Bindra, Ranjit S.

    2014-01-01

    Most cancer therapies involve a component of treatment which inflicts DNA damage in tumor cells, such as double-strand breaks (DSBs), which are considered the most serious threat to genomic integrity. Complex systems have evolved to repair these lesions, and successful DSB repair is essential for tumor cell survival after exposure to ionizing radiation (IR) and other DNA damaging agents. As such, inhibition of DNA repair is a potentially efficacious strategy for chemo- and radio-sensitization. Homologous recombination (HR) and nonhomologous end-joining (NHEJ) represent the two major pathways by DSBs are repaired in mammalian cells. Here, we report the design and execution of a high-throughput, cell-based small molecule screen for novel DSB repair inhibitors. We miniaturized our recently developed dual NHEJ and HR reporter system into a 384-well plate-based format and interrogated a diverse library of 20,000 compounds for molecules which selectively modulate NHEJ and HR repair in tumor cells. We identified a collection of novel hits which potently inhibit DSB repair, and we have validated their functional activity in comprehensive panel of orthogonal secondary assays. A selection of these inhibitors were found to radiosensitize cancer cell lines in vitro, which suggests they may be useful as novel chemo- and radio-sensitizers. Surprisingly, we identified several FDA-approved drugs, including the calcium channel blocker, mibefradil dihydrochloride, which demonstrated activity as DSB repair inhibitors and radiosensitizers. These findings suggest the possibility for repurposing them as tumor cell radiosensitizers in the future. Accordingly, we recently initiated a Phase I clinical trial testing mibefradil as glioma radiosensitizer. PMID:25512618

  10. Sent packing: protein engineering generates a new crystal form of Pseudomonas aeruginosa DsbA1 with increased catalytic surface accessibility

    PubMed Central

    McMahon, Roisin M.; Coinçon, Mathieu; Tay, Stephanie; Heras, Begoña; Morton, Craig J.; Scanlon, Martin J.; Martin, Jennifer L.

    2015-01-01

    Pseudomonas aeruginosa is an opportunistic human pathogen for which new antimicrobial drug options are urgently sought. P. aeruginosa disulfide-bond protein A1 (PaDsbA1) plays a pivotal role in catalyzing the oxidative folding of multiple virulence proteins and as such holds great promise as a drug target. As part of a fragment-based lead discovery approach to PaDsbA1 inhibitor development, the identification of a crystal form of PaDsbA1 that was more suitable for fragment-soaking experiments was sought. A previously identified crystallization condition for this protein was unsuitable, as in this crystal form of PaDsbA1 the active-site surface loops are engaged in the crystal packing, occluding access to the target site. A single residue involved in crystal-packing interactions was substituted with an amino acid commonly found at this position in closely related enzymes, and this variant was successfully used to generate a new crystal form of PaDsbA1 in which the active-site surface is more accessible for soaking experiments. The PaDsbA1 variant displays identical redox character and in vitro activity to wild-type PaDsbA1 and is structurally highly similar. Two crystal structures of the PaDsbA1 variant were determined in complex with small molecules bound to the protein active site. These small molecules (MES, glycerol and ethylene glycol) were derived from the crystallization or cryoprotectant solutions and provide a proof of principle that the reported crystal form will be amenable to co-crystallization and soaking with small molecules designed to target the protein active-site surface. PMID:26627647

  11. Sulfur Denitrosylation by an Engineered Trx-like DsbG Enzyme Identifies Nucleophilic Cysteine Hydrogen Bonds as Key Functional Determinant.

    PubMed

    Lafaye, Céline; Van Molle, Inge; Tamu Dufe, Veronica; Wahni, Khadija; Boudier, Ariane; Leroy, Pierre; Collet, Jean-François; Messens, Joris

    2016-07-15

    Exposure of bacteria to NO results in the nitrosylation of cysteine thiols in proteins and low molecular weight thiols such as GSH. The cells possess enzymatic systems that catalyze the denitrosylation of these modified sulfurs. An important player in these systems is thioredoxin (Trx), a ubiquitous, cytoplasmic oxidoreductase that can denitrosylate proteins in vivo and S-nitrosoglutathione (GSNO) in vitro However, a periplasmic or extracellular denitrosylase has not been identified, raising the question of how extracytoplasmic proteins are repaired after nitrosative damage. In this study, we tested whether DsbG and DsbC, two Trx family proteins that function in reducing pathways in the Escherichia coli periplasm, also possess denitrosylating activity. Both DsbG and DsbC are poorly reactive toward GSNO. Moreover, DsbG is unable to denitrosylate its specific substrate protein, YbiS. Remarkably, by borrowing the CGPC active site of E. coli Trx-1 in combination with a T200M point mutation, we transformed DsbG into an enzyme highly reactive toward GSNO and YbiS. The pKa of the nucleophilic cysteine, as well as the redox and thermodynamic properties of the engineered DsbG are dramatically changed and become similar to those of E. coli Trx-1. X-ray structural insights suggest that this results from a loss of two direct hydrogen bonds to the nucleophilic cysteine sulfur in the DsbG mutant. Our results highlight the plasticity of the Trx structural fold and reveal that the subtle change of the number of hydrogen bonds in the active site of Trx-like proteins is the key factor that thermodynamically controls reactivity toward nitrosylated compounds. PMID:27226614

  12. Rapid MCNP simulation of DNA double strand break (DSB) relative biological effectiveness (RBE) for photons, neutrons, and light ions.

    PubMed

    Stewart, Robert D; Streitmatter, Seth W; Argento, David C; Kirkby, Charles; Goorley, John T; Moffitt, Greg; Jevremovic, Tatjana; Sandison, George A

    2015-11-01

    To account for particle interactions in the extracellular (physical) environment, information from the cell-level Monte Carlo damage simulation (MCDS) for DNA double strand break (DSB) induction has been integrated into the general purpose Monte Carlo N-particle (MCNP) radiation transport code system. The effort to integrate these models is motivated by the need for a computationally efficient model to accurately predict particle relative biological effectiveness (RBE) in cell cultures and in vivo. To illustrate the approach and highlight the impact of the larger scale physical environment (e.g. establishing charged particle equilibrium), we examined the RBE for DSB induction (RBEDSB) of x-rays, (137)Cs γ-rays, neutrons and light ions relative to γ-rays from (60)Co in monolayer cell cultures at various depths in water. Under normoxic conditions, we found that (137)Cs γ-rays are about 1.7% more effective at creating DSB than γ-rays from (60)Co (RBEDSB  =  1.017) whereas 60-250 kV x-rays are 1.1 to 1.25 times more efficient at creating DSB than (60)Co. Under anoxic conditions, kV x-rays may have an RBEDSB up to 1.51 times as large as (60)Co γ-rays. Fission neutrons passing through monolayer cell cultures have an RBEDSB that ranges from 2.6 to 3.0 in normoxic cells, but may be as large as 9.93 for anoxic cells. For proton pencil beams, Monte Carlo simulations suggest an RBEDSB of about 1.2 at the tip of the Bragg peak and up to 1.6 a few mm beyond the Bragg peak. Bragg peak RBEDSB increases with decreasing oxygen concentration, which may create opportunities to apply proton dose painting to help address tumor hypoxia. Modeling of the particle RBE for DSB induction across multiple physical and biological scales has the potential to aid in the interpretation of laboratory experiments and provide useful information to advance the safety and effectiveness of hadron therapy in the treatment of cancer. PMID:26449929

  13. Rapid MCNP simulation of DNA double strand break (DSB) relative biological effectiveness (RBE) for photons, neutrons, and light ions

    NASA Astrophysics Data System (ADS)

    Stewart, Robert D.; Streitmatter, Seth W.; Argento, David C.; Kirkby, Charles; Goorley, John T.; Moffitt, Greg; Jevremovic, Tatjana; Sandison, George A.

    2015-11-01

    To account for particle interactions in the extracellular (physical) environment, information from the cell-level Monte Carlo damage simulation (MCDS) for DNA double strand break (DSB) induction has been integrated into the general purpose Monte Carlo N-particle (MCNP) radiation transport code system. The effort to integrate these models is motivated by the need for a computationally efficient model to accurately predict particle relative biological effectiveness (RBE) in cell cultures and in vivo. To illustrate the approach and highlight the impact of the larger scale physical environment (e.g. establishing charged particle equilibrium), we examined the RBE for DSB induction (RBEDSB) of x-rays, 137Cs γ-rays, neutrons and light ions relative to γ-rays from 60Co in monolayer cell cultures at various depths in water. Under normoxic conditions, we found that 137Cs γ-rays are about 1.7% more effective at creating DSB than γ-rays from 60Co (RBEDSB  =  1.017) whereas 60-250 kV x-rays are 1.1 to 1.25 times more efficient at creating DSB than 60Co. Under anoxic conditions, kV x-rays may have an RBEDSB up to 1.51 times as large as 60Co γ-rays. Fission neutrons passing through monolayer cell cultures have an RBEDSB that ranges from 2.6 to 3.0 in normoxic cells, but may be as large as 9.93 for anoxic cells. For proton pencil beams, Monte Carlo simulations suggest an RBEDSB of about 1.2 at the tip of the Bragg peak and up to 1.6 a few mm beyond the Bragg peak. Bragg peak RBEDSB increases with decreasing oxygen concentration, which may create opportunities to apply proton dose painting to help address tumor hypoxia. Modeling of the particle RBE for DSB induction across multiple physical and biological scales has the potential to aid in the interpretation of laboratory experiments and provide useful information to advance the safety and effectiveness of hadron therapy in the treatment of cancer.

  14. Francisella tularensis type B ΔdsbA mutant protects against type A strain and induces strong inflammatory cytokine and Th1-like antibody response in vivo.

    PubMed

    Straskova, Adela; Spidlova, Petra; Mou, Sherry; Worsham, Patricia; Putzova, Daniela; Pavkova, Ivona; Stulik, Jiri

    2015-11-01

    Francisella tularensis subspecies tularensis is a highly virulent intracellular bacterial pathogen, causing the disease tularemia. However, a safe and effective vaccine for routine application against F. tularensis has not yet been developed. We have recently constructed the deletion mutants for the DsbA homolog protein (ΔdsbA/FSC200) and a hypothetical protein IglH (ΔiglH/FSC200) in the type B F. tularensis subsp. holarctica FSC200 strain, which exerted different protection capacity against parental virulent strain. In this study, we further investigated the immunological correlates for these different levels of protection provided by ΔdsbA/FSC200 and ΔiglH/FSC200 mutants. Our results show that ΔdsbA/FSC200 mutant, but not ΔiglH/FSC200 mutant, induces an early innate inflammatory response leading to strong Th1-like antibody response. Furthermore, vaccination with ΔdsbA/FSC200 mutant, but not with ΔiglH/FSC200, elicited protection against the subsequent challenge with type A SCHU S4 strain in mice. An immunoproteomic approach was used to map a spectrum of antigens targeted by Th1-like specific antibodies, and more than 80 bacterial antigens, including novel ones, were identified. Comparison of tularemic antigens recognized by the ΔdsbA/FSC200 post-vaccination and the SCHU S4 post-challenge sera then revealed the existence of 22 novel SCHU S4 specific antibody clones. PMID:26253078

  15. Four structural subclasses of the antivirulence drug target disulfide oxidoreductase DsbA provide a platform for design of subclass-specific inhibitors.

    PubMed

    McMahon, Róisín M; Premkumar, Lakshmanane; Martin, Jennifer L

    2014-08-01

    By catalyzing oxidative protein folding, the bacterial disulfide bond protein A (DsbA) plays an essential role in the assembly of many virulence factors. Predictably, DsbA disruption affects multiple downstream effector molecules, resulting in pleiotropic effects on the virulence of important human pathogens. These findings mark DsbA as a master regulator of virulence, and identify the enzyme as a target for a new class of antivirulence agents that disarm pathogenic bacteria rather than killing them. The purpose of this article is to discuss and expand upon recent findings on DsbA and to provide additional novel insights into the druggability of this important disulfide oxidoreductase by comparing the structures and properties of 13 well-characterized DsbA enzymes. Our structural analysis involved comparison of the overall fold, the surface properties, the conformations of three loops contributing to the binding surface and the sequence identity of residues contributing to these loops. Two distinct structural classes were identified, classes I and II, which are differentiated by their central β-sheet arrangements and which roughly separate the DsbAs produced by Gram-negative from Gram-positive organisms. The classes can be further subdivided into a total of four subclasses on the basis of surface features. Class Ia is equivalent to the Enterobacteriaceae class that has been defined previously. Bioinformatic analyses support the classification of DsbAs into 3 of the 4 subclasses, but did not pick up the 4th subclass which is only apparent from analysis of DsbA electrostatic surface properties. In the context of inhibitor development, the discrete structural subclasses provide a platform for developing DsbA inhibitory scaffolds with a subclass-wide spectrum of activity. We expect that more DsbA classes are likely to be identified, as enzymes from other pathogens are explored, and we highlight the issues associated with structure-based inhibitor development targeting

  16. DsbA2 (27 kDa Com1-like protein) of Legionella pneumophila catalyses extracytoplasmic disulphide-bond formation in proteins including the Dot/Icm type IV secretion system.

    PubMed

    Jameson-Lee, Max; Garduño, Rafael A; Hoffman, Paul S

    2011-05-01

    In Gram-negative bacteria, thiol oxidoreductases catalyse the formation of disulphide bonds (DSB) in extracytoplasmic proteins. In this study, we sought to identify DSB-forming proteins required for assembly of macromolecular structures in Legionella pneumophila. Here we describe two DSB-forming proteins, one annotated as dsbA1 and the other annotated as a 27 kDa outer membrane protein similar to Com1 of Coxiella burnetii, which we designate as dsbA2. Both proteins are predicted to be periplasmic, and while dsbA1 mutants were readily isolated and without phenotype, dsbA2 mutants were not obtained. To advance studies of DsbA2, a cis-proline residue at position 198 was replaced with threonine that enables formation of stable disulphide-bond complexes with substrate proteins. Expression of DsbA2 P198T mutant protein from an inducible promoter produced dominant-negative effects on DsbA2 function that resulted in loss of infectivity for amoeba and HeLa cells and loss of Dot/Icm T4SS-mediated contact haemolysis of erythrocytes. Analysis of captured DsbA2 P198T-substrate complexes from L. pneumophila by mass spectrometry identified periplasmic and outer membrane proteins that included components of the Dot/Icm T4SS. More broadly, our studies establish a DSB oxidoreductase function for the Com1 lineage of DsbA2-like proteins which appear to be conserved among those bacteria also expressing T4SS. PMID:21375592

  17. DsbA2 (27-kDa Com1-Like Protein) of Legionella pneumophila Catalyses Extracytoplasmic Disulfide-Bond Formation in Proteins Including the Dot/Icm Type IV Secretion System

    PubMed Central

    Jameson-Lee, Max; Garduno, Rafael A.; Hoffman, Paul S.

    2011-01-01

    Summary In Gram negative bacteria, thiol oxidoreductases catalyze the formation of disulfide bonds (DSB) in extracytoplasmic proteins. In this study, we sought to identify DSB-forming proteins required for assembly of macromolecular structures in Legionella pneumophila. Here we describe two DSB forming proteins, one annotated as dsbA1 and the other annotated as a 27-kDa outer membrane protein similar to Com1 of Coxiella burnetii, which we designate as dsbA2. Both proteins are predicted to be periplasmic, and while dsbA1 mutants were readily isolated and without phenotype, dsbA2 mutants were not obtained. To advance studies of DsbA2, a cis-proline residue at position 198 was replaced with threonine that enables formation of stable disulfide-bond complexes with substrate proteins. Expression of DsbA2 P198T-mutant protein from an inducible promoter produced dominant-negative effects on DsbA2 function that resulted in loss of infectivity for amoeba and HeLa cells and loss of Dot/Icm T4SS-mediated contact hemolysis of erythrocytes. Analysis of captured DsbA2 P198T-substrate complexes from L. pneumophila by mass spectrometry identified periplasmic and outer membrane proteins that included components of the Dot/Icm T4SS. More broadly, our studies establish a DSB oxidoreductase function for the Com1 lineage of DsbA2-like proteins which appear to be conserved among those bacteria also expressing T4SS. PMID:21375592

  18. Application of an InGaAsP diode laser to probe photodissociation dynamics - I(asterisk) quantum yields from n- and i-C3F7I and CH3I by laser gain vs absorption spectroscopy

    NASA Technical Reports Server (NTRS)

    Hess, W. P.; Kohler, S. J.; Haugen, H. K.; Leone, S. R.

    1986-01-01

    Initial measurements on I-asterisk yields of alkyl iodides at 266 nm are reported using gain vs. absorption spectroscopy with an InGaAsP diode probe laser. The results are 102 percent + or - 4 percent, 102 percent + or - 7 percent, and 73 percent + or - 4 percent for n-C3F7I, i-C3F7I, and CH3I respectively. Future prospects for the development of diode laser systems and for their use in dynamical studies are discussed.

  19. Sent packing: protein engineering generates a new crystal form of Pseudomonas aeruginosa DsbA1 with increased catalytic surface accessibility

    SciTech Connect

    McMahon, Roisin M. Coinçon, Mathieu; Tay, Stephanie; Heras, Begoña; Morton, Craig J.; Scanlon, Martin J.; Martin, Jennifer L.

    2015-11-26

    The crystal structure of a P. aeruginosa DsbA1 variant is more suitable for fragment-based lead discovery efforts to identify inhibitors of this antimicrobial drug target. In the reported structures the active site of the protein can simultaneously bind multiple ligands introduced in the crystallization solution or via soaking. Pseudomonas aeruginosa is an opportunistic human pathogen for which new antimicrobial drug options are urgently sought. P. aeruginosa disulfide-bond protein A1 (PaDsbA1) plays a pivotal role in catalyzing the oxidative folding of multiple virulence proteins and as such holds great promise as a drug target. As part of a fragment-based lead discovery approach to PaDsbA1 inhibitor development, the identification of a crystal form of PaDsbA1 that was more suitable for fragment-soaking experiments was sought. A previously identified crystallization condition for this protein was unsuitable, as in this crystal form of PaDsbA1 the active-site surface loops are engaged in the crystal packing, occluding access to the target site. A single residue involved in crystal-packing interactions was substituted with an amino acid commonly found at this position in closely related enzymes, and this variant was successfully used to generate a new crystal form of PaDsbA1 in which the active-site surface is more accessible for soaking experiments. The PaDsbA1 variant displays identical redox character and in vitro activity to wild-type PaDsbA1 and is structurally highly similar. Two crystal structures of the PaDsbA1 variant were determined in complex with small molecules bound to the protein active site. These small molecules (MES, glycerol and ethylene glycol) were derived from the crystallization or cryoprotectant solutions and provide a proof of principle that the reported crystal form will be amenable to co-crystallization and soaking with small molecules designed to target the protein active-site surface.

  20. Increased sensitivity of BRCA defective triple negative breast tumors to plumbagin through induction of DNA Double Strand Breaks (DSB).

    PubMed

    Nair, Rakesh Sathish; Kumar, Jerald Mahesh; Jose, Jedy; Somasundaram, Veena; Hemalatha, Sreelatha K; Sengodan, Satheesh Kumar; Nadhan, Revathy; Anilkumar, Thapasimuthu V; Srinivas, Priya

    2016-01-01

    We have earlier shown that Plumbagin (PB) can induce selective cytotoxicity to BRCA1 defective ovarian cancer cells; however, the effect of this molecule in BRCA1 mutated breast cancers has not been analyzed yet. Here, we report that reactive oxygen species (ROS) induced by PB resulted in DNA DSB and activates downstream signaling by ATR/ATM kinases and subsequent apoptosis. PB reduces DNA- dependent protein kinase (DNA-PK) expression and inhibits NHEJ (Non Homologous End Joining) activity in BRCA1 defective breast cancer cells. Also, PB induces apoptosis in two different BRCA1 conditional knock out murine models: MMTV-Cre; BRCA1(Co/Co) and WAP-Cre; BRCA1(Co/Co), at 2 mg/kg body weight, but 32 mg/kg of carboplatin (CN) was needed to induce apoptosis in them. This is the first study where two different tissue specific promoter driven transgenic mice models with BRCA1 exon 11 deletions are used for preclinical drug testing. The apoptosis induced by PB in HR (Homologous Recombination) defective triple negative BRCA1 mutant cell lines and in mouse models occur by inducing ROS mediated DNA DSB. The toxicity profile as compared with CN in transgenic mice provides evidence for PB's safer disposition as a therapeutic lead in breast cancer drug development. PMID:27220670

  1. Increased sensitivity of BRCA defective triple negative breast tumors to plumbagin through induction of DNA Double Strand Breaks (DSB)

    PubMed Central

    Nair, Rakesh Sathish; Kumar, Jerald Mahesh; Jose, Jedy; Somasundaram, Veena; Hemalatha, Sreelatha K.; Sengodan, Satheesh Kumar; Nadhan, Revathy; Anilkumar, Thapasimuthu V.; Srinivas, Priya

    2016-01-01

    We have earlier shown that Plumbagin (PB) can induce selective cytotoxicity to BRCA1 defective ovarian cancer cells; however, the effect of this molecule in BRCA1 mutated breast cancers has not been analyzed yet. Here, we report that reactive oxygen species (ROS) induced by PB resulted in DNA DSB and activates downstream signaling by ATR/ATM kinases and subsequent apoptosis. PB reduces DNA- dependent protein kinase (DNA-PK) expression and inhibits NHEJ (Non Homologous End Joining) activity in BRCA1 defective breast cancer cells. Also, PB induces apoptosis in two different BRCA1 conditional knock out murine models: MMTV-Cre; BRCA1Co/Co and WAP-Cre; BRCA1Co/Co, at 2 mg/kg body weight, but 32 mg/kg of carboplatin (CN) was needed to induce apoptosis in them. This is the first study where two different tissue specific promoter driven transgenic mice models with BRCA1 exon 11 deletions are used for preclinical drug testing. The apoptosis induced by PB in HR (Homologous Recombination) defective triple negative BRCA1 mutant cell lines and in mouse models occur by inducing ROS mediated DNA DSB. The toxicity profile as compared with CN in transgenic mice provides evidence for PB’s safer disposition as a therapeutic lead in breast cancer drug development. PMID:27220670

  2. Co-expression of Dsb proteins enables soluble expression of a single-chain variable fragment (scFv) against human type 1 insulin-like growth factor receptor (IGF-1R) in E. coli.

    PubMed

    Sun, Xue-Wen; Wang, Xiao-Hua; Yao, Yan-Bing

    2014-12-01

    Type 1 insulin-like growth factor receptor (IGF-1R) is a promising therapeutic target for cancer treatment. A single-chain variable fragment (scFv) against human IGF-1R forms inclusion body when expressed in periplasmic space of E. coli routinely. Here, we described that co-expression of appropriate disulfide bonds (Dsb) proteins known to catalyze the formation and isomerization of Dsb can markedly recover the soluble expression of target scFv in E. coli. A 50 % recovery in solubility of the scFv was observed upon co-expression of DsbC alone, and a maximum solubility (80 %) was obtained when DsbA and DsbC were co-expressed in combination. Furthermore, the soluble scFv present full antigen-binding activity with IGF-1R, suggesting its correct folding. This study also suggested that the selection of Dsb proteins should be tested case-by-case if the approach of co-expression of Dsb system is adopted to address the problem of insoluble expression of proteins carrying Dsb. PMID:25256416

  3. miR-3940-5p enhances homologous recombination after DSB in Cr(VI) exposed 16HBE cell.

    PubMed

    Li, Yang; Hu, Guiping; Li, Ping; Tang, Shichuan; Zhang, Ji; Jia, Guang

    2016-02-17

    Hexavalent chromium (Cr(VI)) is a well-recognized human carcinogen, yet the molecular mechanisms by which cause human cancer are still not well understood. MicroRNAs (miRNAs), which are small non-coding RNAs, are involved in carcinogenesis and DNA damage repair. Previous occupational population study showed that hexavalent chromium (Cr(VI)) downregulated plasma miR-3940-5p level, and a low miR-3940-5p level was associated with high XRCC2 expression in lymphocytes, indicating that miR-3940-5p maybe play a protective effect in Cr(VI) induced DNA damage. Here we investigated miR-3940-5p expression and its roles in DNA repair in Cr(VI)-treated 16HBE cells. miR-3940-5p change was detected by qRT-PCR. Rad51 foci formation and double strand break (DSB) were investigated to assess homologous recombination repair (HR) capacity by Immunofluorescent assay and Neutral Comet assay. XRCC2 expression was also evaluated after miRNA oligonucleotides transfection using Western blot. Cr(VI) treatment suppressed miR-3940-5p level in 16HBE cells. miR-3904-5p mimic downregulated XRCC2 expression. As a result, the formation of Rad51-foci was inhibited and DSB repair was prolonged. The results indicate that miR-3940-5p plays a protective effect in Cr(VI) induced DNA damage. PMID:26860703

  4. Novel Biological Approaches for Testing the Contributions of Single DSBs and DSB Clusters to the Biological Effects of High LET Radiation

    PubMed Central

    Mladenova, Veronika; Mladenov, Emil; Iliakis, George

    2016-01-01

    The adverse biological effects of ionizing radiation (IR) are commonly attributed to the generation of DNA double-strand breaks (DSBs). IR-induced DSBs are generated by clusters of ionizations, bear damaged terminal nucleotides, and frequently comprise base damages and single-strand breaks in the vicinity generating a unique DNA damage-clustering effect that increases DSB “complexity.” The number of ionizations in clusters of different radiation modalities increases with increasing linear energy transfer (LET), and is thought to determine the long-known LET-dependence of the relative biological effectiveness (RBE). Multiple ionizations may also lead to the formation of DSB clusters, comprising two or more DSBs that destabilize chromatin further and compromise overall processing. DSB complexity and DSB-cluster formation are increasingly considered in the development of mathematical models of radiation action, which are then “tested” by fitting available experimental data. Despite a plethora of such mathematical models the ultimate goal, i.e., the “a priori” prediction of the radiation effect, has not yet been achieved. The difficulty partly arises from unsurmountable difficulties in testing the fundamental assumptions of such mathematical models in defined biological model systems capable of providing conclusive answers. Recently, revolutionary advances in methods allowing the generation of enzymatic DSBs at random or in well-defined locations in the genome, generate unique testing opportunities for several key assumptions frequently fed into mathematical modeling – including the role of DSB clusters in the overall effect. Here, we review the problematic of DSB-cluster formation in radiation action and present novel biological technologies that promise to revolutionize the way we address the biological consequences of such lesions. We describe new ways of exploiting the I-SceI endonuclease to generate DSB-clusters at random locations in the genome and

  5. Novel Biological Approaches for Testing the Contributions of Single DSBs and DSB Clusters to the Biological Effects of High LET Radiation.

    PubMed

    Mladenova, Veronika; Mladenov, Emil; Iliakis, George

    2016-01-01

    The adverse biological effects of ionizing radiation (IR) are commonly attributed to the generation of DNA double-strand breaks (DSBs). IR-induced DSBs are generated by clusters of ionizations, bear damaged terminal nucleotides, and frequently comprise base damages and single-strand breaks in the vicinity generating a unique DNA damage-clustering effect that increases DSB "complexity." The number of ionizations in clusters of different radiation modalities increases with increasing linear energy transfer (LET), and is thought to determine the long-known LET-dependence of the relative biological effectiveness (RBE). Multiple ionizations may also lead to the formation of DSB clusters, comprising two or more DSBs that destabilize chromatin further and compromise overall processing. DSB complexity and DSB-cluster formation are increasingly considered in the development of mathematical models of radiation action, which are then "tested" by fitting available experimental data. Despite a plethora of such mathematical models the ultimate goal, i.e., the "a priori" prediction of the radiation effect, has not yet been achieved. The difficulty partly arises from unsurmountable difficulties in testing the fundamental assumptions of such mathematical models in defined biological model systems capable of providing conclusive answers. Recently, revolutionary advances in methods allowing the generation of enzymatic DSBs at random or in well-defined locations in the genome, generate unique testing opportunities for several key assumptions frequently fed into mathematical modeling - including the role of DSB clusters in the overall effect. Here, we review the problematic of DSB-cluster formation in radiation action and present novel biological technologies that promise to revolutionize the way we address the biological consequences of such lesions. We describe new ways of exploiting the I-SceI endonuclease to generate DSB-clusters at random locations in the genome and describe the

  6. RADIATION SENSITIVITY & PROCESSING OF DNA DAMAGE FOLLOWING LOW DOSES OF GAMMA-RAY ALPHA PARTICLES & HZE IRRADIATION OF NORMAL DSB REPAIR DEFICIENT CELLS

    SciTech Connect

    O'Neil, Peter

    2009-05-15

    Non-homologous end joining (NHEJ) predominates in the repair of DNA double strand breaks (DSB) over homologous recombination (HR). NHEJ occurs throughout the cell cycle whereas HR occurs in late S/G2 due to the requirement of a sister chromatid (Rothkamm et al, Mol Cell Biol 23 5706-15 [2003]). To date evidence obtained with DSB repair deficient cells using pulsed-field gel electrophoresis has revealed the major pathway throughout all phases of the cell cycle for processing high dose induced DSBs is NHEJ (Wang et al, Oncogene 20 2212-24 (2001); Pluth et al, Cancer Res. 61 2649-55 [2001]). These findings however were obtained at high doses when on average >> 20-30 DSBs are formed per cell. The contribution of the repair pathways (NHEJ and HR) induced in response to DNA damage during the various phases of the cell cycle may depend upon the dose (the level of initial DSBs) especially since low levels of DSBs are induced at low dose. To date, low dose studies using NHEJ and HR deficient mutants have not been carried out to address this important question with radiations of different quality. The work presented here leads us to suggest that HR plays a relatively minor role in the repair of radiation-induced prompt DSBs. SSBs lead to the induction of DSBs which are associated specifically with S-phase cells consistent with the idea that they are formed at stalled replication forks in which HR plays a major role in repair. That DNA-PKcs is in some way involved in the repair of the precursors to replication-induced DSB remains an open question. Persistent non-DSB oxidative damage also leads to an increase in RAD51 positive DSBs. Both simple and complex non-DSB DNA damage may therefore contribute to indirect DSBs induced by ionising radiation at replication forks.

  7. Genistein sensitizes sarcoma cells in vitro and in vivo by enhancing apoptosis and by inhibiting DSB repair pathways

    PubMed Central

    Liu, X.X.; Sun, C.; Jin, X.D.; Li, P.; Zheng, X.G.; Zhao, T.; Li, Q.

    2016-01-01

    The aim of this work was to investigate the radiosensitization effects of genistein on mice sarcoma cells and the corresponding biological mechanisms in vitro and in vivo. Using the non-toxic dosage of 10 μM genistein, the sensitizer enhancement ratios after exposure to X-rays at 50% cell survival (IC50) was 1.45 for S180 cells. For mice cotreated with genistein and X-rays, the excised tumor tissues had reduced blood vessels and decreased size and volume compared with the control and irradiation-only groups. Moreover, a significant increase in apoptosis was accompanied by upregulation of Bax and downregulation of Bcl-2 in the mitochondria, and lots of cytochrome c being transferred to the cytoplasm. Furthermore, X-rays combined with genistein inhibited the activity of DNA-PKcs, so DNA-injured sites were dominated by Ku70/80, leading to incompleteness of homologous recombination (HR) and non-homologous end-joining (NHEJ) repairs and the eventual occurrence of cell apoptosis. Our study, for the first time, demonstrated that genistein sensitized sarcoma cells to X-rays and that this radiosensitizing effect depended on induction of the mitochondrial apoptosis pathway and inhibition of the double-strand break (DSB) repair pathways. PMID:26922091

  8. Genistein sensitizes sarcoma cells in vitro and in vivo by enhancing apoptosis and by inhibiting DSB repair pathways.

    PubMed

    Liu, X X; Sun, C; Jin, X D; Li, P; Zheng, X G; Zhao, T; Li, Q

    2016-06-01

    The aim of this work was to investigate the radiosensitization effects of genistein on mice sarcoma cells and the corresponding biological mechanisms in vitro and in vivo Using the non-toxic dosage of 10 μM genistein, the sensitizer enhancement ratios after exposure to X-rays at 50% cell survival (IC50) was 1.45 for S180 cells. For mice cotreated with genistein and X-rays, the excised tumor tissues had reduced blood vessels and decreased size and volume compared with the control and irradiation-only groups. Moreover, a significant increase in apoptosis was accompanied by upregulation of Bax and downregulation of Bcl-2 in the mitochondria, and lots of cytochrome c being transferred to the cytoplasm. Furthermore, X-rays combined with genistein inhibited the activity of DNA-PKcs, so DNA-injured sites were dominated by Ku70/80, leading to incompleteness of homologous recombination (HR) and non-homologous end-joining (NHEJ) repairs and the eventual occurrence of cell apoptosis. Our study, for the first time, demonstrated that genistein sensitized sarcoma cells to X-rays and that this radiosensitizing effect depended on induction of the mitochondrial apoptosis pathway and inhibition of the double-strand break (DSB) repair pathways. PMID:26922091

  9. Automated resonance assignment of the 21 kDa stereo-array isotope labeled thioldisulfide oxidoreductase DsbA

    NASA Astrophysics Data System (ADS)

    Schmidt, Elena; Ikeya, Teppei; Takeda, Mitsuhiro; Löhr, Frank; Buchner, Lena; Ito, Yutaka; Kainosho, Masatsune; Güntert, Peter

    2014-12-01

    The automated chemical shift assignment algorithm FLYA has been extended for use with stereo-array isotope labeled (SAIL) proteins to determine the sequence-specific resonance assignments of large proteins. Here we present the assignment of the backbone and sidechain chemical shifts of the 21 kDa thioldisulfide oxidoreductase DsbA from Escherichia coli that were determined with the SAIL-FLYA algorithm in conjunction with automated peak picking. No manual corrections of peak lists or assignments were applied. The assignments agreed with manually determined reference assignments in 95.4% of the cases if 16 input spectra were used, 94.1% if only 3D 13C/15N-resolved NOESY, CBCA(CO)NH, and 2D [13C/15N,1H]-HSQC were used, and 86.8% if exclusively 3D 13C/15N-resolved NOESY spectra were used. Considering only the assignments that are classified as reliable by the SAIL-FLYA algorithm, the degrees of agreement increased to 97.5%, 96.5%, and 94.2%, respectively. With our approach it is thus possible to automatically obtain almost complete and correct assignments of proteins larger than 20 kDa.

  10. KARP-1 works as a heterodimer with Ku70, but the function of KARP-1 cannot perfectly replace that of Ku80 in DSB repair

    SciTech Connect

    Koike, Manabu; Yutoku, Yasutomo; Koike, Aki

    2011-10-01

    Ku, the heterodimer of Ku70 and Ku80, plays an essential role in the DNA double-strand break (DSB) repair pathway, i.e., non-homologous end-joining (NHEJ). Two isoforms of Ku80 encoded by the same genes, namely, Ku80 and KARP-1 are expressed and function in primate cells, but not in rodent cells. Ku80 works as a heterodimer with Ku70. However, it is not yet clear whether KARP-1 forms a heterodimer with Ku70 and works as a heterodimer. Although KARP-1 appears to work in NHEJ, its physiological role remains unclear. In this study, we established and characterized EGFP-KARP-1-expressing xrs-6 cell lines, EGFP-KARP-1/xrs-6. We found that nuclear localization signal (NLS) of KARP-1 is localized in the C-terminal region. Our data showed that KARP-1 localizes within the nucleus in NLS-dependent and NLS-independent manner and forms a heterodimer with Ku70, and stabilizes Ku70. On the other hand, EGFP-KARP-1 could not perfectly complement the radiosensitivity and DSB repair activity of Ku80-deficient xrs-6 cells. Furthermore, KARP-1 could not accumulate at DSBs faster than Ku80, although EGFP-KARP-1 accumulates at DSBs. Our data demonstrate that the function of KARP-1 could not perfectly replace that of Ku80 in DSB repair, although KARP-1 has some biochemical properties, which resemble those of Ku80, and works as a heterodimer with Ku70. On the other hand, the number of EGFP-KARP-1-expressing xrs-6 cells showing pan-nuclear {gamma}-H2AX staining significantly increases following X-irradiation, suggesting that KARP-1 may have a novel role in DSB response.

  11. Defective DSB repair correlates with abnormal nuclear morphology and is improved with FTI treatment in Hutchinson-Gilford progeria syndrome fibroblasts

    SciTech Connect

    Constantinescu, Dan; Csoka, Antonei B.; Navara, Christopher S.; Schatten, Gerald P.

    2010-10-15

    Impaired DSB repair has been implicated as a molecular mechanism contributing to the accelerating aging phenotype in Hutchinson-Gilford progeria syndrome (HGPS), but neither the extent nor the cause of the repair deficiency has been fully elucidated. Here we perform a quantitative analysis of the steady-state number of DSBs and the repair kinetics of ionizing radiation (IR)-induced DSBs in HGPS cells. We report an elevated steady-state number of DSBs and impaired repair of IR-induced DSBs, both of which correlated strongly with abnormal nuclear morphology. We recreated the HGPS cellular phenotype in human coronary artery endothelial cells for the first time by lentiviral transduction of GFP-progerin, which also resulted in impaired repair of IR-induced DSBs, and which correlated with abnormal nuclear morphology. Farnesyl transferase inhibitor (FTI) treatment improved the repair of IR-induced DSBs, but only in HGPS cells whose nuclear morphology was also normalized. Interestingly, FTI treatment did not result in a statistically significant reduction in the higher steady-state number of DSBs. We also report a delay in localization of phospho-NBS1 and MRE11, MRN complex repair factors necessary for homologous recombination (HR) repair, to DSBs in HGPS cells. Our results demonstrate a correlation between nuclear structural abnormalities and the DSB repair defect, suggesting a mechanistic link that may involve delayed repair factor localization to DNA damage. Further, our results show that similar to other HGPS phenotypes, FTI treatment has a beneficial effect on DSB repair.

  12. Close, stable homolog juxtaposition during meiosis in budding yeast is dependent on meiotic recombination, occurs independently of synapsis, and is distinct from DSB-independent pairing contacts

    PubMed Central

    Peoples, Tamara L.; Dean, Eric; Gonzalez, Oscar; Lambourne, Lindsey; Burgess, Sean M.

    2002-01-01

    A site-specific recombination system that probes the relative probabilities that pairs of chromosomal loci collide with one another in living cells of budding yeast was used to explore the relative contributions of pairing, recombination, synaptonemal complex formation, and telomere clustering to the close juxtaposition of homologous chromosome pairs during meiosis. The level of Cre-mediated recombination between a pair of loxP sites located at an allelic position on homologous chromosomes was 13-fold greater than that between a pair of loxP sites located at ectopic positions on nonhomologous chromosomes. Mutations affecting meiotic recombination initiation and the processing of DNA double-strand breaks (DSBs) into single-end invasions (SEIs) reduced the levels of allelic Cre-mediated recombination levels by three- to sixfold. The severity of Cre/loxP phenotypes is presented in contrast to relatively weak DSB-independent pairing defects as assayed using fluorescence in situ hybridization for these mutants. Mutations affecting synaptonemal complex (SC) formation or crossover control gave wild-type levels of allelic Cre-mediated recombination. A delay in attaining maximum levels of allelic Cre-mediated recombination was observed for a mutant defective in telomere clustering. None of the mutants affected ectopic levels of recombination. These data suggest that stable, close homolog juxtaposition in yeast is distinct from pre-DSB pairing interactions, requires both DSB and SEI formation, but does not depend on crossovers or SC. PMID:12101126

  13. The UBC Domain Is Required for BRUCE to Promote BRIT1/MCPH1 Function in DSB Signaling and Repair Post Formation of BRUCE-USP8-BRIT1 Complex

    PubMed Central

    Ge, Chunmin; Che, Lixiao; Du, Chunying

    2015-01-01

    BRUCE is implicated in the regulation of DNA double-strand break response to preserve genome stability. It acts as a scaffold to tether USP8 and BRIT1, together they form a nuclear BRUCE-USP8-BRIT1 complex, where BRUCE holds K63-ubiquitinated BRIT1 from access to DSB in unstressed cells. Following DSB induction, BRUCE promotes USP8 mediated deubiquitination of BRIT1, a prerequisite for BRIT1 to be released from the complex and recruited to DSB by binding to γ-H2AX. BRUCE contains UBC and BIR domains, but neither is required for the scaffolding function of BRUCE mentioned above. Therefore, it remains to be determined whether they are required for BRUCE in DSB response. Here we show that the UBC domain, not the BIR domain, is required for BRUCE to promote DNA repair at a step post the formation of BRUCE-USP8-BRIT1 complex. Mutation or deletion of the BRUCE UBC domain did not disrupt the BRUCE-USP8-BRIT1 complex, but impaired deubiquitination and consequent recruitment of BRIT1 to DSB. This leads to impaired chromatin relaxation, decreased accumulation of MDC1, NBS1, pATM and RAD51 at DSB, and compromised homologous recombination repair of DNA DSB. These results demonstrate that in addition to the scaffolding function in complex formation, BRUCE has an E3 ligase function to promote BRIT1 deubiquitination by USP8 leading to accumulation of BRIT1 at DNA double-strand break. These data support a crucial role for BRUCE UBC activity in the early stage of DSB response. PMID:26683461

  14. Proteome Analysis of the Effect of Mucoid Conversion on Global Protein Expression in Pseudomonas aeruginosa Strain PAO1 Shows Induction of the Disulfide Bond Isomerase, DsbA

    PubMed Central

    Malhotra, Sonal; Silo-Suh, Laura A.; Mathee, Kalai; Ohman, Dennis E.

    2000-01-01

    Pseudomonas aeruginosa strains that cause chronic pulmonary infections in cystic fibrosis patients typically undergo mucoid conversion. The mucoid phenotype indicates alginate overproduction and is often due to defects in MucA, an antisigma factor that controls the activity of sigma-22 (AlgT [also called AlgU]), which is required for the activation of genes for alginate biosynthesis. In this study we hypothesized that mucoid conversion may be part of a larger response that activates genes other than those for alginate synthesis. To address this, a two-dimensional (2-D) gel analysis was employed to compare total proteins in strain PAO1 to those of its mucA22 derivative, PDO300, in order to identify protein levels enhanced by mucoid conversion. Six proteins that were clearly more abundant in the mucoid strain were observed. The amino termini of such proteins were determined and used to identify the gene products in the genomic database. Proteins involved in alginate biosynthesis were expected among these, and two (AlgA and AlgD) were identified. This result verified that the 2-D gel approach could identify gene products under sigma-22 control and upregulated by mucA mutation. Two other protein spots were also clearly upregulated in the mucA22 background, and these were identified as porin F (an outer membrane protein) and a homologue of DsbA (a disulfide bond isomerase). Single-copy gene fusions were constructed to test whether these proteins were enhanced in the mucoid strain due to increased transcription. The oprF-lacZ fusion showed little difference in levels of expression in the two strains. However, the dsbA-lacZ fusion showed two- to threefold higher expression in PDO300 than in PAO1, suggesting that its promoter was upregulated by the deregulation of sigma-22 activity. A dsbA-null mutant was constructed in PAO1 and shown to have defects predicted for a cell with reduced disulfide bond isomerase activity, namely, reduction in periplasmic alkaline phosphatase

  15. Novel Image Processing Interface to Relate DSB Spatial Distribution from Immunofluorescence Foci Experiments to the State-of-the-Art Models of DNA Breakage

    NASA Technical Reports Server (NTRS)

    Ponomarev, A. L.; Cucinotta, F. A.

    2004-01-01

    A recently developed software (NASARadiationTrackImage) allows a quick and automatic segmentation of foci that indicate spatial localization of specific proteins that are visualized by immunofluorescence. Of interest are the spatial and temporal distribution of foci such as gammaH2AX, a signal of the phosphorylation of a variant of the histone H2A that has been shown to correspond to DSBs, or proteins involved in DSB processing, such as ATM, Rad51, and p53, following exposures of human cells to high charge and energy (HZE) ion irradiation. Experimental data are recorded as sets of two-dimensional images in color with cells and foci of gammaH2AX, ATM, Rad51 or others shown. Different cells, levels of radiation and timing after radiation were recorded. The software allows us to calculate the number of foci per cell, overall intensity of light in foci and their spatial organization. A simple statistical model allows for testing of foci overlap (eclipse). A more complex statistical model previously known as DNAbreak simulates track structure and random chromosome geometry. It has one adjustable parameter corresponding to an average intensity of DSB creation in cubic micrometers of DNA volume per particle track or unit dose. Its limitation is the low-resolution limit both in physical space and DSB's along DNA. It works adequately on the scale of a cell and provides further insights on how the geometry of tracks and DNA affects genomic damage of the cell and subsequent repair. Future developments of the model for the description of the time evolution of DNA damage response proteins, and more robust track structure models will be discussed.

  16. Asymmetrical TWDM-PON with 4 × 25-Gb/s downstream DSB OFDM and 4 × 10-Gb/s upstream OOK modulations

    NASA Astrophysics Data System (ADS)

    Lin, Bangjiang; Li, Yiwei; Zhang, Shihao; Tang, Xuan

    2015-12-01

    We propose and experimentally demonstrate an asymmetrical time wavelength division multiplexing passive optical network (TWDM-PON) scheme with 100-Gb/s downstream and 40-Gb/s upstream capacity using four pairs of wavelengths. Double sideband (DSB) orthogonal frequency division multiplexing (OFDM) with 16-QAM mapping is used for downstream transmission, while on-off keying (OOK) modulation is used for upstream transmission. The experimental results show that the power budget for bidirectional transmission is more than 35.5-dB. This scheme can be a promising solution for 100-Gb/s TWDM-PON with high power budget.

  17. Biochemical kinetics model of DSB repair and induction of gamma-H2AX foci by non-homologous end joining.

    PubMed

    Cucinotta, Francis A; Pluth, Janice M; Anderson, Jennifer A; Harper, Jane V; O'Neill, Peter

    2008-02-01

    We developed a biochemical kinetics approach to describe the repair of double-strand breaks (DSBs) produced by low-LET radiation by modeling molecular events associated with non-homologous end joining (NHEJ). A system of coupled nonlinear ordinary differential equations describes the induction of DSBs and activation pathways for major NHEJ components including Ku70/80, DNA-PKcs, and the ligase IV-XRCC4 heterodimer. The autophosphorylation of DNA-PKcs and subsequent induction of gamma-H2AX foci observed after ionizing radiation exposure were modeled. A two-step model of regulation of repair by DNA-PKcs was developed with an initial step allowing access of other NHEJ components to breaks and a second step limiting access to ligase IV-XRCC4. Our model assumes that the transition from the first to the second step depends on DSB complexity, with a much slower rate for complex DSBs. The model faithfully reproduced several experimental data sets, including DSB rejoining as measured by pulsed-field gel electrophoresis (PFGE) at 10 min postirradiation or longer and quantification of the induction of gamma-H2AX foci. A process that is independent of DNA-PKcs is required for the model to reproduce experimental data for rejoining before 10 min postirradiation. Predictions are made for the behaviors of NHEJ components at low doses and dose rates, and a steady state is found at dose rates of 0.1 Gy/h or lower. PMID:18220463

  18. The oxidoreductase DsbA plays a key role in the ability of the Crohn's disease-associated adherent-invasive Escherichia coli strain LF82 to resist macrophage killing.

    PubMed

    Bringer, Marie-Agnès; Rolhion, Nathalie; Glasser, Anne-Lise; Darfeuille-Michaud, Arlette

    2007-07-01

    Adherent-invasive Escherichia coli (AIEC) isolated from Crohn's disease patients is able to adhere to and invade intestinal epithelial cells and to replicate in mature phagolysosomes within macrophages. Here, we show that the dsbA gene, encoding a periplasmic oxidoreductase, was required for AIEC strain LF82 to adhere to intestinal epithelial cells and to survive within macrophages. The LF82-DeltadsbA mutant did not express flagella and, probably as a consequence of this, did not express type 1 pili. The role of DsbA in adhesion is restricted to the loss of flagella and type 1 pili, as forced contact between bacteria and cells and induced expression of type 1 pili restored the wild-type phenotype. In contrast, the dsbA gene is essential for AIEC LF82 bacteria to survive within macrophages, irrespective of the loss of flagella and type 1 pilus expression, and the survival ability of LF82-DeltadsbA was as low as that of the nonpathogenic E. coli K-12, which was efficiently killed by macrophages. We also provide evidence that the dsbA gene is needed for LF82 bacteria to grow and survive in an acidic and nutrient-poor medium that partly mimics the harsh environment of the phagocytic vacuole. In addition, under such stress conditions dsbA transcription is highly up-regulated. Finally, the CpxRA signaling pathway does not play a role in regulation of dsbA expression in AIEC LF82 bacteria under conditions similar to those of mature phagolysosomes. PMID:17449627

  19. Induction and Processing of the Radiation-Induced Gamma-H2AX Signal and Its Link to the Underlying Pattern of DSB: A Combined Experimental and Modelling Study

    PubMed Central

    Tommasino, Francesco; Friedrich, Thomas; Jakob, Burkhard; Meyer, Barbara; Durante, Marco; Scholz, Michael

    2015-01-01

    We present here an analysis of DSB induction and processing after irradiation with X-rays in an extended dose range based on the use of the γH2AX assay. The study was performed by quantitative flow cytometry measurements, since the use of foci counting would result in reasonable accuracy only in a limited dose range of a few Gy. The experimental data are complemented by a theoretical analysis based on the GLOBLE model. In fact, original aim of the study was to test GLOBLE predictions against new experimental data, in order to contribute to the validation of the model. Specifically, the γH2AX signal kinetics has been investigated up to 24 h after exposure to increasing photon doses between 2 and 500 Gy. The prolonged persistence of the signal at high doses strongly suggests dose dependence in DSB processing after low LET irradiation. Importantly, in the framework of our modelling analysis, this is related to a gradually increased fraction of DSB clustering at the micrometre scale. The parallel study of γH2AX dose response curves shows the onset of a pronounced saturation in two cell lines at a dose of about 20 Gy. This dose is much lower than expected according to model predictions based on the values usually adopted for the DSB induction yield (≈ 30 DSB/Gy) and for the γH2AX foci extension of approximately 2 Mbp around the DSB. We show and discuss how theoretical predictions and experimental findings can be in principle reconciled by combining an increased DSB induction yield with the assumption of a larger genomic extension for the single phosphorylated regions. As an alternative approach, we also considered in our model the possibility of a 3D spreading-mechanism of the H2AX phosphorylation around the induced DSB, and applied it to the analysis of both the aspects considered. Our results are found to be supportive for the basic assumptions on which GLOBLE is built. Apart from giving new insights into the H2AX phosphorylation process, experiments performed

  20. TrbB from Conjugative Plasmid F Is a Structurally Distinct Disulfide Isomerase That Requires DsbD for Redox State Maintenance ▿ †

    PubMed Central

    Hemmis, Casey W.; Berkmen, Mehmet; Eser, Markus; Schildbach, Joel F.

    2011-01-01

    TrbB, a periplasmic protein encoded by the conjugative plasmid F, has a predicted thioredoxin-like fold and possesses a C-X-X-C redox active site motif. TrbB may function in the conjugative process by serving as a disulfide bond isomerase, facilitating proper folding of a subset of F-plasmid-encoded proteins in the periplasm. Previous studies have demonstrated that a ΔtrbB F plasmid in Escherichia coli lacking DsbCE.coli, its native disulfide bond isomerase, experiences a 10-fold decrease in mating efficiency but have not provided direct evidence for disulfide bond isomerase activity. Here we demonstrate that trbB can partially restore transfer of a variant of the distantly related R27 plasmid when both chromosomal and plasmid genes encoding disulfide bond isomerases have been disrupted. In addition, we show that TrbB displays both disulfide bond isomerase and reductase activities on substrates not involved in the conjugative process. Unlike canonical members of the disulfide bond isomerase family, secondary structure predictions suggest that TrbB lacks both an N-terminal dimerization domain and an α-helical domain found in other disulfide bond isomerases. Phylogenetic analyses support the conclusion that TrbB belongs to a unique family of plasmid-based disulfide isomerases. Interestingly, although TrbB diverges structurally from other disulfide bond isomerases, we show that like those isomerases, TrbB relies on DsbD from E. coli for maintenance of its C-X-X-C redox active site motif. PMID:21742866

  1. Phylogeny of the Vitamin K 2,3-Epoxide Reductase (VKOR) Family and Evolutionary Relationship to the Disulfide Bond Formation Protein B (DsbB) Family

    PubMed Central

    Bevans, Carville G.; Krettler, Christoph; Reinhart, Christoph; Watzka, Matthias; Oldenburg, Johannes

    2015-01-01

    In humans and other vertebrate animals, vitamin K 2,3-epoxide reductase (VKOR) family enzymes are the gatekeepers between nutritionally acquired K vitamins and the vitamin K cycle responsible for posttranslational modifications that confer biological activity upon vitamin K-dependent proteins with crucial roles in hemostasis, bone development and homeostasis, hormonal carbohydrate regulation and fertility. We report a phylogenetic analysis of the VKOR family that identifies five major clades. Combined phylogenetic and site-specific conservation analyses point to clade-specific similarities and differences in structure and function. We discovered a single-site determinant uniquely identifying VKOR homologs belonging to human pathogenic, obligate intracellular prokaryotes and protists. Building on previous work by Sevier et al. (Protein Science 14:1630), we analyzed structural data from both VKOR and prokaryotic disulfide bond formation protein B (DsbB) families and hypothesize an ancient evolutionary relationship between the two families where one family arose from the other through a gene duplication/deletion event. This has resulted in circular permutation of primary sequence threading through the four-helical bundle protein folds of both families. This is the first report of circular permutation relating distant α-helical membrane protein sequences and folds. In conclusion, we suggest a chronology for the evolution of the five extant VKOR clades. PMID:26230708

  2. A human iPSC model of Ligase IV deficiency reveals an important role for NHEJ-mediated-DSB repair in the survival and genomic stability of induced pluripotent stem cells and emerging haematopoietic progenitors

    PubMed Central

    Tilgner, K; Neganova, I; Moreno-Gimeno, I; AL-Aama, J Y; Burks, D; Yung, S; Singhapol, C; Saretzki, G; Evans, J; Gorbunova, V; Gennery, A; Przyborski, S; Stojkovic, M; Armstrong, L; Jeggo, P; Lako, M

    2013-01-01

    DNA double strand breaks (DSBs) are the most common form of DNA damage and are repaired by non-homologous-end-joining (NHEJ) or homologous recombination (HR). Several protein components function in NHEJ, and of these, DNA Ligase IV is essential for performing the final ‘end-joining' step. Mutations in DNA Ligase IV result in LIG4 syndrome, which is characterised by growth defects, microcephaly, reduced number of blood cells, increased predisposition to leukaemia and variable degrees of immunodeficiency. In this manuscript, we report the creation of a human induced pluripotent stem cell (iPSC) model of LIG4 deficiency, which accurately replicates the DSB repair phenotype of LIG4 patients. Our findings demonstrate that impairment of NHEJ-mediated-DSB repair in human iPSC results in accumulation of DSBs and enhanced apoptosis, thus providing new insights into likely mechanisms used by pluripotent stem cells to maintain their genomic integrity. Defects in NHEJ-mediated-DSB repair also led to a significant decrease in reprogramming efficiency of human cells and accumulation of chromosomal abnormalities, suggesting a key role for NHEJ in somatic cell reprogramming and providing insights for future cell based therapies for applications of LIG4-iPSCs. Although haematopoietic specification of LIG4-iPSC is not affected per se, the emerging haematopoietic progenitors show a high accumulation of DSBs and enhanced apoptosis, resulting in reduced numbers of mature haematopoietic cells. Together our findings provide new insights into the role of NHEJ-mediated-DSB repair in the survival and differentiation of progenitor cells, which likely underlies the developmental abnormalities observed in many DNA damage disorders. In addition, our findings are important for understanding how genomic instability arises in pluripotent stem cells and for defining appropriate culture conditions that restrict DNA damage and result in ex vivo expansion of stem cells with intact genomes. PMID

  3. A human iPSC model of Ligase IV deficiency reveals an important role for NHEJ-mediated-DSB repair in the survival and genomic stability of induced pluripotent stem cells and emerging haematopoietic progenitors.

    PubMed

    Tilgner, K; Neganova, I; Moreno-Gimeno, I; Al-Aama, J Y; Burks, D; Yung, S; Singhapol, C; Saretzki, G; Evans, J; Gorbunova, V; Gennery, A; Przyborski, S; Stojkovic, M; Armstrong, L; Jeggo, P; Lako, M

    2013-08-01

    DNA double strand breaks (DSBs) are the most common form of DNA damage and are repaired by non-homologous-end-joining (NHEJ) or homologous recombination (HR). Several protein components function in NHEJ, and of these, DNA Ligase IV is essential for performing the final 'end-joining' step. Mutations in DNA Ligase IV result in LIG4 syndrome, which is characterised by growth defects, microcephaly, reduced number of blood cells, increased predisposition to leukaemia and variable degrees of immunodeficiency. In this manuscript, we report the creation of a human induced pluripotent stem cell (iPSC) model of LIG4 deficiency, which accurately replicates the DSB repair phenotype of LIG4 patients. Our findings demonstrate that impairment of NHEJ-mediated-DSB repair in human iPSC results in accumulation of DSBs and enhanced apoptosis, thus providing new insights into likely mechanisms used by pluripotent stem cells to maintain their genomic integrity. Defects in NHEJ-mediated-DSB repair also led to a significant decrease in reprogramming efficiency of human cells and accumulation of chromosomal abnormalities, suggesting a key role for NHEJ in somatic cell reprogramming and providing insights for future cell based therapies for applications of LIG4-iPSCs. Although haematopoietic specification of LIG4-iPSC is not affected per se, the emerging haematopoietic progenitors show a high accumulation of DSBs and enhanced apoptosis, resulting in reduced numbers of mature haematopoietic cells. Together our findings provide new insights into the role of NHEJ-mediated-DSB repair in the survival and differentiation of progenitor cells, which likely underlies the developmental abnormalities observed in many DNA damage disorders. In addition, our findings are important for understanding how genomic instability arises in pluripotent stem cells and for defining appropriate culture conditions that restrict DNA damage and result in ex vivo expansion of stem cells with intact genomes. PMID

  4. Structural insights into NHEJ: building up an integrated picture of the dynamic DSB repair super complex, one component and interaction at a time

    PubMed Central

    Williams, Gareth J.; Hammel, Michal; Radhakrishnan, Sarvan Kumar; Ramsden, Dale; Lees-Miller, Susan P.; Tainer, John A.

    2014-01-01

    Non-homologous end joining (NHEJ) is the major pathway for repair of DNA double-strand breaks (DSBs) in human cells. NHEJ is also needed for V(D)J recombination and the development of T and B cells in vertebrate immune systems, and acts in both the generation and prevention of non-homologous chromosomal translocations, a hallmark of genomic instability and many human cancers. X-ray crystal structures, cryo-electron microscopy envelopes, and small angle X-ray scattering (SAXS) solution conformations and assemblies are defining most of the core protein components for NHEJ: Ku70/Ku80 heterodimer; the DNA dependent protein kinase catalytic subunit (DNA-PKcs); the structure-specific endonuclease Artemis along with polynucleotide kinase/phosphatase (PNKP), aprataxin and PNKP related protein (APLF); the scaffolding proteins XRCC4 and XLF (XRCC4-like factor); DNA polymerases, and DNA ligase IV (Lig IV). The dynamic assembly of multi-protein NHEJ complexes at DSBs is regulated in part by protein phosphorylation. The basic steps of NHEJ have been biochemically defined to require: 1) DSB detection by the Ku heterodimer with subsequent DNA-PKcs tethering to form the DNA-PKcs-Ku-DNA complex (termed DNA-PK), 2) lesion processing, and 3) DNA end ligation by Lig IV, which functions in complex with XRCC4 and XLF. The current integration of structures by combined methods is resolving puzzles regarding the mechanisms, coordination and regulation of these three basic steps. Overall, structural results suggest the NHEJ system forms a flexing scaffold with the DNA-PKcs HEAT repeats acting as compressible macromolecular springs suitable to store and release conformational energy to apply forces to regulate NHEJ complexes and the DNA substrate for DNA end protection, processing, and ligation. PMID:24656613

  5. Robotics Training.

    ERIC Educational Resources Information Center

    Ettlie, John E.; And Others

    1988-01-01

    Discusses the need for training and education in new skill areas. Points out that the right people often do not get the right training. Too often engineers and skilled workers are trained to the exclusion of supervisors and operators. (JOW)

  6. Acute Normal Tissue Reactions in Head-and-Neck Cancer Patients Treated With IMRT: Influence of Dose and Association With Genetic Polymorphisms in DNA DSB Repair Genes

    SciTech Connect

    Werbrouck, Joke Ruyck, Kim de; Duprez, Frederic; Veldeman, Liv; Claes, Kathleen; Eijkeren, Marc van; Boterberg, Tom; Willems, Petra; Vral, Anne; Neve, Wilfried de; Thierens, Hubert

    2009-03-15

    Purpose: To investigate the association between dose-related parameters and polymorphisms in DNA DSB repair genes XRCC3 (c.-1843A>G, c.562-14A>G, c.722C>T), Rad51 (c.-3429G>C, c.-3392G>T), Lig4 (c.26C>T, c.1704T>C), Ku70 (c.-1310C>G), and Ku80 (c.2110-2408G>A) and the occurrence of acute reactions after radiotherapy. Materials and Methods: The study population consisted of 88 intensity-modulated radiation therapy (IMRT)-treated head-and-neck cancer patients. Mucositis, dermatitis, and dysphagia were scored using the Common Terminology Criteria (CTC) for Adverse Events v.3.0 scale. The population was divided into a CTC0-2 and CTC3+ group for the analysis of each acute effect. The influence of the dose on critical structures was analyzed using dose-volume histograms. Genotypes were determined by polymerase chain reaction (PCR) combined with restriction fragment length polymorphism or PCR-single base extension assays. Results: The mean dose (D{sub mean}) to the oral cavity and constrictor pharyngeus (PC) muscles was significantly associated with the development of mucositis and dysphagia, respectively. These parameters were considered confounding factors in the radiogenomics analyses. The XRCC3c.722CT/TT and Ku70c.-1310CG/GG genotypes were significantly associated with the development of severe dysphagia (CTC3+). No association was found between the investigated polymorphisms and the development of mucositis or dermatitis. A risk analysis model for severe dysphagia, which was developed based on the XRCC3c.722CT/TT and Ku70c.-1310CG/GG genotypes and the PC dose, showed a sensitivity of 78.6% and a specificity of 77.6%. Conclusions: The XRCC3c.722C>T and Ku70c.-1310C>G polymorphisms as well as the D{sub mean} to the PC muscles were highly associated with the development of severe dysphagia after IMRT. The prediction model developed using these parameters showed a high sensitivity and specificity.

  7. Development of a radiation track structure clustering algorithm for the prediction of DNA DSB yields and radiation induced cell death in Eukaryotic cells

    NASA Astrophysics Data System (ADS)

    Douglass, Michael; Bezak, Eva; Penfold, Scott

    2015-04-01

    The preliminary framework of a combined radiobiological model is developed and calibrated in the current work. The model simulates the production of individual cells forming a tumour, the spatial distribution of individual ionization events (using Geant4-DNA) and the stochastic biochemical repair of DNA double strand breaks (DSBs) leading to the prediction of survival or death of individual cells. In the current work, we expand upon a previously developed tumour generation and irradiation model to include a stochastic ionization damage clustering and DNA lesion repair model. The Geant4 code enabled the positions of each ionization event in the cells to be simulated and recorded for analysis. An algorithm was developed to cluster the ionization events in each cell into simple and complex double strand breaks. The two lesion kinetic (TLK) model was then adapted to predict DSB repair kinetics and the resultant cell survival curve. The parameters in the cell survival model were then calibrated using experimental cell survival data of V79 cells after low energy proton irradiation. A monolayer of V79 cells was simulated using the tumour generation code developed previously. The cells were then irradiated by protons with mean energies of 0.76 MeV and 1.9 MeV using a customized version of Geant4. By replicating the experimental parameters of a low energy proton irradiation experiment and calibrating the model with two sets of data, the model is now capable of predicting V79 cell survival after low energy (<2 MeV) proton irradiation for a custom set of input parameters. The novelty of this model is the realistic cellular geometry which can be irradiated using Geant4-DNA and the method in which the double strand breaks are predicted from clustering the spatial distribution of ionisation events. Unlike the original TLK model which calculates a tumour average cell survival probability, the cell survival probability is calculated for each cell in the geometric tumour model

  8. Training Visions

    ERIC Educational Resources Information Center

    Training, 2011

    2011-01-01

    In this article, "Training" asks the 2011 winners to give their predictions for what training--either in general or specifically at their companies--will look like in the next five to 10 years. Perhaps their "training visions" will spark some ideas in one's organization--or at least help prepare for what might be coming in the next decade or so.

  9. [Teacher Training.

    ERIC Educational Resources Information Center

    Palmatier, Robert A., Ed.

    1977-01-01

    This issue collects three articles concerning reading-teacher training. "Language, Failure, and Panda Bears" by Patricia M. Cunningham calls attention to dialect difficulties in the classroom and provides ideas for teacher training programs and for public schools to solve this problem. William H. Rupley, in "Improving Teacher Effectiveness in…

  10. Marketing Training

    NASA Technical Reports Server (NTRS)

    Leonard, Eric

    1998-01-01

    All of our ideas have been field tested and have proven effective in our environment. Our objectives are: We will share our ideas about marketing training and what we've implemented at Michoud Space Systems. You will go away with at least one new idea or insight about how to more effectively market your training.

  11. Project Training

    ERIC Educational Resources Information Center

    Cooper, Hubert

    1970-01-01

    The training manager of the Aero Engine Division of Rolls Royce Limited describes his graduate level program for retaining and developing mechanical engineers, mathematicians, scientists, and other college graduates. Three diagrams. (LY)

  12. Choosing Training?

    ERIC Educational Resources Information Center

    Stephen, Jennifer

    This guide is designed to help the user enter into the job market by making the most of their existing skills and finding additional training. Section 1, Vocations, Occupations, Careers, looks at the assessment tools used by employers and trainers to prepare people for today's job market. It describes how to develop a personal inventory of skills…

  13. Youth Training.

    ERIC Educational Resources Information Center

    General Accounting Office, Washington, DC. Program Evaluation and Methodology Div.

    A descriptive study was made of the roles of training and federal programs in helping youths gain employment in selected high-wage occupations that do not require a four-year college degree. Interviews conducted with federal agency officials and industry representatives found little hard data but elicited officials' views on this issue. The study…

  14. Training Unpolluters

    ERIC Educational Resources Information Center

    Manpower, 1970

    1970-01-01

    The shortage of trained manpower remains a troublesome problem that must be overcome if the national attack on water pollution succeeds. The total number of persons treating waste water in 1967 was 11,300 professionals, 10,300 technicians, 23,500 sewage treatment plant operators, and 50,000 maintenance workers. (BC)

  15. Employment Training.

    ERIC Educational Resources Information Center

    Further Education Unit, London (England).

    This document reviews the progress of the Employment Training (ET) initiative for employed adults in Great Britain. The document begins by explaining the changing context in which the project is operating, which includes a downgrading of the alternatives to ET, increased participation in ET, and increased employer involvement in ET. The response…

  16. Evaluating Training.

    ERIC Educational Resources Information Center

    Brethower, Karen S.; Rummler, Geary A.

    1979-01-01

    Presents general systems models (ballistic system, guided system, and adaptive system) and an evaluation matrix to help in examining training evaluation alternatives and in deciding what evaluation is appropriate. Includes some guidelines for conducting evaluation studies using four designs (control group, reversal, multiple baseline, and "before…

  17. Extreme Training

    ERIC Educational Resources Information Center

    Dolezalek, Holly

    2010-01-01

    Can one learn to lead on the flying trapeze? That's the question prompted by the exotic variations on the theme of experiential learning. By taking employees or executives (or both) out of their work element and putting them through an experience together, training professionals try to create learning that is more meaningful than PowerPoint…

  18. Toppling Trains.

    ERIC Educational Resources Information Center

    Parry, Malcolm

    1998-01-01

    Explains a novel way of approaching centripetal force: theory is used to predict an orbital period at which a toy train will topple from a circular track. The demonstration has elements of prediction (a criterion for a good model) and suspense (a criterion for a good demonstration). The demonstration proved useful in undergraduate physics and…

  19. Interval Training.

    ERIC Educational Resources Information Center

    President's Council on Physical Fitness and Sports, Washington, DC.

    Regardless of the type of physical activity used, interval training is simply repeated periods of physical stress interspersed with recovery periods during which activity of a reduced intensity is performed. During the recovery periods, the individual usually keeps moving and does not completely recover before the next exercise interval (e.g.,…

  20. Power Trains.

    ERIC Educational Resources Information Center

    Kukuk, Marvin; Mathis, Joe

    This curriculum guide is part of a series designed to teach students about diesel engines. The materials in this power trains guide apply to both on-road and off-road vehicles and include information about chain and belt drives used in tractors and combines. These instructional materials, containing nine units, are written in terms of student…

  1. Management Training, Yes! Excellence?

    ERIC Educational Resources Information Center

    Davis, Gary E.

    1990-01-01

    Management training programs are a necessity for transportation supervisors. Basic and advanced training programs are available through associations for business officials and university fleet management training programs. (MLF)

  2. Sports Training

    NASA Technical Reports Server (NTRS)

    1988-01-01

    Practitioners of martial arts have long seen a need for a precise method of measuring the power of a karate kick or a boxer's punch in training and competition. Impax sensor is a piezoelectric film less than one thousandth of an inch thick, yet extremely durable. They give out a voltage impulse when struck, the greater the force of impact, the higher the voltage. The impulse is transmitted to a compact electronics package where voltage is translated into a force-pounds reading shown on a digital display. Impax, manufactured by Impulse Technology, Inc. is used by martial arts instructors, practitioners, U.S. Olympic Committee Training Center, football blocking sleds, and boxers as well as police defensive tactics, providing a means of evaluating the performance of recruits.

  3. Train Analysis

    NASA Technical Reports Server (NTRS)

    1985-01-01

    Running resistance, the combined resistance induced by aerodynamic drag and mechanical friction, absorbs about 16 percent of railroad budget and maintenance costs. In order to study these losses, AT&SF Railroad entered into a joint research project with the Jet Propulsion Laboratory. Various train configurations were tested when the resulting data was computer analyzed. The Coast-Down Technique was estimated at one percent accuracy. AT&SF is evaluating the data further.

  4. Hazmat Training: Recognizing "Gremlins."

    ERIC Educational Resources Information Center

    Cherniak, Michael J.

    1992-01-01

    Presents suggestions and benefits of training programs that extend beyond government regulations for hazardous waste management training procedures. Discusses requirements, environmental concerns, identifying employees for training, administrative details that support training needs, and evaluations. (MCO)

  5. Training for Tech Transitions.

    ERIC Educational Resources Information Center

    InfoWorld, 1984

    1984-01-01

    Discusses nature of training objectives, sources of training materials, and who will direct the training. Also discusses a model training program at Penn Mutual Life Insurance, a customized approach to individual learning, and computer security. (JN)

  6. Education and Training.

    ERIC Educational Resources Information Center

    Social and Labour Bulletin, 1980

    1980-01-01

    Topics discussed include (1) establishment of a national vocational training service in Nicaragua; (2) vocational training in Senegal; (3) national vocational training week in the Ukraine; and (4) inplant training in the U.S.S.R. (SK)

  7. STS-93 Crew Training

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Live footage of the STS-93 crewmembers shows Commander Eileen M. Collins, Pilot Jeffrey S. Ashby, Mission Specialists Steven A. Hawley, Catherine G. Coleman, and Michel Tognini going through various training activities. These activities include Bail Out Training NBL, Emergency Egress Training, Earth Observations Classroom Training, Simulator Training, T-38 Departure from Ellington Field, Chandra Deploy Training, SAREX Shuttle Amateur Radio Experiment, CCT Bail Out Crew Compartment Training, and Southwest Research Ultraviolet Imaging System (SWUIS) Training.

  8. Media Training

    SciTech Connect

    2009-12-11

    With the LHC starting up soon, the world's media are again turning their attention to CERN. We're all likely to be called upon to explain what is happening at CERN to media, friends and neighbours. The seminar will be given by BBC television news journalists Liz Pike and Nadia Marchant, and will deal with the kind of questions we're likely to be confronted with through the restart period. The training is open for everybody. Make sure you arrive early enough to get a seat - there are only 200 seats in the Globe. The session will also be webcast: http://webcast.cern.ch/

  9. Media Training

    ScienceCinema

    None

    2011-10-06

    With the LHC starting up soon, the world's media are again turning their attention to CERN. We're all likely to be called upon to explain what is happening at CERN to media, friends and neighbours. The seminar will be given by BBC television news journalists Liz Pike and Nadia Marchant, and will deal with the kind of questions we're likely to be confronted with through the restart period. The training is open for everybody. Make sure you arrive early enough to get a seat - there are only 200 seats in the Globe. The session will also be webcast: http://webcast.cern.ch/

  10. The Pilot Training Study: Advanced Pilot Training.

    ERIC Educational Resources Information Center

    Kennedy, P. J.

    An overview is presented of advanced pilot training and of the formal advanced pilot training program that constitutes the primary means of providing this training. Section I deals with the various phases of advanced pilot training that a pilot may encounter during his career; Section II deals with the types of aircraft that require some form of…

  11. "Training Floors" and "Training Ceilings": Metonyms for Understanding Training Trends

    ERIC Educational Resources Information Center

    Felstead, Alan; Jewson, Nick

    2014-01-01

    This article outlines a conceptual framework for mapping and understanding training trends. It uses the metonyms of floors and ceilings to distinguish between different types of training configurations. The argument is made that the ups and downs of employer reports of training activity are a crude basis on which to make judgements about the…

  12. Advanced training systems

    NASA Technical Reports Server (NTRS)

    Savely, Robert T.; Loftin, R. Bowen

    1990-01-01

    Training is a major endeavor in all modern societies. Common training methods include training manuals, formal classes, procedural computer programs, simulations, and on-the-job training. NASA's training approach has focussed primarily on on-the-job training in a simulation environment for both crew and ground based personnel. NASA must explore new approaches to training for the 1990's and beyond. Specific autonomous training systems are described which are based on artificial intelligence technology for use by NASA astronauts, flight controllers, and ground based support personnel that show an alternative to current training systems. In addition to these specific systems, the evolution of a general architecture for autonomous intelligent training systems that integrates many of the features of traditional training programs with artificial intelligence techniques is presented. These Intelligent Computer Aided Training (ICAT) systems would provide much of the same experience that could be gained from the best on-the-job training.

  13. Pilot Training Study.

    ERIC Educational Resources Information Center

    Mooz, William E.

    The purpose of the Pilot Training Study is to produce tools with which to analyze the pilot training process of the Air Force in terms of the resources required to train pilots and the cost of pilot training. These tools allow examination of the training courses themselves, and also of the policy factors which drive the need for pilots. The tools…

  14. Management Training in Retailing.

    ERIC Educational Resources Information Center

    Veness, C. Rosina

    Intended for prospective members of the new Distributive Industrial Training Board in Great Britain, this training guide concentrates on managerial functions in retailing; the selection of trainees; the planning of in-company and external training programs; scheduling and continuity of training; roles of training personnel; and the use of various…

  15. 2011 Training Industry Report

    ERIC Educational Resources Information Center

    Training, 2011

    2011-01-01

    This article presents "Training" magazine's exclusive analysis of the U.S. training industry, featuring 2011 training expenditures, budgetary allocations, delivery methods, and training priorities. Now in its 30th year, The Industry Report is recognized as the training industry's most trusted source of data on budgets, staffing, and programs. This…

  16. STS-100 Crew Training

    NASA Technical Reports Server (NTRS)

    2001-01-01

    Footage shows the crew of STS-100, Commander Kent Rominger, Pilot Jeffrey Ashby, and Mission Specialists Chris Hadfield, Scott Parazynski, John Phillips, Umberto Guidoni, and Yuri Valentinovich Lonchakov, during various parts of their training, including the crew photo session, postlanding egress, extravehicular activity (EVA) large tool training, EVA training in the Neutral Buoyancy Laboratory (NBL), secondary payload training, and during VHF training.

  17. Training from the Start.

    ERIC Educational Resources Information Center

    Cocheu, Ted

    1990-01-01

    The time to plan training is in the research and development stage of a new product. Seven major steps are online training, process validation, skills certification, development of manufacturing training instructions, skills verification, transfer training, and offline training and certification. (SK)

  18. Spaced cognitive training promotes training transfer

    PubMed Central

    Wang, Zuowei; Zhou, Renlai; Shah, Priti

    2014-01-01

    Cognitive training studies yield wildly inconsistent results. One dimension on which studies vary is the scheduling of training sessions (Morrison and Chein, 2011). In this study, we systematically address whether or not spacing of practice influences training and transfer. We randomly assigned 115 fifth grade children to an active control group or one of four training groups who received working memory training based on a “running span” task (Zhao et al., 2011). All groups received the same total amount of training: 20 sessions of training with 60 trials for an average of 20 min per session. The training was spread across 2, 5, 10, or 20 days. The active control group received 20-min sessions of math instruction for 20 sessions. Before and after training participants in all five groups performed a single transfer test that assessed fluid intelligence, the Raven's Progressive Matrices Test. Overall, participants in all four training groups improved significantly on the training task (at least partially), as reflected by increased speed. More importantly, the only training group to show significant improvement on the Raven's was the group who had the greatest amount of spacing (20 days group) during training and improvement in this group was significantly higher than that of the control group. PMID:24782744

  19. STS-98 Crew Training

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Footage shows the crew of STS-98 during various phases of their training, including an undocking simulation in the Fixed Bases Shuttle Mission Simulator (SMS), bailout training, and extravehicular activity (EVA) training at the NBL.

  20. TEC Revolutionizes Military Training.

    ERIC Educational Resources Information Center

    Bracco, Donald C.

    1979-01-01

    Training Extension Course (TEC) system, based on individual, performance-oriented achievement, represents a revolution in Army training concepts and practices. It involves using measurable behavioral objectives, criterion-referenced testing, and validated training materials. (JOW)

  1. Dynamic training devices in CRM training

    NASA Technical Reports Server (NTRS)

    Lawver, J.

    1984-01-01

    Pilot training effectiveness and flying safety of a seasonal tour flight company are described. The change from single pilot to two pilot operated twin otters is examined. The use of the ATC 810 training device, its possibilities and training capacity is outlined. Problem areas which may arise, emergency system and pilot/passenger interaction are analyzed.

  2. Train Smart: Perfect Trainings Every Time.

    ERIC Educational Resources Information Center

    Allen, Rich

    This book explains the TrainSmart approach to training, which is based on how the authors believe the brain naturally learns best. Part 1 introduces the five pillars of the TrainSmart Model, which are as follows: (1) engage (engage participants' attention); (2) frame (establish a frame of reference); (3) explore (introduce a conceptual activity);…

  3. STS-92 Crew Training

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Footage shows the crew of STS-92, Commander Brian Duffy, Pilot Pamela A. Melroy, and Mission Specialists Koichi Wakata, Leroy Chiao, Peter J.K. Wisoff, Michael E. Lopez-Alegria, and William S. McArthur during various parts of their training. Clips are seen of the Shuttle bailout training, Shuttle arm and extravehicular activity (EVA) training at the Virtual Reality Lab, EVA training at the Neutral Buoyancy Lab, Shuttle operations training, EVA prep and post training in the Full Fuselage Trainer, ascent and post insertion training in the Guidance Navigation Simulator, and Mission Specialist Wakata in the Shuttle Engineering Dome and training on the Manipulator Development Facility.

  4. A Vocational Training Directory.

    ERIC Educational Resources Information Center

    Heymans, Susan; Hirschowitz, Ros

    This directory describes the training that is taking place in the private sector in the various industries in South Africa, excluding agriculture and domestic service, at a macro level. It focuses on training given or coordinated by industry training boards, regional training centers, certain employer organizations, and certain large employers, as…

  5. Changes in Benchmarked Training.

    ERIC Educational Resources Information Center

    Bassi, Laurie J.; Cheney, Scott

    1996-01-01

    Comparisons of the training practices of large companies confirm that the delivery and financing of training is changing rapidly. Companies in the American Society for Training and Development Benchmarking Forum are delivering less training with permanent staff and more with strategic use of technology, contract staff, and external providers,…

  6. Training Guide: Road Transport.

    ERIC Educational Resources Information Center

    Kogan Page, Ltd., London (England).

    The third in a series of guides to British industrial training, this publication begins with a survey of training issues and tasks confronting the Road Transport Industry Training Board (RTITB). This is followed by information on RTITB policies and provisions; RTITB members, officers, and committees; apprenticeships and other training schemes;…

  7. Assessing Intercultural Training Designs

    ERIC Educational Resources Information Center

    Graf, Andrea

    2004-01-01

    Training designs are critical to the success of intercultural training programmes. A common typology for classifying intercultural training designs distinguishes among the following dimensions: experiential discovery versus didactic expository and culture-specific versus culture-general training. The purpose of this paper is to assess different…

  8. Training Fishermen at Sea.

    ERIC Educational Resources Information Center

    Hermansson, Birgir

    This instructor's manual is designed to serve as a guide for a one-year course for training fishermen on board training vessels of 100 to 200 gross tons. Fourteen chapters are included to deal with the subjects and problems most frequently faced by skippers and other instructors on training vessels. Chapter titles are (1) The Training Vessel, (2)…

  9. Creating Training Miracles.

    ERIC Educational Resources Information Center

    Rylatt, Alastair; Lohan, Kevin

    This book, which is intended as a comprehensive practical reference source for training professionals, consultants, and managers, examines emerging trends in training and explains how to get training results in the workplace by using various educational strategies and advanced training methods. The following are among the topics discussed:…

  10. Training evaluation final report

    NASA Technical Reports Server (NTRS)

    Sepulveda, Jose A.

    1992-01-01

    In the area of management training, 'evaluation' refers both to the specific evaluation instrument used to determine whether a training effort was considered effective, and to the procedures followed to evaluate specific training requests. This report recommends to evaluate new training requests in the same way new procurement or new projects are evaluated. This includes examining training requests from the perspective of KSC goals and objectives, and determining expected ROI of proposed training program (does training result in improved productivity, through savings of time, improved outputs, and/or personnel reduction?). To determine whether a specific training course is effective, a statement of what constitutes 'good performance' is required. The user (NOT the Training Branch) must define what is 'required level of performance'. This 'model' will be the basis for the design and development of an objective, performance-based, training evaluation instrument.

  11. Directory of Evaluation Training Programs.

    ERIC Educational Resources Information Center

    May, Ralph M.; And Others

    1986-01-01

    Descriptions of 48 evaluation training programs offered by colleges and universities are organized into five major divisions: postdoctoral training, doctoral training, master's training, specialist training, and multiple programs. Essential characteristics, placement statistics, admission requirements, and curriculum descriptions are listed. (BS)

  12. STS-102 Crew Training

    NASA Technical Reports Server (NTRS)

    2001-01-01

    Footage shows the crew of STS-102, Commander James D. Wetherbee, Pilot James M. Kelly, and Mission Specialists Andrew S. W. Thomas and Paul Richards, during various parts of their training. Scenes include: (1) neutral buoyancy lab training; (2) undocking/fly-around training in the GNS (Navigation Simulator); (3) crew equipment interface test; (4) Remote Manipulator System (RMS) training in the GNS; and (5) docking training in the GNS.

  13. Crew Strength Training

    NASA Video Gallery

    Train to develop your upper and lower body strength in your muscles and bones by performing body-weight squats and push-ups.The Train Like an Astronaut project uses the excitement of exploration to...

  14. Training European Trade Unionists.

    ERIC Educational Resources Information Center

    Miller, Doug; Stirling, John

    1998-01-01

    A study of trade union education in Germany, Italy, Sweden, and the United Kingdom finds training is being adapted to meet new political and economic conditions. Significant national differences appeared in terms of legislation, funding, training, and accreditation. (SK)

  15. Training Earns Its Keep.

    ERIC Educational Resources Information Center

    Krell, Eric

    2001-01-01

    Training centers are now developing and marketing profitable external learning initiatives. Training programs are strengthening customer satisfaction, contributing to partnership development, enhancing research and development activities, and bolstering the bottom line. (JOW)

  16. Biofeedback Training as Counseling

    ERIC Educational Resources Information Center

    Danskin, David G.; Walters, E. Dale

    1975-01-01

    Encourages professionals in helping relationships to explore and experience biofeedback training for voluntary self-regulation. A sample biofeedback training program is described. Observations of participants in biofeedback programs are presented. (Author/BW)

  17. Expedition 34 Final Training

    NASA Video Gallery

    The Expedition 34 crew members conduct final training at the Gagarin Cosmonaut Training Center before their Dec. 19 launch to the International Space Station. Flight Engineers Chris Hadfield, Roman...

  18. Equating Training to Education.

    ERIC Educational Resources Information Center

    Davis, Lansing J.

    1993-01-01

    Distinguishes between education and employer-sponsored training in terms of process, purpose, and providers. Concludes that work-related training and postsecondary education are cognates within the classification education, and equating their learning outcomes is appropriate. (SK)

  19. Training with the BBC.

    ERIC Educational Resources Information Center

    Anderson, Jane

    1983-01-01

    Describes the special courses run by the British Broadcasting Corporation for overseas broadcasters. These include journalist training, television production, radio production, engineering training courses, and a course for educational broadcasters. (MBR)

  20. Criticality safety training

    SciTech Connect

    Woodruff, S.K.

    1997-06-01

    Criticality safety training is an important element of the Plutonium Facility safety program at Los Alamos National Laboratory. Training consists of student self-study handbooks and hands-on performance-based training in a mock-up laboratory containing gloveboxes, trolley conveyor system, and self-monitoring instruments. A 10-minute video tape and lecture was presented to describe how training in this area is conducted.

  1. Computerized leak training

    SciTech Connect

    Parella, C.; Monroe, A.

    1985-11-01

    Niagara Mohawk Power Corporation's computerized leak detection training system is discussed. The system is able to simulate gas leak situations by means of a computer. The training setup includes actual visual display via slides of houses represented on a plotting board; computer with plotter board in front that simulates an area and various leakage situations; a typical handheld CGI; and a control pad for the computer. The training system has filled a valuable need in the area of emergency training.

  2. On the Job Training.

    ERIC Educational Resources Information Center

    Barron, John M.; And Others

    Using a review of literature and data collected from worker and employer surveys, a study examined on-the-job training and its implications. Among the factors analyzed were the following: (1) on-the-job training as an investment in human capital; (2) measures of on-the-job training; (3) who receives on-the-job training; (4) how well do we measure…

  3. Contract Training for Industry.

    ERIC Educational Resources Information Center

    Averill, Donald F.

    1983-01-01

    Describes contract training whereby industries arrange with community colleges, technical institutes, and vocational schools to prepare employees for specific job assignments. Indicates that industrial training performed under contract with public institutions should be encouraged in favor of expansion of training that industry performs for…

  4. Apprenticeship Training: Gasfitter Program.

    ERIC Educational Resources Information Center

    Alberta Learning, Edmonton. Apprenticeship and Industry Training.

    This document presents information about the apprenticeship training program of Alberta, Canada, in general and the gasfitter program in particular. The first part of the document discusses the following items: Alberta's apprenticeship and industry training system; the apprenticeship and industry training committee structure; local apprenticeship…

  5. Training Top 125

    ERIC Educational Resources Information Center

    Training, 2011

    2011-01-01

    Top companies realize how vital training is to their success and continue to invest in it, even in trying times. This article presents "Training" magazine's 11th annual ranking of the top companies with employee-sponsored workforce training and development. First-time No. 1 winner Farmers Insurance puts such a premium on learning that its new…

  6. Delta Airlines LOFT training

    NASA Technical Reports Server (NTRS)

    Whitehead, J.

    1981-01-01

    A LOFT program was developed as part of the DC-9 training program which serves as a prototype for much of Delta's other aircraft training programs. The LOFT used differs little from the ideology presented in the Advisory Circular. Difficulty and experienced concerns regarding the effectiveness of LOFT as a complete training vehicle are explored.

  7. TRAINING IN INDUSTRY.

    ERIC Educational Resources Information Center

    GLASER, ROBERT

    THIS CHAPTER IN A LARGER WORK ON INDUSTRIAL PSYCHOLOGY DEALS LARGELY WITH THE NEED TO SPECIFY TRAINING OBJECTIVES THROUGH JOB ANALYSIS, USES OF TESTING IN TRAINEE SELECTION, TRAINING VARIABLES AND LEARNING PROCESSES, TRAINING TECHNOLOGY (MAINLY THE CHARACTERISTICS OF PROGRAMED INSTRUCTION), THE EVALUATION OF PROFICIENCY, THE VALUE OF…

  8. Innovations for Training.

    ERIC Educational Resources Information Center

    McFann, Howard H.; And Others

    Four papers on research and innovation in military training within the Army Training system deal with procedures for individualizing training, the Project IMPACT prototype system of computer assisted and programed instructions, student motivation and performance, and prospects for the 1970's and 1980's, and the implications of research in learning…

  9. Human Factors in Training

    NASA Technical Reports Server (NTRS)

    Barshi, Immanuel; Byrne, Vicky; Arsintescu, Lucia; Connell, Erin

    2010-01-01

    Future space missions will be significantly longer than current shuttle missions and new systems will be more complex than current systems. Increasing communication delays between crews and Earth-based support means that astronauts need to be prepared to handle the unexpected on their own. As crews become more autonomous, their potential span of control and required expertise must grow to match their autonomy. It is not possible to train for every eventuality ahead of time on the ground, or to maintain trained skills across long intervals of disuse. To adequately prepare NASA personnel for these challenges, new training approaches, methodologies, and tools are required. This research project aims at developing these training capabilities. By researching established training principles, examining future needs, and by using current practices in space flight training as test beds, both in Flight Controller and Crew Medical domains, this research project is mitigating program risks and generating templates and requirements to meet future training needs. Training efforts in Fiscal Year 09 (FY09) strongly focused on crew medical training, but also began exploring how Space Flight Resource Management training for Mission Operations Directorate (MOD) Flight Controllers could be integrated with systems training for optimal Mission Control Center (MCC) operations. The Training Task addresses Program risks that lie at the intersection of the following three risks identified by the Project: 1) Risk associated with poor task design; 2) Risk of error due to inadequate information; and 3) Risk associated with reduced safety and efficiency due to poor human factors design.

  10. Should HR Control Training?

    ERIC Educational Resources Information Center

    Schettler, Joel

    2002-01-01

    Discusses the debate over whether the training department of a company should be placed within human resources. Offers opinions of training and human resources executives and concludes that a varied structure enables training to remain flexible in response to changing conditions. (JOW)

  11. Human Factors in Training

    NASA Technical Reports Server (NTRS)

    Barshi, Immanuel; Byrne, Vicky; Arsintescu, Lucia; Connell, Erin; Sandor, Aniko

    2009-01-01

    Future space missions will be significantly longer than current shuttle missions and new systems will be more complex than current systems. Increasing communication delays between crews and Earth-based support means that astronauts need to be prepared to handle the unexpected on their own. As crews become more autonomous, their potential span of control and required expertise must grow to match their autonomy. It is not possible to train for every eventuality ahead of time on the ground, or to maintain trained skills across long intervals of disuse. To adequately prepare NASA personnel for these challenges, new training approaches, methodologies, and tools are required. This research project aims at developing these training capabilities. By researching established training principles, examining future needs, and by using current practices in space flight training as test beds, both in Flight Controller and Crew Medical domains, this research project is mitigating program risks and generating templates and requirements to meet future training needs. Training efforts in Fiscal Year 08 (FY08) strongly focused on crew medical training, but also began exploring how Space Flight Resource Management training for Mission Operations Directorate (MOD) Flight Controllers could be integrated with systems training for optimal Mission Control Center (MCC) operations. The Training Task addresses Program risks that lie at the intersection of the following three risks identified by the Project: (1) Risk associated with poor task design (2) Risk of error due to inadequate information (3) Risk associated with reduced safety and efficiency due to poor human factors design

  12. Training for GMPs.

    PubMed

    Levchuk, J W

    1991-01-01

    Training is a dynamic process to assure that personnel are capable of performing their assigned functions. CGMP regulations contain only general expectations, and no FDA guideline regarding training has been issued. Training programs are generally in place in pharmaceutical firms. However, training quality and effectiveness may be inadequate in a number of firms. A pharmaceutical firm should be able to show that its training program consistently meets its training goals as purported, and that each trainee completing an instructional module has acquired the competencies as purported. The proper application of sound principles of instructional design should help firms overcome GMP deficiencies regarding training and personnel qualification. For example, principles of mastery learning, competency-based instruction, performance objectives, a systems approach to instructional design, and the evaluation of instruction as well as the instructional program should help ensure meaningful, relevant training and appropriate, effective instruction. Review of training should be included in the firm's program for managing change. Firms should also ensure adequate training documentation, a positive attitude toward training, and that training is not used inappropriately to remedy performance deviations not resulting from skills deficiencies. PMID:1802982

  13. Functional Training Revisited.

    ERIC Educational Resources Information Center

    Siff, Mel C.

    2002-01-01

    Asserts that though functional training is vital in all sporting preparation, it is only one aspect of the overall process. The paper defines functional training; discusses facets of functionality, functionality and balancing drills, and functional training and periodization; and concludes that functionality is best defined in terms of the outcome…

  14. Adult Vocational Training.

    ERIC Educational Resources Information Center

    Ministry of Labour, Copenhagen (Denmark).

    Danish adult vocational training activities take the form of specifically targeted initial and continued training for employed and unemployed adults. Planning, development, and adaptation of vocational training programs (AMU programs) are characterized by tripartite cooperation among public authorities and organizations of employers and employees.…

  15. Hairstylist Program. Apprenticeship Training.

    ERIC Educational Resources Information Center

    Alberta Learning, Edmonton. Apprenticeship and Industry Training.

    This document presents information about the apprenticeship training program of Alberta, Canada, in general and the hairstylist program in particular. The first part of the document discusses the following items: Alberta's apprenticeship and industry training system; the apprenticeship and industry training committee structure; local…

  16. Cabinetmaker Program. Apprenticeship Training.

    ERIC Educational Resources Information Center

    Alberta Learning, Edmonton. Apprenticeship and Industry Training.

    This document presents information about the apprenticeship training program of Alberta, Canada, in general and the cabinetmaking program in particular. The first part of the document discusses the following items: Alberta's apprenticeship and industry training system; the apprenticeship and industry training committee structure; local…

  17. Cilia Train Spotting.

    PubMed

    Kuhns, Stefanie; Blacque, Oliver E

    2016-06-01

    Cilium formation depends on intraflagellar transport trains that move bidirectionally along ciliary microtubules. Reporting in Science, Stepanek and Pigino (2016) employ correlative light and electron microscopy in algae to determine the ultrastructure of anterograde and retrograde trains and discover that these trains avoid collision by running on B- and A-tubules, respectively. PMID:27270038

  18. Online Training in Australia

    ERIC Educational Resources Information Center

    Kuzic, Joze

    2013-01-01

    On-line training is becoming an interesting phenomenon in Australia and has attracted a lot of interest across many industries and businesses (Chan and Ngai, 2007). The research reported here looks at the use of online training in corporations in Australia. It focuses on two aspects of online training, the factors that "warrant" its…

  19. Training Top 50.

    ERIC Educational Resources Information Center

    Training, 2001

    2001-01-01

    Identifies the top 50 companies in terms of the amount spent on training and development, the number of hours of training per employee, and percentage of payroll spent on training. Profiles the top five plus four additional companies selected by the magazine's editors. (SK)

  20. Women's Job Training Agenda.

    ERIC Educational Resources Information Center

    Women Work! The National Network for Women's Employment, Washington, DC.

    The Coalition on Women and Job Training has developed a women's job training agenda that focuses on nine issues related to women's full participation in training and achievement of economic self-sufficiency. The issues highlighted in the agenda are as follows: (1) the need to make long-term economic self-sufficiency the goal of all employment and…

  1. Strength Training for Girls.

    ERIC Educational Resources Information Center

    Connaughton, Daniel; Connaughton, Angela; Poor, Linda

    2001-01-01

    Strength training can be fun, safe, and appropriate for young girls and women and is an important component of any fitness program when combined with appropriate cardiovascular and flexibility activities. Concerns and misconceptions regarding girls' strength training are discussed, presenting general principles of strength training for children…

  2. Abramovo Counterterrorism Training Center

    SciTech Connect

    Hayes, Christopher M; Ross, Larry; Lingenfelter, Forrest E; Sokolnikov, Pavel I; Kaldenbach, Karen Yvonne; Estigneev, Yuri; Murievav, Andrey

    2011-01-01

    The U.S. government has been assisting the Russian Federation (RF) Ministry of Defense (MOD) for many years with nuclear weapons transportation security (NWTS) through the provision of specialized guard escort railcars and cargo railcars with integrated physical security and communication systems, armored transport vehicles, and armored escort vehicles. As a natural continuation of the NWTS program, a partnership has been formed to construct a training center that will provide counterterrorism training to personnel in all branches of the RF MOD. The Abramovo Counterterrorism Training Center (ACTC) is a multinational, multiagency project with funding from Canada, RF and the U.S. Departments of Defense and Energy. ACTC will be a facility where MOD personnel can conduct basic through advanced training in various security measures to protect Category IA material against the threat of terrorist attack. The training will enhance defense-in-depth principles by integrating MOD guard force personnel into the overall physical protection systems and improving their overall response time and neutralization capabilities. The ACTC project includes infrastructure improvements, renovation of existing buildings, construction of new buildings, construction of new training facilities, and provision of training and other equipment. Classroom training will be conducted in a renovated training building. Basic and intermediate training will be conducted on three different security training areas where various obstacles and static training devices will be constructed. The central element of ACTC, where advanced training will be held, is the 'autodrome,' a 3 km road along which various terrorist events can be staged to challenge MOD personnel in realistic and dynamic nuclear weapons transportation scenarios. This paper will address the ACTC project elements and the vision for training development and integrating this training into actual nuclear weapons transportation operations.

  3. STS-96 Crew Training

    NASA Technical Reports Server (NTRS)

    1999-01-01

    The training for the crew members of the STS-96 Discovery Shuttle is presented. Crew members are Kent Rominger, Commander; Rick Husband, Pilot; Mission Specialists, Tamara Jernigan, Ellen Ochoa, and Daniel Barry; Julie Payette, Mission Specialist (CSA); and Valery Ivanovich Tokarev, Mission Specialist (RSA). Scenes show the crew sitting and talking about the Electrical Power System; actively taking part in virtual training in the EVA Training VR (Virtual Reality) Lab; using the Orbit Space Vision Training System; being dropped in water as a part of the Bail-Out Training Program; and taking part in the crew photo session.

  4. Radiological worker training

    SciTech Connect

    1998-10-01

    This Handbook describes an implementation process for core training as recommended in Implementation Guide G441.12, Radiation Safety Training, and as outlined in the DOE Radiological Control Standard (RCS). The Handbook is meant to assist those individuals within the Department of Energy, Managing and Operating contractors, and Managing and Integrating contractors identified as having responsibility for implementing core training recommended by the RCS. This training is intended for radiological workers to assist in meeting their job-specific training requirements of 10 CFR 835. While this Handbook addresses many requirements of 10 CFR 835 Subpart J, it must be supplemented with facility-specific information to achieve full compliance.

  5. Strength Training and Your Child

    MedlinePlus

    ... Story" 5 Things to Know About Zika & Pregnancy Strength Training and Your Child KidsHealth > For Parents > Strength Training ... help prevent injuries and speed up recovery. About Strength Training Strength training is the practice of using free ...

  6. Human Factors in Training

    NASA Technical Reports Server (NTRS)

    Barshi, Immanuel; Byme, Vicky; Arsintescu, Lucia

    2008-01-01

    Future space missions will be significantly longer than current Shuttle missions and new systems will be more complex than current systems. Increasing communication delays between crews and Earth-based support means that astronauts need to be prepared to handle the unexpected on their own. As crews become more autonomous, their potential span of control and required expertise must grow to match their autonomy. It is not possible to train for every eventuality ahead of time on the ground, or to maintain trained skills across long intervals of disuse. To adequately prepare NASA personnel for these challenges, new training approaches, methodologies, and tools are required. This research project aims at developing these training capabilities. Training efforts in FY07 strongly focused on crew medical training, but also began exploring how Space Flight Resource Management training for Mission Operations Directorate (MOD) Flight Controllers could be integrated with systems training for optimal Mission Control Center operations. Beginning in January 2008, the training research effort will include team training prototypes and tools. The Training Task addresses Program risks that lie at the intersection of the following three risks identified by the Project: 1) Risk associated with poor task design; 2) Risk of error due to inadequate information; 3) Risk associated with reduced safety and efficiency due to poor human factors design.

  7. T-1 Training Area

    SciTech Connect

    2014-11-07

    Another valuable homeland security asset at the NNSS is the T-1 training area, which covers more than 10 acres and includes more than 20 separate training venues. Local, County, and State first responders who train here encounter a variety of realistic disaster scenarios. A crashed 737 airliner lying in pieces across the desert, a helicopter and other small aircraft, trucks, buses, and derailed train cars are all part of the mock incident scene. After formal classroom education, first responders are trained to take immediate decisive action to prevent or mitigate the use of radiological or nuclear devices by terrorists. The Counterterrorism Operations Support Center for Radiological Nuclear Training conducts the courses and exercises providing first responders from across the nation with the tools they need to protect their communities. All of these elements provide a training experience that cannot be duplicated anywhere else in the country.

  8. Intelligent Computerized Training System

    NASA Technical Reports Server (NTRS)

    Wang, Lui; Baffes, Paul; Loftin, R. Bowen; Hua, Grace C.

    1991-01-01

    Intelligent computer-aided training system gives trainees same experience gained from best on-the-job training. Automated system designed to emulate behavior of experienced teacher devoting full time and attention to training novice. Proposes challenging training scenarios, monitors and evaluates trainee's actions, makes meaningful comments in response to errors, reponds to requests for information, gives hints when appropriate, and remembers strengths and weaknesses so it designs suitable exercises. Used to train flight-dynamics officers in deploying satellites from Space Shuttle. Adapted to training for variety of tasks and situations, simply by modifying one or at most two of its five modules. Helps to ensure continuous supply of trained specialists despite scarcity of experienced and skilled human trainers.

  9. Payload training methodology study

    NASA Technical Reports Server (NTRS)

    1990-01-01

    The results of the Payload Training Methodology Study (PTMS) are documented. Methods and procedures are defined for the development of payload training programs to be conducted at the Marshall Space Flight Center Payload Training Complex (PCT) for the Space Station Freedom program. The study outlines the overall training program concept as well as the six methodologies associated with the program implementation. The program concept outlines the entire payload training program from initial identification of training requirements to the development of detailed design specifications for simulators and instructional material. The following six methodologies are defined: (1) The Training and Simulation Needs Assessment Methodology; (2) The Simulation Approach Methodology; (3) The Simulation Definition Analysis Methodology; (4) The Simulator Requirements Standardization Methodology; (5) The Simulator Development Verification Methodology; and (6) The Simulator Validation Methodology.

  10. T-1 Training Area

    ScienceCinema

    None

    2015-01-09

    Another valuable homeland security asset at the NNSS is the T-1 training area, which covers more than 10 acres and includes more than 20 separate training venues. Local, County, and State first responders who train here encounter a variety of realistic disaster scenarios. A crashed 737 airliner lying in pieces across the desert, a helicopter and other small aircraft, trucks, buses, and derailed train cars are all part of the mock incident scene. After formal classroom education, first responders are trained to take immediate decisive action to prevent or mitigate the use of radiological or nuclear devices by terrorists. The Counterterrorism Operations Support Center for Radiological Nuclear Training conducts the courses and exercises providing first responders from across the nation with the tools they need to protect their communities. All of these elements provide a training experience that cannot be duplicated anywhere else in the country.

  11. Training System for the Expiration Training

    NASA Astrophysics Data System (ADS)

    Ogoshi, Yasuhiro; Nakai, Akio; Mitsuhashi, Yoshinori; Araki, Chikahiro

    For a speech therapist, the breathing method is one of the important techniques to train a person who has difficulty in speaking. Insufficient number of therapists makes it difficult to apply these breath training methods widely. Therefore, development of a new breath training method will be mandatory. With this reason, a new training system is developed and described in this paper. In this new method, expiration pattern of a speech therapist analyzing through an air flow sensor is constructed into visual graphic image in order to recognize expiration pattern easily. With experimental use of this system to without handicapped trainees, they could successfully imitate trainer's pronunciation easily. A person who has difficulty in speaking will be able to learn easily how to do expiration for speaking through visual pattern.

  12. STS-105 Crew Training Clip

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The STS-105 crewmembers, Commander Scott Horowitz, Pilot Frederick Sturckow, and Mission Specialists Daniel Barry and Patrick Forrester, are seeing during various stages of their training. Footage includes Post-Insertion Training, Virtual Reality Laboratory Remote Manipulator System Training, Emergency Egress Training in the CCT, Neutral Buoyancy Laboratory Training, Post Egress Landing Training at FFT, Long Rendezvous Training in the GNS (Navigation Simulator), and Post Insertion Operations Training at FFT.

  13. STS-109 Crew Training

    NASA Technical Reports Server (NTRS)

    2002-01-01

    Footage shows the crew of STS-109 (Commander Scott Altman, Pilot Duane Carey, Payload Commander John Grunsfeld, and Mission Specialists Nancy Currie, James Newman, Richard Linnehan, and Michael Massimino) during various parts of their training. Scenes show the crew's photo session, Post Landing Egress practice, training in Dome Simulator, Extravehicular Activity Training in the Neutral Buoyancy Laboratory (NBL), and using the Virtual Reality Laboratory Robotic Arm. The crew is also seen tasting food as they choose their menus for on-orbit meals.

  14. Crew Skills and Training

    NASA Technical Reports Server (NTRS)

    Jones, Thomas; Burbank, Daniel C.; Eppler, Dean; Garrison, Robert; Harvey, Ralph; Hoffman, Paul; Schmitt, Harrison

    1998-01-01

    One of the major focus points for the workshop was the topic of crew skills and training necessary for the Mars surface mission. Discussions centered on the mix of scientific skills necessary to accomplish the proposed scientific goals, and the training environment that can bring the ground and flight teams to readiness. Subsequent discussion resulted in recommendations for specific steps to begin the process of training an experienced Mars exploration team.

  15. World's simplest electric train

    NASA Astrophysics Data System (ADS)

    Criado, C.; Alamo, N.

    2016-01-01

    We analyze the physics of the "world's simplest electric train." The "train" consists of a AA battery with a strong magnet on each end that moves through a helical coil of copper wire. The motion of the train results from the interaction between the magnetic field created by the current in the wire and the magnetic field of the magnets. We calculate the force of this interaction and the terminal velocity of the train due to eddy currents and friction. Our calculations provide a good illustration of Faraday's and Lenz's laws, as well as of the concepts of the Lorentz force and eddy currents.

  16. Correctional Training. Institution Familiarization. Part II: The Training Program.

    ERIC Educational Resources Information Center

    Bureau of Prisons (Dept. of Justice), Washington, DC.

    Designed to assist training coordinators in the initial institution familiarization training for new employees in correctional institutions, this manual consists of two documents: a training coordinator's guide (Part I - CE 017 285) and this document, the training program (Part II). Four training areas are treated: (1) an introduction consisting…

  17. STS-104 Crew Training Clips

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The crewmembers of STS-104, Commander Steven Lindsey, Pilot Charles Hobaugh, and Mission Specialists Michael Gernhardt, James Reilly, and Janet Kavandi, are seen during various stages of their training. Footage shows the following: (1) Water Survival Training at the Neutral Buoyancy Laboratory (NBL); (2) Rendezvous and Docking Training in the Shuttle Mission Simulator; (3) Training in the Space Station Airlock; (4) Training in the Virtual Reality Lab; (5) Post-insertion Operations in the Fixed Base Simulator; (6) Extravehicular Activity Training at the NBL; (7) Crew Stowage Training in the Space Station Mock-up Training Facility; and (8) Water Transfer Training in the Crew Compartment Trainer.

  18. Supporting Learners in Training.

    ERIC Educational Resources Information Center

    Montague, Ann; Hopkins, Linda

    A project was conducted in Australia to investigate the most effective ways whereby learners receive informal and formal support to assist them to successfully complete vocational education and training (VET), either in school or in business job-training programs. Data were collected through a literature review, a case study of 11 organizations,…

  19. Trains across the USA

    ERIC Educational Resources Information Center

    McIntosh, Phyllis

    2013-01-01

    This feature article explores the topic of trains from their early history to recent trends in railroading. A glossary provides related vocabulary, a sidebar called "Romance of the Rails" adds colorful detail about train songs and railroad lore. An additional section, "Classroom Activities," by John Silver, presents three…

  20. Experimenting with Electric Trains

    ERIC Educational Resources Information Center

    Wick, D. P.; Ramsdell, M. W.

    2007-01-01

    A simple experiment can be performed to characterize the relationship between applied voltage and velocity (steady state and transient) for an electric toy train. The results can be used by teams of students to solve a series of challenges in which they attempt to predict the performance of a particular train. Some sample challenges might include…

  1. Training Success Stories.

    ERIC Educational Resources Information Center

    Overmeyer-Day, Leslie; Benson, George

    1996-01-01

    Offers case studies of best training practices in leading companies (AEtna, Andersen Worldwide, Boeing, Digital Equipment, Tektronix, Texas Instruments, Allstate, and Sprint), elected by the Benchmarking Forum of the American Society for Training and Development. The practices are technology based and performance oriented and herald common…

  2. Who's Getting Trained?

    ERIC Educational Resources Information Center

    Froiland, Paul

    1993-01-01

    An annual business/industry survey showed a 15% increase in the number of people receiving formal training. Top five methods were videotape, lecture, one-on-one, role plays, and simulations. Training focused on management, computer, communication, supervisory, and technical skills. Percentage of employers providing remedial education remained…

  3. Failure-Oriented Training.

    ERIC Educational Resources Information Center

    Pickens, Diana; Lorenz, Paul

    This document consists of a number of figures and diagrams suitable for overhead transparencies that illustrate and elaborate on the prnciples of failure-oriented training (a model for improving the effectiveness of instructional analysis). By adding a few simple steps to analysis, the resulting training will be closer to the idealized tutor:…

  4. Evaluating Court Training Programs.

    ERIC Educational Resources Information Center

    Hudzik, John K.; Wakeley, John H.

    1981-01-01

    Careful evaluation of court training programs can use two approaches, program monitoring and training/learning. Four areas of inquiry include: reactions to the program, learning, behavior, and results (behavior measured against organizational goals). The Michigan Judicial Institute program is noted. Journal availability: 200 W. Monroe, Suite 1606,…

  5. Nuclear Medical Technology Training.

    ERIC Educational Resources Information Center

    Simmons, Guy H., Ed.

    This 1-day colloquium, attended by 23 participants representing societies, government agencies, colleges and universities, and other training programs, was conducted for the purpose of reporting on and discussing the curriculums developed at the University of Cincinnati for training nuclear medical technologists. Pilot programs at both the…

  6. Technical Training for Managers.

    ERIC Educational Resources Information Center

    Haverland, Edgar M.

    The question has arisen as to what kind of information a manager without extensive technical training needs to learn to supervise effectively. For example, the Nike Hercules fire control platoon leader, usually an officer in his first active duty assignment, seldom has had extensive technical training. Yet he is responsibile for the…

  7. Rapid Response Skills Training

    ERIC Educational Resources Information Center

    Kelley-Winders, Anna Faye

    2008-01-01

    Mississippi Gulf Coast Community College's (MGCCC) long-term commitment to providing workforce training in a post-Katrina environment became a catalyst for designing short-term flexible educational opportunities. Providing nationally recognized skills training for the recovery/rebuilding of communities challenged the college to develop innovative,…

  8. Training Management Information System

    SciTech Connect

    Rackley, M.P.

    1989-01-01

    The Training Management Information System (TMIS) is an integrated information system for all training related activities. TMIS is at the leading edge of training information systems used in the nuclear industry. The database contains all the necessary records to confirm the department's adherence to accreditation criteria and houses all test questions, student records and information needed to evaluate the training process. The key to the TMIS system is that the impact of any change (i.e., procedure change, new equipment, safety incident in the commercial nuclear industry, etc.) can be tracked throughout the training process. This ensures the best training can be performed that meets the needs of the employees. TMIS is comprised of six functional areas: Job and Task Analysis, Training Materials Design and Development, Exam Management, Student Records/Scheduling, Evaluation, and Commitment Tracking. The system consists of a VAX 6320 Cluster with IBM and MacIntosh computers tied into an ethernet with the VAX. Other peripherals are also tied into the system: Exam Generation Stations to include mark sense readers for test grading, Production PC's for Desk-Top Publishing of Training Material, and PC Image Workstations. 5 figs.

  9. BIBLIOGRAPHY OF TRAINING AIDS.

    ERIC Educational Resources Information Center

    MCKEONE, CHARLES J.

    THIS COMPILATION OF INSTRUCTIONAL AIDS FOR USE IN AIR-CONDITIONING AND REFRIGERATION TRAINING PROGRAMS CONTAINS LISTS OF VISUAL AND AUDIOVISUAL TRAINING AIDS AND GUEST LECTURERS AVAILABLE FROM MEMBER COMPANIES OF THE AIR-CONDITIONING AND REFRIGERATION INSTITUTE AS AN INDUSTRY SERVICE TO SCHOOL OFFICIALS INTERESTED IN CONDUCTING SUCH PROGRAMS. THE…

  10. Training TA's to Teach

    ERIC Educational Resources Information Center

    Stockdale, Dennis L.; Wochok, Zachary S.

    1974-01-01

    Summarizes teaching assistant training programs conducted in 50 graduate schools. Indicates that mose institutions see the teaching assistant as a means of providing undergraduate instruction and of providing financial support for graduate students, not as a means of training future college teachers. (CC)

  11. TAT [Training and Technology.

    ERIC Educational Resources Information Center

    Oak Ridge Associated Universities, TN. Manpower Development Div.

    The Oak Ridge Associated Universities (ORAU) of Tennessee and the Nuclear Division of the Union Carbide Corporation established an industrial training program called Training and Technology (TAT) which was conducted at the Oak Ridge Y-12 plant. TAT instructors were provided by the regular work force of Union Carbide while ORAU provided the…

  12. Interschool Production Training Combines

    ERIC Educational Resources Information Center

    Il'ina, L. A.; Fat'ianov, V. V.

    1977-01-01

    An educational program draws together students from several secondary schools in one region in a labor practicum which combines training shops, classroom work, and laboratory experience. Examples are presented of schools throughout the USSR which are training pupils to be computer and machine operators, construction workers, cooks, automotive…

  13. TRAINING AND DEVELOPMENT HANDBOOK.

    ERIC Educational Resources Information Center

    CRAIG, ROBERT L., ED.; BITTEL, LESTER R., ED.

    TO PROVIDE A BROAD REFERENCE SOURCE, FROM THE VIEWPOINT OF THE EMPLOYER ORGANIZATION, FOR THOSE RESPONSIBLE FOR DEVELOPING HUMAN RESOURCES IN ANY ORGANIZATION, THIS HANDBOOK INCLUDES MATERIALS FOR THE SOPHISTICATED MANAGER OF A LARGE TRAINING STAFF AS WELL AS THE FUNDAMENTALS OF TRAINING FOR THE BEGINNING OR PART-TIME TRAINER. THE LEVELS OF…

  14. Respite Care. Training Manual.

    ERIC Educational Resources Information Center

    Lukenbill, Ron; And Others

    The manual is intended for use in training institutes for persons providing respite care for families of children with developmental disabilities in Montana. Expectations for participants and objectives for training are outlined in preliminary sections. Each chapter contains a brief statement of the chapter's main point or points, a selected…

  15. 2010 Training Industry Report

    ERIC Educational Resources Information Center

    Training, 2010

    2010-01-01

    Now in its 29th year, The Industry Report is recognized as the training industry's most trusted source of data on budgets, staffing, and programs. This year, the study was conducted by an outside research firm in June-August 2010, when members from the "Training" magazine database were e-mailed an invitation to participate in an online survey.…

  16. Systems Approach to Training.

    ERIC Educational Resources Information Center

    British Gas, London (England). Training and Development Dept.

    This pamphlet is intended to assist managers and professional trainers alike in using a systems approach to training. Addressed in the individual sections of the guide are the following topics: identifying the training need (the main job objectives, the conditions under which the job is performed, and the responsibilities it involves); analyzing…

  17. Training On the Job.

    ERIC Educational Resources Information Center

    Walter, Diane

    This book provides training and development professionals with step-by-step guidelines for developing and delivering comprehensive structured the on-the-job training (OJT). Chapter 1 introduces the concept of team-driven structured OJT and discusses its philosophy and basic components in the context of fundamental concepts of human behavior and…

  18. Make Room for Training.

    ERIC Educational Resources Information Center

    Filipczak, Bob

    1991-01-01

    Discusses audiovisual technology and its place in today's state-of-the-art training classroom. Looks at costs, design considerations, and who really uses the technology. Describes Dow Chemical Company's state-of-the-art training center in Midland, Michigan. (JOW)

  19. Training Budgets Boom.

    ERIC Educational Resources Information Center

    Filipczak, Bob

    1993-01-01

    An annual business/industry survey showed 7% increase in spending on formal training in organizations of 100 or more employees, accounted for by 36% of all organizations. Large organizations are either paying trainers more or hiring more trainers; many are using community colleges. Percentage of training dollar per employee category is 26%…

  20. Training Top 125

    ERIC Educational Resources Information Center

    Training, 2012

    2012-01-01

    The best learning and development organizations support business initiatives tactically "and" help drive strategic change. Verizon did just that, earning it the No. 1 spot for the first time on the Training Top 125. Verizon and the other 2012 Top 125 winners continued to invest in training, collectively dedicating a mean of 4.52 percent of their…

  1. Worker-Directed Training.

    ERIC Educational Resources Information Center

    Wagner, Stacey

    2001-01-01

    Describes the training at the Idaho National Engineering and Environmental Laboratory, the foremost nuclear energy and environmental laboratory in the United States. Suggests that the key to assurance is getting workers, most of whom are unionized, involved in their own safety training. (JOW)

  2. Toilet Training Carl.

    ERIC Educational Resources Information Center

    Dunklee, Nancy R.

    1989-01-01

    The process of toilet training a boy with Rubinstein-Taybi Syndrome is addressed. Emphasized are the joint effort of the teacher and parents, the frustration at the seeming lack of progress, the use of rewards and "positive practices," and the ultimate success of the boy's toilet training program. (JDD)

  3. Training in British Industry

    ERIC Educational Resources Information Center

    Jackson, Willis

    2008-01-01

    Purpose: The purpose of this paper is to examine the need for, and development of, further education and practical training for recruits into industry. Design/methodology/approach: The paper discusses how, at the time of writing, many firms were developing and operating training schemes for industrial personnel. Firms benefit themselves from…

  4. Training the Hardcore.

    ERIC Educational Resources Information Center

    Urban Research Corp., Chicago, IL.

    This part of the series on Training the Hardcore covers factors involved in hiring and training the hardcore, abstracts of the 12 volumes included in the whole, and three other documents which will give management further insight into how the employment situation looks to the man in the street. Based on his experience at KLH and Polaroid, Henry M.…

  5. Training at Aerospatiale

    NASA Astrophysics Data System (ADS)

    Chabod, Rene

    A training plan for Aerospatiale is outlined which incorporates goals for both the organization and the individual. Emphasis is placed on the training of staff in multiple skills such as mechanics/electronics. Other areas considered include the development of information tools, the development of commercial approaches, and the learning of new industrial techniques and foreign languages.

  6. Can Trains Really Float?

    ERIC Educational Resources Information Center

    McCartney, Robin Ward; Deroche, Sarah; Pontiff, Danielle

    2008-01-01

    Have you ever heard of a Maglev train? Who would be crazy enough to think that exploring how a high-tech train little known in the United States works with a group of fourth-grade students would yield understandings about the properties of magnetism, force and motion, and inquiry science? Fortunately, the authors--a college methods professor and…

  7. What Price Online Training?

    ERIC Educational Resources Information Center

    Ganzel, Rebecca

    1999-01-01

    Discusses the difficulties involved with pricing an online training course. Looks at such issues as the size of the marketplace, the ability to use the same course many times, the switch from time-centered to content-centered courses, the amount of time needed to complete a specific course, and the need for multimedia training materials. (JOW)

  8. Montana Bicyclist Training Program.

    ERIC Educational Resources Information Center

    DiBrito, Roger, Ed.; And Others

    This guide is designed to train elementary students in the riding skills, hazard identification, and traffic analysis necessary for safe bicycling under nearly all urban traffic and roadway conditions. The training manual is divided into four sections. The Instructor's Manual presents brief introductions, background information, and detailed…

  9. Automotive Power Trains.

    ERIC Educational Resources Information Center

    Marine Corps Inst., Washington, DC.

    This correspondence course, originally developed for the Marine Corps, is designed to provide mechanics with an understanding of the operation, maintenance, and troubleshooting of automotive power trains and certain auxiliary equipment. The course contains six study units covering basic power trains; clutch principles and operations; conventional…

  10. Conducting Effective Simulator Training.

    ERIC Educational Resources Information Center

    Gerling, Kenneth D.

    This paper describes the simulator phase of Commonwealth Edison's program for training and licensing operators of nuclear power stations. Topics covered include (1) preparing the students before starting the simulator phase; (2) the simulator schedule and the number of students that can be trained effectively in a class; (3) format and structure…

  11. TRAINING IN PROBLEM SOLVING.

    ERIC Educational Resources Information Center

    DEVANE, J.R.; RIMOLDI, H.J.A.

    CHANGES WERE STUDIED IN THE PROBLEM-SOLVING BEHAVIOR OF HIGH SCHOOL STUDENTS AS A FUNCTION OF A CAREFULLY DESIGNED TRAINING PROGRAM. TRAINING WAS DEFINED AS THE DEVELOPMENT OF STUDENT AWARENESS OF PROBLEM-SOLVING STRATEGIES USED. INSTRUMENTS WERE DEVELOPED AND REFINED TO MEASURE PROBLEM-SOLVING BEHAVIOR. SPECIFICALLY TESTED WAS THE FOLLOWING…

  12. Medical Training Aid

    NASA Technical Reports Server (NTRS)

    1992-01-01

    Dr. Steven S. Saliterman developed his Dynacath Critical Care Patient Simulator as a training system for medical personnel involved in critical care management and hemodynamic monitoring. The system incorporates NASA simulation technology and allows hospitals and medical manufacturers to conduct training away from the patient's bedside. Dr. Saliterman was formerly employed by Ames Research Center and Johnson Space Center.

  13. Dance Training for Gymnastics.

    ERIC Educational Resources Information Center

    Pica, Rae

    This book views gymnastics as both sport and art and provides gymnasts with an introduction to the types of dance training that can enhance their performances. Dance training routines that incorporate several dance styles are presented to help gymnasts acquire polish, prevent injuries, improve their body alignment, express their creativity, and…

  14. Training Evaluation: Clients' Roles.

    ERIC Educational Resources Information Center

    Hashim, Junaidah

    2001-01-01

    A study was conducted of 262 training providers in Malaysia, where providers must be government approved. Government regulation, client demands for high quality, and an economic downturn that focused attention on training costs have all influenced evaluation in a positive direction. (SK)

  15. Research Diver Training Program.

    ERIC Educational Resources Information Center

    Somers, Lee H.

    This publication provides a listing of the components of a specialized diver training program developed at the University of Michigan. Because of the demand for specialized diver training in the scientific community, a speciality diving course was developed to serve those in the various fields of marine and aquatic science and engineering. This…

  16. Job Instruction Training.

    ERIC Educational Resources Information Center

    Pfau, Richard H.

    Job Instruction Training (JIT) is a step-by-step, relatively simple technique used to train employees on the job. It is especially suitable for teaching manual skills or procedures; the trainer is usually an employee's supervisor but can be a co-worker. The JIT technique consists of a series of steps that a supervisor or other instructor follows…

  17. Industrial Training Research Project.

    ERIC Educational Resources Information Center

    Swanson, R. A.; Sawzin, S. A.

    The study was an experimental comparison of the structured versus unstructed training of semiskilled production workers. The experiment was implemented using the following procedures, which are presented in detail: a representative semiskilled production job was selected, the two training programs were characterized and developed, trainees…

  18. Leader Training Program.

    ERIC Educational Resources Information Center

    Hoehn, Lilburn P., Ed.

    This document describes the Leader Training Program (LTP) component of the Teaching Behavior Improvement Program (TBIP). The LTP was funded under ESEA Title III and was designed to train school-based inservice leaders to implement the TBIP in their districts. The document suggests possible grouping of participants, materials, equipment, physical…

  19. Tax Breaks for Training.

    ERIC Educational Resources Information Center

    Feuer, Dale

    1986-01-01

    Discusses the policy of using tax incentives to stimulate investment in training, which allows the federal government to offer financial support without getting directly involved. The popularity of this policy and the reasons for it are examined. Proposed legislation directed at training needs is described. (CT)

  20. Training for Productivity.

    ERIC Educational Resources Information Center

    Maglen, Leo; Hopkins, Sonnie; Burke, Gerald

    An exploratory study was conducted to evaluate the utility of a method to demonstrate that Australian enterprises that invest in the training of their employees gain a return from that investment through an increase in employee productivity. The method, which compares enterprise expenditure on training of personnel with labor productivity across a…

  1. Peer Leadership Training Manual.

    ERIC Educational Resources Information Center

    Kuhn, Margaret E.; And Others

    Presented are two documents focusing on a peer leadership training program which pairs better students and leaders with younger high-risk students. One document briefly describes the peer leadership training program as a program in which students traveled to an environmental education center for 3 days of competitive and challenging activities,…

  2. Nonformal Education Training Module.

    ERIC Educational Resources Information Center

    Fox, Helen; And Others

    This Peace Corps Nonformal Education (NFE) training module provides training ideas for volunteers. This module contains 10 sessions, each designed to fit into a 3-hour period: (1) What is NFE? (2) adult learning; (3) helping people identify their needs; (4) facilitation skills--part 1; (5) facilitation skills--part 2; (6) NFE materials…

  3. Customized Training Marketing Plan.

    ERIC Educational Resources Information Center

    Lay, Ted

    This report outlines Oregon's Lane Community College's (LCC's) plan for marketing its customized training program for business, community organizations, public agencies, and their employees. Following a mission statement for the customized training program, a brief analysis is provided of the economic environment; of competition from educational…

  4. Do Sectoral Training Funds Stimulate Training?

    ERIC Educational Resources Information Center

    Kamphuis, Pascal; Glebbeek, Arie C.; Van Lieshout, Harm

    2010-01-01

    Sectoral levelling funds are an arrangement aimed at alleviating a well-known theoretical problem of underinvestment in worker training because of free-rider behaviour of firms. In the Netherlands, collective agreements require firms to participate in such funds in a number of sectors. Using a comprehensive dataset of Dutch firms, we attempt to…

  5. Intercultural Training: Six Measures Assessing Training Needs

    ERIC Educational Resources Information Center

    Graf, Andrea; Mertesacker, Marion

    2009-01-01

    Purpose: The purpose of this paper is to develop recommendations for measures assessing intercultural training needs for international human resource management. Based on scientific as well as application-oriented criteria the aim is to select six measures assessing general intercultural competencies and with the help of behaviour ratings in…

  6. Training Public Health Advisors

    PubMed Central

    Meyer, Pamela A.; Brusuelas, Kristin M.; Baden, Daniel J.; Duncan, Heather L.

    2015-01-01

    Federal public health advisors provide guidance and assistance to health departments to improve public health program work. The Centers for Disease Control and Prevention (CDC) prepares them with specialized training in administering public health programs. This article describes the evolving training and is based on internal CDC documents and interviews. The first federal public health advisors worked in health departments to assist with controlling syphilis after World War II. Over time, more CDC prevention programs hired them. To meet emerging needs, 3 major changes occurred: the Public Health Prevention Service, a fellowship program, in 1999; the Public Health Associate Program in 2007; and integration of those programs. Key components of the updated training are competency-based training, field experience, supervision, recruitment and retention, and stakeholder support. The enduring strength of the training has been the experience in a public health agency developing practical skills for program implementation and management. PMID:25564995

  7. Training Public Health Advisors.

    PubMed

    Meyer, Pamela A; Brusuelas, Kristin M; Baden, Daniel J; Duncan, Heather L

    2015-01-01

    Federal public health advisors provide guidance and assistance to health departments to improve public health program work. The Centers for Disease Control and Prevention (CDC) prepares them with specialized training in administering public health programs. This article describes the evolving training and is based on internal CDC documents and interviews. The first federal public health advisors worked in health departments to assist with controlling syphilis after World War II. Over time, more CDC prevention programs hired them. To meet emerging needs, 3 major changes occurred: the Public Health Prevention Service, a fellowship program, in 1999; the Public Health Associate Program in 2007; and integration of those programs. Key components of the updated training are competency-based training, field experience, supervision, recruitment and retention, and stakeholder support. The enduring strength of the training has been the experience in a public health agency developing practical skills for program implementation and management. PMID:25564995

  8. 38 CFR 21.4265 - Practical training approved as institutional training or on-job training.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... approved as institutional training or on-job training. 21.4265 Section 21.4265 Pensions, Bonuses, and... training or on-job training. (a) Medical-dental internships and residencies. (1) Medical residencies (other...) of this section. If the course is not so accredited such practical or on-the-job training...

  9. A True Training Needs Analysis.

    ERIC Educational Resources Information Center

    Nowack, Kenneth M.

    1991-01-01

    Employees often want training in areas that are irrelevant to their jobs or inconsistent with organizational objectives. A well-designed questionnaire can weed out training wants to uncover an employee's true training needs. (Author)

  10. Training by simulation

    SciTech Connect

    Angelini, E.

    1984-12-01

    Effective training by simulation is a matter of 1. accurate definition of the training need, 2. selecting the right hardware/software package to do the job cost effectively, 3. careful, methodical building of the training program and 4. an appropriate setting, such as a well-equipped training center. Several points seem clear: 1. ''Computer shock'' or computer phobia -- that is, fear of the simulator, can be a factor with some of the older operators. But it quickly disappears once they're into the program, and learn the versatility and the exciting dynamics of simulators. 2. Operator training is becoming more sophisticated throughout the HPI. Among other things, operators should get out into the plant to learn more about how their actions in the control room impact the plant. Operators are becoming more ''big picture'' oriented. 3. When inexperienced training instructors are used, they should attend a good ''train the trainer'' course emphasizing platform skills and instructional technology. 4. Operators need as much ''hands on'' experience with the plant's actual operating equipment as possible. The cognitive ''linkage'' between that equipment and what they see on the CRT is vitally important.

  11. Conducting effective tailgate trainings.

    PubMed

    Harrington, David; Materna, Barbara; Vannoy, Jim; Scholz, Peter

    2009-07-01

    The California Department of Health Services' Occupational Health Branch and others have identified the construction industry as being at high risk for injuries, illnesses, and fatalities. Effective tailgate trainings (brief job site safety meetings) can be a powerful tool to promote hazard awareness and safe work practices. The authors found that many contractors and supervisors conducted ineffective tailgate trainings. They developed the BuildSafe California Project to assist contractors to have more effective programs by holding 25 training-of-trainers sessions reaching 1,525 participants. The needs assessment, intervention, and evaluation results from the first 18 trainings are presented. Eighty-six percent of the participants found the program "very helpful." Participants used the materials and made improvements in the quality and frequency of trainings. Supervisors must be skilled at conducting tailgate trainings as part of their responsibilities. There is a serious need to provide more culturally appropriate safety training in a workforce increasingly made up of Latino workers. PMID:18372431

  12. How shall we train?

    PubMed

    De Lorenzo, Robert A

    2005-10-01

    The prosecution of modern war and the competing missions of peacekeeping, humanitarian missions, and beneficiary care place great demands on the military medical system. Meeting the military medical training challenges of the new millennium requires the best trained and most experienced medical personnel possible. Various strategies for initial and sustainment (continuing) medical training are available to ensure that the medical force is ready for the next mission. Accredited programs both in and out of the military are the mainstay of training for both enlisted personnel and officers, with professional certification serving as the standard for competency. Clinical sustainment training can take place in military medical treatment facilities, civilian institutions, or a combination of the two. When direct patient care opportunities cannot provide the proper mixture of experiences to maintain certain skills, short courses, distance education, and patient simulators can play important roles. Because each training strategy offers certain advantages in different settings, it is likely that military medical departments will need to use all of them. An optimal training environment benefits from all strategies used, in combination or separately. PMID:16435752

  13. Space Age Training

    NASA Technical Reports Server (NTRS)

    1996-01-01

    Teledyne Brown developed a computer-based interactive multimedia training system for use with the Crystal Growth Furnace in the U.S. Microgravity Laboratory-2 mission on the Space Shuttle. Teledyne Brown commercialized the system and customized it for PPG Industries Aircraft Products. The system challenges learners with role-playing scenarios and software-driven simulations engaging all the senses using text, video, animation, voice, sounds and music. The transfer of this technology to commercial industrial process training has resulted in significant improvements in effectiveness, standardization, and quality control, as well as cost reductions over the usual classroom and on-the- job training approaches.

  14. Training: Bridging the gap

    SciTech Connect

    Not Available

    1991-07-01

    These proceedings contain abstracts and papers of the general subject of training for the US DOE by Oak Ridge Associated Universities (ORAU). ORAU is a consortium of colleges and universities that explore opportunities, solve problems, and seek to make increasingly positive contributions to society through science and technology. ORAU operates in four major areas:medical sciences, science and engineering education, training and management systems, and energy and environment systems. Training programs at the Y-12 plant, WMCO, the Mound Laboratory, Sandia laboratories, Hanford, Rocky Flats plant, Los Alamos National Laboratory, and other DOE contractor facilities are presented. Individual papers are indexed separately.

  15. Conducting Elite Performance Training.

    PubMed

    Silverman, Elliott; Tucker, Scott A; Imsdahl, Solveig; Charles, Justin A; Stellato, Mallory A; Wagner, Mercy D; Brown, Kimberly M

    2015-08-01

    Training to excellence in the conduct of surgical procedures has many similarities to the acquisition and mastery of technical skills in elite-level music and sports. By using coaching techniques and strategies gleaned from analysis of professional music ensembles and athletic training, surgical educators can set conditions that increase the success rate of training to elite performance. This article describes techniques and strategies used in both music and athletic coaching, and it discusses how they can be applied and integrated into surgical simulation and education. PMID:26210975

  16. Factory Skills Training at Honeywell.

    ERIC Educational Resources Information Center

    Fey, Carol L.

    1981-01-01

    Because of increased training needs, caused by complex products and employee turnover, Honeywell provides trainers with training packages which facilitate more effective teaching of new production workers. (JOW)

  17. TAP 1: Training Program Manual

    SciTech Connect

    Not Available

    1993-08-01

    The Training Accreditation Program (TAP) was established by the Department of Energy (DOE) to assist in achieving excellence in the development and implementation of performance-based nuclear facility training programs. The TAP establishes the objectives and criteria against which DOE nuclear facility training is evaluated for accreditation. The TAP Staff provides assistance to contractors, develops training guidelines, and evaluates the quality and effectiveness of facility training. This manual describes the accreditation process, provides functional descriptions for positions which require accredited training programs, provides a brief discussion of performance-based training, contains the objectives and criteria that must be addressed in training programs subject to accreditation, and includes a glossary.

  18. Meeting training needs through operator training simulators

    SciTech Connect

    Sendelbach, S.P.; Stewart, W.B.; Morgan, S.W.

    1994-12-31

    An Operator Training Simulator (OTS) can be used to train operators how to safely operate their process plant. Of particular concern are the most critical plant operations and upsets which occur very infrequently during the course of normal every day operations. In that context, the OTS is similar in concept to federally mandated flight simulators which are used to certify commercial airline pilots in take-off, landing and emergency conditions. Of the 170 largest industry losses, 127 occurred during normal operation, while the remaining 43 occurred during other activities. When these figures are combined with the fact that the industry average on-stream factor is 94%, an accident is over five times more likely to occur during other than normal plant operation. The costs associated with atypical plant operation accidents (start-up, shutdown, severe upsets, etc.) are 50% higher than those which occur during normal operation. These statistics combined with new OSHA training requirements suggest that operating companies must now take additional measures to ensure that their operating staff are totally familiar with abnormal process operations, particularly handling of process upsets and emergency operating procedures.

  19. [Government and training demands].

    PubMed

    van der Klaauw, J M

    1998-06-01

    The availability of adequately trained professionals in sufficient numbers is an essential condition for guaranteeing good quality of health care. The Individual Health Care Professions Act fosters the quality by defining the educational requirements and the area of professional competence. The proposal of the working group 'Competition, Deregulation and Quality of Legislation in the health care' to make the dental hygienist free accessible for the patient is in accordance with the policy of the government to foster the efficiency by task delegation. The replacement of sectorial EU-directives by general directives for the free movement of professionals within the European Economic Area has more drawbacks than advantages. Most EU-memberstates are not a proponent for a vocational year after the basic dental training. Deficiencies in the basic dental training should have been met by lengthening the basic training. PMID:12132401

  20. Captains' training at Swissair

    NASA Technical Reports Server (NTRS)

    Grob, N.

    1980-01-01

    The objectives, crucial points, and problems of the individual phase of the captain's basic training are examined. The prerequisites to pilot selection are given as well as a generalized outline of pilot career progression.

  1. Does CRM training work?

    NASA Technical Reports Server (NTRS)

    Helmreich, R. L.

    1991-01-01

    Formal cockpit resource management training in crew coordination concepts increases the percentage of crews rated as above average in performance and decreases the percentage of crews rated as below average.

  2. The Future Training Room.

    ERIC Educational Resources Information Center

    Barbian, Jeff

    2001-01-01

    Looks at some of the electronic learning technology that has already been developed and will become common for training, including robots, lucid dreaming, tele-immersion, human interface technology, among others. (JOW)

  3. Sales Training Center

    ERIC Educational Resources Information Center

    Training in Business and Industry, 1971

    1971-01-01

    After a year of planning, the American Republic Insurance Company has opened a new training facility which occupies a complete floor of the National Headquarters building in Des Moines. Pictures of the facilities are shown. (EB)

  4. Computerized training management system

    DOEpatents

    Rice, H.B.; McNair, R.C.; White, K.; Maugeri, T.

    1998-08-04

    A Computerized Training Management System (CTMS) is disclosed for providing a procedurally defined process that is employed to develop accreditable performance based training programs for job classifications that are sensitive to documented regulations and technical information. CTMS is a database that links information needed to maintain a five-phase approach to training-analysis, design, development, implementation, and evaluation independent of training program design. CTMS is designed using R-Base{trademark}, an-SQL compliant software platform. Information is logically entered and linked in CTMS. Each task is linked directly to a performance objective, which, in turn, is linked directly to a learning objective; then, each enabling objective is linked to its respective test items. In addition, tasks, performance objectives, enabling objectives, and test items are linked to their associated reference documents. CTMS keeps all information up to date since it automatically sorts, files and links all data; CTMS includes key word and reference document searches. 18 figs.

  5. Computerized training management system

    DOEpatents

    Rice, Harold B.; McNair, Robert C.; White, Kenneth; Maugeri, Terry

    1998-08-04

    A Computerized Training Management System (CTMS) for providing a procedurally defined process that is employed to develop accreditable performance based training programs for job classifications that are sensitive to documented regulations and technical information. CTMS is a database that links information needed to maintain a five-phase approach to training-analysis, design, development, implementation, and evaluation independent of training program design. CTMS is designed using R-Base.RTM., an-SQL compliant software platform. Information is logically entered and linked in CTMS. Each task is linked directly to a performance objective, which, in turn, is linked directly to a learning objective; then, each enabling objective is linked to its respective test items. In addition, tasks, performance objectives, enabling objectives, and test items are linked to their associated reference documents. CTMS keeps all information up to date since it automatically sorts, files and links all data; CTMS includes key word and reference document searches.

  6. United Airlines LOFT training

    NASA Technical Reports Server (NTRS)

    Cavanagh, D.; Traub, B.

    1981-01-01

    Line oriented training is used in a broader, more generic sense that as a specific program under FAR 12.1409 and AC 120-35. A company policy was adopted more than twenty years ago requiring that all pilot checks and recurrent training be conducted with a full crew occupying the seats they occupy on the line. Permission was obtained to reschedule the hours for recurrent proficiency training to include one and one-half hours of LOFT flight. The number of emergencies and abnormal procedures which could be undertaken were considered and the introduction of an a occasional incapacitation revealed which person is the most difficult to replace on the widebodies. By using the LOFT concept, every training period can be structured like a typical line flight. The use of LOFT in simulator syllabus development and problems that need to be refined are discussed.

  7. Television Training in India

    ERIC Educational Resources Information Center

    Malik, Iqbal

    1973-01-01

    A general discussion of training programs which resulted from India's decision to expand television as a nationwide network and a vastly expanded use of educational technology within the educational system. (Author/HB)

  8. Training Visual Attention

    ERIC Educational Resources Information Center

    Mulholland, Thomas B.

    1974-01-01

    The effects of brain waves and alpha rhythms on attentiveness to visual stimuli are discussed, and preliminary research findings and research needs are considered in connection with measuring and training for attention. (LH)

  9. Cockpit resource management training

    NASA Technical Reports Server (NTRS)

    White, Lawson C.

    1987-01-01

    The 6th General Flight Crew Training Meeting held in Montreal in May, 1984 was for most IATA member airlines the first time they had been exposed to what was then a relatively new aspect of flight crew training-resource management training. In reviewing the results of this meeting the IATA Flight Crew Training SubCommittee (FCTSC), which had been responsible for the agenda and the meeting itself, concluded that because very few airlines had implemented a program or even appeared to understand the term resource management, a member airline survey should be conducted and the results analyzed. This presentation shows the results of that survey in a form which can be related to the topics of the workshop.

  10. Boiler Operator Training.

    ERIC Educational Resources Information Center

    Miller, Stanton S.

    1978-01-01

    This program, developed by the Nalco Chemical Company, helps with energy conservation in industrial plants. The program takes four to six weeks to complete. The training sessions last for about two hours. (BB)

  11. The Training Guage

    ERIC Educational Resources Information Center

    Palmer, Charles F.

    1970-01-01

    A marketing trainer discusses implications of differing training function concepts and attitudes ranging from completely negative ( None") to firm commitment to upgrading the performance of all members of the organization ( Maximization"). (LY)

  12. Training in Pennsylvania.

    ERIC Educational Resources Information Center

    West, Leo R.

    1983-01-01

    A train trip through Pennsylvania is an education in the history and current activities and industries of the state. One can observe the old railroad depots, landscapes, railroad work crews, strip mines, and Amish communities. (KC)

  13. Swimmer-Training Institutions

    ERIC Educational Resources Information Center

    O'Donnell, R. W.

    1972-01-01

    This satirical essay proposes an institution of higher learning that would prepare students to become swimmers" and swimming instructors. Curriculum, teaching methods, student selection and evaluation are modelled on certain contemporary teacher-training institutes. (PD)

  14. Safety analysts training

    SciTech Connect

    Bolton, P.

    2000-10-01

    The purpose of this task was to support ESH-3 in providing Airborne Release Fraction and Respirable Fraction training to safety analysts at LANL who perform accident analysis, hazard analysis, safety analysis, and/or risk assessments at nuclear facilities. The task included preparation of materials for and the conduct of two 3-day training courses covering the following topics: safety analysis process; calculation model; aerosol physic concepts for safety analysis; and overview of empirically derived airborne release fractions and respirable fractions.

  15. Take the "C" Train

    ERIC Educational Resources Information Center

    Lawton, Rebecca

    2008-01-01

    In this essay, the author recalls several of her experiences in which she successfully pulled her boats out of river holes by throwing herself to the water as a sea-anchor. She learned this trick from her senior guides at a spring training. Her guides told her, "When you're stuck in a hole, take the "C" train."" "Meaning?" The author asked her…

  16. The Training Decision: Training in the Private Sector.

    ERIC Educational Resources Information Center

    Ontario Ministry of Skills Development, Toronto.

    A study was undertaken to examine the factors that influenced the decision of firms to provide training for their employees. It also identified the types of training provided, how the training function was organized and administered within firms, and the extent to which firms provided formal training programs for their employees. The study was…

  17. Training Costs with Reference to the Industrial Training Act.

    ERIC Educational Resources Information Center

    Garbutt, Douglas

    Provisions and implications of the British Industrial Training Act of 1964 (including the system of training grants and levies) are set forth. Procedures for accounting and budgeting for training costs, routines for collecting training information, documents (budgets, cost sheets, control statements) for collecting and controlling costs, means of…

  18. Updated Guidelines for Training Package Developers. Australia's National Training Framework.

    ERIC Educational Resources Information Center

    Australian National Training Authority, Melbourne.

    This document contains the latest guidelines for developing nationally endorsed training packages for use in Australia's system of vocational education and training (VET). Discussed in section A are the following aspects of the context in which the training packages emerged: development of the National Training Framework and associated new…

  19. Research on Synthetic Training: Device Evaluation and Training Program Development.

    ERIC Educational Resources Information Center

    Caro, Paul W.; And Others

    Two studies were conducted to evaluate a fixed-wing instrument procedures training device and to develop a training program for use with it. In the first, a group of trainees who received synthetic instrument flight training with the new device were compared with a control group who did not. Men trained with the device performed more…

  20. Train Right or Don't Train at All.

    ERIC Educational Resources Information Center

    Napier, Nancy K.; Deller, John

    1985-01-01

    A program to train bank operations supervisors to conduct quarterly informal performance appraisals involved three modes: content-only training, content-plus-procedure training, and no training. While content-plus-procedure was predictably the most satisfactory, content-only, because it lacked a practice component, was less effective than no…

  1. Monitoring spike train synchrony.

    PubMed

    Kreuz, Thomas; Chicharro, Daniel; Houghton, Conor; Andrzejak, Ralph G; Mormann, Florian

    2013-03-01

    Recently, the SPIKE-distance has been proposed as a parameter-free and timescale-independent measure of spike train synchrony. This measure is time resolved since it relies on instantaneous estimates of spike train dissimilarity. However, its original definition led to spuriously high instantaneous values for eventlike firing patterns. Here we present a substantial improvement of this measure that eliminates this shortcoming. The reliability gained allows us to track changes in instantaneous clustering, i.e., time-localized patterns of (dis)similarity among multiple spike trains. Additional new features include selective and triggered temporal averaging as well as the instantaneous comparison of spike train groups. In a second step, a causal SPIKE-distance is defined such that the instantaneous values of dissimilarity rely on past information only so that time-resolved spike train synchrony can be estimated in real time. We demonstrate that these methods are capable of extracting valuable information from field data by monitoring the synchrony between neuronal spike trains during an epileptic seizure. Finally, the applicability of both the regular and the real-time SPIKE-distance to continuous data is illustrated on model electroencephalographic (EEG) recordings. PMID:23221419

  2. Good Clinical Practice Training

    PubMed Central

    Arango, Jaime; Chuck, Tina; Ellenberg, Susan S.; Foltz, Bridget; Gorman, Colleen; Hinrichs, Heidi; McHale, Susan; Merchant, Kunal; Shapley, Stephanie; Wild, Gretchen

    2016-01-01

    Good Clinical Practice (GCP) is an international standard for the design, conduct, performance, monitoring, auditing, recording, analyses, and reporting of clinical trials. The goal of GCP is to ensure the protection of the rights, integrity, and confidentiality of clinical trial participants and to ensure the credibility and accuracy of data and reported results. In the United States, trial sponsors generally require investigators to complete GCP training prior to participating in each clinical trial to foster GCP and as a method to meet regulatory expectations (ie, sponsor’s responsibility to select qualified investigators per 21 CFR 312.50 and 312.53(a) for drugs and biologics and 21 CFR 812.40 and 812.43(a) for medical devices). This training requirement is often extended to investigative site staff, as deemed relevant by the sponsor, institution, or investigator. Those who participate in multiple clinical trials are often required by sponsors to complete repeated GCP training, which is unnecessarily burdensome. The Clinical Trials Transformation Initiative convened a multidisciplinary project team involving partners from academia, industry, other researchers and research staff, and government to develop recommendations for streamlining current GCP training practices. Recommendations drafted by the project team, including the minimum key training elements, frequency, format, and evidence of training completion, were presented to a broad group of experts to foster discussion of the current issues and to seek consensus on proposed solutions. PMID:27390628

  3. The alchemy of training.

    PubMed

    Egger-Biniores, Deborah

    2007-04-01

    Training in Zurich has its own special character but also is marked by the very fact that it is in Zurich. Zurich radiates its own distinct energy and carries a specific historical significance in the world of analytical psychology. This, like all things with psychic energy, has a spectrum of meaning. This spectrum, as well as the 'spirit' of the place, will be critically examined, taking into account the 'blessings' and 'curses' of such genius loci. Training in Zurich is experientially based and is first and foremost an initiation: an initiation into symbolic life, or rather life where symbol plays an important role. Training is understood to involve a transformation of one's self, much like the 8th century alchemist Morienus Romanus understood the opus as a 'human transformation system'. It is not merely an education. The requirement of 'immersion' is core to the experience of becoming an analyst in Zurich and this sets up a valuable discomfort between rational intellectual learning and intuitive experience, between knowing and not-knowing. How does this dis-serve the making of an analyst? What is implicit in this immersion and its discomfort? Does it have a role in today's emphasis on clinical and empirical training? Does Zurich still offer something unique and valuable in the world of training, or is it passé? From these questions, the dichotomy of what is 'urgent' and 'essential' in training will be examined. PMID:17403211

  4. Certification of training

    NASA Technical Reports Server (NTRS)

    Gibson, Richard S.

    1994-01-01

    Training has been around as an informal process for countless years. Most higher order animals require some level of training in hunting, social skills, or other survival related skills to continue their existence beyond early infancy. Much of the training is accomplished through imitation, trial and error, and good luck. In some ways the essentials of training in aviation have not deviated from this original formula a great deal. One of the major changes in aviation and other technical areas is that more complex response chains based on a broader base of knowledge are now required. 'To certify' means many things according to the American Heritage dictionary of the English Language. These meanings range from 'to guarantee as meeting a standard' to 'to declare legally insane'. For this discussion, we will use the definition 'an action taken by some authoritative body that essentially guarantees that the instruction meets some defined standard'. In order to make this certification, the responsible body subjects the educational process, training, training device, or simulator to some type of examination to determine its adequacy or validity.

  5. Annual training manual for security training: Protective force

    SciTech Connect

    Not Available

    1990-10-01

    Westinghouse is committed to high quality training relevant to the need of the Protective Forces at the Waste Isolation Pilot Plant (WIPP). The training programs at WIPP are designed to qualify Security personnel to perform WIPP security missions in a professional and responsible manner. The program consists of basic as well as sustainment training, as further described in this plan. This plan documents the WIPP Security training program for security personnel for calendar year 1990. The programs detailed in this plan are designed to adequately train persons to ensure the uninterrupted continuity of Department of Energy (DOE)/Westinghouse operations. The Security Training Program consists of four basic elements. These elements are (1) basic level training; (2) on-the-job training; (3) refresher training; and (4) in-service training.

  6. Job Training Coordination Committee Report.

    ERIC Educational Resources Information Center

    Royall, Robert V., Jr.

    A governor-appointed committee examined South Carolina's job training systems and developed recommendations on providing more effective and efficient job training. A consensus was first developed on conditions that currently exist in the areas of job training, job training's relationship to economic development, and services to target groups.…

  7. Aircraft Simulators and Pilot Training.

    ERIC Educational Resources Information Center

    Caro, Paul W.

    Flight simulators are built as realistically as possible, presumably to enhance their training value. Yet, their training value is determined by the way they are used. Traditionally, simulators have been less important for training than have aircraft, but they are currently emerging as primary pilot training vehicles. This new emphasis is an…

  8. Development Prospects of Vocational Training.

    ERIC Educational Resources Information Center

    Alex, Laszlo

    In the Federal Republic of Germany, apprenticeship is the preponderant form of vocational training. The basis of the training is a private legal contract between the trainer and the trainee. The practical part of the training takes place in the training firm from 3 to 4 days per week, and the theoretical part takes place in a vocational school 1…

  9. Results of Computer Based Training.

    ERIC Educational Resources Information Center

    1978

    This report compares the projected savings of using computer based training to conduct training for newly hired pilots to the results of that application. New Hire training, one of a number of programs conducted continuously at the United Airline Flight Operations Training Center, is designed to assure that any newly hired pilot will be able to…

  10. A Study of Training Program Characteristics and Training Program Effectiveness among Organizations Receiving Training Services from External Training Providers

    ERIC Educational Resources Information Center

    Paek, Jeeyon; Hawley, Joshua D.

    2006-01-01

    The purpose of this study is to examine the impact of training program characteristics on training effectiveness among organizations receiving training services from external training providers. Two surveys were sent to HRD managers and senior managers per company. The results showed that the operational margin of the programs where private…

  11. Model Training Guide. Firefighter I.

    ERIC Educational Resources Information Center

    Hagevig, William A.; Gallagher, Leigh S.

    This firefighter training guide for a 180-hour course was developed to assist training officers in planning training with emphasis on conformance to recommended National Fire Protection Association (NFPA 1001) standards. The material in the guide is referenced to current editions of the International Fire Service Training Association manuals and…

  12. Customer Satisfaction with Training Programs.

    ERIC Educational Resources Information Center

    Mulder, Martin

    2001-01-01

    A model for evaluating customer satisfaction with training programs was tested with training purchasers. The model confirmed two types of projects: training aimed at achieving learning results and at changing job performance. The model did not fit for training intended to support organizational change. (Contains 31 references.) (SK)

  13. The First Aid Training Picture.

    ERIC Educational Resources Information Center

    Moore, Ian

    2000-01-01

    Discusses the history of first aid training provisions in the United Kingdom with respect to the outdoor industry, what to look for in a first aid training provider, an experiential model of first aid training, and the current National Governing Body requirements for first aid training for various types of coaches and instructors. (TD)

  14. Sustainable Library Development Training Package

    ERIC Educational Resources Information Center

    Peace Corps, 2012

    2012-01-01

    This Sustainable Library Development Training Package supports Peace Corps' Focus In/Train Up strategy, which was implemented following the 2010 Comprehensive Agency Assessment. Sustainable Library Development is a technical training package in Peace Corps programming within the Education sector. The training package addresses the Volunteer…

  15. Analysis of Commercial Contract Training.

    ERIC Educational Resources Information Center

    Copeland, D. Robert; And Others

    The report describes the Phase I findings of a two-phase study to determine the applications of commercial contract training to the Navy Training System. The objective of the study is to determine if commercial sources would be cost and training effective alternatives for current and peak Navy skill training requirements. The study was conducted…

  16. ESCAP mobile training scheme.

    PubMed

    Yasas, F M

    1977-01-01

    In response to a United Nations resolution, the Mobile Training Scheme (MTS) was set up to provide training to the trainers of national cadres engaged in frontline and supervisory tasks in social welfare and rural development. The training is innovative in its being based on an analysis of field realities. The MTS team consisted of a leader, an expert on teaching methods and materials, and an expert on action research and evaluation. The country's trainers from different departments were sent to villages to work for a short period and to report their problems in fulfilling their roles. From these grass roots experiences, they made an analysis of the job, determining what knowledge, attitude and skills it required. Analysis of daily incidents and problems were used to produce indigenous teaching materials drawn from actual field practice. How to consider the problems encountered through government structures for policy making and decisions was also learned. Tasks of the students were to identify the skills needed for role performance by job analysis, daily diaries and project histories; to analyze the particular community by village profiles; to produce indigenous teaching materials; and to practice the role skills by actual role performance. The MTS scheme was tried in Nepal in 1974-75; 3 training programs trained 25 trainers and 51 frontline workers; indigenous teaching materials were created; technical papers written; and consultations were provided. In Afghanistan the scheme was used in 1975-76; 45 participants completed the training; seminars were held; and an ongoing Council was created. It is hoped that the training program will be expanded to other countries. PMID:12265562

  17. Psychoanalytic training today.

    PubMed

    Morris, J

    1992-01-01

    Psychoanalytic education in the United States faces multiple challenges as we enter the last decade of this century. (1) Changing interest and career path patterns for psychiatrists have resulted in fewer medical applications for psychoanalytic training. (2) Increased opportunities for full psychoanalytic training of nonphysicians have resulted in increased applications from highly skilled clinicians who often have more clinical experience than their medical colleagues. (3) Increased enrollment of women candidates has required rethinking of progression requirements, in light of their combined careers as professionals and mothers. (4) Independent institutes not accredited by the American Psychoanalytic Association compete for applicants while maintaining training standards that require less time and immersion in psychoanalytic theory and practice. (5) Economic factors increasingly influence the desirability of prolonged psychoanalytic training and the availability of suitable analysands for control analyses. (6) Evolution of theory and practice and the emergence of "new schools" of psychoanalytic thought have rendered the previous psychoanalytic landscape dominated by drive theory and ego psychology more multifaceted and less uniform. The American Psychoanalytic Association and its institutes attempt to understand these changing patterns and take them into consideration in the design and implementation of psychoanalytic training programs. Only one aspect of this complex situation will be described in this work, the current state of psychoanalytic training in the 28 institutes accredited by the American Psychoanalytic Association. Although the data available at this time leave unanswered many important questions about the philosophies that organize the content and emphases of the curriculum in different institutes, much has been learned about the overall structure of psychoanalytic training programs. PMID:1430764

  18. Microneurosurgical Skills Training.

    PubMed

    Yadav, Yad Ram; Parihar, Vijay; Ratre, Shailendra; Kher, Yatin; Iqbal, Mohmed

    2016-03-01

    Microneurosurgical operations differ from other surgery. Longer operative time, narrow and deep-seated operative corridors, hand-eye coordination, fine manipulation, and physiologic tremor present special problems. Proper understanding of visual feedback, control of physiologic tremor, better instrument design, and development of surgical skills with better precision is important for optimal surgical results. Using the pen-type precision grip with well-supported arm, wrist, hand, and fingers avoids fatigue and improves precision. Proper instrument design, patient positioning, hemostasis techniques, tilting operative table, good operative microscope, an adjustable chair, careful use of suction tube, bipolar forceps, and brain retraction play important roles in microneurosurgery. Sufficient clinical case volume or opportunity during routine operative hours may not be available in the beginning for young neurosurgeons; microsurgical training using various models can enable them to gain experience. Training models using deep-seated and narrow operative corridors, drilling, knot-tying technique, and anastomosis using fine sutures under high magnification can be practiced for skill improvement. Training laboratory and simulation modules can be useful for resident training and skill acquisition. Indigenously made inexpensive models and comparatively less expensive microscopes can be used in resource-constrained situations. The maintenance of microsurgical ability should be preserved by staying active in operative practice. The knowledge of ergonomics, proper training, observing hand movements of skillful surgeons, and the use of operative videos can improve skill. Endoscopic assistance, computer-assisted robot hand technique, and microtechnology can provide access to the smallest areas of the body. PMID:25915501

  19. LEAH interdisciplinary training program.

    PubMed

    Robbins, Cynthia L; Rickert, Vaughn D

    2016-08-01

    This article describes the Leadership Education in Adolescent Health (LEAH) interdisciplinary training program in the United States. The Maternal and Child Health Bureau authorized by legislation provides monies to train leaders in adolescent health through a competitive grant process. Currently, seven academic medical centers have funding to provide leadership in adolescent health (LEAH) training in five core disciplines: medicine, nursing, psychology, social work and nutrition. LEAH training programs both ensure high clinical competence in core disciplines serving adolescents and prepare trainees for leadership positions in adolescent health and public health care realms. Together, these programs trained almost 1000 long-term trainees across these five disciplines, and graduates from these programs are working in 45 of the 50 states within the United States. About 90% of these graduates are working with maternal and child/adolescent health populations, and almost all have held leadership positions in the areas of public health, advocacy, public policy, academic medical centers and/or clinical care settings. PMID:26115496

  20. Optimal Physical Training During Military Basic Training Period.

    PubMed

    Santtila, Matti; Pihlainen, Kai; Viskari, Jarmo; Kyröläinen, Heikki

    2015-11-01

    The goal for military basic training (BT) is to create a foundation for physical fitness and military skills of soldiers. Thereafter, more advanced military training can safely take place. Large differences in the initial physical performance of conscripts or recruits have led military units to develop more safe and effective training programs. The purpose of this review article was to describe the limiting factors of optimal physical training during the BT period. This review revealed that the high volume of low-intensity physical activity combined with endurance-type military training (like combat training, prolonged physical activity, and field shooting) during BT interferes with optimal development of maximal oxygen uptake and muscle strength of the soldiers. Therefore, more progressive, periodized, and individualized training programs are needed. In conclusion, optimal training programs lead to higher training responses and lower risks for injuries and overloading. PMID:26506180

  1. [Dichoptic training for amblyopia].

    PubMed

    Bach, M

    2016-04-01

    Dichoptic training is a promising new therapeutic approach to amblyopia, which employs simultaneous and separate stimulation of both eyes (thus dichoptic). The contrast for the good eye is reduced thus aiming at a balance with the amblyopic eye. In contrast to monocular patching, binocular vision is trained by video game tasks that can only be solved binocularly. To date the average gain in visual acuity achieved in currently available studies is only 0.20 ± 0.07 logMAR and is not significantly better than competing treatment options. This article explains the basic approach of dichoptic training, summarizes pertinent studies, names unsolved problems and closes with a personal critical assessment. PMID:27059985

  2. Airline Crew Training

    NASA Technical Reports Server (NTRS)

    1989-01-01

    The discovery that human error has caused many more airline crashes than mechanical malfunctions led to an increased emphasis on teamwork and coordination in airline flight training programs. Human factors research at Ames Research Center has produced two crew training programs directed toward more effective operations. Cockpit Resource Management (CRM) defines areas like decision making, workload distribution, communication skills, etc. as essential in addressing human error problems. In 1979, a workshop led to the implementation of the CRM program by United Airlines, and later other airlines. In Line Oriented Flight Training (LOFT), crews fly missions in realistic simulators while instructors induce emergency situations requiring crew coordination. This is followed by a self critique. Ames Research Center continues its involvement with these programs.

  3. Alliance-focused training.

    PubMed

    Eubanks-Carter, Catherine; Muran, J Christopher; Safran, Jeremy D

    2015-06-01

    Alliance-focused training (AFT) aims to increase therapists' ability to recognize, tolerate, and negotiate alliance ruptures by increasing the therapeutic skills of self-awareness, affect regulation, and interpersonal sensitivity. In AFT, therapists are encouraged to draw on these skills when metacommunicating about ruptures with patients. In this article, we present the 3 main supervisory tasks of AFT: videotape analysis of rupture moments, awareness-oriented role-plays, and mindfulness training. We describe the theoretical and empirical support for each supervisory task, provide examples based on actual supervision sessions, and present feedback about the usefulness of the techniques from trainees in our program. We also note some of the challenges involved in conducting AFT and the importance of maintaining a strong supervisory alliance when using this training approach. PMID:25150677

  4. Training SVMs without offset

    SciTech Connect

    Steinwart, Ingo; Hush, Don; Scovel, Clint

    2009-01-01

    We develop, analyze, and test a training algorithm for support vector machine cla.'>sifiers without offset. Key features of this algorithm are a new stopping criterion and a set of working set selection strategies that, although inexpensive, do not lead to substantially more iterations than the optimal working set selection strategy. For these working set strategies, we establish convergence rates that coincide with the best known rates for SYMs with offset. We further conduct various experiments that investigate both the run time behavior and the performed iterations of the new training algorithm. It turns out, that the new algorithm needs less iterations and run-time than standard training algorithms for SYMs with offset.

  5. Training for telemedicine.

    PubMed

    Blignault, I; Kennedy, C

    1999-01-01

    All technology requires training. Simply installing videoconferencing equipment in a hospital or health centre, and leaving the manufacturer's manual nearby, is not sufficient to encourage or maintain its use for telemedicine. Presenting clinicians and other users with thick policy and procedure documents to be read and understood is also not helpful. Following initial awareness raising about telemedicine and its potential, staff must be properly trained: not just in how to turn on the equipment but in how to use it effectively for consultation, education and administrative purposes. Training needs to be continual, especially where staff turnover is high. It should be practical, pitched at different levels, incorporated into mainstream activities such as orientation courses and staff development days wherever possible, and supplemented with clear, straightforward protocols and user-friendly instruction manuals. PMID:10534864

  6. Financing Residency Training Redesign

    PubMed Central

    Carney, Patricia A.; Waller, Elaine; Green, Larry A.; Crane, Steven; Garvin, Roger D.; Pugno, Perry A.; Kozakowski, Stanley M.; Douglass, Alan B.; Jones, Samuel; Eiff, M. Patrice

    2014-01-01

    Background Redesign in the health care delivery system creates a need to reorganize resident education. How residency programs fund these redesign efforts is not known. Methods Family medicine residency program directors participating in the Preparing Personal Physicians for Practice (P4) project were surveyed between 2006 and 2011 on revenues and expenses associated with training redesign. Results A total of 6 university-based programs in the study collectively received $5,240,516 over the entire study period, compared with $4,718,943 received by 8 community-based programs. Most of the funding for both settings came from grants, which accounted for 57.8% and 86.9% of funding for each setting, respectively. Department revenue represented 3.4% of university-based support and 13.1% of community-based support. The total average revenue (all years combined) per program for university-based programs was just under $875,000, and the average was nearly $590,000 for community programs. The vast majority of funds were dedicated to salary support (64.8% in university settings versus 79.3% in community-based settings). Based on the estimated ratio of new funding relative to the annual costs of training using national data for a 3-year program with 7 residents per year, training redesign added 3% to budgets for university-based programs and about 2% to budgets for community-based programs. Conclusions Residencies undergoing training redesign used a variety of approaches to fund these changes. The costs of innovations marginally increased the estimated costs of training. Federal and local funding sources were most common, and costs were primarily salary related. More research is needed on the costs of transforming residency training. PMID:26140119

  7. Astronaut training manual

    NASA Technical Reports Server (NTRS)

    Coleman, E. A.

    1980-01-01

    Scientific information from previous space flights, space medicine, exercise physiology, and sports medicine was used to prepare a physical fitness manual suitable for use by members of the NASA astronaut population. A variety of scientifically valid exercise programs and activities suitable for the development of physical fitness are provided. Programs, activities, and supportive scientific data are presented in a concise, easy to read format so as to permit the user to select his or her mode of training with confidence and devote time previously spent experimenting with training routines to preparation for space flight. The programs and activities included were tested and shown to be effective and enjoyable.

  8. Accrediting industrial safety training programs

    SciTech Connect

    Beitel, L.

    1992-12-31

    There are job-specific training requirements established by regulations that Impose stringent training requirements on a contractor, for example, the Occupational Safety & Health Act (OSHA). Failure to comply with OSHA training requirements can result in severe penalties being levied against a company. Although an accredited training program is expensive, it is a possible solution for minimizing risks associated with job-specific training requirements for employees. Operating DOE contractors direct approximately 10 percent of the operating funds toward training activities. Training needs for contractors span a broad range, from requirements awareness training for managers, to general training required on a one-time basis for all employees, to highly specialized training programs for employees involved In clean-up operations at hazardous waste sites. With this kind of an investment in training, it is logical to maximize the most return on an investment of training funds and to limit exposure to liability suits whenever possible. This presentation will provide an overview of accredited industrial safety programs. The criteria for accredited industrial safety programs will be defined. The question of whether accredited training programs are necessary will be examined. Finally, advantages and disadvantages will be identified for accrediting industrial safety training programs.

  9. Accrediting industrial safety training programs

    SciTech Connect

    Beitel, L.

    1992-01-01

    There are job-specific training requirements established by regulations that Impose stringent training requirements on a contractor, for example, the Occupational Safety Health Act (OSHA). Failure to comply with OSHA training requirements can result in severe penalties being levied against a company. Although an accredited training program is expensive, it is a possible solution for minimizing risks associated with job-specific training requirements for employees. Operating DOE contractors direct approximately 10 percent of the operating funds toward training activities. Training needs for contractors span a broad range, from requirements awareness training for managers, to general training required on a one-time basis for all employees, to highly specialized training programs for employees involved In clean-up operations at hazardous waste sites. With this kind of an investment in training, it is logical to maximize the most return on an investment of training funds and to limit exposure to liability suits whenever possible. This presentation will provide an overview of accredited industrial safety programs. The criteria for accredited industrial safety programs will be defined. The question of whether accredited training programs are necessary will be examined. Finally, advantages and disadvantages will be identified for accrediting industrial safety training programs.

  10. [Surgical training with simulators in training centers].

    PubMed

    Rodríguez-García, José Ignacio; Turienzo-Santos, Estrella; Vigal-Brey, Guillermo; Brea-Pastor, Agustín

    2006-06-01

    The development of new endoscopic procedures and minimally-invasive surgical interventions has led the methodology used to date to be questioned. Greater demand for safety by patients, the growth of the health budget and the reduced time available for training have led to the proliferation of centers with accredited personnel in which the knowledge and surgical skills necessary for the controlled incorporation of these techniques can be acquired. Simulators are available for the learning of both digestive endoscopy and laparoscopic techniques. These simulators are more or less dynamic, virtual, with viscera or mixed; even live animals can be used. Thus, the various techniques can be incorporated into clinical practice safely and effectively and at a reasonable cost. Simulators also allow evaluation and follow-up of the skills acquired. PMID:16768997

  11. Automating the training development process

    NASA Technical Reports Server (NTRS)

    Scott, Carol J.

    1993-01-01

    The Automated Training Development System (ATDS) was developed as a training tool for the JPL training environment. ATDS is based on the standard for military training programs and is designed to develop training from a system perspective, focusing on components in terms of the whole process. The principal feature of ATDS is data base maintainability. Everything is contained and maintained within the data base, and, if properly developed, it could be a training component of a software delivery and provided to CM as a controlled item. The analysis, development, design, presentation, and reporting phases in the ATDS instructional design method are illustrated.

  12. Intensive training in young athletes.

    PubMed Central

    Maffulli, N; Pintore, E

    1990-01-01

    An increasing number of children take part in organized sporting activities, undergoing intensive training and high level competition from an early age. Although intensive training in children may foster health benefits, many are injured as a result of training, often quite seriously. This paper reviews some of the areas of research dealing with intensively trained young athletes, and focuses on physical, cardiovascular and muscular effects, sports injuries and psychological effects of intensive training. It is concluded that measures should be taken to modify present training and competition schemes to avoid the deleterious effects of intensive physical activity on these children. PMID:2097019

  13. Boating Safety Training Manual.

    ERIC Educational Resources Information Center

    Coast Guard, Washington, DC.

    The training manual serves as the text for the Coast Guard's boating safety 32-hour course and for the D-8 Qualification Code Recertification Course. The manual is designed for self-study or for use with an instructor-led course. Each chapter concludes with a quiz to be used as a review of chaper content. Opening chapters review the use of the…

  14. Two Centuries of Training

    ERIC Educational Resources Information Center

    Greenwood, Frank; And Others

    1976-01-01

    Some principles which have guided the U.S. Coast Guard in selecting, training, and developing capable managers in the past 200 years include: quality organizational executives, a selection process based on past performance, management of time, management experience, high ethical level, organizational "belonging," and immediate and serious…

  15. Basic Science Training Program.

    ERIC Educational Resources Information Center

    Brummel, Clete

    These six learning modules were developed for Lake Michigan College's Basic Science Training Program, a workshop to develop good study skills while reviewing basic science. The first module, which was designed to provide students with the necessary skills to study efficiently, covers the following topics: time management; an overview of a study…

  16. Training at Kodak

    ERIC Educational Resources Information Center

    Training in Business and Industry, 1971

    1971-01-01

    Article discusses the vast number of educational programs which the Kodak company offers to employees, supervisors, workers in related fields and to the community by way of educational training programs for inner city and underprivileged youth. Discussed are the ways in which Kodak enhances its own position in the industry by keeping its employees…

  17. Virtual Safety Training.

    ERIC Educational Resources Information Center

    Fuller, Scott; Davis, Jason

    2003-01-01

    The Multimedia Tool Box Talk is a web-based quick reference safety guide and training tool for construction personnel. An intended outcome of this effort was to provide an efficient and effective way to locate and interpret crucial safety information while at the job site. The tool includes information from the Occupational Safety and Health…

  18. Training on Demand

    ERIC Educational Resources Information Center

    Maliszewski, Casey; Nespoli, Lawrence; Rosa, Robert

    2013-01-01

    Nearly a decade ago officials at New Jersey's 19 community colleges sought a way for the state's two-year career and technical college system to work together to more efficiently serve the workforce development needs of the state. The goal was to say to businesses, "If you have a training need, we can help by delivering the "combined" training…

  19. Music Training in Germany.

    ERIC Educational Resources Information Center

    Tapia, Ivan, Ed.

    1994-01-01

    This special-issue volume examines music education in the two Germanies and how music has had a great influence in the culture of the nations. The presentation is a professional and objective portrayal of music training and cultivation in Germany in the last decade of the present century. The articles attempt to outline the problems and tasks that…

  20. Transfer of Training.

    ERIC Educational Resources Information Center

    1996

    This document consists of three papers presented at a symposium on transfer of training moderated by Gene Roth at the 1996 conference of the Academy of Human Resource Development (AHRD). "Validation of a Transfer Climate Instrument" (Reid A. Bates et al.) reports a study that attempted to validate Rouiller and Goldstein's (1993) eight-factor…

  1. Train better by computer

    SciTech Connect

    Furgang, S.R.

    1989-01-01

    Computer-based training (CBT) is used in a new approach to training refinery operators. The system was developed at Mobil's U.S. headquarters, with field help. In the early 1980s the authors evaluated the mainframe Plato system from Control Data, considered by most the ''granddad'' of CBT systems. A couple of years later, when the CPM-based micro computers became popular, they traded in Plato for stand-alone desktop that offered most of the same features without the mainframe's cost and connectivity limitations. Their evaluation continued into the mid-1980s after IBM established its PC line which became the corporate hardware standard. CBT is both right for the times and particularly suited to the hydrocarbon processing industry's needs. Mobil has taken significant steps to benefit from all that CBT offers by developing two curricula for operator training. The first, the Fundamentals Series, provides the background needed for any operations assignment. The second curriculum, the Advanced Series, covers specific process unit job training.

  2. Training Career Practitioners

    ERIC Educational Resources Information Center

    Niles, Spencer G.; Engels, Dennis; Lenz, Janet

    2009-01-01

    This article summarizes 8 presentations from Group 8 on the theme "Training of Researchers and Practitioners," which were part of the 2007 joint symposium of the International Association for Educational and Vocational Guidance, Society for Vocational Psychology, and National Career Development Association held in Padua, Italy. Three themes…

  3. FIRE SERVICE TRAINING.

    ERIC Educational Resources Information Center

    BERNDT, WILLIAM M.; AND OTHERS

    STUDENTS MAY USE THIS REVISED MANUAL IN FIRE STATION OR TRAINING CENTER EXTENSION PROGRAMS FOR IMPROVING THE COMPETENCIES AND SKILLS OF LOCAL FIRE PERSONNEL IN THE SPECIALIZED FIELD OF FIRE SERVICE. IT WAS DEVELOPED BY A STATEWIDE COMMITTEE OF FIRE-FIGHTING CONSULTANTS AND ADVISORY GROUPS. THE 26 CHAPTERS PROVIDE BOTH BASIC AND ADVANCED TECHNICAL…

  4. Generalization and Training.

    ERIC Educational Resources Information Center

    Jenkins, W. O.

    This paper contends that human organisms are not qualitatively different from infra-human ones. The same principles apply to the acquisition, maintenance and weakening of complex verbal skills as mathematics as to rats pressing bars or pigeons pecking windows. Two aspects of child research are assessment and training. Assessment involves…

  5. Model Cities Training Program.

    ERIC Educational Resources Information Center

    Tennessee Univ., Chattanooga.

    The Model Cities Training Program, the first in the country, is a 10-session course to be conducted in seminar form under the direction of the University of Tennessee at Chattanooga. The objective is to enable the 50 members of the Community Development Administration Board of Directors to: acquire knowledge of the structure of the Model Cities…

  6. Lennox - Student Training Equipment.

    ERIC Educational Resources Information Center

    Lennox Industries, Inc., Marshalltown, IA.

    Presents a series of demonstration units designed by Lennox Industries for the purpose of training students to become familiar with Lennox mechanical equipment. Demonstrators are designed to present technical information in a clear simplified manner thus reducing frustration for the beginning trainee. The following demonstrators are available--(1)…

  7. Recruitment and Training. Symposium.

    ERIC Educational Resources Information Center

    2002

    This document contains three papers from a symposium on recruitment and training. "College Choice: The State of Marketing and Effective Student Recruitment Strategies" (Fredrick Muyia Nafukho, Michael F. Burnett) reports on a study of the recruitment strategies used by Louisiana State University's admissions office and College of Agriculture that…

  8. Training for Nonviolent Action.

    ERIC Educational Resources Information Center

    Olson, Theodore W.; Shivers, Lynne

    The theory and practice of nonviolent action training as it exists to date are reviewed in this pamphlet. A response to a renewal of interest in alternative forms of social action, the pamphlet results specifically from an international seminar of experienced organizers and trainers held at Preston Patrick, Westmorland, England, June 27 - July 2,…

  9. Rx for Tutor Training.

    ERIC Educational Resources Information Center

    Rubenstein, Ilene; And Others

    Tutor training programs in composition which emphasize interpersonal skills while offering concentrated correctness doses of mechanics and grammar are inherently limiting. While interpersonal skills are important, they only superficially address the complex situation of tutoring. A prescription for a healthy tutor program, one which would allow…

  10. Tips on Training.

    ERIC Educational Resources Information Center

    DePutte, Toni Van, Ed.; And Others

    The articles in this collection are addressed to athletic coaches or ancillary health personnel practitioners who treat athletes' injuries. Emphasis is upon proper immediate care, recognition and referral of serious injuries, and preventive measures to reduce injuries. Papers include: (1) "Planning a High School Athletic Training Program" (Stanley…

  11. Career Guidance Training.

    ERIC Educational Resources Information Center

    Young, John E.

    The 1972 Jet Propulsion Laboratory (JPL) Career Training Program was aimed at placing counselors in actual work situations (as new employees) to enable them to experience that which they must describe to students if they are to do an effective job in career counseling. The overall purpose was to give counselors or teachers and administrators an…

  12. Adjustment to Recruit Training.

    ERIC Educational Resources Information Center

    Anderson, Betty S.

    The thesis examines problems of adjustment encountered by new recruits entering the military services. Factors affecting adjustment are discussed: the recruit training staff and environment, recruit background characteristics, the military's image, the changing values and motivations of today's youth, and the recruiting process. Sources of…

  13. Training of Trainers.

    ERIC Educational Resources Information Center

    Gormley, Wilma J.; Austin, John H.

    1985-01-01

    Discusses specific training methods and common characteristics of participants in workshops sponsored by Agency for International Development Water and Sanitation for Health Project for extension agents, who will act as trainers in transfer of sanitation technology in developing nations. Recommendations for conducting such workshops in…

  14. Training of Penitentiary Agents

    ERIC Educational Resources Information Center

    Zaharia, Victor

    2009-01-01

    The training of penitentiary staff directly influences the efficiency of a prison system and the observance of human rights in places of detention. On the one hand, there are no "perfect" detainees. The causes that led to imprisonment could be diverse. On the other hand, the possibilities and resources available to a prison system could be limited…

  15. Training Circuit Boards

    NASA Technical Reports Server (NTRS)

    1986-01-01

    Young Electronics Company's new product is a testboard developed by the Jet Propulsion Laboratory as a tool for training and qualifying personnel in board assembly and in the art of soldering components without damaging boards or components. The boards are used for pre- employment testing and employee requalification.

  16. LCDs Revolutionize Group Training.

    ERIC Educational Resources Information Center

    Mandell, Mel

    1987-01-01

    Describes a screen projector based on liquid crystal display (LCD) that duplicates the monitor of a microcomputer and may be used in group training sessions for demonstration purposes. Suggestions of what features to look for and a buyer's guide are provided. (CLB)

  17. INDUSTRIAL TRAINING RESEARCH REGISTER.

    ERIC Educational Resources Information Center

    Ministry of Labour, London (England).

    IN THIS CLASSIFIED REGISTER OF CURRENT AND RECENTLY COMPLETED STUDIES OF INDUSTRIAL TRAINING IN GREAT BRITAIN, INDIVIDUAL PROJECTS ARE ARRANGED BY THE ITEM NUMBER JUDGED MOST IMPORTANT, AND THE NUMBERS OF OTHER RELEVANT INDUSTRIAL RESEARCH PROJECTS ARE INSERTED AT THE END OF EACH SECTION TO PROVIDE CROSS REFERENCES. DESCRIPTIONS INCLUDE THE TITLE…

  18. Training in the Clouds

    ERIC Educational Resources Information Center

    Pretlow, Cassi; Jayroe, Tina

    2010-01-01

    In this article, the authors share how cloud-based applications, such as Google Calendar, Wikidot, Google Docs, Google Sites, YouTube, and Craigslist, played a big part in the success of their plan of implementing a technology training program for customers and employees. A few years ago the Denver Public Library, where the authors work, developed…

  19. Occupational Training in Industry.

    ERIC Educational Resources Information Center

    Stromsdorfer, Ernst W.; Barclay, Suzanne

    A significant amount of on-the-job occupational training is occurring in the private sector, though the data on its extent and nature are extremely sketchy. Estimates of total economic costs in the 1974-75 period range from a crude measure of 100 billion dollars to one that is somewhat more reliable of about 40 to 50 billion dollars. Most of this…

  20. The Training Audit.

    ERIC Educational Resources Information Center

    Buckley, Roger; Caple, Jim

    1984-01-01

    This article discusses the role of the auditor and describes a seven-stage training audit model currently being employed by a multinational United Kingdom bank. Stages include familiarization, auditing the client, auditing the program, auditing the program organizer, auditing the consumer, presenting the report, and tasking action agencies. (CT)

  1. Assertive Training in Groups

    ERIC Educational Resources Information Center

    Sansbury, David L.

    1974-01-01

    This article describes a group approach to helping the nonassertive client. After describing the group composition and goals, he presents a session by session description for conducting the assertive training group. In addition, he presents suggestions based on experiences in leading the group. (Author)

  2. Training Public Health Assistants.

    ERIC Educational Resources Information Center

    Quinlan, Paul; And Others

    Funded by the Office of Economic Opportunity and carried out in Springfield, Massachusetts, during 1965-67, this training project sought to meet employment needs of disadvantaged high school graduates, the shortage of health professionals, and the need to improve and coordinate professional public health services. It combined a half-time,…

  3. Microcomputer Videogame Based Training.

    ERIC Educational Resources Information Center

    Driskell, James E.; Dwyer, Daniel J.

    1984-01-01

    Examines advantages of using videogame format for training, including practicality, relatively inexpensive hardware, ability to use multi-purpose software modules, motivational advantages of increased student interest and instructional effectiveness, and instructional features such as the adaptive interactive nature of videogames. An example of a…

  4. [Training can be fun!].

    PubMed

    Piano, Julie; Robert, Philippe; Foulon, Pierre

    2015-05-01

    Serious games are becoming ever more innovative and increasingly popular within the healthcare profession. Developed as a remote training tool, they support health professionals in a fun way, enabling them to update their knowledge and aptitude. This article looks at a serious game developed for nursing homes. PMID:26145695

  5. Slow Train Coming

    ERIC Educational Resources Information Center

    McNair, Stephen

    2011-01-01

    The battle for an adequate careers guidance service for adults has been a long one. A succession of governments have been formally committed to lifelong learning, to broadening choice and diversity in education and training, and to asking individuals to take more control of (and pay more for) their education, but only in the last few years have…

  6. Old Train, New Track.

    ERIC Educational Resources Information Center

    Prokopeak, Andrew W.

    1984-01-01

    Presents ideas for using model trains as a teaching tool and/or minicourse in junior high school science classes. Students investigate such topics as electric motor operation, electric potential, resistance, electromagnets, transformers, switches, centripetal force, cam mechanism, circuitry and wiring techniques, and ammeters. Directions for…

  7. Vocational Education and Training.

    ERIC Educational Resources Information Center

    van Leeuwen, Fred, Ed.

    2000-01-01

    This issue of the quarterly Education International focuses on vocational education and training (VET). The editorial, "Education and the Wealth of Nations" (Fred van Leeuwen), focuses on provision of quality education for all. "Education International's (EI's) First Joint Worldwide Action on Education Issues" (Elie Jouen) describes the Global…

  8. Internet Training Needs Inventory.

    ERIC Educational Resources Information Center

    Weglarz, Shirley

    The Office of Institutional Research at Johnson County Community College (JCCC), Kansas, conducted a survey to help determine the level of interest in Internet training courses. Surveys were mailed in November 1998 to 9,982 individuals selected from six lists representing the JCCC Business and Industry Division's current and potential clients. A…

  9. Children's Collective Training Manual.

    ERIC Educational Resources Information Center

    Kimbrough, Jackie; And Others

    This children's collective manual describes a training program for parents and teachers of preschool children which is designed to encourage cooperative, community-oriented styles of group interaction in black preschool children. Developed by the Children's Collective of the Coordinated Child Care Council of South Los Angeles, the program is based…

  10. Mir Training Facility

    NASA Technical Reports Server (NTRS)

    1995-01-01

    A full-scale mockup of Russia's Space Station with the core module called Mir in the center. Other modules connected to the core include Kvant, Kvant II and Kristall. The mockup at the Gagarin Cosmonaut Training Center in Star City, Russia is used for cos

  11. Mir Training Facility

    NASA Technical Reports Server (NTRS)

    1995-01-01

    A full-scale mockup of Russia's Space Station serves as one of the several training aids for cosmonaut flights aboard the orbiting laboratory. The core module - called Mir, for world of space - was launched in February 1986 and now serves as the main livi

  12. Training Health Care Paraprofessionals

    ERIC Educational Resources Information Center

    Linton, Corinne B.

    1977-01-01

    This review of the allied health occupations training programs offered by Brevard Community College (Cocoa, Florida) covers organization of the division, objectives, selection and admission process, instructional delivery system, clinical facilities, advisory committees, high school relations, continuing education programs, and program success.…

  13. Assessing Technical Training Needs.

    ERIC Educational Resources Information Center

    Schwaller, Anthony E.; Slipy, Dave

    1985-01-01

    Describes the results of a joint project of St. Cloud State University (Minnesota) and DeZURIK Corporation (a manufacturer and distributor of industrial valves) which developed and implemented a technical training needs questionnaire for use with the company's employees. Student involvement in the process is noted. (MBR)

  14. Explicating Individual Training Decisions

    ERIC Educational Resources Information Center

    Walter, Marcel; Mueller, Normann

    2015-01-01

    In this paper, we explicate individual training decisions. For this purpose, we propose a framework based on instrumentality theory, a psychological theory of motivation that has frequently been applied to individual occupational behavior. To test this framework, we employ novel German individual data and estimate the effect of subjective expected…

  15. Training's Annual Salary Survey.

    ERIC Educational Resources Information Center

    Goldwasser, Donna

    2000-01-01

    A survey of trainers' (n=2,003) salaries compared salaries based on size of companies, geographic location, level of education, experience, gender, and age. At $60,794, the average training salary is only 2.8 percent higher than last year, compared to a 4.6 percent increase for the average salaried employee. (JOW)

  16. Bus Training Handbook.

    ERIC Educational Resources Information Center

    Dorny, Audrea; Cole, ChiKay

    This manual presents guidelines for teaching students with disabilities necessary skills for safe and independent travel on public buses. Six guidelines for teachers include: (1) participate in bus training; (2) use wise and intelligent judgment; (3) utilize the bus checklist; (4) know and teach bus rules; (5) know bus routes; and (6) know bus…

  17. Hiring and Training Workers.

    ERIC Educational Resources Information Center

    Bishop, John, Ed.

    This report is one of a series of papers analyzing how employers select and train employees and the implications of their behavior for schools. Data for the study were gathered in telephone interviews with more than 3,500 employees conducted by the Gallup Organization. This report is organized in three chapters. In the first chapter, John Barron…

  18. Teacher-Training Projects

    ERIC Educational Resources Information Center

    Greer, Leslie

    1977-01-01

    The Sociedade Brasileira de Cultura Inglesa of Sao Paolo, Brazil, is an English teaching center which also runs an introductory course to train teachers of English. This article describes some of the projects completed by prospective teachers; they include language games, pictures, cartoons, role-playing and writing creative dialogue. (CHK)

  19. Functional Communication Training

    ERIC Educational Resources Information Center

    Durand, V. Mark; Moskowitz, Lauren

    2015-01-01

    Thirty years ago, the first experimental demonstration was published showing that educators could improve significant challenging behavior in children with disabilities by replacing these behaviors with forms of communication that served the same purpose, a procedure called functional communication training (FCT). Since the publication of that…

  20. Rhetorical Types and "Training."

    ERIC Educational Resources Information Center

    Palmerton, Patricia R.

    The speech communications professionals who are involved in training and consulting need to conceptualize and talk about their activities as communication educators--not as trainers. Clients should be taught to make interpretive choices appropriate to their rhetorical environment, in addition to learning basic skills. As educators in the corporate…

  1. Vocational Training in Norway.

    ERIC Educational Resources Information Center

    Mortensen, Gunnar; Persson, Sven

    The purpose of this publication is to present Norway's industrial status, industrial manpower profile, and philosophy of vocational training. The relation between general and vocational education is discussed and Norway's educational system is outlined. Occupational areas receiving detailed discussion include (1) Agriculture, (2) Maritime…

  2. Training Student Organizers Curriculum.

    ERIC Educational Resources Information Center

    Zamm, Michael; Hurtado, Denise

    The purpose of this curriculum is to help teachers and field supervisors at the college, high school, and advanced junior high school level train students to organize environmental improvement projects. It can also be used by graduate/undergraduate students who are supervising secondary school students. The curriculum may be started at any point…

  3. Continuing Education and Training.

    ERIC Educational Resources Information Center

    Alger, Ian

    1993-01-01

    Discusses history of first training program in Marriage and Family Counseling (MFC) at Hofstra University established in 1982. Explains context of MFC graduate programs and philosophy and model of MFC programs. Describes Marriage and Family Counseling Club, mentoring program, Marriage and Family Counseling Newsletter, and marriage and family…

  4. Circulation Aide Training Manual.

    ERIC Educational Resources Information Center

    Bergeson, Alan O.

    This training manual provides instruction on shelving and other duties for student assistants in the learning resources center at the College of Dupage, located in Illinois. It is noted that prospective student circulation aides are required to read the manual and pass a written test on policies and procedures before they are allowed to shelve…

  5. Dental Training Films.

    ERIC Educational Resources Information Center

    Veterans Administration Medical Center, Washington, DC.

    This dental training films catalog is organized into two sections. Section I is a category listing of the films by number and title, indexed according to generalized headings; categories are as follow: anatomy, articulator systems, complete dentures, dental assisting, dental laboratory technology, dental materials, dental office emergencies,…

  6. Training 2000. Publication 1480.

    ERIC Educational Resources Information Center

    Internal Revenue Service (Dept. of Treasury), Washington, DC.

    This document reports the results of an examination of the external and internal environments of the Internal Revenue Service (IRS) and the most promising technological tools and methods to create the training system capabilities the IRS will need in the future. Following an executive summary, chapter 1 describes how the document was developed.…

  7. Training Guidelines: Bricks Operatives.

    ERIC Educational Resources Information Center

    Ceramics, Glass, and Mineral Products Industry Training Board, Harrow (England).

    This manual offers guidelines for training of personnel involved in the manufacture of bricks, including employment practices; handling and preparation of raw materials; making, drying, firing, sorting, packing, and loading of bricks. A major emphasis is placed on industrial safety. (MF)

  8. Exercise training in asthma.

    PubMed

    Satta, A

    2000-12-01

    Asthma is a chronic disease that is often limiting the exercise capacity. Rehabilitation programs are recommended and widely applied in asthmatic patients, and exercise prescription is a keystone of these programs. The impairment of exercise performance in asthmatics, the role of exercise training in such patients, the mechanisms of its beneficial effects and the suggested programs are discussed in a review, accordingly to the current evidence and available data in scientific literature. Exercise performance is impaired in most asthmatics. There is no conclusive evidence that asthma may involve a ventilatory limitation to exercise. The lesser fitness in asthmatics seems mainly due to inactivity and sedentary lifestyle. Exercise induced asthma (EIA) is a significant problem, and the best approach to minimise its effects on exercise capacity is prevention. Exercise training has been proved to have health-related benefits and to improve the quality of life. There is substantial evidence that exercise training increases exercise performance and fitness in asthmatics. It is still unclear whether physical training improves pulmonary function and bronchial responsiveness. Since asthma ranges widely, exercise prescription varies for each patient. The proper selection of the patients and the choice of exercise programs are the steps required. Accordingly with the severity of the disease, exercise strategies may range from sports activities to, when the disease is severe, inpatient hospital programs that overlap with COPD rehabilitation. Further research to clarify some aspects (effects on pulmonary function and EIA, outcomes, cost-benefit relationship) is necessary. PMID:11296996

  9. Drive-Through Training

    ERIC Educational Resources Information Center

    Carter, Margie

    2010-01-01

    In this article, the author discusses how the early childhood field's approach to staff training reflects the drive-through, fast-food culture. Year after year directors send their teachers to workshops to get some quick refresher techniques. The author suggests that rather than focusing professional development on topics, focus on observing…

  10. Vocational Training in Ireland.

    ERIC Educational Resources Information Center

    Dooney, Roy; Dunne, Paul

    This monograph, one of a series of studies of vocational education in the countries of the European Communities, describes the vocational training system in Ireland. The study was compiled from existing statistics and descriptions, and most figures cited refer to 1984. The report is organized in eight chapters. Chapter 1 covers population,…

  11. Vocational Training in Belgium.

    ERIC Educational Resources Information Center

    Debaty, Pol

    This document on vocational training in Belgium contains eight chapters, a list of abbreviations, and a nine-item list of main texts consulted. Chapter 1 describes Belgium and its inhabitants, including population, age structures, migration, the working population, public and private sectors, unemployment, and measures to combat unemployment.…

  12. Training for Hospitality.

    ERIC Educational Resources Information Center

    Herman, Francine A.; Eller, Martha E.

    1991-01-01

    The labor problems of hotels and restaurants are being addressed with partnership training initiatives. Examples are the Stratford Chef's School in Ontario, the University of Hawaii's Tourism Industry Management Program, and an English-as-a-Second-Language program sponsored by labor and management in New York City hotels. (SK)

  13. Waterworks Operator Training Manual.

    ERIC Educational Resources Information Center

    Missouri Univ., Columbia. Instructional Materials Lab.

    Sixteen self-study waterworks operators training modules are provided. Module titles are the following: basic mathematics, basic chemistry, analysis procedures, microbiology, basic electricity, hydraulics, chlorination, plant operation, surface water, ground water, pumps, cross connections, distribution systems, safety, public relations, and…

  14. Recruitment Selection and Training.

    ERIC Educational Resources Information Center

    National Economic Development Office, London (England).

    This booklet outlines facts that should be considered during a new employee's initial weeks with a firm, in this case a clothing manufacturer; particular emphasis is placed on the principles of good selection and training. Various sources are also given from which to get help in implementing the basic principles described. The following points are…

  15. Training for Leadership.

    ERIC Educational Resources Information Center

    Adair, John

    An indepth study of how to select and train for effective leadership is based on the belief that though innate character traits play a role of leadership, leadership potential can be developed. The author's theory known as "functional leadership," stresses that leadership is an interaction among leader, group members, and situation. The good…

  16. TNT: Teams Need Training.

    ERIC Educational Resources Information Center

    Centre County Vocational-Technical School, Pleasant Gap, PA. CIU 10 Bi-County Development Center for Adults.

    This document includes a final report and curriculum manual from a project to help adult educators teach team training by developing a curriculum for use in teaching teamwork skills in work force literacy programs and by providing two half-day seminars to assist adult educators with effectively using the curriculum. The manual for work force…

  17. Hospitalwide Education and Training.

    ERIC Educational Resources Information Center

    Munk, Robert J.; Lovett, Marc

    Findings and recommendations of a five-year demonstration project designed to clarify the role and organized structure of employee education and training in health care institutions are presented. In the first section of the book, hospitalwide education functions that were implemented at sixteen individual hospitals are examined. Identified are…

  18. Change and Athletic Training

    ERIC Educational Resources Information Center

    Ray, Richard

    2006-01-01

    Dr. R. Richard Ray, Jr. is a Professor and Chair of Kinesiology at Hope College in Holland, Michigan. Since 1982 he has served as the Program Director for the Athletic Training Education Program at Hope College. Dr. Ray is the author of several books and over 40 articles. In this article he begins by laying a foundation for the reasons why change…

  19. Conducting Leadership Training Workshops.

    ERIC Educational Resources Information Center

    New York State Education Dept., Albany.

    The purpose of this publication is to provide informational and organizational aid to students, teachers and administrators interested in developing and strengthening student leadership through local and regional student leadership training workshops. Divided into two major parts, the first part discusses the improvement of leadership training…

  20. BA Sales Training.

    ERIC Educational Resources Information Center

    Coutts-Clay, Jennifer

    1979-01-01

    Describes the programs and courses of the sales training department at British Airways from induction to management level. The staff uses one of the largest commercial computer facilities in the world to provide seat reservations, fare quotations and tickets, hotel reservations, and tours. (MF)

  1. Experiential Training: Practical Guidelines.

    ERIC Educational Resources Information Center

    Hobbs, Tony, Ed.

    This book examines the use of an experiential (participative) learning approach in training people in areas that are particularly emotionally challenging. Each of the nine articles focuses on the design and conduct of workshops in which the principles of experiential learning are used to help trainees acquire knowledge related to and cope with…

  2. The Top Training Priorities for 2003.

    ERIC Educational Resources Information Center

    Hall, Brandon

    2003-01-01

    A survey of 222 training professionals identified current training priorities: soft skills training; technical training; enhancing the quality of training; business skills; business alignment, business impact, and return on investment; online learning; sales training; safety and compliance training; performance management; and human…

  3. Payload Crew Training Complex (PCTC) utilization and training plan

    NASA Technical Reports Server (NTRS)

    Self, M. R.

    1980-01-01

    The physical facilities that comprise the payload crew training complex (PCTC) are described including the host simulator; experiment simulators; Spacelab aft flight deck, experiment pallet, and experiment rack mockups; the simulation director's console; payload operations control center; classrooms; and supporting soft- and hardware. The parameters of a training philosophy for payload crew training at the PCTC are established. Finally the development of the training plan is addressed including discussions of preassessment, and evaluation options.

  4. Behavioral Vision Training for Myopia: Stimulus Specificity of Training Effects.

    ERIC Educational Resources Information Center

    Leung, Jin-Pang

    1988-01-01

    The study assessed transfer of visual training for myopia using two different training stimuli and a single subject A-B-C-A design with a male student volunteer. A procedure including stimulus fading and reinforcement (positive verbal feedback) was used to effectively improve performance on both behavioral acuity tests during the training phases…

  5. Sales Training: Effects of Spaced Practice on Training Transfer

    ERIC Educational Resources Information Center

    Kauffeld, Simone; Lehmann-Willenbrock, Nale

    2010-01-01

    Purpose: The benefits of spaced training over massed training practice are well established in the laboratory setting. In a field study design with sales trainings, the purpose of this paper is to investigate the effects of spaced compared with massed practice on transfer quantity and quality, sales competence, and key figures.…

  6. Transfer of Instrument Training and the Synthetic Flight Training System.

    ERIC Educational Resources Information Center

    Caro, Paul W.

    One phase of an innovative flight training program, its development, and initial administration is described in this paper. The operational suitability test activities related to a determination of the transfer of instrument training value of the Army's Synthetic Flight Training System (SFTS) Device 2B24. Sixteen active Army members of an Officer…

  7. Training and Technology: A Systems Approach for Industrial Training.

    ERIC Educational Resources Information Center

    Myers, William E.

    The document analyzes the Training and Technology (TAT) Industrial Skill and Technical Training (ISTT) program and describes the basic relationships between various training components and their linkages to certain aspects of program structure and organization. The TAT ISTT program operations are presented within the conceptual framework of an…

  8. Peace Corps Aquaculture Training Manual. Training Manual T0057.

    ERIC Educational Resources Information Center

    Peace Corps, Washington, DC. Information Collection and Exchange Div.

    This Peace Corps training manual was developed from two existing manuals to provide a comprehensive training program in fish production for Peace Corps volunteers. The manual encompasses the essential elements of the University of Oklahoma program that has been training volunteers in aquaculture for 25 years. The 22 chapters of the manual are…

  9. Training Analysis Digest for Technical and Military Training, No. 5.

    ERIC Educational Resources Information Center

    Air Training Command, Randolph AFB, TX.

    The Training Analysis Digest is a compilation of training improvement studies and projects which have contributed to better instruction and the refinement of course content. It provides a system for reporting the efforts of individual training centers at the Chanute, Keesler, Lackland, Lowry, and Sheppard Air Force Bases in improving enlisted men…

  10. Making Training Core Business: Enterprise Registered Training Organisations in Australia

    ERIC Educational Resources Information Center

    Smith, Erica; Smith, Andy

    2009-01-01

    This paper examines the Australian phenomenon of Enterprise Registered Training Organizations (RTOs). These are organizations that do not have training as their main business but that are accredited to deliver training and award qualifications, primarily to their own workers. Although Enterprise RTOs have been in existence in one form or another…

  11. Smart Training: The Manager's Guide to Training for Improved Performance.

    ERIC Educational Resources Information Center

    Carr, Clay

    This book describes and advocates "smart training," an approach to on-the-job training that helps employees gain competence and empowers them to act to satisfy customers. The book is organized in 16 chapters grouped into 4 parts. Part 1 outlines the basics of smart training, including its performance base, the need for initial analysis, and the…

  12. The Efficacy of Stuttering Measurement Training: Evaluating Two Training Programs

    ERIC Educational Resources Information Center

    Bainbridge, Lauren A.; Stavros, Candace; Ebrahimian, Mineh; Wang, Yuedong; Ingham, Roger J.

    2015-01-01

    Purpose: Two stuttering measurement training programs currently used for training clinicians were evaluated for their efficacy in improving the accuracy of total stuttering event counting. Method: Four groups, each with 12 randomly allocated participants, completed a pretest-posttest design training study. They were evaluated by their counts of…

  13. A Parent Training Model for Toilet Training Children with Autism

    ERIC Educational Resources Information Center

    Kroeger, K.; Sorensen, R.

    2010-01-01

    Background: Azrin & Foxx pioneered an intensive toilet training protocol for individuals with intellectual disability living in a residential setting. Since the development of the Rapid Toilet Training (RTT) protocol, many have replicated the efficacy, most notably in educational and outpatient treatment settings, but often training over longer…

  14. Administrators' Roles in Training Programs and Training Transfer

    ERIC Educational Resources Information Center

    Ismail, Azman; Hua, Ng Kueh; Ismail, Yusof; Samah, Ainon Jauhariah Abu; Bakar, Rixal Abu; Ibrahim, Nurshahira

    2015-01-01

    An administrator plays a vital role in the growth and development of his/her subordinates. Despite this notion, the role of an administrator in the context of training programs and transfer of training is not well studied. Therefore, this study is set to examine the relationship between administrator's role in training programs and training…

  15. Training Requirements in OSHA Standards and Training Guidelines. Revised.

    ERIC Educational Resources Information Center

    Occupational Safety and Health Administration, Washington, DC.

    This guide provides an overview of Occupational Safety and Health Act (OSHA) standards and training guidelines for various industries. The first section introduces the concept of voluntary training guidelines, explaining that the guidelines are designed to help employers determine whether a worksite problem can be solved by training, what training…

  16. Pre-Employment Laboratory Training

    ERIC Educational Resources Information Center

    Vela, Rene H.; Correa, Jose

    1976-01-01

    The article describes the development of a pre-employment laboratory training program in meat processing and its successful use in conjunction with a cooperative training program in a high school agriculture curriculum. (MS)

  17. Innovative approaches to recurrent training

    NASA Technical Reports Server (NTRS)

    Noon, H.; Murphy, M.

    1984-01-01

    Innovative approaches to recurrent training for regional airline aircrews are explored. Guidelines for recurrent training programs which include in corporation of cockpit resource management are discussed. B.W.

  18. Training Personnel for Computer Conversions.

    ERIC Educational Resources Information Center

    Rulffes, Walt

    1986-01-01

    A computer training model for school personnel can be divided into several phases that meet a district's goal of decentralizing the computerized administrative and instructional support system. Training involves such considerations as cost and job description revisions. (CJH)

  19. Creating a Toilet Training Plan

    MedlinePlus

    ... Size Email Print Share Creating a Toilet Training Plan Page Content Article Body These are the tools ... will need to create your own toilet-training plan and implement it at the best time for ...

  20. Theories Supporting Transfer of Training.

    ERIC Educational Resources Information Center

    Yamnill, Siriporn; McLean, Gary N.

    2001-01-01

    Reviews theories about factors affecting the transfer of training, including theories on motivation (expectancy, equity, goal setting), training transfer design (identical elements, principle, near and far), and transfer climate (organizational). (Contains 36 references.) (SK)

  1. Enterprise Training: This Changes Everything.

    ERIC Educational Resources Information Center

    Stamps, David

    1999-01-01

    In many companies, training management and knowledge management are merging. Enterprise-resource-planning software products promise seamless integration of all information flowing through a company. Training then becomes part of a process, not a course or event. (SK)

  2. Vocational Training in Latin America.

    ERIC Educational Resources Information Center

    Wallenborn, Manfred

    2001-01-01

    Explores historical development and weaknesses and change efforts in vocational training institutions in Chile, Colombia, Costa Rica, and Peru. Identifies future directions for the training enterprise in Latin America. (SK)

  3. Pelvic floor muscle training exercises

    MedlinePlus

    Pelvic floor muscle training exercises are a series of exercises designed to strengthen the muscles of the pelvic floor. ... Pelvic floor muscle training exercises are recommended for: Women ... Men with urinary stress incontinence after prostate surgery ...

  4. Payload IVA training and simulation

    NASA Technical Reports Server (NTRS)

    Monsees, J. H.

    1982-01-01

    The development of a training program for the intravehicular operation of space shuttle payloads is discussed. The priorities for the program are compliance with established training standards, and accommodating changes. Simulation devices are also reviewed.

  5. Training of component reading skills.

    PubMed

    Fiedorowicz, C A

    1986-01-01

    Fifteen reading-disabled boys, classified according to reading subskill deficits, formed three subgroups: Oral Reading, Associative, and Sequential. On the rationale that training procedures emphasizing accuracy and speed of response to letters, syllables, and words would improve reading skills, the children were trained daily for two and one-half months. Two schedules of testing and training allowed for an untrained control group and a follow-up group. The results indicated that the computer-assisted training procedures were not only effective in improving component reading skills, but in addition there was a transfer of training to achievement measures of reading word recognition. This study lends support to the hypothesis that training according to subgroup classification, using training procedures which incorporate an application of the automaticity theory and a combination of task-analytic and process-oriented models, is an effective training approach for reading-disabled children. PMID:24243467

  6. Prepubescent Strength Training. Some Considerations.

    ERIC Educational Resources Information Center

    Priest, Joe W.; Holshouser, Richard S.

    1987-01-01

    Under the careful supervision of a trained fitness professional, the benefits of prepubescent strength training (improved strength, power, muscular endurance, bone density) outweigh the risks (acute and chronic musculoskeletal injuries). (CB)

  7. Operator Training: Who Is Responsible?

    ERIC Educational Resources Information Center

    Wubbena, Robert L.

    1979-01-01

    Summarized are the findings of a study to identify and correct water pollution control operator training deficiencies. Several models are presented to aid in developing a coordinated delivery system for operator training and certification. (CS)

  8. Radiological training for tritium facilities

    SciTech Connect

    1996-12-01

    This program management guide describes a recommended implementation standard for core training as outlined in the DOE Radiological Control Manual (RCM). The standard is to assist those individuals, both within DOE and Managing and Operating contractors, identified as having responsibility for implementing the core training recommended by the RCM. This training may also be given to radiological workers using tritium to assist in meeting their job specific training requirements of 10 CFR 835.

  9. Training: Plugging the information gaps

    NASA Technical Reports Server (NTRS)

    Scott, Carol J.

    1993-01-01

    Training is commonly viewed as an add-on function to the development cycle. It is imperative that this view be changed. The training developer needs to be a colleague in the system development process, contributing and learning along with the other development participants. Training developers can make contributions to design concepts that favor end-users. Early involvement will enhance the likelihood of training availability concurrent with software delivery. End-users will benefit and cost-savings will be realized.

  10. Water Resources Division training catalog

    USGS Publications Warehouse

    Hotchkiss, W.R.; Foxhoven, L.A.

    1984-01-01

    The National Training Center provides technical and management sessions nesessary for the conductance of the U.S. Geological Survey 's training programs. This catalog describes the facilities and staff at the Lakewood Training Center and describes Water Resources Division training courses available through the center. In addition, the catalog describes the procedures for gaining admission, formulas for calculating fees, and discussion of course evaluations. (USGS)

  11. Constellation Training Facility Support

    NASA Technical Reports Server (NTRS)

    Flores, Jose M.

    2008-01-01

    The National Aeronautics and Space Administration is developing the next set of vehicles that will take men back to the moon under the Constellation Program. The Constellation Training Facility (CxTF) is a project in development that will be used to train astronauts, instructors, and flight controllers on the operation of Constellation Program vehicles. It will also be used for procedure verification and validation of flight software and console tools. The CxTF will have simulations for the Crew Exploration Vehicle (CEV), Crew Module (CM), CEV Service Module (SM), Launch Abort System (LAS), Spacecraft Adapter (SA), Crew Launch Vehicle (CLV), Pressurized Cargo Variant CM, Pressurized Cargo Variant SM, Cargo Launch Vehicle, Earth Departure Stage (EDS), and the Lunar Surface Access Module (LSAM). The Facility will consist of part-task and full-task trainers, each with a specific set of mission training capabilities. Part task trainers will be used for focused training on a single vehicle system or set of related systems. Full task trainers will be used for training on complete vehicles and all of its subsystems. Support was provided in both software development and project planning areas of the CxTF project. Simulation software was developed for the hydraulic system of the Thrust Vector Control (TVC) of the ARES I launch vehicle. The TVC system is in charge of the actuation of the nozzle gimbals for navigation control of the upper stage of the ARES I rocket. Also, software was developed using C standards to send and receive data to and from hand controllers to be used in CxTF cockpit simulations. The hand controllers provided movement in all six rotational and translational axes. Under Project Planning & Control, support was provided to the development and maintenance of integrated schedules for both the Constellation Training Facility and Missions Operations Facilities Division. These schedules maintain communication between projects in different levels. The Cx

  12. Directions in Center Director Training

    ERIC Educational Resources Information Center

    Bloom, Paula Jorde; Vinci, Yasmina; Rafanello, Donna; Donohue, Chip

    2011-01-01

    Exchange invited some of the leading trend watchers in the arena of director training to share their insights on the current state and future directions in this country. This article presents the authors' insights on the directions in center director training. They also share their views on whether the amount of and quality of training out there…

  13. Vocational Training in the GDR.

    ERIC Educational Resources Information Center

    Panorama DDR, 1981

    1981-01-01

    Vocational training forms part of the integrated educational system in the German Democratic Republic (GDR). About 85 percent of young people attend vocational training courses after completion of 10-year general schooling. Skilled worker training is offered in 291 trades with opportunities for specialization in 500 fields. In 98 trades,…

  14. Formal Training and Labor Productivity.

    ERIC Educational Resources Information Center

    Medoff, James L.

    The impact of certain types of training on labor productivity or performance in different settings was assessed. In the manufacturing sector, it was found that the extent of in-house, employer-sponsored, job-related training has a positive relationship with output per labor hour. The extent of this type of training received out-of-house did not…

  15. ISD Designed Medical Specialist Training.

    ERIC Educational Resources Information Center

    Rock, Samuel K., Jr.; Chagalis, George P.

    The Basic Medical Specialist course has one of the largest enrollments of the U.S. Army's Academy of Health Sciences; 11,000 soldiers were trained in this course in 1977 and 1978. Training encompasses both emergency first aid (for field medics) and basic nursing skills. A task force working to improve Army training developed this course, in…

  16. Alumni Training by the Book.

    ERIC Educational Resources Information Center

    Zelno, Robert P.

    1992-01-01

    Recommendations for planning a successful training workshop for college alumni volunteers include assuring the association is organized and the program well timed; making sure everyone agrees to budget and/or financial commitments; using seasoned volunteers to conduct a feasibility study and determine training needs; making training enjoyable; and…

  17. Instructor Training on British Railways

    ERIC Educational Resources Information Center

    Gibson, J. D.

    1970-01-01

    The value of instructor training was recognized by British Railways as early as 1950 with the setting up of a training center at Darlington. This article shows the results of this continuous training experience in the benefits to be obtained from re-appraisal techniques and practical work. (Author/EB)

  18. Forecasting the Value of Training

    ERIC Educational Resources Information Center

    Basarab, Dave

    2011-01-01

    The Predictive Evaluation (PE) model is a training and evaluation approach with the element of prediction. PE allows trainers and business leaders to predict the results, value, intention, adoption, and impact of training, allowing them to make smarter, more strategic training and evaluation investments. PE is invaluable for companies that…

  19. Prepubescent Strength Training Gains Support.

    ERIC Educational Resources Information Center

    Duda, Marty

    1986-01-01

    Recent studies have stimulated greater support for prepubescent weight training. There seems to be general agreement that strength and weight training, when practiced under properly controlled conditions, is safe and efficacious for prepubescents. Weight lifting is not supported. Recommendations for weight training are made, and reservations are…

  20. Training within the Accounting Firm.

    ERIC Educational Resources Information Center

    Finch, Beth; And Others

    1991-01-01

    A survey received 509 responses from 2,000 randomly selected accounting employees about which training topics are receiving the most attention and who is receiving the training. Results prove that training has become an integral part of a certified public accountant's job; topics most often covered were tax related--individual and corporate income…

  1. Warehousing Competency Profile. Apprenticeship Training.

    ERIC Educational Resources Information Center

    Alberta Learning, Edmonton. Apprenticeship and Industry Training.

    This document presents information about the apprenticeship training program of Alberta, Canada, in general and the warehousing program in particular. The first part of the document discusses the following items: Alberta's apprenticeship and industry training system; the apprenticeship and industry training committee structure; local…

  2. The Training-Waste Conspiracy.

    ERIC Educational Resources Information Center

    Shaw, Edward

    1995-01-01

    Half of the $50 billion spent on training each year is wasted. The training industry is responsible for at least half of that waste because the field is not fully professionalized and too many programs are not based on research on human psychology and behavior. The training industry must improve both the quality of its product and the quality of…

  3. Sales Training for Small Companies

    ERIC Educational Resources Information Center

    Kinder, James F.

    1974-01-01

    The author suggests methods a sales manager in a small company can use to train effective salesmen; he defines the sales manager's role, and cites internal and external training resources. The absence of sales training courses in metropolitan Canada is noted. (AJ)

  4. TAT Welding Technology Training Program.

    ERIC Educational Resources Information Center

    Rogers, Everette N.; Cook, Jerry L.

    The Training and Technology (TAT) Welding Technology Training Program is an intensive industrial training program conducted by Oak Ridge Associated Universities and Union Carbide Corporation designed to upgrade the skills of unemployed and underemployed individuals so they can command good jobs in industry. The document provides an introduction…

  5. Expedition 3 Crew Training Clips

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The Expedition 3 crewmembers, Frank Culbertson, Jr., Mikhail Turin, and Vladimir Dezhurov, are seen during various stages of their training. Footage includes Extravehicular Activity (EVA) Training at the Neutral Buoyancy Laboratory (NBL), EVA Preparation and Post Training in the International Space Station Airlock Mock-up, in the NBL Space Station Remote Manipulator System Workstation, and during the T-38 flight at Ellington Field.

  6. Technical Training: A Systematic Approach.

    ERIC Educational Resources Information Center

    Gammuto, John J.

    1980-01-01

    To employ scarce personnel and money to best advantage, company management must diagnose critical training needs and assign them priorities. A systems approach is recommended, which gives focus to training design while accommodating most organization training goals. Included are suggestions for system design, facility management, and cost…

  7. A Handbook for Training Strategy.

    ERIC Educational Resources Information Center

    Sloman, Martyn

    This handbook introduces a new training model that reflects the complexity of organizational life, changes in the human resources (HR) function, and the need to involve line management. Part I outlines the new context in which training must be delivered. It discusses the changing organizational requirements for training and the changing position…

  8. Apprenticeship Training: Communication Technician Program.

    ERIC Educational Resources Information Center

    Alberta Learning, Edmonton. Apprenticeship and Industry Training.

    This document presents information about the apprenticeship training program of Alberta, Canada, in general and the communication technician program in particular. The first part of the document discusses the following items: Alberta's apprenticeship and industry training system; the apprenticeship and industry training committee structure; local…

  9. Anthropometric Training Project. Final Report.

    ERIC Educational Resources Information Center

    San Diego State Coll., CA.

    The Anthropometric Training Project was aimed at providing a training program for select students to develop research competency in an area relating body type, composition, anthropometric assessment, and physical performance measures. The program involves interdisciplinary cooperation in training through seminars, laboratory practice, and…

  10. Targeted Training: An Integrated Initiative.

    ERIC Educational Resources Information Center

    Valvasori, Joe

    The Learning Enrichment Foundation (LEF) in Toronto, Ontario (Canada) offers 12 targeted training programs that have successfully helped "hard-to-serve" clients return to the workforce. Compared with traditional training, targeted training has a much narrower focus and adapts quickly to industry trends to meet employers' changing demands. LEF's…

  11. The Training Evaluation Inventory (TEI)--Evaluation of Training Design and Measurement of Training Outcomes for Predicting Training Success

    ERIC Educational Resources Information Center

    Ritzmann, Sandrina; Hagemann, Vera; Kluge, Annette

    2014-01-01

    Training evaluation in research and organisational contexts is vital to ensure informed decisions regarding the value of training. The present study describes the development of a valid and reliable training evaluation inventory (TEI), as it does not exist so far. The objectives were a) to construct an instrument that is theoretically and…

  12. Radiological Worker Computer Based Training

    Energy Science and Technology Software Center (ESTSC)

    2003-02-06

    Argonne National Laboratory has developed an interactive computer based training (CBT) version of the standardized DOE Radiological Worker training program. This CD-ROM based program utilizes graphics, animation, photographs, sound and video to train users in ten topical areas: radiological fundamentals, biological effects, dose limits, ALARA, personnel monitoring, controls and postings, emergency response, contamination controls, high radiation areas, and lessons learned.

  13. Research Training--Present & Future.

    ERIC Educational Resources Information Center

    Organisation for Economic Cooperation and Development, Paris (France).

    In 10 papers by independent experts, this volume explores the trends in and prospects for research training in member nations of the Organization for Economic Cooperation and Development. "Problems and Prospects of Research Training in the 1990s" (Stuart Blume) looks at trends in national policy toward research training and issues of quality.…

  14. Concrete Finisher Program. Apprenticeship Training.

    ERIC Educational Resources Information Center

    Alberta Learning, Edmonton. Apprenticeship and Industry Training.

    This document presents information about the apprenticeship training program of Alberta, Canada, in general and the concrete finishing program in particular. The first part of the document discusses the following items: Alberta's apprenticeship and industry training system; the apprenticeship and industry training committee structure; local…

  15. The Context Oriented Training Method.

    ERIC Educational Resources Information Center

    Cavrini, Andrea

    The Context Oriented Training (COT) method is introduced and explored in this paper. COT is a means of improving the training process, beginning with the observation and analysis of current corporate experiences in the field. The learning context lies between the development of professional competencies in training and the operational side in the…

  16. Weight Training for Wheelchair Sports.

    ERIC Educational Resources Information Center

    Practical Pointers, 1978

    1978-01-01

    The article examines weight lifting training procedures for persons involved in wheelchair sports. Popular myths about weight training are countered, and guidelines for a safe and sound weight or resistance training program are given. Diagrams and descriptions follow for specific weightlifting activities: regular or standing press, military press,…

  17. The 2002 Training Top 100.

    ERIC Educational Resources Information Center

    Training, 2002

    2002-01-01

    Identifies the top 100 companies in terms of the amount spent on training and development, the number of hours of training per employee, percentage of payroll spent on training, and tuition allotment per employee. Describes best practices in succession planning and leadership development, mentoring, and job shadowing and provides a detailed…

  18. The 2003 Training Top 100.

    ERIC Educational Resources Information Center

    Galvin, Tammy

    2003-01-01

    Identifies the top 100 U.S. companies in terms of the amount spent on training and development, the number of hours of training per employee, percentage of payroll spent on training, and tuition allotment per employee. Describes best practices in succession planning and leadership development, mentoring, and job shadowing and provides a detailed…

  19. Guidance for training program evaluation

    SciTech Connect

    Not Available

    1984-01-01

    An increased concern about the training of nuclear reactor operators resulted from the incident at TMI-2 in 1979. Purpose of this guide is to provide a general framework for the systematic evaluation of training programs for DOE Category-A reactors. The primary goal of such evaluations is to promote continuing quality improvements in the selection, training and qualification programs.

  20. Strength Training and Children's Health.

    ERIC Educational Resources Information Center

    Faigenbaum, Avery D.

    2001-01-01

    Provides an overview of the potential health benefits of strength training for children, discussing the role of strength training in preventing sports-related injuries and highlighting design considerations for such programs. The focus is on musculoskeletal adaptations to strength training that are observable in healthy children. Guidelines for…

  1. A systematic approach to training: A training needs assessment

    NASA Technical Reports Server (NTRS)

    Manning, Margaret H.

    1989-01-01

    In an effort to determine the gap between the actual performance and the necessary performance of employees for the effective and efficient accomplishment of an organization's mission and goals, an organization-wide Training Needs Assessment must be conducted. The purpose of this work was to conduct a training needs analysis and prepare a NASA Langley Catalog of On-Site Training programs. The work included developing a Training Needs Assessment Survey, implementing the survey, analyzing and researching the training needs, identifying the courses to meet the needs, and preparing and designing an On-Site Training Catalog. This needs analysis attempted to identify performance weaknesses and deficits; seek out and provide opportunities for improved performance; anticipate and avoid future problems; enhance and create new strengths. The end product is a user-friendly catalog of on-site training available. The results include: top-down approach to needs assessment; improved communication with training coordinators; 98 percent return rate of the Training Needs Assessment survey; complete, newly designed, user-friendly catalog; 167 catalog descriptions advertised; 82 new courses advertised; training logo; and request for the training application form.

  2. The Efficacy of Stuttering Measurement Training: Evaluating Two Training Programs

    PubMed Central

    Bainbridge, Lauren A.; Stavros, Candace; Ebrahimian, Mineh; Wang, Yuedong

    2015-01-01

    Purpose Two stuttering measurement training programs currently used for training clinicians were evaluated for their efficacy in improving the accuracy of total stuttering event counting. Method Four groups, each with 12 randomly allocated participants, completed a pretest–posttest design training study. They were evaluated by their counts of stuttering events on eight 3-min audiovisual speech samples from adults and children who stutter. Stuttering judgment training involved use of either the Stuttering Measurement System (SMS), Stuttering Measurement Assessment and Training (SMAAT) programs, or no training. To test for the reliability of any training effect, SMS training was repeated with the 4th group. Results Both SMS-trained groups produced approximately 34% improvement, significantly better than no training or the SMAAT program. The SMAAT program produced a mixed result. Conclusions The SMS program was shown to produce a “medium” effect size improvement in the accuracy of stuttering event counts, and this improvement was almost perfectly replicated in a 2nd group. Half of the SMAAT judges produced a 36% improvement in accuracy, but the other half showed no improvement. Additional studies are needed to demonstrate the durability of the reported improvements, but these positive effects justify the importance of stuttering measurement training. PMID:25629956

  3. Resistance training and sarcopenia.

    PubMed

    Giallauria, Francesco; Cittadini, Antonio; Smart, Neil Andrew; Vigorito, Carlo

    2016-01-01

    Aging is inexorably accompanied by a progressive decline of muscle mass, quality and strength. The resulting condition has been termed sarcopenia. Age-related sarcopenia can be accelerated by a variety of factors including changes in the hormonal milieu, inactivity, poor nutrition, chronic illness, and loss of integrity and function in the peripheral and central nervous systems. The downstream mechanisms by which these risk factors cause sarcopenia are not completely understood. Exercise training (particularly resistance training) has long been identified as the most promising method for increasing muscle mass and strength among older people. New interventions aimed at preventing muscle atrophy, promoting muscle growth and ultimately, maintaining muscle functions during aging are discussed. Understanding how age affects muscle-related gene expression, protein recycling and resynthesis, post-translational modification and turnover will be crucial to identify new treatment options. PMID:27374049

  4. Women Scientists in Training

    NASA Technical Reports Server (NTRS)

    1975-01-01

    Women scientists in training at Marshall Space Flight Center, (top to bottom) Carolyn Griner, Ann Whitaker, and Dr. Mary Johnston, are shown simulating weightlessness while undergoing training in the Neutral Buoyancy Simulator. These women were part of a special program dedicated to gaining a better understanding of problems involved in performing experiments in space. The three were engaged in designing and developing experiments for space, such as materials processing for Spacelabs. Dr. Johnston specialized in metallurgical Engineering, Dr. Whitaker in lubrication and surface physics, and Dr. Griner in material science. Dr. Griner went on to become Acting Center Director at Marshall Space Flight Center from January to September 1998. She was the first woman to serve

  5. NASA Headquarters training catalog

    NASA Technical Reports Server (NTRS)

    1990-01-01

    The NASA Headquarters training catalog is a comprehensive listing of all educational and employee development programs. This course catalog contains descriptions of course content, objectives, target audience, prerequisites, length of course, approximate number of times the course is offered per year, and cost of the course. Curriculum areas include graduate and undergraduate academic study; professional development program; and executive management, senior management, and supervisory development programs. Secretarial/clerical and general computer skills programs are also included.

  6. Strangers on a Train

    ERIC Educational Resources Information Center

    Davidson, Cathy N.

    2011-01-01

    Early on in her first tenure-track job at Michigan State University, in the late 1970s, the author happened to be riding the same train from Lansing, Michigan, to Chicago as the department chair who hired her, Alan Hollingsworth, who had since become dean of the College of Arts and Sciences. It had taken her three years to land a tenure-track…

  7. Effect of Exercise Training and +Gz Acceleration Training on Men

    NASA Technical Reports Server (NTRS)

    Greenleaf, John E.; Simonson, Shawn R.; Stocks, Jodie M.; Evans, Joyce; Knapp, Charles F.; Cowell, Stephenie A.; Pemberton, Kendra N.; Wilson, Heather W.; Vener, Jamie M.; Evetts, Simon N.

    2001-01-01

    Countermeasures for reduction in work capacity (maximal oxygen uptake and strength) during spaceflight and enhanced orthostatic intolerance during re-entry, landing and egress from the return vehicle are continuing problems. The purpose for this study was to test the hypothesis that passive-acceleration training; supine, interval, exercise plus acceleration training and exercise combined with acceleration training would improve orthostatic tolerance in ambulatory men; and that addition of the aerobic exercise conditioning would not alter this improved tolerance from that of passive-acceleration training. Seven men (24-38 yr) underwent "Passive" training on the Ames human-powered centrifuge (HPC) for 30 min, "Exercise" training on the cycle ergometer with constant +Gz acceleration; and "Combined" exercise training at 40% to 90% of the HPC +Gz(max) exercise level. Maximal supine exercise loads increased significant (P<0.05) by 8.3% (Passive), 12.6% (Exercise), and by 15.4% (Combined) after training, but their post-training maximal oxygen uptakes and maximal heart rates were unchanged. Maximal time to fatigue (endurance) was unchanged with Passive was increased (P<0.05) with Exercise and Combined training. Thus, the exercise in the Exercise and Combined training Phases resulted in greater maximal loads and endurance without effect on maximal oxygen uptake or heart rate. There was a 4% to 6% increase (P<0.05) in all four quadriceps muscle volumes (right and left) after post-Combined training. Resting pre-tilt heart rate was elevated by 12.9% (P<0.05) only after Passive training suggesting that the exercise training attenuated the HR response. Plasma volume (% Delta) was uniformly decreased by 8% to 14% (P<0.05) at tilt-tolerance pre- vs. post-training indicating essentially no effect of training on the level of hypovolemia. Post-training tilt-tolerance time and heart rate were increased (P<0.05) only with Passive training by 37.8% and by 29.1%, respectively. Thus

  8. Capsule-train stability

    NASA Astrophysics Data System (ADS)

    Bryngelson, Spencer H.; Freund, Jonathan B.

    2016-07-01

    Elastic capsules flowing in small enough tubes, such as red blood cells in capillaries, are well known to line up into regular single-file trains. The stability of such trains in somewhat wider channels, where this organization is not observed, is studied in a two-dimensional model system that includes full coupling between the viscous flow and suspended capsules. A diverse set of linearly amplifying disturbances, both long-time asymptotic (modal) and transient (nonmodal) perturbations, is identified and analyzed. These have a range of amplification rates and their corresponding forms are wavelike, typically dominated by one of five principal perturbation classes: longitudinal and transverse translations, tilts, and symmetric and asymmetric shape distortions. Finite-amplitude transiently amplifying perturbations are shown to provide a mechanism that can bypass slower asymptotic modal linear growth and precipitate the onset of nonlinear effects. Direct numerical simulations are used to verify the linear analysis and track the subsequent transition of the regular capsule trains into an apparently chaotic flow.

  9. Clinical toxinology specialty training.

    PubMed

    White, Julian

    2013-07-01

    Clinical toxinology is the medical discipline dealing with the diagnosis, treatment and prevention of toxin diseases caused by exposure to venomous animals and poisonous animals, plants and mushrooms. Currently there is no national or international organisation accrediting or training doctors in this discipline, but the role of the IST in this area is the subject of a recently approved revised Constitution. A few courses covering some aspects of clinical toxinology exist, either with limited curricula, or with only a minor clinical focus, or with a very regional, non-global focus. The only comprehensive clinical toxinology course is the one provided in Adelaide, Australia, running regularly since 1997. This course may form the nucleus from which IST can develop a global accredited training scheme in clinical toxinology. Such a scheme will require input from diverse global regions and will be far more comprehensive and over a much longer time than the current Short Course, though may incorporate the Short Course in some way, or a derivative of it. Accreditation of medical expertise in clinical toxinology will be required at the national level and this might be accomplished by the IST working with existing national medical specialty organisations and governments, with the IST supervising the training and accreditation requirements and the national organisations providing the framework for registration of medical expertise at the local level. PMID:23524067

  10. Training a quantum optimizer

    NASA Astrophysics Data System (ADS)

    Wecker, Dave; Hastings, Matthew B.; Troyer, Matthias

    2016-08-01

    We study a variant of the quantum approximate optimization algorithm [E. Farhi, J. Goldstone, and S. Gutmann, arXiv:1411.4028] with a slightly different parametrization and a different objective: rather than looking for a state which approximately solves an optimization problem, our goal is to find a quantum algorithm that, given an instance of the maximum 2-satisfiability problem (MAX-2-SAT), will produce a state with high overlap with the optimal state. Using a machine learning approach, we chose a "training set" of instances and optimized the parameters to produce a large overlap for the training set. We then tested these optimized parameters on a larger instance set. As a training set, we used a subset of the hard instances studied by Crosson, Farhi, C. Y.-Y. Lin, H.-H. Lin, and P. Shor (CFLLS) (arXiv:1401.7320). When tested, on the full set, the parameters that we find produce a significantly larger overlap than the optimized annealing times of CFLLS. Testing on other random instances from 20 to 28 bits continues to show improvement over annealing, with the improvement being most notable on the hardest instances. Further tests on instances of MAX-3-SAT also showed improvement on the hardest instances. This algorithm may be a possible application for near-term quantum computers with limited coherence times.

  11. Training and Simulation in Otolaryngology

    PubMed Central

    Wiet, Gregory J.; Stredney, Don; Wan, Dinah

    2011-01-01

    This article focuses on key issues surrounding the needs and application of simulation technologies for technical skills training in otolaryngology. The discussion includes an overview of key topics in training and learning, the application of these issues in simulation environments, and the subsequent applications of these simulation environments to the field of otolaryngology. Examples of past applications are presented, with discussion of how the interplay of cultural changes in surgical training in general, along with the rapid advancements in technology have shaped and influenced their adoption and adaptation. The authors conclude with emerging trends and potential influences advanced simulation and training will have on technical skills training in otolaryngology. PMID:22032486

  12. PECO Energy Company training experience

    SciTech Connect

    Stankiewicz, J.J.

    1994-12-31

    Line managers at Peach Bottom atomic power station are now accountable for and committed to improving the effectiveness of training at the station. Assignments of new personnel to key management positions have resulted in an improved working relationship between the line and training organizations. Training and line managers are now working closely with the station vice president to ensure that the root causes of the recurring management issues are corrected and that processes are institutionalized that will preclude the recurrence of the noted deficiencies. Managers now believe that training is a tool used to improve station performance. Senior station management will continue to monitor and direct further enhancements to improve training effectiveness.

  13. 25 CFR 26.30 - Does the Job Training Program provide part-time training or short-term training?

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 25 Indians 1 2011-04-01 2011-04-01 false Does the Job Training Program provide part-time training or short-term training? 26.30 Section 26.30 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE... Program provide part-time training or short-term training? Yes, part-time and short-term training...

  14. Examining the Effectiveness of a Train-the-Trainer Model: Training Teachers to Use Pivotal Response Training

    ERIC Educational Resources Information Center

    Suhrheinrich, Jessica

    2011-01-01

    The train-the-trainer (TTT) model, which has also been called pyramidal training, triadic training, and helper model training, focuses on initially training a person or people who, in turn, train other people at their home agency. The TTT model has promise of being both efficient and cost-effective. The TTT model may be especially useful in…

  15. Cooking Up World-Class Training.

    ERIC Educational Resources Information Center

    Zemke, Ron

    1997-01-01

    The Culinary Institute of America (CIA), a training ground for aspiring chefs, is a sophisticated training organization that practices many philosophies and techniques, including team learning, training the whole cook, and training the trainer. (JOW)

  16. Training the Auxiliary Health Workers; An Analysis of Functions, Training Content, Training Costs, and Facilities.

    ERIC Educational Resources Information Center

    Health Services and Mental Health Administration (DHEW), Bethesda, MD.

    The booklet describes what each type of worker is allowed to do and presents an overview of the substantive content of the training, length of training, training costs, and kinds of facilities and staff needed. The types of workers include community health aide, homemaker-home health aide, social worker aide, food service supervisor, physical…

  17. TAP 1, Training Program Manual

    SciTech Connect

    Not Available

    1991-01-01

    Training programs at DOE nuclear facilities should provide well-trained, qualified personnel to safely and efficiently operate the facilities in accordance with DOE requirements. A need has been identified for guidance regarding analysis, design, development, implementation, and evaluation of consistent and reliable performance-based training programs. Accreditation of training programs at Category A reactors and high-hazard and selected moderate-hazard nonreactor nuclear facilities will assure consistent, appropriate, and cost-effective training of personnel responsible for the operation, maintenance, and technical support of these facilities. Training programs that are designed and based on systematically determined job requirements, instead of subjective estimation of trainee needs, yield training activities that are consistent and develop or improve knowledge, skills, and abilities that can be directly related to the work setting. Because the training is job-related, the content of these programs more efficiently meets the needs of the employee. Besides a better trained work force, a greater level of operational reactor safety can be realized. This manual is intended to provide an overview of the accreditation process and a brief description of the elements necessary to construct and maintain training programs that are based on the requirements of the job. Two companion manuals provide additional information to assist contractors in their efforts to accredit training programs.

  18. Change Detection: Training and Transfer

    PubMed Central

    Gaspar, John G.; Neider, Mark B.; Simons, Daniel J.; McCarley, Jason S.; Kramer, Arthur F.

    2013-01-01

    Observers often fail to notice even dramatic changes to their environment, a phenomenon known as change blindness. If training could enhance change detection performance in general, then it might help to remedy some real-world consequences of change blindness (e.g. failing to detect hazards while driving). We examined whether adaptive training on a simple change detection task could improve the ability to detect changes in untrained tasks for young and older adults. Consistent with an effective training procedure, both young and older adults were better able to detect changes to trained objects following training. However, neither group showed differential improvement on untrained change detection tasks when compared to active control groups. Change detection training led to improvements on the trained task but did not generalize to other change detection tasks. PMID:23840775

  19. Stroboscopic Training Enhances Anticipatory Timing

    PubMed Central

    SMITH, TREVOR Q.; MITROFF, STEPHEN R.

    2012-01-01

    The dynamic aspects of sports often place heavy demands on visual processing. As such, an important goal for sports training should be to enhance visual abilities. Recent research has suggested that training in a stroboscopic environment, where visual experiences alternate between visible and obscured, may provide a means of improving attentional and visual abilities. The current study explored whether stroboscopic training could impact anticipatory timing – the ability to predict where a moving stimulus will be at a specific point in time. Anticipatory timing is a critical skill for both sports and non-sports activities, and thus finding training improvements could have broad impacts. Participants completed a pre-training assessment that used a Bassin Anticipation Timer to measure their abilities to accurately predict the timing of a moving visual stimulus. Immediately after this initial assessment, the participants completed training trials, but in one of two conditions. Those in the Control condition proceeded as before with no change. Those in the Strobe condition completed the training trials while wearing specialized eyewear that had lenses that alternated between transparent and opaque (rate of 100ms visible to 150ms opaque). Post-training assessments were administered immediately after training, 10-minutes after training, and 10-days after training. Compared to the Control group, the Strobe group was significantly more accurate immediately after training, was more likely to respond early than to respond late immediately after training and 10 minutes later, and was more consistent in their timing estimates immediately after training and 10 minutes later. PMID:27182391

  20. 48 CFR 752.7019 - Participant training.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Participant training. 752... Participant training. For use in any USAID direct contract involving training of USAID participants. Participant Training (JAN 1999) (a) Definitions. (1) Participant training is the training of any...

  1. 20 CFR 632.78 - Training activities.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... EMPLOYMENT AND TRAINING PROGRAMS Program Design and Management § 632.78 Training activities. Native American... following: (a) Classroom training. This program activity is any training of the type normally conducted in... classroom training, employment and training services, or supportive services, costs for which the...

  2. 20 CFR 632.78 - Training activities.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... EMPLOYMENT AND TRAINING PROGRAMS Program Design and Management § 632.78 Training activities. Native American... following: (a) Classroom training. This program activity is any training of the type normally conducted in... classroom training, employment and training services, or supportive services, costs for which the...

  3. Analysis of chromatin structure at meiotic DSB sites in yeasts.

    PubMed

    Hirota, Kouji; Fukuda, Tomoyuki; Yamada, Takatomi; Ohta, Kunihiro

    2009-01-01

    One of the major features of meiosis is a high frequency of homologous recombination that not only confers genetic diversity to a successive generation but also ensures proper segregation of chromosomes. Meiotic recombination is initiated by DNA double-strand breaks that require many proteins including the catalytic core, Spo11. In this regard, like transcription and repair, etc., recombination is hindered by a compacted chromatin structure because trans-acting factors cannot easily access the DNA. Such inhibitory effects must be alleviated prior to recombination initiation. Indeed, a number of groups showed that chromatin around recombination hotspots is less condensed, by using nucleases as a probe to assess local DNA accessibility. Here we describe a method to analyze chromatin structure of a recombination hotspot in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. This method, combining micrococcal nuclease (MNase) digestion ofchromatin DNA and subsequent Southern blotting, is expected to provide information as to chromatin context around a hotspot. Moreover, by virtue of MNase preferentially targeting linker DNA, positions of several nucleosomes surrounding a hotspot can also be determined. Our protocol is a very powerful way to analyze several-kb regions of interest and can be applied to other purposes. PMID:19799187

  4. Biochemical mechanism of DSB end resection and its regulation.

    PubMed

    Daley, James M; Niu, Hengyao; Miller, Adam S; Sung, Patrick

    2015-08-01

    DNA double-strand breaks (DSBs) in cells can undergo nucleolytic degradation to generate long 3' single-stranded DNA tails. This process is termed DNA end resection, and its occurrence effectively commits to break repair via homologous recombination, which entails the acquisition of genetic information from an intact, homologous donor DNA sequence. Recent advances, prompted by the identification of the nucleases that catalyze resection, have revealed intricate layers of functional redundancy, interconnectedness, and regulation. Here, we review the current state of the field with an emphasis on the major questions that remain to be answered. Topics addressed will include how resection initiates via the introduction of an endonucleolytic incision close to the break end, the molecular mechanism of the conserved MRE11 complex in conjunction with Sae2/CtIP within such a model, the role of BRCA1 and 53BP1 in regulating resection initiation in mammalian cells, the influence of chromatin in the resection process, and potential roles of novel factors. PMID:25956866

  5. AAV Vectorization of DSB-mediated Gene Editing Technologies.

    PubMed

    Moser, Rachel J; Hirsch, Matthew L

    2016-01-01

    Recent work both at the bench and the bedside demonstrate zinc-finger nucleases (ZFNs), CRISPR/Cas9, and other programmable site-specific endonuclease technologies are being successfully utilized within and alongside AAV vectors to induce therapeutically relevant levels of directed gene editing within the human chromosome. Studies from past decades acknowledge that AAV vector genomes are enhanced substrates for homology-directed repair in the presence or absence of targeted DNA damage within the host genome. Additionally, AAV vectors are currently the most efficient format for in vivo gene delivery with no vector related complications in >100 clinical trials for diverse diseases. At the same time, advancements in the design of custom-engineered site-specific endonucleases and the utilization of elucidated endonuclease formats have resulted in efficient and facile genetic engineering for basic science and for clinical therapies. AAV vectors and gene editing technologies are an obvious marriage, using AAV for the delivery of repair substrate and/or a gene encoding a designer endonuclease; however, while efficient delivery and enhanced gene targeting by vector genomes are advantageous, other attributes of AAV vectors are less desirable for gene editing technologies. This review summarizes the various roles that AAV vectors play in gene editing technologies and provides insight into its trending applications for the treatment of genetic diseases. PMID:27280971

  6. Trainee and training issues.

    PubMed

    Redfern, Nancy; Bartley, Catherine

    2006-12-01

    This chapter deals with the obligations of trainers and trainees to each other, the responsibilities of the programme and the conflicts of providing a service while training. Management of trainees with differing needs, such as those working part-time or returning to training after sickness, is reviewed. Assessment of performance and the obligation of consultants to identify, manage and support struggling trainees are discussed. Ethical discussion is based on the principles of autonomy, non-maleficence, beneficence, and justice to which fidelity is added. Case studies illustrating the application of ethical principles to work and decision-making are presented to stimulate debate. Opinions vary as to which principle carries more weight in individual cases, and how best to balance the conflicting requirements of the parties involved (patient, trainee,.trainer, employer, society). For all healthcare practitioners, the needs of patients remain our first concern. Acting in a consequentialist way, we must "maximise the good" and minimise the attendant harms in training. However, deontology states that certain sacrosanct rules and principles should never be breached. Doctors must abide by the duties of a doctor described in Good Medical Practice, maintaining standards in a way that ensures professional qualifications are respected. For the patient, there are advantages and disadvantages to receiving care in an educational setting. A 'teaching environment' tends to encourage and maintain high standards of practice from senior clinicians, but it also exposes patients to new learners, who are less efficient and polished and perhaps more prone to make errors. Learning has to fit round and complement the clinical and emotional needs of patients. PMID:17219945

  7. Endurance training at altitude.

    PubMed

    Saunders, Philo U; Pyne, David B; Gore, Christopher J

    2009-01-01

    Since the 1968 Olympic Games when the effects of altitude on endurance performance became evident, moderate altitude training ( approximately 2000 to 3000 m) has become popular to improve competition performance both at altitude and sea level. When endurance athletes are exposed acutely to moderate altitude, a number of physiological responses occur that can comprise performance at altitude; these include increased ventilation, increased heart rate, decreased stroke volume, reduced plasma volume, and lower maximal aerobic power ((.)Vo(2max)) by approximately 15% to 20%. Over a period of several weeks, one primary acclimatization response is an increase in the volume of red blood cells and consequently of (.)Vo(2max). Altitudes > approximately 2000 m for >3 weeks and adequate iron stores are required to elicit these responses. However, the primacy of more red blood cells for superior sea-level performance is not clear-cut since the best endurance athletes in the world, from Ethiopia (approximately 2000 to 3000 m), have only marginally elevated hemoglobin concentrations. The substantial reduction in (.)Vo(2max) of athletes at moderate altitude implies that their training should include adequate short-duration (approximately 1 to 2 min), high-intensity efforts with long recoveries to avoid a reduction in race-specific fitness. At the elite level, athlete performance is not dependent solely on (.)Vo(2max), and the "smallest worthwhile change" in performance for improving race results is as little as 0.5%. Consequently, contemporary statistical approaches that utilize the concept of the smallest worthwhile change are likely to be more appropriate than conventional statistical methods when attempting to understand the potential benefits and mechanisms of altitude training. PMID:19519223

  8. Basic Instructor Training Course

    SciTech Connect

    Morgan, M.P.

    1987-01-01

    Originally, the program intent was to establish foundation for instructors in the UNC Training Department to become certified in accordance to nuclear industry-accepted standards. However, personnel from other organizations, along with DOE representatives and United States government contractors, have utilized the program as a learning base for planning, developing, and implementing instructional presentations. The course content presents a wide variety of material relative to learning behaviors, educational theory, and instructional methodology. The content is presented on a team-teaching basis with usually three or more different instructors. This provides students opportunity to observe a variety of instructional styles, mannerisms, and personalities, lending interest and student introspection.

  9. Training high performance skills using above real-time training

    NASA Technical Reports Server (NTRS)

    Guckenberger, Dutch; Uliano, Kevin C.; Lane, Norman E.

    1993-01-01

    The Above Real-Time Training (ARTT) concept is a unique approach to training high performance skills. ARTT refers to a training paradigm that places the operator in a simulated environment that functions at faster than normal time. Such a training paradigm represents a departure from the intuitive, but not often supported, feeling that the best practice is determined by the training environment with the highest fidelity. This approach is hypothesized to provide greater 'transfer value' per simulation trial, by incorporating training techniques and instructional features into the simulator. These techniques allow individuals to acquire these critical skills faster and with greater retention. ARTT also allows an individual trained in 'fast time' to operate at what appears to be a more confident state, when the same task is performed in a real-time environment. Two related experiments are discussed. The findings appear to be consistent with previous findings that show positive effects of task variation during training. Moreover, ARTT has merit in improving or maintaining transfer with sharp reductions in training time. There are indications that the effectiveness of ARTT varies as a function of task content and possibly task difficulty. Other implications for ARTT are discussed along with future research directions.

  10. 25 CFR 26.30 - Does the Job Training Program provide part-time training or short-term training?

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 25 Indians 1 2014-04-01 2014-04-01 false Does the Job Training Program provide part-time training or short-term training? 26.30 Section 26.30 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR HUMAN SERVICES JOB PLACEMENT AND TRAINING PROGRAM Training Services § 26.30 Does the Job Training Program provide part-time training...

  11. 25 CFR 26.30 - Does the Job Training Program provide part-time training or short-term training?

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 25 Indians 1 2012-04-01 2011-04-01 true Does the Job Training Program provide part-time training or short-term training? 26.30 Section 26.30 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR HUMAN SERVICES JOB PLACEMENT AND TRAINING PROGRAM Training Services § 26.30 Does the Job Training Program provide part-time training or...

  12. 25 CFR 26.30 - Does the Job Training Program provide part-time training or short-term training?

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 25 Indians 1 2013-04-01 2013-04-01 false Does the Job Training Program provide part-time training or short-term training? 26.30 Section 26.30 Indians BUREAU OF INDIAN AFFAIRS, DEPARTMENT OF THE INTERIOR HUMAN SERVICES JOB PLACEMENT AND TRAINING PROGRAM Training Services § 26.30 Does the Job Training Program provide part-time training...

  13. Robotic surgical training.

    PubMed

    Ben-Or, Sharon; Nifong, L Wiley; Chitwood, W Randolph

    2013-01-01

    In July 2000, the da Vinci Surgical System (Intuitive Surgical, Inc) received Food and Drug Administration approval for intracardiac applications, and the first mitral valve repair was done at the East Carolina Heart Institute in May 2000. The system is now approved and used in many surgical specialties. With this disruptive technology and accepted use, surgeons and hospitals are seeking the most efficacious training pathway leading to safe use and responsible credentialing.One of the most important issues related to safe use is assembling the appropriate team of professionals involved with patient care. Moreover, proper patient selection and setting obtainable goals are also important.Creation and maintenance of a successful program are discussed in the article focusing on realistic goals. This begins with a partnership between surgeon leaders, hospital administrators, and industry support. Through this partnership, an appropriate training pathway and clinical pathway for success can be outlined. A timeline can then be created with periods of data analysis and adjustments as necessary. A successful program is attainable by following this pathway and attending to every detail along the journey. PMID:23528718

  14. EVA Skills Training

    NASA Technical Reports Server (NTRS)

    Parazynski, Scott

    2012-01-01

    Dr. Parazynski and a colleague from Extravehicular Activity (EVA), Robotics, & Crew Systems Operations (DX) worked closely to build the EVA Skills Training Program, and for the first time, defined the gold standards of EVA performance, allowing crewmembers to increase their performance significantly. As part of the program, individuals had the opportunity to learn at their own rate, taking additional water time as required, to achieve that level of performance. This focus on training to one's strengths and weaknesses to bolster them enabled the Crew Office and DX to field a much larger group of spacewalkers for the daunting "wall of EVA" required for the building and maintenance of the ISS. Parazynski also stressed the need for designers to understand the capabilities and the limitations of a human in a spacesuit, as well as opportunities to improve future generations of space. He shared lessons learned (how the Crew Office engaged in these endeavors) and illustrated the need to work as a team to develop these complex systems.

  15. Crew Selection and Training

    NASA Technical Reports Server (NTRS)

    Helmreich, Robert L.

    1996-01-01

    This research addressed a number of issues relevant to the performance of teams in demanding environments. Initial work, conducted in the aviation analog environment, focused on developing new measures of performance related attitudes and behaviors. The attitude measures were used to assess acceptance of concepts related to effective teamwork and personal capabilities under stress. The behavioral measures were used to evaluate the effectiveness of flight crews operating in commercial aviation. Assessment of team issues in aviation led further to the evaluation and development of training to enhance team performance. Much of the work addressed evaluation of the effectiveness of such training, which has become known as Crew Resource Management (CRM). A second line of investigation was into personality characteristics that predict performance in challenging environments such as aviation and space. A third line of investigation of team performance grew out of the study of flight crews in different organizations. This led to the development of a theoretical model of crew performance that included not only individual attributes such as personality and ability, but also organizational and national culture. A final line of investigation involved beginning to assess whether the methodologies and measures developed for the aviation analog could be applied to another domain -- the performance of medical teams working in the operating room.

  16. Virtual reality for emergency training

    SciTech Connect

    Altinkemer, K.

    1995-12-31

    Virtual reality is a sequence of scenes generated by a computer as a response to the five different senses. These senses are sight, sound, taste, touch, smell. Other senses that can be used in virtual reality include balance, pheromonal, and immunological senses. Many application areas include: leisure and entertainment, medicine, architecture, engineering, manufacturing, and training. Virtual reality is especially important when it is used for emergency training and management of natural disasters including earthquakes, floods, tornados and other situations which are hard to emulate. Classical training methods for these extraordinary environments lack the realistic surroundings that virtual reality can provide. In order for virtual reality to be a successful training tool the design needs to include certain aspects; such as how real virtual reality should be and how much fixed cost is entailed in setting up the virtual reality trainer. There are also pricing questions regarding the price per training session on virtual reality trainer, and the appropriate training time length(s).

  17. Enhancing performance in professional water polo players: dryland training, in-water training, and combined training.

    PubMed

    Sáez de Villarreal, Eduardo; Suarez-Arrones, Luis; Requena, Bernardo; Haff, G Gregory; Ramos Veliz, Rafael

    2015-04-01

    We compared the effects of 6 weeks of dryland, in-water-specific strength training and plyometric training combined with a water polo (WP) training program on 7 sport-specific performance parameters. Thirty professional players were randomly assigned to 3 experimental groups: combined training (CG), in-water-specific strength (WSG), and plyometrics (PG). The program included 3 weekly strength training sessions and 5 days of WP training per week for a total of 6 weeks during the preseason. The 10-m T-agility test, 20-m maximal sprint swim, maximal dynamic strength (1 repetition maximum [1RM], bench press [BP] and full squat [FS]), in-water boost, countermovement jump (CMJ) and throwing speed (ThS) were measured before and after the 6-week training period. There were no significant differences between the groups for any of the tested variables before the initiation of the 6-week training period. After 6 weeks of training, significant improvements (p ≤ 0.001) were found in the PG group for the CMJ (6.1%) and in all groups for the in-water boost (4.4-5.1%) test. The 1RM BP (7.6-12.6%) and FS (11.5-14.6%) significantly (p ≤ 0.05) increased in all groups. Additionally, ThS significantly increased in all groups (11.4-17.5%), whereas the agility test was significantly decreased (-7.3%) in only the CG group. Combined, in-water-specific strength and plyometric training produced medium to large effects on most WP-specific performance parameters. Therefore, we propose preseason WP training should include a combined training program that contains dryland and in-water-specific strength and plyometric training to optimize the WP preparation for competition. PMID:25259469

  18. Stockpile Dismantlement Database Training Materials

    SciTech Connect

    Not Available

    1993-11-01

    This document, the Stockpile Dismantlement Database (SDDB) training materials is designed to familiarize the user with the SDDB windowing system and the data entry steps for Component Characterization for Disposition. The foundation of information required for every part is depicted by using numbered graphic and text steps. The individual entering data is lead step by step through generic and specific examples. These training materials are intended to be supplements to individual on-the-job training.

  19. Evaluating cockpit resource management training

    NASA Technical Reports Server (NTRS)

    Helmreich, Robert L.; Wilhelm, John A.

    1987-01-01

    The determinants of effective or ineffective cockpit resurce management and the difficulties these multiple factors pose for validation of the effectiveness of cockpit resource management (CRM) training are discussed. A model of an evaluation design that may be applied to this type of training is presented. Concept validation is discussed as well as criteria for judging crew proficiency. Attention is given to accidents and proficiency checks, incidents and repeated maneuvers, attitude measuremet, and self-report evauation of training.

  20. COPD: benefits of exercise training.

    PubMed

    2016-03-01

    In patients with stable, moderate or severe chronic obstructive pulmonary disease (COPD), general exercise training, including limb exercises, provides sustained improvement in various quality of life domains, compared with care without pulmonary rehabilitation. After a COPD exacerbation, exercise training appears to reduce the risk of hospitalisation in the following months by at least half. Few studies have evaluated the adverse effects of exercise training in COPD, but based on the data available in 2015, its harm-benefit balance appears favourable. PMID:27152405