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Sample records for affect nonhomologous end-joining

  1. The mechanism of non-homologous end-joining: a synopsis of synapsis.

    Science.gov (United States)

    Weterings, Eric; van Gent, Dik C

    2004-11-02

    Repair of DNA double-strand breaks (DSBs) by non-homologous end-joining (NHEJ) is required for resistance to genotoxic agents, such as ionizing radiation, but also for proper development of the vertebrate immune system. Much progress has been made in identifying the factors that are involved in this repair pathway. We are now entering the phase in which we begin to understand basic concepts of the reaction mechanism and regulation of non-homologous end-joining. This review concentrates on novel insights into damage recognition and subsequent tethering, processing and joining of DNA ends.

  2. Modeling Non-homologous End Joining

    Science.gov (United States)

    Li, Yongfeng

    2013-01-01

    Non-homologous end joining (NHEJ) is the dominant DNA double strand break (DSB) repair pathway and involves several NHEJ proteins such as Ku, DNA-PKcs, XRCC4, Ligase IV and so on. Once DSBs are generated, Ku is first recruited to the DNA end, followed by other NHEJ proteins for DNA end processing and ligation. Because of the direct ligation of break ends without the need for a homologous template, NHEJ turns out to be an error-prone but efficient repair pathway. Some mechanisms have been proposed of how the efficiency of NHEJ repair is affected. The type of DNA damage is an important factor of NHEJ repair. For instance, the length of DNA fragment may determine the recruitment efficiency of NHEJ protein such as Ku [1], or the complexity of the DNA breaks [2] is accounted for the choice of NHEJ proteins and subpathway of NHEJ repair. On the other hand, the chromatin structure also plays a role of the accessibility of NHEJ protein to the DNA damage site. In this talk, some mathematical models of NHEJ, that consist of series of biochemical reactions complying with the laws of chemical reaction (e.g. mass action, etc.), will be introduced. By mathematical and numerical analysis and parameter estimation, the models are able to capture the qualitative biological features and show good agreement with experimental data. As conclusions, from the viewpoint of modeling, how the NHEJ proteins are recruited will be first discussed for connection between the classical sequential model [4] and recently proposed two-phase model [5]. Then how the NHEJ repair pathway is affected, by the length of DNA fragment [6], the complexity of DNA damage [7] and the chromatin structure [8], will be addressed

  3. Nonhomologous DNA End Joining in Cell-Free Extracts

    Directory of Open Access Journals (Sweden)

    Sheetal Sharma

    2010-01-01

    Full Text Available Among various DNA damages, double-strand breaks (DSBs are considered as most deleterious, as they may lead to chromosomal rearrangements and cancer when unrepaired. Nonhomologous DNA end joining (NHEJ is one of the major DSB repair pathways in higher organisms. A large number of studies on NHEJ are based on in vitro systems using cell-free extracts. In this paper, we summarize the studies on NHEJ performed by various groups in different cell-free repair systems.

  4. Detection and Repair of Ionizing Radiation-Induced DNA Double Strand Breaks: New Developments in Nonhomologous End Joining

    International Nuclear Information System (INIS)

    Wang, Chen; Lees-Miller, Susan P.

    2013-01-01

    DNA damage can occur as a result of endogenous metabolic reactions and replication stress or from exogenous sources such as radiation therapy and chemotherapy. DNA double strand breaks are the most cytotoxic form of DNA damage, and defects in their repair can result in genome instability, a hallmark of cancer. The major pathway for the repair of ionizing radiation-induced DSBs in human cells is nonhomologous end joining. Here we review recent advances on the mechanism of nonhomologous end joining, as well as new findings on its component proteins and regulation

  5. Detection and Repair of Ionizing Radiation-Induced DNA Double Strand Breaks: New Developments in Nonhomologous End Joining

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Chen [Departments of Biochemistry and Molecular Biology and Oncology, and Southern Alberta Cancer Research Institute, University of Calgary, Calgary (Canada); Lees-Miller, Susan P., E-mail: leesmill@ucalgary.ca [Departments of Biochemistry and Molecular Biology and Oncology, and Southern Alberta Cancer Research Institute, University of Calgary, Calgary (Canada)

    2013-07-01

    DNA damage can occur as a result of endogenous metabolic reactions and replication stress or from exogenous sources such as radiation therapy and chemotherapy. DNA double strand breaks are the most cytotoxic form of DNA damage, and defects in their repair can result in genome instability, a hallmark of cancer. The major pathway for the repair of ionizing radiation-induced DSBs in human cells is nonhomologous end joining. Here we review recent advances on the mechanism of nonhomologous end joining, as well as new findings on its component proteins and regulation.

  6. The endless tale of non-homologous end-joining.

    Science.gov (United States)

    Weterings, Eric; Chen, David J

    2008-01-01

    DNA double-strand breaks (DSBs) are introduced in cells by ionizing radiation and reactive oxygen species. In addition, they are commonly generated during V(D)J recombination, an essential aspect of the developing immune system. Failure to effectively repair these DSBs can result in chromosome breakage, cell death, onset of cancer, and defects in the immune system of higher vertebrates. Fortunately, all mammalian cells possess two enzymatic pathways that mediate the repair of DSBs: homologous recombination and non-homologous end-joining (NHEJ). The NHEJ process utilizes enzymes that capture both ends of the broken DNA molecule, bring them together in a synaptic DNA-protein complex, and finally repair the DNA break. In this review, all the known enzymes that play a role in the NHEJ process are discussed and a working model for the co-operation of these enzymes during DSB repair is presented.

  7. Distinct DNA repair pathways involving RecA and nonhomologous end joining in Mycobacterium smegmatis.

    OpenAIRE

    Korycka-Machala, M; Brzostek, A; Rozalska, S; Rumijowska-Galewicz, A; Dziedzic, R; Bowater, R; Dziadek, J

    2006-01-01

    Mycobacterium smegmatis was used to study the relationship between DNA repair processes involving RecA and nonhomologous end joining (NHEJ). The effect of gene deletions in recA and/or in two genes involved in NHEJ (ku and ligD) was tested on the ability of bacteria to join breaks in plasmids transformed into them and in their response to chemicals that damage DNA. The results provide in vivo evidence that only NHEJ is required for the repair of noncompatible DNA ends. By contrast, the respon...

  8. Distinct DNA repair pathways involving RecA and nonhomologous end joining in Mycobacterium smegmatis.

    Science.gov (United States)

    Korycka-Machala, Malgorzata; Brzostek, Anna; Rozalska, Sylwia; Rumijowska-Galewicz, Anna; Dziedzic, Renata; Bowater, Richard; Dziadek, Jaroslaw

    2006-05-01

    Mycobacterium smegmatis was used to study the relationship between DNA repair processes involving RecA and nonhomologous end joining (NHEJ). The effect of gene deletions in recA and/or in two genes involved in NHEJ (ku and ligD) was tested on the ability of bacteria to join breaks in plasmids transformed into them and in their response to chemicals that damage DNA. The results provide in vivo evidence that only NHEJ is required for the repair of noncompatible DNA ends. By contrast, the response of mycobacteria to mitomycin C preferentially involved a RecA-dependent pathway.

  9. Non-homologous end-joining genes are not inactivated in human radiation-induced sarcomas with genomic instability

    International Nuclear Information System (INIS)

    Lefevre, S.H.; Coquelle, A.; Gonin-Laurent, N.

    2005-01-01

    DNA double-strand break (DSB) repair pathways are implicated in the maintenance of genomic stability. However the alterations of these pathways, as may occur in human tumor cells with strong genomic instability, remain poorly characterized. We analyzed the loss of heterozygosity (LOH) and the presence of mutations for a series of genes implicated in DSB repair by non-homologous end-joining in five radiation-induced sarcomas devoid of both active Tp53 and Rb1. LOH was recurrently observed for 8 of the 9 studied genes (KU70, KU80, XRCC4, LIG4, Artemis, MRE11, RAD50, NBS1) but not for DNA-PKcs. No mutation was found in the remaining allele of the genes with LOH and the mRNA expression did not correlate with the allelic status. Our findings suggest that non-homologous end-joining repair pathway alteration is unlikely to be involved in the high genomic instability observed in these tumors. (author)

  10. Coincident In Vitro Analysis of DNA-PK-Dependent and -Independent Nonhomologous End Joining

    Directory of Open Access Journals (Sweden)

    Cynthia L. Hendrickson

    2010-01-01

    Full Text Available In mammalian cells, DNA double-strand breaks (DSBs are primarily repaired by nonhomologous end joining (NHEJ. The current model suggests that the Ku 70/80 heterodimer binds to DSB ends and recruits DNA-PKcs to form the active DNA-dependent protein kinase, DNA-PK. Subsequently, XRCC4, DNA ligase IV, XLF and most likely, other unidentified components participate in the final DSB ligation step. Therefore, DNA-PK plays a key role in NHEJ due to its structural and regulatory functions that mediate DSB end joining. However, recent studies show that additional DNA-PK-independent NHEJ pathways also exist. Unfortunately, the presence of DNA-PKcs appears to inhibit DNA-PK-independent NHEJ, and in vitro analysis of DNA-PK-independent NHEJ in the presence of the DNA-PKcs protein remains problematic. We have developed an in vitro assay that is preferentially active for DNA-PK-independent DSB repair based solely on its reaction conditions, facilitating coincident differential biochemical analysis of the two pathways. The results indicate the biochemically distinct nature of the end-joining mechanisms represented by the DNA-PK-dependent and -independent NHEJ assays as well as functional differences between the two pathways.

  11. Requirement for XLF/Cernunnos in alignment-based gap filling by DNA polymerases lambda and mu for nonhomologous end joining in human whole-cell extracts.

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    Akopiants, Konstantin; Zhou, Rui-Zhe; Mohapatra, Susovan; Valerie, Kristoffer; Lees-Miller, Susan P; Lee, Kyung-Jong; Chen, David J; Revy, Patrick; de Villartay, Jean-Pierre; Povirk, Lawrence F

    2009-07-01

    XLF/Cernunnos is a core protein of the nonhomologous end-joining pathway of DNA double-strand break repair. To better define the role of Cernunnos in end joining, whole-cell extracts were prepared from Cernunnos-deficient human cells. These extracts effected little joining of DNA ends with cohesive 5' or 3' overhangs, and no joining at all of partially complementary 3' overhangs that required gap filling prior to ligation. Assays in which gap-filled but unligated intermediates were trapped using dideoxynucleotides revealed that there was no gap filling on aligned DSB ends in the Cernunnos-deficient extracts. Recombinant Cernunnos protein restored gap filling and end joining of partially complementary overhangs, and stimulated joining of cohesive ends more than twentyfold. XLF-dependent gap filling was nearly eliminated by immunodepletion of DNA polymerase lambda, but was restored by addition of either polymerase lambda or polymerase mu. Thus, Cernunnos is essential for gap filling by either polymerase during nonhomologous end joining, suggesting that it plays a major role in aligning the two DNA ends in the repair complex.

  12. Contribution of canonical nonhomologous end joining to chromosomal rearrangements is enhanced by ATM kinase deficiency.

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    Bhargava, Ragini; Carson, Caree R; Lee, Gabriella; Stark, Jeremy M

    2017-01-24

    A likely mechanism of chromosomal rearrangement formation involves joining the ends from two different chromosomal double-strand breaks (DSBs). These events could potentially be mediated by either of two end-joining (EJ) repair pathways [canonical nonhomologous end joining (C-NHEJ) or alternative end joining (ALT-EJ)], which cause distinct rearrangement junction patterns. The relative role of these EJ pathways during rearrangement formation has remained controversial. Along these lines, we have tested whether the DNA damage response mediated by the Ataxia Telangiectasia Mutated (ATM) kinase may affect the relative influence of C-NHEJ vs. ALT-EJ on rearrangement formation. We developed a reporter in mouse cells for a 0.4-Mbp deletion rearrangement that is formed by EJ between two DSBs induced by the Cas9 endonuclease. We found that disruption of the ATM kinase causes an increase in the frequency of the rearrangement as well as a shift toward rearrangement junctions that show hallmarks of C-NHEJ. Furthermore, ATM suppresses rearrangement formation in an experimental condition, in which C-NHEJ is the predominant EJ repair event (i.e., expression of the 3' exonuclease Trex2). Finally, several C-NHEJ factors are required for the increase in rearrangement frequency caused by inhibition of the ATM kinase. We also examined ATM effectors and found that H2AX shows a similar influence as ATM, whereas the influence of ATM on this rearrangement seems independent of 53BP1. We suggest that the contribution of the C-NHEJ pathway to the formation of a 0.4-Mbp deletion rearrangement is enhanced in ATM-deficient cells.

  13. Ectopic expression of RNF168 and 53BP1 increases mutagenic but not physiological non-homologous end joining

    DEFF Research Database (Denmark)

    Zong, Dali; Callén, Elsa; Pegoraro, Gianluca

    2015-01-01

    DNA double strand breaks (DSBs) formed during S phase are preferentially repaired by homologous recombination (HR), whereas G1 DSBs, such as those occurring during immunoglobulin class switch recombination (CSR), are repaired by non-homologous end joining (NHEJ). The DNA damage response proteins ...

  14. Requirement for XLF/Cernunnos in alignment-based gap filling by DNA polymerases ? and ? for nonhomologous end joining in human whole-cell extracts

    OpenAIRE

    Akopiants, Konstantin; Zhou, Rui-Zhe; Mohapatra, Susovan; Valerie, Kristoffer; Lees-Miller, Susan P.; Lee, Kyung-Jong; Chen, David J.; Revy, Patrick; de Villartay, Jean-Pierre; Povirk, Lawrence F.

    2009-01-01

    XLF/Cernunnos is a core protein of the nonhomologous end-joining pathway of DNA double-strand break repair. To better define the role of Cernunnos in end joining, whole-cell extracts were prepared from Cernunnos-deficient human cells. These extracts effected little joining of DNA ends with cohesive 5? or 3? overhangs, and no joining at all of partially complementary 3? overhangs that required gap filling prior to ligation. Assays in which gap-filled but unligated intermediates were trapped us...

  15. Positive selection on the nonhomologous end-joining factor Cernunnos-XLF in the human lineage

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    Jurka Jerzy

    2006-06-01

    Full Text Available Abstract Background Cernunnos-XLF is a nonhomologous end-joining factor that is mutated in patients with a rare immunodeficiency with microcephaly. Several other microcephaly-associated genes such as ASPM and microcephalin experienced recent adaptive evolution apparently linked to brain size expansion in humans. In this study we investigated whether Cernunnos-XLF experienced similar positive selection during human evolution. Results We obtained or reconstructed full-length coding sequences of chimpanzee, rhesus macaque, canine, and bovine Cernunnos-XLF orthologs from sequence databases and sequence trace archives. Comparison of coding sequences revealed an excess of nonsynonymous substitutions consistent with positive selection on Cernunnos-XLF in the human lineage. The hotspots of adaptive evolution are concentrated around a specific structural domain, whose analogue in the structurally similar XRCC4 protein is involved in binding of another nonhomologous end-joining factor, DNA ligase IV. Conclusion Cernunnos-XLF is a microcephaly-associated locus newly identified to be under adaptive evolution in humans, and possibly played a role in human brain expansion. We speculate that Cernunnos-XLF may have contributed to the increased number of brain cells in humans by efficient double strand break repair, which helps to prevent frequent apoptosis of neuronal progenitors and aids mitotic cell cycle progression. Reviewers This article was reviewed by Chris Ponting and Richard Emes (nominated by Chris Ponting, Kateryna Makova, Gáspár Jékely and Eugene V. Koonin.

  16. In vivo blunt-end cloning through CRISPR/Cas9-facilitated non-homologous end-joining

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    Geisinger, Jonathan M.; Turan, Sören; Hernandez, Sophia; Spector, Laura P.; Calos, Michele P.

    2016-01-01

    The CRISPR/Cas9 system facilitates precise DNA modifications by generating RNA-guided blunt-ended double-strand breaks. We demonstrate that guide RNA pairs generate deletions that are repaired with a high level of precision by non-homologous end-joining in mammalian cells. We present a method called knock-in blunt ligation for exploiting these breaks to insert exogenous PCR-generated sequences in a homology-independent manner without loss of additional nucleotides. This method is useful for making precise additions to the genome such as insertions of marker gene cassettes or functional elements, without the need for homology arms. We successfully utilized this method in human and mouse cells to insert fluorescent protein cassettes into various loci, with efficiencies up to 36% in HEK293 cells without selection. We also created versions of Cas9 fused to the FKBP12-L106P destabilization domain in an effort to improve Cas9 performance. Our in vivo blunt-end cloning method and destabilization-domain-fused Cas9 variant increase the repertoire of precision genome engineering approaches. PMID:26762978

  17. Atomic Structure and Nonhomologous End-Joining Function of the Polymerase Component of Bacterial DNA Ligase D

    Energy Technology Data Exchange (ETDEWEB)

    Zhu,H.; Nandakumar, J.; Aniukwu, J.; Wang, L.; Glickman, M.; Lima, C.; Shuman, S.

    2006-01-01

    DNA ligase D (LigD) is a large polyfunctional protein that participates in a recently discovered pathway of nonhomologous end-joining in bacteria. LigD consists of an ATP-dependent ligase domain fused to a polymerase domain (Pol) and a phosphoesterase module. The Pol activity is remarkable for its dependence on manganese, its ability to perform templated and nontemplated primer extension reactions, and its preference for adding ribonucleotides to blunt DNA ends. Here we report the 1.5- Angstroms crystal structure of the Pol domain of Pseudomonas LigD and its complexes with manganese and ATP-dATP substrates, which reveal a minimized polymerase with a two-metal mechanism and a fold similar to that of archaeal DNA primase. Mutational analysis highlights the functionally relevant atomic contacts in the active site. Although distinct nucleoside conformations and contacts for ATP versus dATP are observed in the cocrystals, the functional analysis suggests that the ATP-binding mode is the productive conformation for dNMP and rNMP incorporation. We find that a mutation of Mycobacterium LigD that uniquely ablates the polymerase activity results in increased fidelity of blunt-end double-strand break repair in vivo by virtue of eliminating nucleotide insertions at the recombination junctions. Thus, LigD Pol is a direct catalyst of mutagenic nonhomologous end-joining in vivo. Our studies underscore a previously uncharacterized role for the primase-like polymerase family in DNA repair.

  18. Processing of DNA double strand breaks by alternative non-homologous end-joining in hyperacetylated chromatin.

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    Manova, Vasilissa; Singh, Satyendra K; Iliakis, George

    2012-08-22

    Mammalian cells employ at least two subpathways of non-homologous end-joining for the repair of ionizing radiation induced DNA double strand breaks: The canonical DNA-PK-dependent form of non-homologous end-joining (D-NHEJ) and an alternative, slowly operating, error-prone backup pathway (B-NHEJ). In contrast to D-NHEJ, which operates with similar efficiency throughout the cell cycle, B-NHEJ operates more efficiently in G2-phase. Notably, B-NHEJ also shows strong and as of yet unexplained dependency on growth activity and is markedly compromised in serum-deprived cells, or in cells that enter the plateau-phase of growth. The molecular mechanisms underpinning this response remain unknown. Since chromatin structure or changes in chromatin structure are prime candidate-B-NHEJ-modulators, we study here the role of chromatin hyperacetylation, either by HDAC2 knockdown or treatment with the HDAC inhibitor TSA, on the repair by B-NHEJ of IR-induced DSBs. siRNA-mediated knockdown of HDAC2 fails to provoke histone hyperacetylation in Lig4-/- MEFs and has no detectable effect on B-NHEJ function. Treatment with TSA that inhibits multiple HDACs causes efficient, reversible chromatin hyperacetylation in Lig4-/- MEFs, as well as in human HCT116 Lig4-/- cells and the human glioma cell line M059K. The IR yield of DSBs in TSA-treated cells remains similar to that of untreated cells despite the expected chromatin relaxation. In addition, chromatin hyperacetylation leaves unchanged repair of DSBs by B-NHEJ in irradiated exponentially growing, or plateau-phase cells. Notably, under the experimental conditions employed here, chromatin hyperacetylation fails to detectably modulate B-NHEJ in M059K cells as well. In summary, the results show that chromatin acetylation or deacetylation does not affect the kinetics of alternative NHEJ in all types of cells examined both in exponentially growing and serum deprived cultures. We conclude that parameters beyond chromatin acetylation determine B

  19. Double Strand Break Repair, one mechanism can hide another: Alternative non-homologous end joining

    International Nuclear Information System (INIS)

    Rass, E.; Grabarz, A.; Bertrand, P.; Lopez, B.S.

    2012-01-01

    DNA double strand breaks are major cytotoxic lesions encountered by the cells. They can be induced by ionizing radiation or endogenous stress and can lead to genetic instability. Two mechanisms compete for the repair of DNA double strand breaks: homologous recombination and non-homologous end joining (NHEJ). Homologous recombination requires DNA sequences homology and is initiated by single strand resection. Recently, advances have been made concerning the major steps and proteins involved in resection. NHEJ, in contrast, does not require sequence homology. The existence of a DNA double strand break repair mechanism, independent of KU and ligase IV, the key proteins of the canonical non homologous end joining pathway, has been revealed lately and named alternative non homologous end joining. The hallmarks of this highly mutagenic pathway are deletions at repair junctions and frequent use of distal micro-homologies. This mechanism is also initiated by a single strand resection of the break. The aim of this review is firstly to present recent data on single strand resection, and secondly the alternative NHEJ pathway, including a discussion on the fidelity of NHEJ. Based on current knowledge, canonical NHEJ does not appear as an intrinsically mutagenic mechanism, but in contrast, as a conservative one. The structure of broken DNA ends actually dictates the quality repair of the alternative NHEJ and seems the actual responsible for the mutagenesis attributed beforehand to the canonical NHEJ. The existence of this novel DNA double strand breaks repair mechanism needs to be taken into account in the development of radiosensitizing strategies in order to optimise the efficiency of radiotherapy. (authors)

  20. Altered kinetics of nonhomologous end joining and class switch recombination in ligase IV-deficient B cells.

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    Han, Li; Yu, Kefei

    2008-11-24

    Immunoglobulin heavy chain class switch recombination (CSR) is believed to occur through the generation and repair of DNA double-strand breaks (DSBs) in the long and repetitive switch regions. Although implied, the role of the major vertebrate DSB repair pathway, nonhomologous end joining (NHEJ), in CSR has been controversial. By somatic gene targeting of DNA ligase IV (Lig4; a key component of NHEJ) in a B cell line (CH12F3) capable of highly efficient CSR in vitro, we found that NHEJ is required for efficient CSR. Disruption of the Lig4 gene in CH12F3 cells severely inhibits the initial rate of CSR and causes a late cell proliferation defect under cytokine stimulation. However, unlike V(D)J recombination, which absolutely requires NHEJ, CSR accumulates to a substantial level in Lig4-null cells. The data revealed a fast-acting NHEJ and a slow-acting alterative end joining of switch region breaks during CSR.

  1. Environmental Stress Induces Trinucleotide Repeat Mutagenesis in Human Cells by Alt-Nonhomologous End Joining Repair.

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    Chatterjee, Nimrat; Lin, Yunfu; Yotnda, Patricia; Wilson, John H

    2016-07-31

    Multiple pathways modulate the dynamic mutability of trinucleotide repeats (TNRs), which are implicated in neurodegenerative disease and evolution. Recently, we reported that environmental stresses induce TNR mutagenesis via stress responses and rereplication, with more than 50% of mutants carrying deletions or insertions-molecular signatures of DNA double-strand break repair. We now show that knockdown of alt-nonhomologous end joining (alt-NHEJ) components-XRCC1, LIG3, and PARP1-suppresses stress-induced TNR mutagenesis, in contrast to the components of homologous recombination and NHEJ, which have no effect. Thus, alt-NHEJ, which contributes to genetic mutability in cancer cells, also plays a novel role in environmental stress-induced TNR mutagenesis. Published by Elsevier Ltd.

  2. XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes.

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    Saribasak, Huseyin; Maul, Robert W; Cao, Zheng; McClure, Rhonda L; Yang, William; McNeill, Daniel R; Wilson, David M; Gearhart, Patricia J

    2011-10-24

    Activation-induced deaminase (AID) deaminates cytosine to uracil in immunoglobulin genes. Uracils in DNA can be recognized by uracil DNA glycosylase and abasic endonuclease to produce single-strand breaks. The breaks are repaired either faithfully by DNA base excision repair (BER) or mutagenically to produce somatic hypermutation (SHM) and class switch recombination (CSR). To unravel the interplay between repair and mutagenesis, we decreased the level of x-ray cross-complementing 1 (XRCC1), a scaffold protein involved in BER. Mice heterozygous for XRCC1 showed a significant increase in the frequencies of SHM in Igh variable regions in Peyer's patch cells, and of double-strand breaks in the switch regions during CSR. Although the frequency of CSR was normal in Xrcc1(+/-) splenic B cells, the length of microhomology at the switch junctions decreased, suggesting that XRCC1 also participates in alternative nonhomologous end joining. Furthermore, Xrcc1(+/-) B cells had reduced Igh/c-myc translocations during CSR, supporting a role for XRCC1 in microhomology-mediated joining. Our results imply that AID-induced single-strand breaks in Igh variable and switch regions become substrates simultaneously for BER and mutagenesis pathways.

  3. The Ku70/80 ring in Non-Homologous End-Joining: easy to slip on, hard to remove.

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    Kragelund, Birthe B; Weterings, Eric; Hartmann-Petersen, Rasmus; Keijzers, Guido

    2016-01-01

    Non-homologous end-joining (NHEJ) is an essential DNA double strand break repair pathway during all cell cycle stages. Deficiency in NHEJ factors can lead to accumulation of unrepaired DNA breaks or faulty DNA repair, which may ultimately result in cell death, senescence or carcinogenesis. The Ku70/80 heterodimer is a key-player in the NHEJ pathway and binds to DNA termini with high affinity, where it helps to protect DNA ends from degradation and to recruit other NHEJ factors required for repair. The mechanism of Ku70/80 detachment from the DNA helix after completion of DNA repair is incompletely understood. Some data suggest that certain DNA repair factors are ubiquitylated and targeted for proteasomal degradation after repair. Recent studies suggest that Ku80 is conjugated to lysine48-linked ubiquitin chains by the Skp1-Cullin-F-box (SCF) complex and/or the RING finger protein 8 (RNF8) ubiquitin-protein ligases, followed by rapid proteasomal degradation. In this review we address the structure and function of the Ku70/80 heterodimer and how ubiquitylation may affect the release of Ku70/80 from chromatin and its subsequent degradation via the ubiquitin-proteasome system.

  4. Lithium chloride protects retinal neurocytes from nutrient deprivation by promoting DNA non-homologous end-joining

    International Nuclear Information System (INIS)

    Zhuang Jing; Li Fan; Liu Xuan; Liu Zhiping; Lin Jianxian; Ge Yihong; Kaminski, Joseph M.; Summers, James Bradley; Wang Zhichong; Ge Jian; Yu Keming

    2009-01-01

    Lithium chloride is a therapeutic agent for treatment of bipolar affective disorders. Increasing numbers of studies have indicated that lithium has neuroprotective effects. However, the molecular mechanisms underlying the actions of lithium have not been fully elucidated. This study aimed to investigate whether lithium chloride produces neuroprotective function by improving DNA repair pathway in retinal neurocyte. In vitro, the primary cultured retinal neurocytes (85.7% are MAP-2 positive cells) were treated with lithium chloride, then cultured with serum-free media to simulate the nutrient deprived state resulting from ischemic insult. The neurite outgrowth of the cultured cells increased significantly in a dose-dependent manner when exposed to different levels of lithium chloride. Genomic DNA electrophoresis demonstrated greater DNA integrity of retinal neurocytes when treated with lithium chloride as compared to the control. Moreover, mRNA and protein levels of Ligase IV (involved in DNA non-homologous end-joining (NHEJ) pathway) in retinal neurocytes increased with lithium chloride. The end joining activity assay was performed to determine the role of lithium on NHEJ in the presence of extract from retinal neurocytes. The rejoining levels in retinal neurocytes treated with lithium were significantly increased as compared to the control. Furthermore, XRCC4, the Ligase IV partner, and the transcriptional factor, CREB and CTCF, were up-regulated in retinal cells after treating with 1.0 mM lithium chloride. Therefore, our data suggest that lithium chloride protects the retinal neural cells from nutrient deprivation in vitro, which may be similar to the mechanism of cell death in glaucoma. The improvement in DNA repair pathway involving in Ligase IV might have an important role in lithium neuroprotection. This study provides new insights into the neural protective mechanisms of lithium chloride.

  5. Direct Involvement of Retinoblastoma Family Proteins in DNA Repair by Non-homologous End-Joining

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    Rebecca Cook

    2015-03-01

    Full Text Available Deficiencies in DNA double-strand break (DSB repair lead to genetic instability, a recognized cause of cancer initiation and evolution. We report that the retinoblastoma tumor suppressor protein (RB1 is required for DNA DSB repair by canonical non-homologous end-joining (cNHEJ. Support of cNHEJ involves a mechanism independent of RB1’s cell-cycle function and depends on its amino terminal domain with which it binds to NHEJ components XRCC5 and XRCC6. Cells with engineered loss of RB family function as well as cancer-derived cells with mutational RB1 loss show substantially reduced levels of cNHEJ. RB1 variants disabled for the interaction with XRCC5 and XRCC6, including a cancer-associated variant, are unable to support cNHEJ despite being able to confer cell-cycle control. Our data identify RB1 loss as a candidate driver of structural genomic instability and a causative factor for cancer somatic heterogeneity and evolution.

  6. Hyperactivation of PARP triggers nonhomologous end-joining in repair-deficient mouse fibroblasts.

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    Natalie R Gassman

    Full Text Available Regulation of poly(ADP-ribose (PAR synthesis and turnover is critical to determining cell fate after genotoxic stress. Hyperactivation of PAR synthesis by poly(ADP-ribose polymerase-1 (PARP-1 occurs when cells deficient in DNA repair are exposed to genotoxic agents; however, the function of this hyperactivation has not been adequately explained. Here, we examine PAR synthesis in mouse fibroblasts deficient in the base excision repair enzyme DNA polymerase β (pol β. The extent and duration of PARP-1 activation was measured after exposure to either the DNA alkylating agent, methyl methanesulfonate (MMS, or to low energy laser-induced DNA damage. There was strong DNA damage-induced hyperactivation of PARP-1 in pol β nullcells, but not in wild-type cells. In the case of MMS treatment, PAR synthesis did not lead to cell death in the pol β null cells, but instead resulted in increased PARylation of the nonhomologous end-joining (NHEJ protein Ku70 and increased association of Ku70 with PARP-1. Inhibition of the NHEJ factor DNA-PK, under conditions of MMS-induced PARP-1 hyperactivation, enhanced necrotic cell death. These data suggest that PARP-1 hyperactivation is a protective mechanism triggering the classical-NHEJ DNA repair pathway when the primary alkylated base damage repair pathway is compromised.

  7. Development of versatile non-homologous end joining-based knock-in module for genome editing.

    Science.gov (United States)

    Sawatsubashi, Shun; Joko, Yudai; Fukumoto, Seiji; Matsumoto, Toshio; Sugano, Shigeo S

    2018-01-12

    CRISPR/Cas9-based genome editing has dramatically accelerated genome engineering. An important aspect of genome engineering is efficient knock-in technology. For improved knock-in efficiency, the non-homologous end joining (NHEJ) repair pathway has been used over the homology-dependent repair pathway, but there remains a need to reduce the complexity of the preparation of donor vectors. We developed the versatile NHEJ-based knock-in module for genome editing (VIKING). Using the consensus sequence of the time-honored pUC vector to cut donor vectors, any vector with a pUC backbone could be used as the donor vector without customization. Conditions required to minimize random integration rates of the donor vector were also investigated. We attempted to isolate null lines of the VDR gene in human HaCaT keratinocytes using knock-in/knock-out with a selection marker cassette, and found 75% of clones isolated were successfully knocked-in. Although HaCaT cells have hypotetraploid genome composition, the results suggest multiple clones have VDR null phenotypes. VIKING modules enabled highly efficient knock-in of any vectors harboring pUC vectors. Users now can insert various existing vectors into an arbitrary locus in the genome. VIKING will contribute to low-cost genome engineering.

  8. De novo CNV formation in mouse embryonic stem cells occurs in the absence of Xrcc4-dependent nonhomologous end joining.

    Directory of Open Access Journals (Sweden)

    Martin F Arlt

    2012-09-01

    Full Text Available Spontaneous copy number variant (CNV mutations are an important factor in genomic structural variation, genomic disorders, and cancer. A major class of CNVs, termed nonrecurrent CNVs, is thought to arise by nonhomologous DNA repair mechanisms due to the presence of short microhomologies, blunt ends, or short insertions at junctions of normal and de novo pathogenic CNVs, features recapitulated in experimental systems in which CNVs are induced by exogenous replication stress. To test whether the canonical nonhomologous end joining (NHEJ pathway of double-strand break (DSB repair is involved in the formation of this class of CNVs, chromosome integrity was monitored in NHEJ-deficient Xrcc4(-/- mouse embryonic stem (ES cells following treatment with low doses of aphidicolin, a DNA replicative polymerase inhibitor. Mouse ES cells exhibited replication stress-induced CNV formation in the same manner as human fibroblasts, including the existence of syntenic hotspot regions, such as in the Auts2 and Wwox loci. The frequency and location of spontaneous and aphidicolin-induced CNV formation were not altered by loss of Xrcc4, as would be expected if canonical NHEJ were the predominant pathway of CNV formation. Moreover, de novo CNV junctions displayed a typical pattern of microhomology and blunt end use that did not change in the absence of Xrcc4. A number of complex CNVs were detected in both wild-type and Xrcc4(-/- cells, including an example of a catastrophic, chromothripsis event. These results establish that nonrecurrent CNVs can be, and frequently are, formed by mechanisms other than Xrcc4-dependent NHEJ.

  9. Non-homologous end joining mediated DNA repair is impaired in the NUP98-HOXD13 mouse model for myelodysplastic syndrome.

    Science.gov (United States)

    Puthiyaveetil, Abdul Gafoor; Reilly, Christopher M; Pardee, Timothy S; Caudell, David L

    2013-01-01

    Chromosomal translocations typically impair cell differentiation and often require secondary mutations for malignant transformation. However, the role of a primary translocation in the development of collaborating mutations is debatable. To delineate the role of leukemic translocation NUP98-HOXD13 (NHD13) in secondary mutagenesis, DNA break and repair mechanisms in stimulated mouse B lymphocytes expressing NHD13 were analyzed. Our results showed significantly reduced expression of non-homologous end joining (NHEJ)-mediated DNA repair genes, DNA Pkcs, DNA ligase4, and Xrcc4 leading to cell cycle arrest at G2/M phase. Our results showed that expression of NHD13 fusion gene resulted in impaired NHEJ-mediated DNA break repair. Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. Mycobacterium tuberculosis and Mycobacterium marinum non-homologous end-joining proteins can function together to join DNA ends in Escherichia coli.

    Science.gov (United States)

    Wright, Douglas G; Castore, Reneau; Shi, Runhua; Mallick, Amrita; Ennis, Don G; Harrison, Lynn

    2017-03-01

    Mycobacterium tuberculosis and Mycobacterium smegmatis express a Ku protein and a DNA ligase D and are able to repair DNA double strand breaks (DSBs) by non-homologous end-joining (NHEJ). This pathway protects against DNA damage when bacteria are in stationary phase. Mycobacterium marinum is a member of this mycobacterium family and like M. tuberculosis is pathogenic. M. marinum lives in water, forms biofilms and infects fish and frogs. M. marinum is a biosafety level 2 (BSL2) organism as it can infect humans, although infections are limited to the skin. M. marinum is accepted as a model to study mycobacterial pathogenesis, as M. marinum and M. tuberculosis are genetically closely related and have similar mechanisms of survival and persistence inside macrophage. The aim of this study was to determine whether M. marinum could be used as a model to understand M. tuberculosis NHEJ repair. We identified and cloned the M. marinum genes encoding NHEJ proteins and generated E. coli strains that express the M. marinum Ku (Mm-Ku) and ligase D (Mm-Lig) individually or together (LHmKumLig strain) from expression vectors integrated at phage attachment sites in the genome. We demonstrated that Mm-Ku and Mm-Lig are both required to re-circularize Cla I-linearized plasmid DNA in E. coli. We compared repair of strain LHmKumLig with that of an E. coli strain (BWKuLig#2) expressing the M. tuberculosis Ku (Mt-Ku) and ligase D (Mt-Lig), and found that LHmKumLig performed 3.5 times more repair and repair was more accurate than BWKuLig#2. By expressing the Mm-Ku with the Mt-Lig, or the Mt-Ku with the Mm-Lig in E. coli, we have shown that the NHEJ proteins from M. marinum and M. tuberculosis can function together to join DNA DSBs. NHEJ repair is therefore conserved between the two species. Consequently, M. marinum is a good model to study NHEJ repair during mycobacterial pathogenesis. © The Author 2016. Published by Oxford University Press on behalf of the UK Environmental Mutagen

  11. Mycobacterial nonhomologous end joining mediates mutagenic repair of chromosomal double-strand DNA breaks.

    Science.gov (United States)

    Stephanou, Nicolas C; Gao, Feng; Bongiorno, Paola; Ehrt, Sabine; Schnappinger, Dirk; Shuman, Stewart; Glickman, Michael S

    2007-07-01

    Bacterial nonhomologous end joining (NHEJ) is a recently described DNA repair pathway best characterized in mycobacteria. Bacterial NHEJ proteins LigD and Ku have been analyzed biochemically, and their roles in linear plasmid repair in vivo have been verified genetically; yet the contributions of NHEJ to repair of chromosomal DNA damage are unknown. Here we use an extensive set of NHEJ- and homologous recombination (HR)-deficient Mycobacterium smegmatis strains to probe the importance of HR and NHEJ in repairing diverse types of chromosomal DNA damage. An M. smegmatis Delta recA Delta ku double mutant has no apparent growth defect in vitro. Loss of the NHEJ components Ku and LigD had no effect on sensitivity to UV radiation, methyl methanesulfonate, or quinolone antibiotics. NHEJ deficiency had no effect on sensitivity to ionizing radiation in logarithmic- or early-stationary-phase cells but was required for ionizing radiation resistance in late stationary phase in 7H9 but not LB medium. In addition, NHEJ components were required for repair of I-SceI mediated chromosomal double-strand breaks (DSBs), and in the absence of HR, the NHEJ pathway rapidly mutates the chromosomal break site. The molecular outcomes of NHEJ-mediated chromosomal DSB repair involve predominantly single-nucleotide insertions at the break site, similar to previous findings using plasmid substrates. These findings demonstrate that prokaryotic NHEJ is specifically required for DSB repair in late stationary phase and can mediate mutagenic repair of homing endonuclease-generated chromosomal DSBs.

  12. Induction and repair of DNA double strand breaks: The increasing spectrum of non-homologous end joining pathways

    International Nuclear Information System (INIS)

    Mladenov, Emil; Iliakis, George

    2011-01-01

    A defining characteristic of damage induced in the DNA by ionizing radiation (IR) is its clustered character that leads to the formation of complex lesions challenging the cellular repair mechanisms. The most widely investigated such complex lesion is the DNA double strand break (DSB). DSBs undermine chromatin stability and challenge the repair machinery because an intact template strand is lacking to assist restoration of integrity and sequence in the DNA molecule. Therefore, cells have evolved a sophisticated machinery to detect DSBs and coordinate a response on the basis of inputs from various sources. A central function of cellular responses to DSBs is the coordination of DSB repair. Two conceptually different mechanisms can in principle remove DSBs from the genome of cells of higher eukaryotes. Homologous recombination repair (HRR) uses as template a homologous DNA molecule and is therefore error-free; it functions preferentially in the S and G2 phases. Non-homologous end joining (NHEJ), on the other hand, simply restores DNA integrity by joining the two ends, is error prone as sequence is only fortuitously preserved and active throughout the cell cycle. The basis of DSB repair pathway choice remains unknown, but cells of higher eukaryotes appear programmed to utilize preferentially NHEJ. Recent work suggests that when the canonical DNA-PK dependent pathway of NHEJ (D-NHEJ), becomes compromised an alternative NHEJ pathway and not HRR substitutes in a quasi-backup function (B-NHEJ). Here, we outline aspects of DSB induction by IR and review the mechanisms of their processing in cells of higher eukaryotes. We place particular emphasis on backup pathways of NHEJ and summarize their increasing significance in various cellular processes, as well as their potential contribution to carcinogenesis.

  13. Garcinol, a Histone Acetyltransferase Inhibitor, Radiosensitizes Cancer Cells by Inhibiting Non-Homologous End Joining

    Energy Technology Data Exchange (ETDEWEB)

    Oike, Takahiro [Division of Multistep Carcinogenesis, National Cancer Center Research Institute, Chuo-ku, Tokyo (Japan); Division of Genome Biology, National Cancer Center Research Institute, Chuo-ku, Tokyo (Japan); Department of Radiation Oncology, Gunma University Graduate School of Medicine, Maebashi, Gunma (Japan); Ogiwara, Hideaki [Division of Genome Biology, National Cancer Center Research Institute, Chuo-ku, Tokyo (Japan); Torikai, Kohta [Gunma University Heavy Ion Medical Center, Maebashi, Gunma (Japan); Nakano, Takashi [Department of Radiation Oncology, Gunma University Graduate School of Medicine, Maebashi, Gunma (Japan); Yokota, Jun [Division of Multistep Carcinogenesis, National Cancer Center Research Institute, Chuo-ku, Tokyo (Japan); Kohno, Takashi, E-mail: tkkohno@ncc.go.jp [Division of Genome Biology, National Cancer Center Research Institute, Chuo-ku, Tokyo (Japan)

    2012-11-01

    Purpose: Non-homologous end joining (NHEJ), a major pathway used to repair DNA double-strand breaks (DSBs) generated by ionizing radiation (IR), requires chromatin remodeling at DSB sites through the acetylation of histones by histone acetyltransferases (HATs). However, the effect of compounds with HAT inhibitory activities on the DNA damage response (DDR), including the NHEJ and cell cycle checkpoint, as well as on the radiosensitivity of cancer cells, remains largely unclear. Here, we investigated whether garcinol, a HAT inhibitor found in the rinds of Garcinia indica fruit (called mangosteens), has effects on DDR, and whether it can be used for radiosensitization. Methods and Materials: The following assays were used to examine the effect of garcinol on the inhibition of DSB repair, including the following: a conventional neutral comet assay; a cell-based assay recently developed by us, in which NHEJ repair of DSBs on chromosomal DNA was evaluated; the micrococcal nuclease sensitivity assay; and immunoblotting for autophosphorylation of DNA-dependent protein kinase catalytic subunit (DNA-PKcs). We assessed the effect of garcinol on the cell cycle checkpoint after IR treatment by analyzing the phosphorylation levels of checkpoint kinases CHK1 and CHK2 and histone H3, and by cell cycle profile analysis using flow cytometry. The radiosensitizing effect of garcinol was assessed by a clonogenic survival assay, whereas its effects on apoptosis and senescence were examined by annexin V and senescence-associated {beta}-galactosidase (SA-{beta}-Gal) staining, respectively. Results: We found that garcinol inhibits DSB repair, including NHEJ, without affecting cell cycle checkpoint. Garcinol radiosensitized A549 lung and HeLa cervical carcinoma cells with dose enhancement ratios (at 10% surviving fraction) of 1.6 and 1.5, respectively. Cellular senescence induced by IR was enhanced by garcinol. Conclusion: These results suggest that garcinol is a radiosensitizer that

  14. Human induced pluripotent cells resemble embryonic stem cells demonstrating enhanced levels of DNA repair and efficacy of nonhomologous end-joining

    Energy Technology Data Exchange (ETDEWEB)

    Fan Jinshui; Robert, Carine [Department of Radiation Oncology, University of Maryland School of Medicine, 655 West Baltimore Street, BRB 7-023A, Baltimore, MD 21201 (United States); Jang, Yoon-Young; Liu Hua; Sharkis, Saul; Baylin, Stephen Bruce [Johns Hopkins University School of Medicine, Department of Oncology, Baltimore, MD 21231-1000 (United States); Rassool, Feyruz Virgilia, E-mail: frassool@som.umaryland.edu [Department of Radiation Oncology, University of Maryland School of Medicine, 655 West Baltimore Street, BRB 7-023A, Baltimore, MD 21201 (United States)

    2011-08-01

    Highlights: {yields} iPSC and hESC demonstrate a similar cell cycle profile, with increased S phase cells and decreased G0/G1. {yields} iPSC and hESC increased ROS and decreased DSBs, compared with differentiated parental cells. {yields} iPSC and hESC demonstrate elevated DSB repair activity, including nonhomologous end-joining, compared with differentiated parental cells. {yields} iPSC however show a partial apoptotic response to DNA damage, compared to hESC. {yields} DNA damage responses may constitute important markers for the efficacy of iPSC reprogramming. - Abstract: To maintain the integrity of the organism, embryonic stem cells (ESC) need to maintain their genomic integrity in response to DNA damage. DNA double strand breaks (DSBs) are one of the most lethal forms of DNA damage and can have disastrous consequences if not repaired correctly, leading to cell death, genomic instability and cancer. How human ESC (hESC) maintain genomic integrity in response to agents that cause DSBs is relatively unclear. Adult somatic cells can be induced to 'dedifferentiate' into induced pluripotent stem cells (iPSC) and reprogram into cells of all three germ layers. Whether iPSC have reprogrammed the DNA damage response is a critical question in regenerative medicine. Here, we show that hESC demonstrate high levels of endogenous reactive oxygen species (ROS) which can contribute to DNA damage and may arise from high levels of metabolic activity. To potentially counter genomic instability caused by DNA damage, we find that hESC employ two strategies: First, these cells have enhanced levels of DNA repair proteins, including those involved in repair of DSBs, and they demonstrate elevated nonhomologous end-joining (NHEJ) activity and repair efficacy, one of the main pathways for repairing DSBs. Second, they are hypersensitive to DNA damaging agents, as evidenced by a high level of apoptosis upon irradiation. Importantly, iPSC, unlike the parent cells they are derived

  15. Human induced pluripotent cells resemble embryonic stem cells demonstrating enhanced levels of DNA repair and efficacy of nonhomologous end-joining

    International Nuclear Information System (INIS)

    Fan Jinshui; Robert, Carine; Jang, Yoon-Young; Liu Hua; Sharkis, Saul; Baylin, Stephen Bruce; Rassool, Feyruz Virgilia

    2011-01-01

    Highlights: → iPSC and hESC demonstrate a similar cell cycle profile, with increased S phase cells and decreased G0/G1. → iPSC and hESC increased ROS and decreased DSBs, compared with differentiated parental cells. → iPSC and hESC demonstrate elevated DSB repair activity, including nonhomologous end-joining, compared with differentiated parental cells. → iPSC however show a partial apoptotic response to DNA damage, compared to hESC. → DNA damage responses may constitute important markers for the efficacy of iPSC reprogramming. - Abstract: To maintain the integrity of the organism, embryonic stem cells (ESC) need to maintain their genomic integrity in response to DNA damage. DNA double strand breaks (DSBs) are one of the most lethal forms of DNA damage and can have disastrous consequences if not repaired correctly, leading to cell death, genomic instability and cancer. How human ESC (hESC) maintain genomic integrity in response to agents that cause DSBs is relatively unclear. Adult somatic cells can be induced to 'dedifferentiate' into induced pluripotent stem cells (iPSC) and reprogram into cells of all three germ layers. Whether iPSC have reprogrammed the DNA damage response is a critical question in regenerative medicine. Here, we show that hESC demonstrate high levels of endogenous reactive oxygen species (ROS) which can contribute to DNA damage and may arise from high levels of metabolic activity. To potentially counter genomic instability caused by DNA damage, we find that hESC employ two strategies: First, these cells have enhanced levels of DNA repair proteins, including those involved in repair of DSBs, and they demonstrate elevated nonhomologous end-joining (NHEJ) activity and repair efficacy, one of the main pathways for repairing DSBs. Second, they are hypersensitive to DNA damaging agents, as evidenced by a high level of apoptosis upon irradiation. Importantly, iPSC, unlike the parent cells they are derived from, mimic hESC in their ROS levels

  16. Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

    Energy Technology Data Exchange (ETDEWEB)

    Henrique Barreta, Marcos [Universidade Federal de Santa Catarina, Campus Universitario de Curitibanos, Curitibanos, SC (Brazil); Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Garziera Gasperin, Bernardo; Braga Rissi, Vitor; Cesaro, Matheus Pedrotti de [Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Ferreira, Rogerio [Centro de Educacao Superior do Oeste-Universidade do Estado de Santa Catarina, Chapeco, SC (Brazil); Oliveira, Joao Francisco de; Goncalves, Paulo Bayard Dias [Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Bordignon, Vilceu, E-mail: vilceu.bordignon@mcgill.ca [Department of Animal Science, McGill University, Ste-Anne-De-Bellevue, QC (Canada)

    2012-10-01

    This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.

  17. Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

    International Nuclear Information System (INIS)

    Henrique Barreta, Marcos; Garziera Gasperin, Bernardo; Braga Rissi, Vitor; Cesaro, Matheus Pedrotti de; Ferreira, Rogério; Oliveira, João Francisco de; Gonçalves, Paulo Bayard Dias; Bordignon, Vilceu

    2012-01-01

    This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.

  18. Alternative end-joining pathway(s): bricolage at DNA breaks.

    Science.gov (United States)

    Frit, Philippe; Barboule, Nadia; Yuan, Ying; Gomez, Dennis; Calsou, Patrick

    2014-05-01

    To cope with DNA double strand break (DSB) genotoxicity, cells have evolved two main repair pathways: homologous recombination which uses homologous DNA sequences as repair templates, and non-homologous Ku-dependent end-joining involving direct sealing of DSB ends by DNA ligase IV (Lig4). During the last two decades a third player most commonly named alternative end-joining (A-EJ) has emerged, which is defined as any Ku- or Lig4-independent end-joining process. A-EJ increasingly appears as a highly error-prone bricolage on DSBs and despite expanding exploration, it still escapes full characterization. In the present review, we discuss the mechanism and regulation of A-EJ as well as its biological relevance under physiological and pathological situations, with a particular emphasis on chromosomal instability and cancer. Whether or not it is a genuine DSB repair pathway, A-EJ is emerging as an important cellular process and understanding A-EJ will certainly be a major challenge for the coming years. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  19. DNA ligase C1 mediates the LigD-independent nonhomologous end-joining pathway of Mycobacterium smegmatis.

    Science.gov (United States)

    Bhattarai, Hitesh; Gupta, Richa; Glickman, Michael S

    2014-10-01

    Nonhomologous end joining (NHEJ) is a recently described bacterial DNA double-strand break (DSB) repair pathway that has been best characterized for mycobacteria. NHEJ can religate transformed linear plasmids, repair ionizing radiation (IR)-induced DSBs in nonreplicating cells, and seal I-SceI-induced chromosomal DSBs. The core components of the mycobacterial NHEJ machinery are the DNA end binding protein Ku and the polyfunctional DNA ligase LigD. LigD has three autonomous enzymatic modules: ATP-dependent DNA ligase (LIG), DNA/RNA polymerase (POL), and 3' phosphoesterase (PE). Although genetic ablation of ku or ligD abolishes NHEJ and sensitizes nonreplicating cells to ionizing radiation, selective ablation of the ligase activity of LigD in vivo only mildly impairs NHEJ of linearized plasmids, indicating that an additional DNA ligase can support NHEJ. Additionally, the in vivo role of the POL and PE domains in NHEJ is unclear. Here we define a LigD ligase-independent NHEJ pathway in Mycobacterium smegmatis that requires the ATP-dependent DNA ligase LigC1 and the POL domain of LigD. Mycobacterium tuberculosis LigC can also support this backup NHEJ pathway. We also demonstrate that, although dispensable for efficient plasmid NHEJ, the activities of the POL and PE domains are required for repair of IR-induced DSBs in nonreplicating cells. These findings define the genetic requirements for a LigD-independent NHEJ pathway in mycobacteria and demonstrate that all enzymatic functions of the LigD protein participate in NHEJ in vivo. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  20. Either non-homologous ends joining or homologous recombination is required to repair double-strand breaks in the genome of macrophage-internalized Mycobacterium tuberculosis.

    Science.gov (United States)

    Brzostek, Anna; Szulc, Izabela; Klink, Magdalena; Brzezinska, Marta; Sulowska, Zofia; Dziadek, Jaroslaw

    2014-01-01

    The intracellular pathogen Mycobacterium tuberculosis (Mtb) is constantly exposed to a multitude of hostile conditions and is confronted by a variety of potentially DNA-damaging assaults in vivo, primarily from host-generated antimicrobial toxic radicals. Exposure to reactive nitrogen species and/or reactive oxygen species causes different types of DNA damage, including oxidation, depurination, methylation and deamination, that can result in single- or double-strand breaks (DSBs). These breaks affect the integrity of the whole genome and, when left unrepaired, can lead to cell death. Here, we investigated the role of the DSB repair pathways, homologous recombination (HR) and non-homologous ends joining (NHEJ), in the survival of Mtb inside macrophages. To this end, we constructed Mtb strains defective for HR (ΔrecA), NHEJ [Δ(ku,ligD)], or both DSB repair systems [Δ(ku,ligD,recA)]. Experiments using these strains revealed that either HR or NHEJ is sufficient for the survival and propagation of tubercle bacilli inside macrophages. Inhibition of nitric oxide or superoxide anion production with L-NIL or apocynin, respectively, enabled the Δ(ku,ligD,recA) mutant strain lacking both systems to survive intracellularly. Complementation of the Δ(ku,ligD,recA) mutant with an intact recA or ku-ligD rescued the ability of Mtb to propagate inside macrophages.

  1. Either non-homologous ends joining or homologous recombination is required to repair double-strand breaks in the genome of macrophage-internalized Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Anna Brzostek

    Full Text Available The intracellular pathogen Mycobacterium tuberculosis (Mtb is constantly exposed to a multitude of hostile conditions and is confronted by a variety of potentially DNA-damaging assaults in vivo, primarily from host-generated antimicrobial toxic radicals. Exposure to reactive nitrogen species and/or reactive oxygen species causes different types of DNA damage, including oxidation, depurination, methylation and deamination, that can result in single- or double-strand breaks (DSBs. These breaks affect the integrity of the whole genome and, when left unrepaired, can lead to cell death. Here, we investigated the role of the DSB repair pathways, homologous recombination (HR and non-homologous ends joining (NHEJ, in the survival of Mtb inside macrophages. To this end, we constructed Mtb strains defective for HR (ΔrecA, NHEJ [Δ(ku,ligD], or both DSB repair systems [Δ(ku,ligD,recA]. Experiments using these strains revealed that either HR or NHEJ is sufficient for the survival and propagation of tubercle bacilli inside macrophages. Inhibition of nitric oxide or superoxide anion production with L-NIL or apocynin, respectively, enabled the Δ(ku,ligD,recA mutant strain lacking both systems to survive intracellularly. Complementation of the Δ(ku,ligD,recA mutant with an intact recA or ku-ligD rescued the ability of Mtb to propagate inside macrophages.

  2. Lung Basal Stem Cells Rapidly Repair DNA Damage Using the Error-Prone Nonhomologous End-Joining Pathway

    Science.gov (United States)

    Weeden, Clare E.; Chen, Yunshun; Ma, Stephen B.; Hu, Yifang; Ramm, Georg; Sutherland, Kate D.; Smyth, Gordon K.

    2017-01-01

    Lung squamous cell carcinoma (SqCC), the second most common subtype of lung cancer, is strongly associated with tobacco smoking and exhibits genomic instability. The cellular origins and molecular processes that contribute to SqCC formation are largely unexplored. Here we show that human basal stem cells (BSCs) isolated from heavy smokers proliferate extensively, whereas their alveolar progenitor cell counterparts have limited colony-forming capacity. We demonstrate that this difference arises in part because of the ability of BSCs to repair their DNA more efficiently than alveolar cells following ionizing radiation or chemical-induced DNA damage. Analysis of mice harbouring a mutation in the DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a key enzyme in DNA damage repair by nonhomologous end joining (NHEJ), indicated that BSCs preferentially repair their DNA by this error-prone process. Interestingly, polyploidy, a phenomenon associated with genetically unstable cells, was only observed in the human BSC subset. Expression signature analysis indicated that BSCs are the likely cells of origin of human SqCC and that high levels of NHEJ genes in SqCC are correlated with increasing genomic instability. Hence, our results favour a model in which heavy smoking promotes proliferation of BSCs, and their predilection for error-prone NHEJ could lead to the high mutagenic burden that culminates in SqCC. Targeting DNA repair processes may therefore have a role in the prevention and therapy of SqCC. PMID:28125611

  3. Molecular mechanism of protein assembly on DNA double-strand breaks in the non-homologous end-joining pathway

    International Nuclear Information System (INIS)

    Yano, Ken-ichi; Morotomi-Yano, Keiko; Adachi, Noritaka; Akiyama, Hidenori

    2009-01-01

    Non-homologous end-joining (NHEJ) is the major repair pathway for DNA double-strand breaks (DSBs) in mammalian species. Upon DSB induction, a living cell quickly activates the NHEJ pathway comprising of multiple molecular events. However, it has been difficult to analyze the initial phase of DSB responses in living cells, primarily due to technical limitations. Recent advances in real-time imaging and site-directed DSB induction using laser microbeam allow us to monitor the spatiotemporal dynamics of NHEJ factors in the immediate-early phase after DSB induction. These new approaches, together with the use of cell lines deficient in each essential NHEJ factor, provide novel mechanistic insights into DSB recognition and protein assembly on DSBs in the NHEJ pathway. In this review, we provide an overview of recent progresses in the imaging analyses of the NHEJ core factors. These studies strongly suggest that the NHEJ core factors are pre-assembled into a large complex on DSBs prior to the progression of the biochemical reactions in the NHEJ pathway. Instead of the traditional step-by-step assembly model from the static view of NHEJ, a novel model for dynamic protein assembly in the NHEJ pathway is proposed. This new model provides important mechanistic insights into the protein assembly at DSBs and the regulation of DSB repair. (author)

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

    Science.gov (United States)

    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.

  5. The Ku70/80 ring in Non-Homologous End-Joining

    DEFF Research Database (Denmark)

    Kragelund, Birthe Brandt; Weterings, Eric; Hartmann-Petersen, Rasmus

    2016-01-01

    /80 heterodimer is a key-player in the NHEJ pathway and binds to DNA termini with high affinity, where it helps to protect DNA ends from degradation and to recruit other NHEJ factors required for repair. The mechanism of Ku70/80 detachment from the DNA helix after completion of DNA repair is incompletely......Non-homologous end-joining (NHEJ) is an essential DNA double strand break repair pathway during all cell cycle stages. Deficiency in NHEJ factors can lead to accumulation of unrepaired DNA breaks or faulty DNA repair, which may ultimately result in cell death, senescence or carcinogenesis. The Ku70...... understood. Some data suggest that certain DNA repair factors are ubiquitylated and targeted for proteasomal degradation after repair. Recent studies suggest that Ku80 is conjugated to lysine48-linked ubiquitin chains by the Skp1-Cullin-F-box (SCF) complex and/or the RING finger protein 8 (RNF8) ubiquitin-protein...

  6. The Ku80 carboxy terminus stimulates joining and artemis-mediated processing of DNA ends

    DEFF Research Database (Denmark)

    Weterings, Eric; Verkaik, Nicole S; Keijzers, Guido

    2008-01-01

    Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA-PK(CS)) an......Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA......-PK(CS)) and the XRCC4/ligase IV complex. Activation of the DNA-PK(CS) serine/threonine kinase requires an interaction with Ku70/80 and is essential for NHEJ-mediated DSB repair. In contrast to previous models, we found that the carboxy terminus of Ku80 is not absolutely required for the recruitment and activation...... was phosphorylated to normal levels. This resulted in severely reduced levels of Artemis nuclease activity in vivo and in vitro. We therefore conclude that the Ku80 carboxy terminus is important to support DNA-PK(CS) autophosphorylation at specific sites, which facilitates DNA end processing by the Artemis...

  7. CRISPR/Cas-mediated knock-in via non-homologous end-joining in the crustacean Daphnia magna.

    Science.gov (United States)

    Kumagai, Hitoshi; Nakanishi, Takashi; Matsuura, Tomoaki; Kato, Yasuhiko; Watanabe, Hajime

    2017-01-01

    The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system (Cas) is widely used for mediating the knock-in of foreign DNA into the genomes of various organisms. Here, we report a process of CRISPR/Cas-mediated knock-in via non-homologous end joining by the direct injection of Cas9/gRNA ribonucleoproteins (RNPs) in the crustacean Daphnia magna, which is a model organism for studies on toxicology, ecology, and evolution. First, we confirmed the cleavage activity of Cas9 RNPs comprising purified Cas9 proteins and gRNAs in D. magna. We used a gRNA that targets exon 10 of the eyeless gene. Cas9 proteins were incubated with the gRNAs and the resulting Cas9 RNPs were injected into D. magna eggs, which led to a typical phenotype of the eyeless mutant, i.e., eye deformity. The somatic and heritable mutagenesis efficiencies were up to 96% and 40%, respectively. Second, we tested the CRISPR/Cas-mediated knock-in of a plasmid by the injection of Cas9 RNPs. The donor DNA plasmid harboring the fluorescent reporter gene was designed to contain the gRNA recognition site. The co-injection of Cas9 RNPs together with the donor DNAs resulted in generation of one founder animal that produced fluorescent progenies. This transgenic Daphnia had donor DNA at the targeted genomic site, which suggested the concurrent cleavage of the injected plasmid DNA and genomic DNA. Owing to its simplicity and ease of experimental design, we suggest that the CRISPR/Cas-mediated knock-in method represents a promising tool for studying functional genomics in D. magna.

  8. BCR/ABL downregulates DNA-PK(CS)-dependent and upregulates backup non-homologous end joining in leukemic cells.

    Science.gov (United States)

    Poplawski, Tomasz; Blasiak, Janusz

    2010-06-01

    Non-homologous end joining (NHEJ) and homologous recombination repair (HRR) are the main mechanisms involved in the processing of DNA double strand breaks (DSBs) in humans. We showed previously that the oncogenic tyrosine kinase BCR/ABL stimulated DSBs repair by HRR. To evaluate the role of BCR/ABL in DSBs repair by NHEJ we examined the ability of leukemic BCR/ABL-expressing cell line BV173 to repair DNA damage induced by two DNA topoisomerase II inhibitors: etoposide and sobuzoxane. DNA lesions induced by sobuzoxane are repaired by a NHEJ pathway which is dependent on the catalytic subunit of protein kinase dependent on DNA (DNA-PK(CS); D-NHEJ), whereas damage evoked by etoposide are repaired by two distinct NHEJ pathways, dependent on or independent of DNA-PK(CS) (backup NHEJ, B-NHEJ). Cells incubated with STI571, a highly specific inhibitor of BCR/ABL, displayed resistance to these agents associated with an accelerated kinetics of DSBs repair, as measured by the neutral comet assay and pulsed field gel electrophoresis. However, in a functional NHEJ assay, cells preincubated with STI571 repaired DSBs induced by a restriction enzyme with a lower efficacy than without the preincubation and addition of wortmannin, a specific inhibitor of DNA-PK(CS), did not change efficacy of the NHEJ reaction. We suggest that BCR/ABL switch on B-NHEJ which is more error-prone then D-NHEJ and in such manner contribute to the increase of the genomic instability of leukemic cells.

  9. Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration[W

    Science.gov (United States)

    Vaghchhipawala, Zarir E.; Vasudevan, Balaji; Lee, Seonghee; Morsy, Mustafa R.; Mysore, Kirankumar S.

    2012-01-01

    Agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (T-DNA) into the plant genome. Several plant factors that play a role in Agrobacterium attachment to plant cells and transport of T-DNA to the nucleus have been identified, but the T-DNA integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (NHEJ)–mediated double-strand DNA break (DSB) repair. Here, we report a negative role of X-RAY CROSS COMPLEMENTATION GROUP4 (XRCC4), one of the key proteins required for NHEJ, in Agrobacterium T-DNA integration. Downregulation of XRCC4 in Arabidopsis and Nicotiana benthamiana increased stable transformation due to increased T-DNA integration. Overexpression of XRCC4 in Arabidopsis decreased stable transformation due to decreased T-DNA integration. Interestingly, XRCC4 directly interacted with Agrobacterium protein VirE2 in a yeast two-hybrid system and in planta. VirE2-expressing Arabidopsis plants were more susceptible to the DNA damaging chemical bleomycin and showed increased stable transformation. We hypothesize that VirE2 titrates or excludes active XRCC4 protein available for DSB repair, thus delaying the closure of DSBs in the chromosome, providing greater opportunity for T-DNA to integrate. PMID:23064322

  10. AAV-Mediated Gene Targeting Is Significantly Enhanced by Transient Inhibition of Nonhomologous End Joining or the Proteasome In Vivo

    Science.gov (United States)

    Paulk, Nicole K.; Loza, Laura Marquez; Finegold, Milton J.

    2012-01-01

    Abstract Recombinant adeno-associated virus (rAAV) vectors have clear potential for use in gene targeting but low correction efficiencies remain the primary drawback. One approach to enhancing efficiency is a block of undesired repair pathways like nonhomologous end joining (NHEJ) to promote the use of homologous recombination. The natural product vanillin acts as a potent inhibitor of NHEJ by inhibiting DNA-dependent protein kinase (DNA-PK). Using a homology containing rAAV vector, we previously demonstrated in vivo gene repair frequencies of up to 0.1% in a model of liver disease hereditary tyrosinemia type I. To increase targeting frequencies, we administered vanillin in combination with rAAV. Gene targeting frequencies increased up to 10-fold over AAV alone, approaching 1%. Fah−/−Ku70−/− double knockout mice also had increased gene repair frequencies, genetically confirming the beneficial effects of blocking NHEJ. A second strategy, transient proteasomal inhibition, also increased gene-targeting frequencies but was not additive to NHEJ inhibition. This study establishes the benefit of transient NHEJ inhibition with vanillin, or proteasome blockage with bortezomib, for increasing hepatic gene targeting with rAAV. Functional metabolic correction of a clinically relevant disease model was demonstrated and provided evidence for the feasibility of gene targeting as a therapeutic strategy. PMID:22486314

  11. Non-homologous end joining dependency of {gamma}-irradiation-induced adaptive frameshift mutation formation in cell cycle-arrested yeast cells

    Energy Technology Data Exchange (ETDEWEB)

    Heidenreich, Erich [Institute of Cancer Research, Division of Molecular Genetics, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria)]. E-mail: erich.heidenreich@meduniwien.ac.at; Eisler, Herfried [Institute of Cancer Research, Division of Molecular Genetics, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria)

    2004-11-22

    There is a strong selective pressure favoring adaptive mutations which relieve proliferation-limiting adverse living conditions. Due to their importance for evolution and pathogenesis, we are interested in the mechanisms responsible for the formation of such adaptive, gain-of-fitness mutations in stationary-phase cells. During previous studies on the occurrence of spontaneous reversions of an auxotrophy-causing frameshift allele in the yeast Saccharomyces cerevisiae, we noticed that about 50% of the adaptive reversions depended on a functional non-homologous end joining (NHEJ) pathway of DNA double-strand break (DSB) repair. Here, we show that the occasional NHEJ component Pol4, which is the yeast ortholog of mammalian DNA polymerase lambda, is not required for adaptive mutagenesis. An artificially imposed excess of DSBs by {gamma}-irradiation resulted in a dramatic increase in the incidence of adaptive, cell cycle arrest-releasing frameshift reversions. By the use of DNA ligase IV-deficient strains we detected that the majority of the {gamma}-induced adaptive mutations were also dependent on a functional NHEJ pathway. This suggests that the same mutagenic NHEJ mechanism acts on spontaneously arising as well as on ionizing radiation-induced DSBs. Inaccuracy of the NHEJ repair pathway may extensively contribute to the incidence of frameshift mutations in resting (non-dividing) eukaryotic cells, and thus act as a driving force in tumor development.

  12. Non-homologous end joining dependency of γ-irradiation-induced adaptive frameshift mutation formation in cell cycle-arrested yeast cells

    International Nuclear Information System (INIS)

    Heidenreich, Erich; Eisler, Herfried

    2004-01-01

    There is a strong selective pressure favoring adaptive mutations which relieve proliferation-limiting adverse living conditions. Due to their importance for evolution and pathogenesis, we are interested in the mechanisms responsible for the formation of such adaptive, gain-of-fitness mutations in stationary-phase cells. During previous studies on the occurrence of spontaneous reversions of an auxotrophy-causing frameshift allele in the yeast Saccharomyces cerevisiae, we noticed that about 50% of the adaptive reversions depended on a functional non-homologous end joining (NHEJ) pathway of DNA double-strand break (DSB) repair. Here, we show that the occasional NHEJ component Pol4, which is the yeast ortholog of mammalian DNA polymerase lambda, is not required for adaptive mutagenesis. An artificially imposed excess of DSBs by γ-irradiation resulted in a dramatic increase in the incidence of adaptive, cell cycle arrest-releasing frameshift reversions. By the use of DNA ligase IV-deficient strains we detected that the majority of the γ-induced adaptive mutations were also dependent on a functional NHEJ pathway. This suggests that the same mutagenic NHEJ mechanism acts on spontaneously arising as well as on ionizing radiation-induced DSBs. Inaccuracy of the NHEJ repair pathway may extensively contribute to the incidence of frameshift mutations in resting (non-dividing) eukaryotic cells, and thus act as a driving force in tumor development

  13. SV40 Utilizes ATM Kinase Activity to Prevent Non-homologous End Joining of Broken Viral DNA Replication Products

    Science.gov (United States)

    Sowd, Gregory A.; Mody, Dviti; Eggold, Joshua; Cortez, David; Friedman, Katherine L.; Fanning, Ellen

    2014-01-01

    Simian virus 40 (SV40) and cellular DNA replication rely on host ATM and ATR DNA damage signaling kinases to facilitate DNA repair and elicit cell cycle arrest following DNA damage. During SV40 DNA replication, ATM kinase activity prevents concatemerization of the viral genome whereas ATR activity prevents accumulation of aberrant genomes resulting from breakage of a moving replication fork as it converges with a stalled fork. However, the repair pathways that ATM and ATR orchestrate to prevent these aberrant SV40 DNA replication products are unclear. Using two-dimensional gel electrophoresis and Southern blotting, we show that ATR kinase activity, but not DNA-PKcs kinase activity, facilitates some aspects of double strand break (DSB) repair when ATM is inhibited during SV40 infection. To clarify which repair factors associate with viral DNA replication centers, we examined the localization of DSB repair proteins in response to SV40 infection. Under normal conditions, viral replication centers exclusively associate with homology-directed repair (HDR) and do not colocalize with non-homologous end joining (NHEJ) factors. Following ATM inhibition, but not ATR inhibition, activated DNA-PKcs and KU70/80 accumulate at the viral replication centers while CtIP and BLM, proteins that initiate 5′ to 3′ end resection during HDR, become undetectable. Similar to what has been observed during cellular DSB repair in S phase, these data suggest that ATM kinase influences DSB repair pathway choice by preventing the recruitment of NHEJ factors to replicating viral DNA. These data may explain how ATM prevents concatemerization of the viral genome and promotes viral propagation. We suggest that inhibitors of DNA damage signaling and DNA repair could be used during infection to disrupt productive viral DNA replication. PMID:25474690

  14. SV40 utilizes ATM kinase activity to prevent non-homologous end joining of broken viral DNA replication products.

    Directory of Open Access Journals (Sweden)

    Gregory A Sowd

    2014-12-01

    Full Text Available Simian virus 40 (SV40 and cellular DNA replication rely on host ATM and ATR DNA damage signaling kinases to facilitate DNA repair and elicit cell cycle arrest following DNA damage. During SV40 DNA replication, ATM kinase activity prevents concatemerization of the viral genome whereas ATR activity prevents accumulation of aberrant genomes resulting from breakage of a moving replication fork as it converges with a stalled fork. However, the repair pathways that ATM and ATR orchestrate to prevent these aberrant SV40 DNA replication products are unclear. Using two-dimensional gel electrophoresis and Southern blotting, we show that ATR kinase activity, but not DNA-PK(cs kinase activity, facilitates some aspects of double strand break (DSB repair when ATM is inhibited during SV40 infection. To clarify which repair factors associate with viral DNA replication centers, we examined the localization of DSB repair proteins in response to SV40 infection. Under normal conditions, viral replication centers exclusively associate with homology-directed repair (HDR and do not colocalize with non-homologous end joining (NHEJ factors. Following ATM inhibition, but not ATR inhibition, activated DNA-PK(cs and KU70/80 accumulate at the viral replication centers while CtIP and BLM, proteins that initiate 5' to 3' end resection during HDR, become undetectable. Similar to what has been observed during cellular DSB repair in S phase, these data suggest that ATM kinase influences DSB repair pathway choice by preventing the recruitment of NHEJ factors to replicating viral DNA. These data may explain how ATM prevents concatemerization of the viral genome and promotes viral propagation. We suggest that inhibitors of DNA damage signaling and DNA repair could be used during infection to disrupt productive viral DNA replication.

  15. Unsolved mystery: the role of BRCA1 in DNA end-joining

    International Nuclear Information System (INIS)

    Saha, Janapriya; Davis, Anthony J.

    2016-01-01

    Heritable mutations in the tumor suppressor gene BRCA1 increase a woman's lifetime risk of developing breast and ovarian cancer. BRCA1's tumor suppressor function is directly linked to its myriad of functions in the cellular response to DNA double-strand breaks (DSBs). BRCA1 interacts with an extensive array of DNA damage responsive proteins and plays important roles in DSB repair, mediated by the homologous recombination pathway, and in the activation of cell cycle checkpoints. However, the role of BRCA1 in the other two DSB repair pathways, classical non-homologous end-joining (C-NHEJ) and alternative NHEJ (A-NHEJ), remains unclear. In this review, we will discuss the current literature on BRCA1's potential role(s) in modulating both C-NHEJ and A-NHEJ. We also present a model showing that BRCA1 contributes to genomic maintenance by promoting precise DNA repair across all cell cycle phases via the direct modulation of DNA end-joining

  16. Human DNA ligase III bridges two DNA ends to promote specific intermolecular DNA end joining

    Science.gov (United States)

    Kukshal, Vandna; Kim, In-Kwon; Hura, Gregory L.; Tomkinson, Alan E.; Tainer, John A.; Ellenberger, Tom

    2015-01-01

    Mammalian DNA ligase III (LigIII) functions in both nuclear and mitochondrial DNA metabolism. In the nucleus, LigIII has functional redundancy with DNA ligase I whereas LigIII is the only mitochondrial DNA ligase and is essential for the survival of cells dependent upon oxidative respiration. The unique LigIII zinc finger (ZnF) domain is not required for catalytic activity but senses DNA strand breaks and stimulates intermolecular ligation of two DNAs by an unknown mechanism. Consistent with this activity, LigIII acts in an alternative pathway of DNA double strand break repair that buttresses canonical non-homologous end joining (NHEJ) and is manifest in NHEJ-defective cancer cells, but how LigIII acts in joining intermolecular DNA ends versus nick ligation is unclear. To investigate how LigIII efficiently joins two DNAs, we developed a real-time, fluorescence-based assay of DNA bridging suitable for high-throughput screening. On a nicked duplex DNA substrate, the results reveal binding competition between the ZnF and the oligonucleotide/oligosaccharide-binding domain, one of three domains constituting the LigIII catalytic core. In contrast, these domains collaborate and are essential for formation of a DNA-bridging intermediate by adenylated LigIII that positions a pair of blunt-ended duplex DNAs for efficient and specific intermolecular ligation. PMID:26130724

  17. Non-homologous end joining pathway is the major route of protection against 4β-hydroxywithanolide E-induced DNA damage in MCF-7 cells.

    Science.gov (United States)

    You, B-J; Wu, Y-C; Lee, C-L; Lee, H-Z

    2014-03-01

    4β-Hydroxywithanolide E is a bioactive withanolide extracted from Physalis peruviana. 4β-Hydroxywithanolide E caused reactive oxygen species production and cell apoptosis in human breast cancer MCF-7 cells. We further found that 4β-hydroxywithanolide E induced DNA damage and regulated the DNA damage signaling in MCF-7 cells. The DNA damage sensors and repair proteins act promptly to remove DNA lesions by 4β-hydroxywithanolide E. The ataxia-telangiectasia mutated protein (ATM)-dependent DNA damage signaling pathway is involved in 4β-hydroxywithanolide E-induced apoptosis of MCF-7 cells. Non-homologous end joining pathway, but not homologous recombination, is the major route of protection of MCF-7 cells against 4β-hydroxywithanolide E-induced DNA damage. 4β-Hydroxywithanolide E had no significant impact on the base excision repair pathway. In this study, we examined the 4β-hydroxywithanolide E-induced DNA damage as a research tool in project investigating the DNA repair signaling in breast cancer cells. We also suggest that 4β-hydroxywithanolide E assert its anti-tumor activity in carcinogenic progression and develop into a dietary chemopreventive agent. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. The Ku80 carboxy terminus stimulates joining and artemis-mediated processing of DNA ends.

    Science.gov (United States)

    Weterings, Eric; Verkaik, Nicole S; Keijzers, Guido; Florea, Bogdan I; Wang, Shih-Ya; Ortega, Laura G; Uematsu, Naoya; Chen, David J; van Gent, Dik C

    2009-03-01

    Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA-PK(CS)) and the XRCC4/ligase IV complex. Activation of the DNA-PK(CS) serine/threonine kinase requires an interaction with Ku70/80 and is essential for NHEJ-mediated DSB repair. In contrast to previous models, we found that the carboxy terminus of Ku80 is not absolutely required for the recruitment and activation of DNA-PK(CS) at DSBs, although cells that harbored a carboxy-terminal deletion in the Ku80 gene were sensitive to ionizing radiation and showed reduced end-joining capacity. More detailed analysis of this repair defect showed that DNA-PK(CS) autophosphorylation at Thr2647 was diminished, while Ser2056 was phosphorylated to normal levels. This resulted in severely reduced levels of Artemis nuclease activity in vivo and in vitro. We therefore conclude that the Ku80 carboxy terminus is important to support DNA-PK(CS) autophosphorylation at specific sites, which facilitates DNA end processing by the Artemis endonuclease and the subsequent joining reaction.

  19. Identification of auxotrophic mutants of the yeast Kluyveromyces marxianus by non-homologous end joining-mediated integrative transformation with genes from Saccharomyces cerevisiae.

    Science.gov (United States)

    Yarimizu, Tohru; Nonklang, Sanom; Nakamura, Junpei; Tokuda, Shuya; Nakagawa, Takaaki; Lorreungsil, Sasithorn; Sutthikhumpha, Surasit; Pukahuta, Charida; Kitagawa, Takao; Nakamura, Mikiko; Cha-Aim, Kamonchai; Limtong, Savitree; Hoshida, Hisashi; Akada, Rinji

    2013-12-01

    The isolation and application of auxotrophic mutants for gene manipulations, such as genetic transformation, mating selection and tetrad analysis, form the basis of yeast genetics. For the development of these genetic methods in the thermotolerant fermentative yeast Kluyveromyces marxianus, we isolated a series of auxotrophic mutants with defects in amino acid or nucleic acid metabolism. To identify the mutated genes, linear DNA fragments of nutrient biosynthetic pathway genes were amplified from Saccharomyces cerevisiae chromosomal DNA and used to directly transform the K. marxianus auxotrophic mutants by random integration into chromosomes through non-homologous end joining (NHEJ). The appearance of transformant colonies indicated that the specific S. cerevisiae gene complemented the K. marxianus mutant. Using this interspecific complementation approach with linear PCR-amplified DNA, we identified auxotrophic mutations of ADE2, ADE5,7, ADE6, HIS2, HIS3, HIS4, HIS5, HIS6, HIS7, LYS1, LYS2, LYS4, LYS9, LEU1, LEU2, MET2, MET6, MET17, TRP3, TRP4 and TRP5 without the labour-intensive requirement of plasmid construction. Mating, sporulation and tetrad analysis techniques for K. marxianus were also established. With the identified auxotrophic mutant strains and S. cerevisiae genes as selective markers, NHEJ-mediated integrative transformation with PCR-amplified DNA is an attractive system for facilitating genetic analyses in the yeast K. marxianus. Copyright © 2013 John Wiley & Sons, Ltd.

  20. Cadmium delays non-homologous end joining (NHEJ) repair via inhibition of DNA-PKcs phosphorylation and downregulation of XRCC4 and Ligase IV

    Energy Technology Data Exchange (ETDEWEB)

    Li, Weiwei; Gu, Xueyan; Zhang, Xiaoning; Kong, Jinxin [Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou 730000 (China); Ding, Nan [Gansu Key laboratory of Space Radiobiology, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Qi, Yongmei; Zhang, Yingmei [Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou 730000 (China); Wang, Jufang [Gansu Key laboratory of Space Radiobiology, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Huang, Dejun, E-mail: huangdj@lzu.edu.cn [Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou 730000 (China)

    2015-09-15

    Highlights: • Cadmium (Cd) exposure delayed the repair of DNA damage induced by X-ray. • Cd exposure altered the phosphorylation of DNA-PKcs on Thr-2609 and Ser-2056 sites. • Cd impaired the formation of XRCC4 and Ligase IV foci, and down-regulated their protein expression. • Zinc mitigated the effects of Cd on DDR by regulating pDNA-PKcs (Thr-2609), XRCC4 and Ligase IV. - Abstract: Although studies have shown that cadmium (Cd) interfered with DNA damage repair (DDR), whether Cd could affect non-homologous end joining (NHEJ) repair remains elusive. To further understand the effect of Cd on DDR, we used X-ray irradiation of Hela cells as an in vitro model system, along with γH2AX and 53BP1 as markers for DNA damage. Results showed that X-ray significantly increased γH2AX and 53BP1 foci in Hela cells (p < 0.01), all of which are characteristic of accrued DNA damage. The number of foci declined rapidly over time (1–8 h postirradiation), indicating an initiation of NHEJ process. However, the disappearance of γH2AX and 53BP1 foci was remarkably slowed by Cd pretreatment (p < 0.01), suggesting that Cd reduced the efficiency of NHEJ. To further elucidate the mechanisms of Cd toxicity, several markers of NHEJ pathway including Ku70, DNA-PKcs, XRCC4 and Ligase IV were examined. Our data showed that Cd altered the phosphorylation of DNA-PKcs, and reduced the expression of both XRCC4 and Ligase IV in irradiated cells. These observations are indicative of the impairment of NHEJ-dependent DNA repair pathways. In addition, zinc (Zn) mitigated the effects of Cd on NHEJ, suggesting that the Cd-induced NHEJ alteration may partly result from the displacement of Zn or from an interference with the normal function of Zn-containing proteins by Cd. Our findings provide a new insight into the toxicity of Cd on NHEJ repair and its underlying mechanisms in human cells.

  1. Lipofection and nucleofection of substrate plasmid can generate widely different readings of DNA end-joining efficiency in different cell lines.

    Science.gov (United States)

    Magin, Simon; Saha, Janapriya; Wang, Minli; Mladenova, Veronika; Coym, Nadine; Iliakis, George

    2013-02-01

    In vivo plasmid end-joining assays are valuable tools for dissecting important qualitative and quantitative aspects of non-homologous end-joining (NHEJ)--a key mechanism for the repair of DNA double-strand breaks (DSBs) in higher eukaryotes. They enable the use of defined DNA ends as substrates for end-joining and the analysis by sequencing of the resulting junctions to identify the repair pathways engaged. Yet, plasmid assays have generated divergent results of end-joining capacity in the same DSB repair mutants when used under different conditions, which implies contributions from undefined and therefore uncontrolled parameters. To help standardize these assays, we searched for parameters underpinning these variations and identified transfection method as an important determinant. Here, we compare a lipid-based transfection method, lipofection, with an electroporation method, nucleofection, and find large, unanticipated and cell line-dependent differences in percent end-joining without recognizable trends. For example, in rodent cells, transfection using lipofection gives nearly WT end-joining in DNA-PKcs mutants and only mildly inhibited end-joining in Lig4 and Ku mutants. In contrast, transfection using nucleofection shows marked end-joining inhibition in all NHEJ mutants tested as compared to the WT. In human HCT116 cells, end-joining after nucleofection is strongly suppressed even in the WT and the differences to the mutants are small. After lipofection, in contrast, end-joining is high in WT cells and markedly suppressed in the mutants. We conclude that better understanding and control of the physicochemical/biological and analytical parameters underpinning these differences will be required to generate with plasmid assays results with quantitative power comparable to that of well-established methods of DSB analysis such as pulsed-field gel electrophoresis or γ-H2AX foci scoring. Until then, caution is needed in the interpretation of the results obtained

  2. DNA double-strand break response factors influence end-joining features of IgH class switch and general translocation junctions.

    Science.gov (United States)

    Panchakshari, Rohit A; Zhang, Xuefei; Kumar, Vipul; Du, Zhou; Wei, Pei-Chi; Kao, Jennifer; Dong, Junchao; Alt, Frederick W

    2018-01-23

    Ig heavy chain (IgH) class switch recombination (CSR) in B lymphocytes switches IgH constant regions to change antibody functions. CSR is initiated by DNA double-strand breaks (DSBs) within a donor IgH switch (S) region and a downstream acceptor S region. CSR is completed by fusing donor and acceptor S region DSB ends by classical nonhomologous end-joining (C-NHEJ) and, in its absence, by alternative end-joining that is more biased to use longer junctional microhomologies (MHs). Deficiency for DSB response (DSBR) factors, including ataxia telangiectasia-mutated (ATM) and 53BP1, variably impair CSR end-joining, with 53BP1 deficiency having the greatest impact. However, studies of potential impact of DSBR factor deficiencies on MH-mediated CSR end-joining have been technically limited. We now use a robust DSB joining assay to elucidate impacts of deficiencies for DSBR factors on CSR and chromosomal translocation junctions in primary mouse B cells and CH12F3 B-lymphoma cells. Compared with wild-type, CSR and c-myc to S region translocation junctions in the absence of 53BP1, and, to a lesser extent, other DSBR factors, have increased MH utilization; indeed, 53BP1-deficient MH profiles resemble those associated with C-NHEJ deficiency. However, translocation junctions between c-myc DSB and general DSBs genome-wide are not MH-biased in ATM-deficient versus wild-type CH12F3 cells and are less biased in 53BP1- and C-NHEJ-deficient cells than CSR junctions or c-myc to S region translocation junctions. We discuss potential roles of DSBR factors in suppressing increased MH-mediated DSB end-joining and features of S regions that may render their DSBs prone to MH-biased end-joining in the absence of DSBR factors.

  3. Redundant function of DNA ligase 1 and 3 in alternative end-joining during immunoglobulin class switch recombination.

    Science.gov (United States)

    Masani, Shahnaz; Han, Li; Meek, Katheryn; Yu, Kefei

    2016-02-02

    Nonhomologous end-joining (NHEJ) is the major DNA double-strand break (DSB) repair pathway in mammals and resolves the DSBs generated during both V(D)J recombination in developing lymphocytes and class switch recombination (CSR) in antigen-stimulated B cells. In contrast to the absolute requirement for NHEJ to resolve DSBs associated with V(D)J recombination, DSBs associated with CSR can be resolved in NHEJ-deficient cells (albeit at a reduced level) by a poorly defined alternative end-joining (A-EJ) pathway. Deletion of DNA ligase IV (Lig4), a core component of the NHEJ pathway, reduces CSR efficiency in a mouse B-cell line capable of robust cytokine-stimulated CSR in cell culture. Here, we report that CSR levels are not further reduced by deletion of either of the two remaining DNA ligases (Lig1 and nuclear Lig3) in Lig4(-/-) cells. We conclude that in the absence of Lig4, Lig1, and Lig3 function in a redundant manner in resolving switch region DSBs during CSR.

  4. Relative contribution of homologous recombination and non-homologous end-joining to DNA double-strand break repair after oxidative stress in Saccharomyces cerevisiae.

    Science.gov (United States)

    Letavayová, Lucia; Marková, Eva; Hermanská, Katarína; Vlcková, Viera; Vlasáková, Danusa; Chovanec, Miroslav; Brozmanová, Jela

    2006-05-10

    Oxidative damage to DNA seems to be an important factor in developing many human diseases including cancer. It involves base and sugar damage, base-free sites, DNA-protein cross-links and DNA single-strand (SSB) and double-strand (DSB) breaks. Oxidative DSB can be formed in various ways such as their direct induction by the drug or their generation either through attempted and aborted repair of primary DNA lesions or through DNA replication-dependent conversion of SSB. In general, two main pathways are responsible for repairing DSB, homologous recombination (HR) and non-homologous end-joining (NHEJ), with both of them being potential candidates for the repair of oxidative DSB. We have examined relative contribution of HR and NHEJ to cellular response after oxidative stress in Saccharomyces cerevisiae. Therefore, cell survival, mutagenesis and DSB induction and repair in the rad52, yku70 and rad52 yku70 mutants after hydrogen peroxide (H(2)O(2)), menadione (MD) or bleomycin (BLM) exposure were compared to those obtained for the corresponding wild type. We show that MD exposure does not lead to observable DSB induction in yeast, suggesting that the toxic effects of this agent are mediated by other types of DNA damage. Although H(2)O(2) treatment generates some DSB, their yield is relatively low and hence DSB may only partially be responsible for toxicity of H(2)O(2), particularly at high doses of the agent. On the other hand, the basis of the BLM toxicity resides primarily in DSB induction. Both HR and NHEJ act on BLM-induced DSB, although their relative participation in the process is not equal. Based on our results we suggest that the complexity and/or the quality of the BLM-induced DSB might represent an obstacle for the NHEJ pathway.

  5. An efficient system for deletion of large DNA fragments in Escherichia coli via introduction of both Cas9 and the non-homologous end joining system from Mycobacterium smegmatis.

    Science.gov (United States)

    Zheng, Xuan; Li, Shi-Yuan; Zhao, Guo-Ping; Wang, Jin

    2017-04-15

    Accompanied with the internal non-homologous end joining (NHEJ) system, Cas9 can be used to easily inactivate a gene or delete a fragment through introduction of DNA double-stranded breaks (DSBs) in eukaryotic cells. While in most prokaryotes (e.g. Escherichia coli), due to the lack of NHEJ, homologous recombination (HR) is required for repair of DSBs, which is less convenient. Here, a markerless system was developed for rapid gene inactivation or fragment deletion in E. coli via introduction of both Cas9 and a bacterial NHEJ system. Three bacterial NHEJ systems, i.e. Mycobacterium smegmatis (Msm), Mycobacterium tuberculosis (Mtb) and Bacillus subtilis (Bs), were tested in E. coli, and the MsmNHEJ system showed the best efficiency. With the employment of Cas9 and MsmNHEJ, we efficiently mutated lacZ gene, deleted glnALG operon and two large DNA fragments (67 kb and 123 kb) in E. coli, respectively. Moreover, the system was further designed to allow for continuous inactivation of genes or deletion of DNA fragments in E. coli. We envision this system can be extended to other bacteria, especially those with low HR efficiency. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Non-homologous end joining is the responsible pathway for the repair of fludarabine-induced DNA double strand breaks in mammalian cells

    International Nuclear Information System (INIS)

    Campos-Nebel, Marcelo de; Larripa, Irene; Gonzalez-Cid, Marcela

    2008-01-01

    Fludarabine (FLU), an analogue of adenosine, interferes with DNA synthesis and inhibits the chain elongation leading to replication arrest and DNA double strand break (DSB) formation. Mammalian cells use two main pathways of DSB repair to maintain genomic stability: homologous recombination (HR) and non-homologous end joining (NHEJ). The aim of the present work was to evaluate the repair pathways employed in the restoration of DSB formed following replication arrest induced by FLU in mammalian cells. Replication inhibition was induced in human lymphocytes and fibroblasts by FLU. DSB occurred in a dose-dependent manner on early/middle S-phase cells, as detected by γH2AX foci formation. To test whether conservative HR participates in FLU-induced DSB repair, we measured the kinetics of Rad51 nuclear foci formation in human fibroblasts. There was no significant induction of Rad51 foci after FLU treatment. To further confirm these results, we analyzed the frequency of sister chromatid exchanges (SCE) in both human cells. We did not find increased frequencies of SCE after FLU treatment. To assess the participation of NHEJ pathway in the repair of FLU-induced damage, we used two chemical inhibitors of the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), vanillin and wortmannin. Human fibroblasts pretreated with DNA-PKcs inhibitors showed increased levels of chromosome breakages and became more sensitive to cell death. An active role of NHEJ pathway was also suggested from the analysis of Chinese hamster cell lines. XR-C1 (DNA-PKcs-deficient) and XR-V15B (Ku80-deficient) cells showed hypersensitivity to FLU as evidenced by the increased frequency of chromosome aberrations, decreased mitotic index and impaired survival rates. In contrast, CL-V4B (Rad51C-deficient) and V-C8 (Brca2-deficient) cell lines displayed a FLU-resistant phenotype. Together, our results suggest a major role for NHEJ repair in the preservation of genome integrity against FLU-induced DSB

  7. Non-homologous end joining is the responsible pathway for the repair of fludarabine-induced DNA double strand breaks in mammalian cells

    Energy Technology Data Exchange (ETDEWEB)

    Campos-Nebel, Marcelo de [Departamento de Genetica, Instituto de Investigaciones Hematologicas Mariano R. Castex, Academia Nacional de Medicina, Buenos Aires (Argentina)], E-mail: mnebel@hematologia.anm.edu.ar; Larripa, Irene; Gonzalez-Cid, Marcela [Departamento de Genetica, Instituto de Investigaciones Hematologicas Mariano R. Castex, Academia Nacional de Medicina, Buenos Aires (Argentina)

    2008-11-10

    Fludarabine (FLU), an analogue of adenosine, interferes with DNA synthesis and inhibits the chain elongation leading to replication arrest and DNA double strand break (DSB) formation. Mammalian cells use two main pathways of DSB repair to maintain genomic stability: homologous recombination (HR) and non-homologous end joining (NHEJ). The aim of the present work was to evaluate the repair pathways employed in the restoration of DSB formed following replication arrest induced by FLU in mammalian cells. Replication inhibition was induced in human lymphocytes and fibroblasts by FLU. DSB occurred in a dose-dependent manner on early/middle S-phase cells, as detected by {gamma}H2AX foci formation. To test whether conservative HR participates in FLU-induced DSB repair, we measured the kinetics of Rad51 nuclear foci formation in human fibroblasts. There was no significant induction of Rad51 foci after FLU treatment. To further confirm these results, we analyzed the frequency of sister chromatid exchanges (SCE) in both human cells. We did not find increased frequencies of SCE after FLU treatment. To assess the participation of NHEJ pathway in the repair of FLU-induced damage, we used two chemical inhibitors of the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), vanillin and wortmannin. Human fibroblasts pretreated with DNA-PKcs inhibitors showed increased levels of chromosome breakages and became more sensitive to cell death. An active role of NHEJ pathway was also suggested from the analysis of Chinese hamster cell lines. XR-C1 (DNA-PKcs-deficient) and XR-V15B (Ku80-deficient) cells showed hypersensitivity to FLU as evidenced by the increased frequency of chromosome aberrations, decreased mitotic index and impaired survival rates. In contrast, CL-V4B (Rad51C-deficient) and V-C8 (Brca2-deficient) cell lines displayed a FLU-resistant phenotype. Together, our results suggest a major role for NHEJ repair in the preservation of genome integrity against FLU

  8. KIN17, XPC, DNA-PKCS and XRCC4 proteins in the cellular response to DNA damages. Relations between nucleotide excision repair and non-homologous end joining in a human syn-genic model

    International Nuclear Information System (INIS)

    Despras, Emmanuelle

    2006-01-01

    The response to genotoxic stress involves many cellular factors in a complex network of mechanisms that aim to preserve the genetic integrity of the organism. These mechanisms enclose the detection and repair of DNA lesions, the regulation of transcription and replication and, eventually, the setting of cell death. Among the nuclear proteins involved in this response, kin17 proteins are zinc-finger proteins conserved through evolution and activated by ultraviolet (UV) or ionizing radiations (IR). We showed that human kin17 protein (HSAkin17) is found in the cell under a soluble form and a form tightly anchored to nuclear structures. A fraction of HSAkin17 protein is directly associated with chromatin. HSAkin17 protein is recruited to nuclear structures 24 hours after treatment with various agents inducing DNA double-strand breaks (DSB) and/or replication forks blockage. Moreover, the reduction of total HSAkin17 protein level sensitizes RKO cells to IR. We also present evidence for the involvement of HSAkin17 protein in DNA replication. This hypothesis was further confirmed by the biochemical demonstration of its belonging to the replication complex. HSAkin17 protein could link DNA replication and DNA repair, a defect in the HSAkin17 pathway leading to an increased radiosensitivity. In a second part, we studied the interactions between two DNA repair mechanisms: nucleotide excision repair (NER) and non-homologous end joining (NHEJ). NER repairs a wide variety of lesions inducing a distortion of the DNA double helix including UV-induced pyrimidine dimers. NHEJ allows the repair of DSB by direct joining of DNA ends. We used a syn-genic model for DNA repair defects based on RNA interference developed in the laboratory. Epstein-Barr virus-derived vectors (pEBV) allow long-term expression of siRNA and specific extinction of the targeted gene. The reduction of the expression of genes involved in NER (XPA and XPC) or NHEJ (DNA-PKcs and XRCC4) leads to the expected

  9. Highly precise and developmentally programmed genome assembly in Paramecium requires ligase IV-dependent end joining.

    Directory of Open Access Journals (Sweden)

    Aurélie Kapusta

    2011-04-01

    Full Text Available During the sexual cycle of the ciliate Paramecium, assembly of the somatic genome includes the precise excision of tens of thousands of short, non-coding germline sequences (Internal Eliminated Sequences or IESs, each one flanked by two TA dinucleotides. It has been reported previously that these genome rearrangements are initiated by the introduction of developmentally programmed DNA double-strand breaks (DSBs, which depend on the domesticated transposase PiggyMac. These DSBs all exhibit a characteristic geometry, with 4-base 5' overhangs centered on the conserved TA, and may readily align and undergo ligation with minimal processing. However, the molecular steps and actors involved in the final and precise assembly of somatic genes have remained unknown. We demonstrate here that Ligase IV and Xrcc4p, core components of the non-homologous end-joining pathway (NHEJ, are required both for the repair of IES excision sites and for the circularization of excised IESs. The transcription of LIG4 and XRCC4 is induced early during the sexual cycle and a Lig4p-GFP fusion protein accumulates in the developing somatic nucleus by the time IES excision takes place. RNAi-mediated silencing of either gene results in the persistence of free broken DNA ends, apparently protected against extensive resection. At the nucleotide level, controlled removal of the 5'-terminal nucleotide occurs normally in LIG4-silenced cells, while nucleotide addition to the 3' ends of the breaks is blocked, together with the final joining step, indicative of a coupling between NHEJ polymerase and ligase activities. Taken together, our data indicate that IES excision is a "cut-and-close" mechanism, which involves the introduction of initiating double-strand cleavages at both ends of each IES, followed by DSB repair via highly precise end joining. This work broadens our current view on how the cellular NHEJ pathway has cooperated with domesticated transposases for the emergence of new

  10. Highly precise and developmentally programmed genome assembly in Paramecium requires ligase IV-dependent end joining.

    Science.gov (United States)

    Kapusta, Aurélie; Matsuda, Atsushi; Marmignon, Antoine; Ku, Michael; Silve, Aude; Meyer, Eric; Forney, James D; Malinsky, Sophie; Bétermier, Mireille

    2011-04-01

    During the sexual cycle of the ciliate Paramecium, assembly of the somatic genome includes the precise excision of tens of thousands of short, non-coding germline sequences (Internal Eliminated Sequences or IESs), each one flanked by two TA dinucleotides. It has been reported previously that these genome rearrangements are initiated by the introduction of developmentally programmed DNA double-strand breaks (DSBs), which depend on the domesticated transposase PiggyMac. These DSBs all exhibit a characteristic geometry, with 4-base 5' overhangs centered on the conserved TA, and may readily align and undergo ligation with minimal processing. However, the molecular steps and actors involved in the final and precise assembly of somatic genes have remained unknown. We demonstrate here that Ligase IV and Xrcc4p, core components of the non-homologous end-joining pathway (NHEJ), are required both for the repair of IES excision sites and for the circularization of excised IESs. The transcription of LIG4 and XRCC4 is induced early during the sexual cycle and a Lig4p-GFP fusion protein accumulates in the developing somatic nucleus by the time IES excision takes place. RNAi-mediated silencing of either gene results in the persistence of free broken DNA ends, apparently protected against extensive resection. At the nucleotide level, controlled removal of the 5'-terminal nucleotide occurs normally in LIG4-silenced cells, while nucleotide addition to the 3' ends of the breaks is blocked, together with the final joining step, indicative of a coupling between NHEJ polymerase and ligase activities. Taken together, our data indicate that IES excision is a "cut-and-close" mechanism, which involves the introduction of initiating double-strand cleavages at both ends of each IES, followed by DSB repair via highly precise end joining. This work broadens our current view on how the cellular NHEJ pathway has cooperated with domesticated transposases for the emergence of new mechanisms

  11. Sirt6 Promotes DNA End Joining in iPSCs Derived from Old Mice

    Directory of Open Access Journals (Sweden)

    Wen Chen

    2017-03-01

    Full Text Available Induced pluripotent stem cells (iPSCs have great potential for treating age-related diseases, but the genome integrity of iPSCs is critically important. Here, we demonstrate that non-homologous end joining (NHEJ, rather than homologous recombination (HR, is less efficient in iPSCs from old mice than young mice. We further find that Sirt6 is downregulated in iPSCs from old mice. Sirt6 directly binds to Ku80 and facilitates the Ku80/DNA-PKcs interaction, thus promoting DNA-PKcs phosphorylation at residue S2056, leading to efficient NHEJ. Rescue experiments show that introducing a combination of Sirt6 and the Yamanaka factors during reprogramming significantly promotes DNA double-strand break (DSB repair by activating NHEJ in iPSCs derived from old mice. Thus, our study suggests a strategy to improve the quality of iPSCs derived from old donors by activating NHEJ and stabilizing the genome.

  12. Alternative end-joining catalyzes robust IgH locus deletions and translocations in the combined absence of ligase 4 and Ku70.

    Science.gov (United States)

    Boboila, Cristian; Jankovic, Mila; Yan, Catherine T; Wang, Jing H; Wesemann, Duane R; Zhang, Tingting; Fazeli, Alex; Feldman, Lauren; Nussenzweig, Andre; Nussenzweig, Michel; Alt, Frederick W

    2010-02-16

    Class switch recombination (CSR) in B lymphocytes is initiated by introduction of multiple DNA double-strand breaks (DSBs) into switch (S) regions that flank immunoglobulin heavy chain (IgH) constant region exons. CSR is completed by joining a DSB in the donor S mu to a DSB in a downstream acceptor S region (e.g., S gamma1) by end-joining. In normal cells, many CSR junctions are mediated by classical nonhomologous end-joining (C-NHEJ), which employs the Ku70/80 complex for DSB recognition and XRCC4/DNA ligase 4 for ligation. Alternative end-joining (A-EJ) mediates CSR, at reduced levels, in the absence of C-NHEJ, even in combined absence of Ku70 and ligase 4, demonstrating an A-EJ pathway totally distinct from C-NHEJ. Multiple DSBs are introduced into S mu during CSR, with some being rejoined or joined to each other to generate internal switch deletions (ISDs). In addition, S-region DSBs can be joined to other chromosomes to generate translocations, the level of which is increased by absence of a single C-NHEJ component (e.g., XRCC4). We asked whether ISD and S-region translocations occur in the complete absence of C-NHEJ (e.g., in Ku70/ligase 4 double-deficient B cells). We found, unexpectedly, that B-cell activation for CSR generates substantial ISD in both S mu and S gamma1 and that ISD in both is greatly increased by the absence of C-NHEJ. IgH chromosomal translocations to the c-myc oncogene also are augmented in the combined absence of Ku70 and ligase 4. We discuss the implications of these findings for A-EJ in normal and abnormal DSB repair.

  13. End-joining inhibition at telomeres requires the translocase and polySUMO-dependent ubiquitin ligase Uls1.

    Science.gov (United States)

    Lescasse, Rachel; Pobiega, Sabrina; Callebaut, Isabelle; Marcand, Stéphane

    2013-03-20

    In eukaryotes, permanent inhibition of the non-homologous end joining (NHEJ) repair pathway at telomeres ensures that chromosome ends do not fuse. In budding yeast, binding of Rap1 to telomere repeats establishes NHEJ inhibition. Here, we show that the Uls1 protein is required for the maintenance of NHEJ inhibition at telomeres. Uls1 protein is a non-essential Swi2/Snf2-related translocase and a Small Ubiquitin-related Modifier (SUMO)-Targeted Ubiquitin Ligase (STUbL) with unknown targets. Loss of Uls1 results in telomere-telomere fusions. Uls1 requirement is alleviated by the absence of poly-SUMO chains and by rap1 alleles lacking SUMOylation sites. Furthermore, Uls1 limits the accumulation of Rap1 poly-SUMO conjugates. We propose that one of Uls1 functions is to clear non-functional poly-SUMOylated Rap1 molecules from telomeres to ensure the continuous efficiency of NHEJ inhibition. Since Uls1 is the only known STUbL with a translocase activity, it can be the general molecular sweeper for the clearance of poly-SUMOylated proteins on DNA in eukaryotes.

  14. Expression of Mycobacterium tuberculosis Ku and Ligase D in Escherichia coli results in RecA and RecB-independent DNA end-joining at regions of microhomology.

    Science.gov (United States)

    Malyarchuk, Svitlana; Wright, Douglas; Castore, Reneau; Klepper, Emily; Weiss, Bernard; Doherty, Aidan J; Harrison, Lynn

    2007-10-01

    Unlike Escherichia coli, Mycobacterium tuberculosis (Mt) expresses a Ku-like protein and an ATP-dependent DNA ligase that can perform non-homologous end-joining (NHEJ). We have expressed the Mt-Ku and Mt-Ligase D in E. coli using an arabinose-inducible promoter and expression vectors that integrate into specific sites in the E. coli chromosome. E. coli strains have been generated that express the Mt-Ku and Mt-Ligase D on a genetic background that is wild-type for repair, or deficient in either the RecA or RecB protein. Transformation of these strains with linearized plasmid DNA containing a 2bp overhang has demonstrated that expression of both the Mt-Ku and Mt-Ligase D is required for DNA end-joining and that loss of RecA does not prevent this double-strand break repair. Analysis of the re-joined plasmid has shown that repair is predominantly inaccurate and results in the deletion of sequences. Loss of RecB did not prevent the formation of large deletions, but did increase the amount of end-joining. Sequencing the junctions has revealed that the majority of the ligations occurred at regions of microhomology (1-4bps), eliminating one copy of the homologous sequence at the junction. The Mt-Ku and Mt-Ligase D can therefore function in E. coli to re-circularize linear plasmid.

  15. Interplay between Ku and Replication Protein A in the Restriction of Exo1-mediated DNA Break End Resection*

    Science.gov (United States)

    Krasner, Danielle S.; Daley, James M.; Sung, Patrick; Niu, Hengyao

    2015-01-01

    DNA double-strand breaks can be eliminated via non-homologous end joining or homologous recombination. Non-homologous end joining is initiated by the association of Ku with DNA ends. In contrast, homologous recombination entails nucleolytic resection of the 5′-strands, forming 3′-ssDNA tails that become coated with replication protein A (RPA). Ku restricts end access by the resection nuclease Exo1. It is unclear how partial resection might affect Ku engagement and Exo1 restriction. Here, we addressed these questions in a reconstituted system with yeast proteins. With blunt-ended DNA, Ku protected against Exo1 in a manner that required its DNA end-binding activity. Despite binding poorly to ssDNA, Ku could nonetheless engage a 5′-recessed DNA end with a 40-nucleotide (nt) ssDNA overhang, where it localized to the ssDNA-dsDNA junction and efficiently blocked resection by Exo1. Interestingly, RPA could exclude Ku from a partially resected structure with a 22-nt ssDNA tail and thus restored processing by Exo1. However, at a 40-nt tail, Ku remained stably associated at the ssDNA-dsDNA junction, and RPA simultaneously engaged the ssDNA region. We discuss a model in which the dynamic equilibrium between Ku and RPA binding to a partially resected DNA end influences the timing and efficiency of the resection process. PMID:26067273

  16. Agrobacterium tumefaciens T-DNA Integration and Gene Targeting in Arabidopsis thaliana Non-Homologous End-Joining Mutants

    Directory of Open Access Journals (Sweden)

    Qi Jia

    2012-01-01

    Full Text Available In order to study the role of AtKu70 and AtKu80 in Agrobacterium-mediated transformation and gene targeting, plant lines with a T-DNA insertion in AtKu80 or AtKu70 genes were functionally characterized. Such plant lines lacked both subunits, indicating that heterodimer formation between AtKu70 and AtKu80 is needed for the stability of the proteins. Homozygous mutants were phenotypically indistinguishable from wild-type plants and were fertile. However, they were hypersensitive to the genotoxic agent bleomycin, resulting in more DSBs as quantified in comet assays. They had lower end-joining efficiency, suggesting that NHEJ is a critical pathway for DSB repair in plants. Both Atku mutants and a previously isolated Atmre11 mutant were impaired in Agrobacterium T-DNA integration via floral dip transformation, indicating that AtKu70, AtKu80, and AtMre11 play an important role in T-DNA integration in Arabidopsis. The frequency of gene targeting was not significantly increased in the Atku80 and Atku70 mutants, but it was increased at least 10-fold in the Atmre11 mutant compared with the wild type.

  17. Role for Artemis nuclease in the repair of radiation-induced DNA double strand breaks by alternative end joining.

    Science.gov (United States)

    Moscariello, Mario; Wieloch, Radi; Kurosawa, Aya; Li, Fanghua; Adachi, Noritaka; Mladenov, Emil; Iliakis, George

    2015-07-01

    Exposure of cells to ionizing radiation or radiomimetic drugs generates DNA double-strand breaks that are processed either by homologous recombination repair (HRR), or by canonical, DNA-PKcs-dependent non-homologous end-joining (C-NHEJ). Chemical or genetic inactivation of factors involved in C-NHEJ or HRR, but also their local failure in repair proficient cells, promotes an alternative, error-prone end-joining pathway that serves as backup (A-EJ). There is evidence for the involvement of Artemis endonuclease, a protein deficient in a human radiosensitivity syndrome associated with severe immunodeficiency (RS-SCID), in the processing of subsets of DSBs by HRR or C-NHEJ. It is thought that within HRR or C-NHEJ Artemis processes DNA termini at complex DSBs. Whether Artemis has a role in A-EJ remains unknown. Here, we analyze using pulsed-field gel electrophoresis (PFGE) and specialized reporter assays, DSB repair in wild-type pre-B NALM-6 lymphocytes, as well as in their Artemis(-/-), DNA ligase 4(-/-) (LIG4(-/-)), and LIG4(-/-)/Artemis(-/-) double mutant counterparts, under conditions allowing evaluation of A-EJ. Our results substantiate the suggested roles of Artemis in C-NHEJ and HRR, but also demonstrate a role for the protein in A-EJ that is confirmed in Artemis deficient normal human fibroblasts. We conclude that Artemis is a nuclease participating in DSB repair by all major repair pathways. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Environmentally friendly joining of tubes by their ends

    DEFF Research Database (Denmark)

    Silva, Carlos M.A.; Nielsen, Chris Valentin; Alves, Luis M.

    2015-01-01

    This paper proposes an environmentally friendly joining process for connecting tubes by their ends that has the potential to replace current solutions based on fastened, crimped, welded, brazed or adhesive bonded joints. The process is based on a new type of tubular lap joint produced by local...... of deformation and the process feasibility window, and destructive testing to establish the working limits of tubular lap joints under different type of loading conditions. Results demonstrate that the proposed joining process is a flexible and cost-effective technology for connecting tubes by their ends...

  19. The role of DNA dependent protein kinase in synapsis of DNA ends

    NARCIS (Netherlands)

    E.P.W.C. Weterings (Eric); N.S. Verkaik (Nicole); H.T. Brüggenwirth (Hennie); D.C. van Gent (Dik); J.H.J. Hoeijmakers (Jan)

    2003-01-01

    textabstractDNA dependent protein kinase (DNA-PK) plays a central role in the non-homologous end-joining pathway of DNA double strand break repair. Its catalytic subunit (DNA-PK(CS)) functions as a serine/threonine protein kinase. We show that DNA-PK forms a stable complex at DNA termini that blocks

  20. A Role for BLM in Double-Strand Break Repair Pathway Choice: Prevention of CtIP/Mre11-Mediated Alternative Nonhomologous End-Joining

    DEFF Research Database (Denmark)

    Grabarz, Anastazja; Guirouilh-Barbat, Josée; Barascu, Aurelia

    2013-01-01

    The choice of the appropriate double-strand break (DSB) repair pathway is essential for the maintenance of genomic stability. Here, we show that the Bloom syndrome gene product, BLM, counteracts CtIP/MRE11-dependent long-range deletions (>200 bp) generated by alternative end-joining (A-EJ). BLM...... represses A-EJ in an epistatic manner with 53BP1 and RIF1 and is required for ionizing-radiation-induced 53BP1 focus assembly. Conversely, in the absence of 53BP1 or RIF1, BLM promotes formation of A-EJ long deletions, consistent with a role for BLM in DSB end resection. These data highlight a dual role...... for BLM that influences the DSB repair pathway choice: (1) protection against CtIP/MRE11 long-range deletions associated with A-EJ and (2) promotion of DNA resection. These antagonist roles can be regulated, according to cell-cycle stage, by interacting partners such as 53BP1 and TopIII, to avoid...

  1. The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends▿

    Science.gov (United States)

    Weterings, Eric; Verkaik, Nicole S.; Keijzers, Guido; Florea, Bogdan I.; Wang, Shih-Ya; Ortega, Laura G.; Uematsu, Naoya; Chen, David J.; van Gent, Dik C.

    2009-01-01

    Repair of DNA double-strand breaks (DSBs) is predominantly mediated by nonhomologous end joining (NHEJ) in mammalian cells. NHEJ requires binding of the Ku70-Ku80 heterodimer (Ku70/80) to the DNA ends and subsequent recruitment of the DNA-dependent protein kinase catalytic subunit (DNA-PKCS) and the XRCC4/ligase IV complex. Activation of the DNA-PKCS serine/threonine kinase requires an interaction with Ku70/80 and is essential for NHEJ-mediated DSB repair. In contrast to previous models, we found that the carboxy terminus of Ku80 is not absolutely required for the recruitment and activation of DNA-PKCS at DSBs, although cells that harbored a carboxy-terminal deletion in the Ku80 gene were sensitive to ionizing radiation and showed reduced end-joining capacity. More detailed analysis of this repair defect showed that DNA-PKCS autophosphorylation at Thr2647 was diminished, while Ser2056 was phosphorylated to normal levels. This resulted in severely reduced levels of Artemis nuclease activity in vivo and in vitro. We therefore conclude that the Ku80 carboxy terminus is important to support DNA-PKCS autophosphorylation at specific sites, which facilitates DNA end processing by the Artemis endonuclease and the subsequent joining reaction. PMID:19103741

  2. DNA-dependent protein kinase in nonhomologous end joining: a lock with multiple keys?

    Science.gov (United States)

    Weterings, Eric; Chen, David J

    2007-10-22

    The DNA-dependent protein kinase (DNA-PK) is one of the central enzymes involved in DNA double-strand break (DSB) repair. It facilitates proper alignment of the two ends of the broken DNA molecule and coordinates access of other factors to the repair complex. We discuss the latest findings on DNA-PK phosphorylation and offer a working model for the regulation of DNA-PK during DSB repair.

  3. Double strand break rejoining by the Ku-dependent mechanism of non-homologous end-joining

    International Nuclear Information System (INIS)

    Jeggo, P.; Singleton, B.; Beamish, H.; Priestley, A.

    1999-01-01

    The DNA-dependent protein kinase functions in the repair of DNA double strand breaks (DSBs) and in V(D)J recombination. To gain insight into the function of DNA-PK in this process we have carried out a mutation analysis of Ku80 and DNA-PKcs. Mutations at multiple sites within the N-terminal two thirds of Ku80 result in loss of Ku70/80 interaction, loss of DNA end-binding activity and inability to complement Ku80 defective cell fines. In contrast, mutations in the carboxy terminal region of the protein do not impair DNA end-binding activity but decrease the ability of Ku to activate DNA-PK. To gain insight into important functional domains within DNA-PKcs, we have analysed defective mutants, including the mouse scid cell line, and the rodent mutants, ire-20 and V-3. Mutational changes in the carboxy terminal region have been identified in all cases. Our results strongly suggest that the C-terminus of DNA-PKcs is required for kinase activity. (author)

  4. Constitutional chromothripsis rearrangements involve clustered double-stranded DNA breaks and nonhomologous repair mechanisms.

    Science.gov (United States)

    Kloosterman, Wigard P; Tavakoli-Yaraki, Masoumeh; van Roosmalen, Markus J; van Binsbergen, Ellen; Renkens, Ivo; Duran, Karen; Ballarati, Lucia; Vergult, Sarah; Giardino, Daniela; Hansson, Kerstin; Ruivenkamp, Claudia A L; Jager, Myrthe; van Haeringen, Arie; Ippel, Elly F; Haaf, Thomas; Passarge, Eberhard; Hochstenbach, Ron; Menten, Björn; Larizza, Lidia; Guryev, Victor; Poot, Martin; Cuppen, Edwin

    2012-06-28

    Chromothripsis represents a novel phenomenon in the structural variation landscape of cancer genomes. Here, we analyze the genomes of ten patients with congenital disease who were preselected to carry complex chromosomal rearrangements with more than two breakpoints. The rearrangements displayed unanticipated complexity resembling chromothripsis. We find that eight of them contain hallmarks of multiple clustered double-stranded DNA breaks (DSBs) on one or more chromosomes. In addition, nucleotide resolution analysis of 98 breakpoint junctions indicates that break repair involves nonhomologous or microhomology-mediated end joining. We observed that these eight rearrangements are balanced or contain sporadic deletions ranging in size between a few hundred base pairs and several megabases. The two remaining complex rearrangements did not display signs of DSBs and contain duplications, indicative of rearrangement processes involving template switching. Our work provides detailed insight into the characteristics of chromothripsis and supports a role for clustered DSBs driving some constitutional chromothripsis rearrangements. Copyright © 2012 The Authors. Published by Elsevier Inc. All rights reserved.

  5. Switch junction sequences in PMS2-deficient mice reveal a microhomology-mediated mechanism of Ig class switch recombination

    Science.gov (United States)

    Ehrenstein, Michael R.; Rada, Cristina; Jones, Anne-Marie; Milstein, César; Neuberger, Michael S.

    2001-01-01

    Isotype switching involves a region-specific, nonhomologous recombinational deletion that has been suggested to occur by nonhomologous joining of broken DNA ends. Here, we find increased donor/acceptor homology at switch junctions from PMS2-deficient mice and propose that class switching can occur by microhomology-mediated end-joining. Interestingly, although isotype switching and somatic hypermutation show many parallels, we confirm that PMS2 deficiency has no major effect on the pattern of nucleotide substitutions generated during somatic hypermutation. This finding is in contrast to MSH2 deficiency. With MSH2, the altered pattern of switch recombination and hypermutation suggests parallels in the mechanics of the two processes, whereas the fact that PMS2 deficiency affects only switch recombination may reflect differences in the pathways of break resolution. PMID:11717399

  6. Analysis of DNA double-strand break repair pathways in mice

    International Nuclear Information System (INIS)

    Brugmans, Linda; Kanaar, Roland; Essers, Jeroen

    2007-01-01

    During the last years significant new insights have been gained into the mechanism and biological relevance of DNA double-strand break (DSB) repair in relation to genome stability. DSBs are a highly toxic DNA lesion, because they can lead to chromosome fragmentation, loss and translocations, eventually resulting in cancer. DSBs can be induced by cellular processes such as V(D)J recombination or DNA replication. They can also be introduced by exogenous agents DNA damaging agents such as ionizing radiation or mitomycin C. During evolution several pathways have evolved for the repair of these DSBs. The most important DSB repair mechanisms in mammalian cells are nonhomologous end-joining and homologous recombination. By using an undamaged repair template, homologous recombination ensures accurate DSB repair, whereas the untemplated nonhomologous end-joining pathway does not. Although both pathways are active in mammals, the relative contribution of the two repair pathways to genome stability differs in the different cell types. Given the potential differences in repair fidelity, it is of interest to determine the relative contribution of homologous recombination and nonhomologous end-joining to DSB repair. In this review, we focus on the biological relevance of DSB repair in mammalian cells and the potential overlap between nonhomologous end-joining and homologous recombination in different tissues

  7. Influence of reduced glutathione on end-joining of DNA double-strand breaks: Cytogenetical and molecular approach

    Energy Technology Data Exchange (ETDEWEB)

    Ghoshal, Nitin [Molecular Genetics Laboratory, Department of Biotechnology & Bioinformatics, North-Eastern Hill University, Shillong, Meghalaya-793022 (India); Sharma, Sheetal [Department of Biochemistry, Indian Institute of Science, Bangalore, 560 012 (India); Banerjee, Atanu; Kurkalang, Sillarine [Molecular Genetics Laboratory, Department of Biotechnology & Bioinformatics, North-Eastern Hill University, Shillong, Meghalaya-793022 (India); Raghavan, Sathees C. [Department of Biochemistry, Indian Institute of Science, Bangalore, 560 012 (India); Chatterjee, Anupam, E-mail: chatterjeeanupam@hotmail.com [Molecular Genetics Laboratory, Department of Biotechnology & Bioinformatics, North-Eastern Hill University, Shillong, Meghalaya-793022 (India)

    2017-01-15

    Highlights: • DNA lesions induced by Blem and radiation interact well and form higher frequency of exchange aberrations. • Cellular level of glutathione does influence such interaction of DNA lesions. • Oligomer-based cell-free assay system demonstrated better end-joining efficiency at higher level of endogenous GSH. - Abstract: Radiation induced DNA double-strand breaks (DSB) are the major initial lesions whose misrejoining may lead to exchange aberrations. However, the role of glutathione (GSH), a major cellular thiol, in regulating cell’s sensitivity to DNA damaging agents is not well understood. Influence of endogenous GSH on the efficiency of X-rays and bleomycin (Blem) induced DNA DSBs end-joining has been tested here cytogenetically, in human lymphocytes and Hct116 cells. In another approach, oligomeric DNA (75 bp) containing 5′-compatible and non-compatible overhangs mimicking the endogenous DSB were for rejoining in presence of cell-free extracts from cells having different endogenous GSH levels. Frequency of aberrations, particularly exchange aberrations, was significantly increased when Blem was combined with radiation. The exchange aberration frequency was further enhanced when combined treatment was given at 4 °C since DNA lesions are poorly repaired at 4 °C so that a higher number of DNA breaks persist and interact when shifted from 4 °C to 37 °C. The exchange aberrations increased further when the combined treatment was given to Glutathione-ester (GE) pre-treated cells, indicating more frequent rejoining of DNA lesions in presence of higher cellular GSH. This is further supported by the drastic reduction in frequency of exchange aberrations but significant increase in incidences of deletions when combined treatment was given to GSH-depleted cells. End-joining efficiency of DNA DSBs with compatible ends was better than for non-compatible ends. End-joining efficiency of testicular and MCF7 cell extracts was better than that of lungs and

  8. Using Negotiated Joining to Construct and Fill Open-ended Roles in Elite Culinary Groups.

    Science.gov (United States)

    Tan, Vaughn

    2015-03-01

    This qualitative study examines membership processes in groups operating in an uncertain environment that prevents them from fully predefining new members' roles. I describe how nine elite high-end, cutting-edge culinary groups in the U.S. and Europe, ranging from innovative restaurants to culinary R&D groups, use negotiated joining-a previously undocumented process-to systematically construct and fill these emergent, open-ended roles. I show that negotiated joining is a consistently patterned, iterative process that begins with a role that both aspirant and target group explicitly understand to be provisional. This provisional role is then jointly modified and constructed by the aspirant and target group through repeated iterations of proposition, validation through trial and evaluation, and selective integration of validated role components. The initially provisional role stabilizes and the aspirant achieves membership if enough role components are validated; otherwise the negotiated joining process is abandoned. Negotiated joining allows the aspirant and target group to learn if a mutually desirable role is likely and, if so, to construct such a role. In addition, the provisional roles in negotiated joining can support absorptive capacity by allowing novel role components to enter target groups through aspirants' efforts to construct stable roles for themselves, while the internal adjustment involved in integrating newly validated role components can have the unintended side effect of supporting adaptation by providing opportunities for the groups to use these novel role components to modify their role structure and goals to suit a changing and uncertain environment. Negotiated joining thus reveals role ambiguity's hitherto unexamined beneficial consequences and provides a foundation for a contingency theory of new-member acquisition.

  9. The role of DNA dependent protein kinase in synapsis of DNA ends.

    Science.gov (United States)

    Weterings, Eric; Verkaik, Nicole S; Brüggenwirth, Hennie T; Hoeijmakers, Jan H J; van Gent, Dik C

    2003-12-15

    DNA dependent protein kinase (DNA-PK) plays a central role in the non-homologous end-joining pathway of DNA double strand break repair. Its catalytic subunit (DNA-PK(CS)) functions as a serine/threonine protein kinase. We show that DNA-PK forms a stable complex at DNA termini that blocks the action of exonucleases and ligases. The DNA termini become accessible after autophosphorylation of DNA-PK(CS), which we demonstrate to require synapsis of DNA ends. Interestingly, the presence of DNA-PK prevents ligation of the two synapsed termini, but allows ligation to another DNA molecule. This alteration of the ligation route is independent of the type of ligase that we used, indicating that the intrinsic architecture of the DNA-PK complex itself is not able to support ligation of the synapsed DNA termini. We present a working model in which DNA-PK creates a stable molecular bridge between two DNA ends that is remodeled after DNA-PK autophosphorylation in such a way that the extreme termini become accessible without disrupting synapsis. We infer that joining of synapsed DNA termini would require an additional protein factor.

  10. DNA apoptosis and stability in B-cell chronic lymphoid leukaemia: implication of the DNA double-strand breaks repair system by non homologous recombination

    International Nuclear Information System (INIS)

    Deriano, L.

    2005-01-01

    After an introduction presenting the diagnosis and treatment of chronic lymphoid leukaemia, its molecular and genetic characteristics, and its cellular origin and clonal evolution, this research thesis describes the apoptosis (definition and characteristics, cancer and chemotherapy, apoptotic ways induced by gamma irradiation), the genotoxic stresses, the different repair mechanisms for different damages, and the DNA repair processes. It reports how human chronic lymphocytic leukaemia B cells can escape DNA damage-induced apoptosis through the non-homologous end-joining DNA repair pathway, and presents non-homologous end-joining DNA repair as a potent mutagenic process in human chronic lymphocytic leukaemia B cells

  11. RECQ HELICASE RECQL4 PARTICIPATES IN NON-HOMOLOGOUS END JOINING AND INTERACTS WITH THE KU COMPLEX

    DEFF Research Database (Denmark)

    Shamanna, Raghavendra A; Singh, Dharmendra Kumar; Lu, Huiming

    2014-01-01

    -irradiation and resulted in accumulation of 53BP1 foci after irradiation, indicating defects in the processing of DSB. We find that RECQL4 interacts with the Ku70/Ku80 heterodimer, part of the DNA-dependent protein kinase (DNA-PK) complex, via its N-terminal domain. Further, RECQL4 stimulates higher order DNA binding...... of Ku70/Ku80 to a blunt end DNA substrate. Taken together, these results implicate that RECQL4 participates in the NHEJ pathway of DSB repair via a functional interaction with the Ku70/Ku80 complex. This is the first study to provide both in vitro and in vivo evidence for a role of a RecQ helicase...

  12. GANP regulates the choice of DNA repair pathway by DNA-PKcs interaction in AID-dependent IgV region diversification.

    Science.gov (United States)

    Eid, Mohammed Mansour Abbas; Maeda, Kazuhiko; Almofty, Sarah Ameen; Singh, Shailendra Kumar; Shimoda, Mayuko; Sakaguchi, Nobuo

    2014-06-15

    RNA export factor germinal center-associated nuclear protein (GANP) interacts with activation-induced cytidine deaminase (AID) and shepherds it from the cytoplasm to the nucleus and toward the IgV region loci in B cells. In this study, we demonstrate a role for GANP in the repair of AID-initiated DNA damage in chicken DT40 B cells to generate IgV region diversity by gene conversion and somatic hypermutation. GANP plays a positive role in IgV region diversification of DT40 B cells in a nonhomologous end joining-proficient state. DNA-PKcs physically interacts with GANP, and this interaction is dissociated by dsDNA breaks induced by a topoisomerase II inhibitor, etoposide, or AID overexpression. GANP affects the choice of DNA repair mechanism in B cells toward homologous recombination rather than nonhomologous end joining repair. Thus, GANP presumably plays a critical role in protection of the rearranged IgV loci by favoring homologous recombination of the DNA breaks under accelerated AID recruitment. Copyright © 2014 by The American Association of Immunologists, Inc.

  13. Genome Editing in Escherichia coli with Cas9 and synthetic CRISPRs

    Energy Technology Data Exchange (ETDEWEB)

    Peng, Ze; Richardson, Sarah; Robinson, David; Deutsch, Samuel; Cheng, Jan-Fang

    2014-03-14

    Recently, the Cas9-CRISPR system has proven to be a useful tool for genome editing in eukaryotes, which repair the double stranded breaks made by Cas9 with non-homologous end joining or homologous recombination. Escherichia coli lacks non-homologous end joining and has a very low homologous recombination rate, effectively rendering targeted Cas9 activity lethal. We have developed a heat curable, serializable, plasmid based system for selectionless Cas9 editing in arbitrary E. coli strains that uses synthetic CRISPRs for targeting and -red to effect repairs of double stranded breaks. We have demonstrated insertions, substitutions, and multi-target deletions with our system, which we have tested in several strains.

  14. Sibling rivalry: competition between Pol X family members in V(D)J recombination and general double strand break repair.

    Science.gov (United States)

    Nick McElhinny, Stephanie A; Ramsden, Dale A

    2004-08-01

    The nonhomologous end-joining pathway is a major means for repairing double-strand breaks (DSBs) in all mitotic cell types. This repair pathway is also the only efficient means for resolving DSB intermediates in V(D)J recombination, a lymphocyte-specific genome rearrangement required for assembly of antigen receptors. A role for polymerases in end-joining has been well established. They are a major factor in determining the character of repair junctions but, in contrast to 'core' end-joining factors, typically appear to have a subtle impact on the efficiency of end-joining. Recent work implicates several members of the Pol X family in end-joining and suggests surprising complexity in the control of how these different polymerases are employed in this pathway.

  15. DNA-Dependent Protein Kinase in Non-Homologous End-Joining: Guarding Strategic Positions

    OpenAIRE

    Weterings, Eric

    2005-01-01

    markdownabstract__Abstract__ Careful maintenance of genetic information throughout generations is of vital importance to all living creatures. A battery of both endogenous and exogenous factors continuously threatens genetic integrity by altering the DNA chemistry. As a consequence, DNA damage types are as diverse as their causes. DNA doublestrand breaks (DSBs) are among the most deleterious lesions, since they introduce chromosomal breakage or translocation and are able to trigger carcinogen...

  16. TDP2 suppresses chromosomal translocations induced by DNA topoisomerase II during gene transcription.

    Science.gov (United States)

    Gómez-Herreros, Fernando; Zagnoli-Vieira, Guido; Ntai, Ioanna; Martínez-Macías, María Isabel; Anderson, Rhona M; Herrero-Ruíz, Andrés; Caldecott, Keith W

    2017-08-10

    DNA double-strand breaks (DSBs) induced by abortive topoisomerase II (TOP2) activity are a potential source of genome instability and chromosome translocation. TOP2-induced DNA double-strand breaks are rejoined in part by tyrosyl-DNA phosphodiesterase 2 (TDP2)-dependent non-homologous end-joining (NHEJ), but whether this process suppresses or promotes TOP2-induced translocations is unclear. Here, we show that TDP2 rejoins DSBs induced during transcription-dependent TOP2 activity in breast cancer cells and at the translocation 'hotspot', MLL. Moreover, we find that TDP2 suppresses chromosome rearrangements induced by TOP2 and reduces TOP2-induced chromosome translocations that arise during gene transcription. Interestingly, however, we implicate TDP2-dependent NHEJ in the formation of a rare subclass of translocations associated previously with therapy-related leukemia and characterized by junction sequences with 4-bp of perfect homology. Collectively, these data highlight the threat posed by TOP2-induced DSBs during transcription and demonstrate the importance of TDP2-dependent non-homologous end-joining in protecting both gene transcription and genome stability.DNA double-strand breaks (DSBs) induced by topoisomerase II (TOP2) are rejoined by TDP2-dependent non-homologous end-joining (NHEJ) but whether this promotes or suppresses translocations is not clear. Here the authors show that TDP2 suppresses chromosome translocations from DSBs introduced during gene transcription.

  17. Inactivation of Pol θ and C-NHEJ eliminates off-target integration of exogenous DNA.

    Science.gov (United States)

    Zelensky, Alex N; Schimmel, Joost; Kool, Hanneke; Kanaar, Roland; Tijsterman, Marcel

    2017-07-07

    Off-target or random integration of exogenous DNA hampers precise genomic engineering and presents a safety risk in clinical gene therapy strategies. Genetic definition of random integration has been lacking for decades. Here, we show that the A-family DNA polymerase θ (Pol θ) promotes random integration, while canonical non-homologous DNA end joining plays a secondary role; cells double deficient for polymerase θ and canonical non-homologous DNA end joining are devoid of any integration events, demonstrating that these two mechanisms define random integration. In contrast, homologous recombination is not reduced in these cells and gene targeting is improved to 100% efficiency. Such complete reversal of integration outcome, from predominately random integration to exclusively gene targeting, provides a rational way forward to improve the efficacy and safety of DNA delivery and gene correction approaches.Random off-target integration events can impair precise gene targeting and poses a safety risk for gene therapy. Here the authors show that repression of polymerase θ and classical non-homologous recombination eliminates random integration.

  18. Comparative analysis of the end-joining activity of several DNA ligases.

    Directory of Open Access Journals (Sweden)

    Robert J Bauer

    Full Text Available DNA ligases catalyze the repair of phosphate backbone breaks in DNA, acting with highest activity on breaks in one strand of duplex DNA. Some DNA ligases have also been observed to ligate two DNA fragments with short complementary overhangs or blunt-ended termini. In this study, several wild-type DNA ligases (phage T3, T4, and T7 DNA ligases, Paramecium bursaria chlorella virus 1 (PBCV1 DNA ligase, human DNA ligase 3, and Escherichia coli DNA ligase were tested for their ability to ligate DNA fragments with several difficult to ligate end structures (blunt-ended termini, 3'- and 5'- single base overhangs, and 5'-two base overhangs. This analysis revealed that T4 DNA ligase, the most common enzyme utilized for in vitro ligation, had its greatest activity on blunt- and 2-base overhangs, and poorest on 5'-single base overhangs. Other ligases had different substrate specificity: T3 DNA ligase ligated only blunt ends well; PBCV1 DNA ligase joined 3'-single base overhangs and 2-base overhangs effectively with little blunt or 5'- single base overhang activity; and human ligase 3 had highest activity on blunt ends and 5'-single base overhangs. There is no correlation of activity among ligases on blunt DNA ends with their activity on single base overhangs. In addition, DNA binding domains (Sso7d, hLig3 zinc finger, and T4 DNA ligase N-terminal domain were fused to PBCV1 DNA ligase to explore whether modified binding to DNA would lead to greater activity on these difficult to ligate substrates. These engineered ligases showed both an increased binding affinity for DNA and increased activity, but did not alter the relative substrate preferences of PBCV1 DNA ligase, indicating active site structure plays a role in determining substrate preference.

  19. Mycobacterium smegmatis Ku binds DNA without free ends.

    Science.gov (United States)

    Kushwaha, Ambuj K; Grove, Anne

    2013-12-01

    Ku is central to the non-homologous end-joining pathway of double-strand-break repair in all three major domains of life, with eukaryotic homologues being associated with more diversified roles compared with prokaryotic and archaeal homologues. Ku has a conserved central 'ring-shaped' core domain. While prokaryotic homologues lack the N- and C-terminal domains that impart functional diversity to eukaryotic Ku, analyses of Ku from certain prokaryotes such as Pseudomonas aeruginosa and Mycobacterium smegmatis have revealed the presence of distinct C-terminal extensions that modulate DNA-binding properties. We report in the present paper that the lysine-rich C-terminal extension of M. smegmatis Ku contacts the core protein domain as evidenced by an increase in DNA-binding affinity and a decrease in thermal stability and intrinsic tryptophan fluorescence upon its deletion. Ku deleted for this C-terminus requires free DNA ends for binding, but translocates to internal DNA sites. In contrast, full-length Ku can directly bind DNA without free ends, suggesting that this property is conferred by its C-terminus. Such binding to internal DNA sites may facilitate recruitment to sites of DNA damage. The results of the present study also suggest that extensions beyond the shared core domain may have independently evolved to expand Ku function.

  20. Fumarase is involved in DNA double-strand break resection through a functional interaction with Sae2

    DEFF Research Database (Denmark)

    Leshets, Michael; Ramamurthy, Dharanidharan; Lisby, Michael

    2018-01-01

    One of the most severe forms of DNA damage is the double-strand break (DSB). Failure to properly repair the damage can cause mutation, gross chromosomal rearrangements and lead to the development of cancer. In eukaryotes, homologous recombination (HR) and non-homologous end joining (NHEJ) are the......One of the most severe forms of DNA damage is the double-strand break (DSB). Failure to properly repair the damage can cause mutation, gross chromosomal rearrangements and lead to the development of cancer. In eukaryotes, homologous recombination (HR) and non-homologous end joining (NHEJ......) are the main DSB repair pathways. Fumarase is a mitochondrial enzyme which functions in the tricarboxylic acid cycle. Intriguingly, the enzyme can be readily detected in the cytosolic compartment of all organisms examined, and we have shown that cytosolic fumarase participates in the DNA damage response...

  1. The use of TALENs for nonhomologous end joining mutagenesis in silkworm and fruitly

    Czech Academy of Sciences Publication Activity Database

    Takasu, Y.; Tamura, T.; Sajwan, Suresh; Kobayashi, I.; Žurovec, Michal

    2014-01-01

    Roč. 69, č. 1 (2014), s. 46-57 ISSN 1046-2023 R&D Projects: GA ČR GAP305/10/2406 Grant - others:Japan Society for the Promotion of Science(JP) 23580083 Institutional support: RVO:60077344 Keywords : Bombyx mori * Drosophila melanogaster * pBlue-TAL Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.645, year: 2014 http://www.sciencedirect.com/science/article/pii/S1046202314000449

  2. Ionizing radiation sensitivity of DNA polymerase lambda-deficient cells.

    NARCIS (Netherlands)

    Vermeulen, C.; Bertocci, B.; Begg, A.C.; Vens, C.

    2007-01-01

    Ionizing radiation induces a diverse spectrum of DNA lesions, including strand breaks and oxidized bases. In mammalian cells, ionizing radiation-induced lesions are targets of non-homologous end joining, homologous recombination, and base excision repair. In vitro assays show a potential involvement

  3. Screening of Pesticides with the Potential of Inducing DSB and Successive Recombinational Repair

    Directory of Open Access Journals (Sweden)

    Karen Suárez-Larios

    2017-01-01

    Full Text Available A study was realized to ascertain whether eight selected pesticides would induce double strand breaks (DSB in lymphocyte cultures and whether this damage would induce greater levels of proteins Rad51 participating in homologous recombination or of p-Ku80 participating in nonhomologous end joining. Only five pesticides were found to induce DSB of which only glyphosate and paraoxon induced a significant increase of p-Ku80 protein, indicating that nonhomologous end joining recombinational DNA repair system would be activated. The type of gamma-H2AX foci observed was comparable to that induced by etoposide at similar concentrations. These results are of importance since these effects occurred at low concentrations in the micromolar range, in acute treatments to the cells. Effects over longer exposures in actual environmental settings are expected to produce cumulative damage if repeated events of recombination take place over time.

  4. A robust network of double-strand break repair pathways governs genome integrity during C. elegans development.

    NARCIS (Netherlands)

    Pontier, D.B.; Tijsterman, M.

    2009-01-01

    To preserve genomic integrity, various mechanisms have evolved to repair DNA double-strand breaks (DSBs). Depending on cell type or cell cycle phase, DSBs can be repaired error-free, by homologous recombination, or with concomitant loss of sequence information, via nonhomologous end-joining (NHEJ)

  5. Crypt base columnar stem cells in small intestines of mice are radioresistant

    NARCIS (Netherlands)

    Hua, G.; Thin, T.H.; Feldman, R.; Haimovitz-Friedman, A.; Clevers, H.; Fuks, Z.; Kolesnick, R.

    2012-01-01

    BACKGROUND & AIMS: Adult stem cells have been proposed to be quiescent and radiation resistant, repairing DNA double-strand breaks by nonhomologous end joining. However, the population of putative small intestinal stem cells (ISCs) at position +4 from the crypt base contradicts this model, in that

  6. Physical and biological parameters affecting DNA double strand break misrejoining in mammalian cells

    International Nuclear Information System (INIS)

    Kuehne, M.; Rothkamm, K.; Loebrich, M.

    2002-01-01

    In an attempt to investigate the effect of radiation quality, dose and specific repair pathways on correct and erroneous rejoining of DNA double strand breaks (DSBs), an assay was applied that allows the identification and quantification of incorrectly rejoined DSB ends produced by ionising radiation. While substantial misrejoining occurs in mammalian cells after high acute irradiation doses, decreasing misrejoining frequencies were observed in dose fractionation experiments with X rays. In line with this finding, continuous irradiation with gamma rays at low dose rate leads to non detectable misrejoining. This indicates that the probability for a DSB to be misrejoined decreases drastically when DSBs are separated in time and space. The same dose fractionation approach was applied to determine DSB misrejoining after a particle exposure. In contrast to the results with X rays, there was no significant decrease in DSB misrejoining with increasing fractionation. This suggests that DSB misrejoining after a irradiation is not significantly affected by a separation of particle tracks. To identify the enzymatic pathways that are involved in DSB misrejoining, cell lines deficient in non-homologous end-joining (NHEJ) were examined. After high X ray doses, DSB misrejoining is considerable reduced in NHEJ mutants. Low dose rate experiments show elevated DSB misrejoining in NHEJ mutants compared with wild-type cells. The authors propose that NHEJ serves as an efficient pathway for rejoining correct break ends in situations of separated breaks but generates genomic rearrangements if DSBs are close in time and space. (author)

  7. Pharmacodynamic Assay Panel for Monitoring Phospho-Signaling Networks | Office of Cancer Clinical Proteomics Research

    Science.gov (United States)

    The DNA damage response (DDR) is a highly regulated signal transduction network that orchestrates the temporal and spatial organization of protein complexes required to repair (or tolerate) DNA damage (e.g., nucleotide excision repair, base excision repair, homologous recombination, non-homologous end joining, post-replication repair).

  8. Non-homologous isofunctional enzymes: a systematic analysis of alternative solutions in enzyme evolution.

    Science.gov (United States)

    Omelchenko, Marina V; Galperin, Michael Y; Wolf, Yuri I; Koonin, Eugene V

    2010-04-30

    Evolutionarily unrelated proteins that catalyze the same biochemical reactions are often referred to as analogous - as opposed to homologous - enzymes. The existence of numerous alternative, non-homologous enzyme isoforms presents an interesting evolutionary problem; it also complicates genome-based reconstruction of the metabolic pathways in a variety of organisms. In 1998, a systematic search for analogous enzymes resulted in the identification of 105 Enzyme Commission (EC) numbers that included two or more proteins without detectable sequence similarity to each other, including 34 EC nodes where proteins were known (or predicted) to have distinct structural folds, indicating independent evolutionary origins. In the past 12 years, many putative non-homologous isofunctional enzymes were identified in newly sequenced genomes. In addition, efforts in structural genomics resulted in a vastly improved structural coverage of proteomes, providing for definitive assessment of (non)homologous relationships between proteins. We report the results of a comprehensive search for non-homologous isofunctional enzymes (NISE) that yielded 185 EC nodes with two or more experimentally characterized - or predicted - structurally unrelated proteins. Of these NISE sets, only 74 were from the original 1998 list. Structural assignments of the NISE show over-representation of proteins with the TIM barrel fold and the nucleotide-binding Rossmann fold. From the functional perspective, the set of NISE is enriched in hydrolases, particularly carbohydrate hydrolases, and in enzymes involved in defense against oxidative stress. These results indicate that at least some of the non-homologous isofunctional enzymes were recruited relatively recently from enzyme families that are active against related substrates and are sufficiently flexible to accommodate changes in substrate specificity.

  9. Deletion of individual Ku subunits in mice causes an NHEJ-independent phenotype potentially by altering apurinic/apyrimidinic site repair

    NARCIS (Netherlands)

    Y.J. Choi (Yong Jun); H. Li (Han); M.Y. Son (Mi Young); X.-H. Wang (Xiao-Hong); J.L. Fornsaglio (Jamie L.); R.W. Sobol (Robert W.); M. Lee (Moonsook); J. Vijg (Jan); S. Imholz (Sandra); M.E.T. Dollé (Martijn); H. van Steeg (Harry); E. Reiling (Erwin); P. Hasty (Paul)

    2014-01-01

    textabstractKu70 and Ku80 form a heterodimer called Ku that forms a holoenzyme with DNA dependent-protein kinase catalytic subunit (DNA-PKCS) to repair DNA double strand breaks (DSBs) through the nonhomologous end joining (NHEJ) pathway. As expected mutating these genes in mice caused a similar DSB

  10. Chromosome End Repair and Genome Stability in Plasmodium falciparum

    Directory of Open Access Journals (Sweden)

    Susannah F. Calhoun

    2017-08-01

    Full Text Available The human malaria parasite Plasmodium falciparum replicates within circulating red blood cells, where it is subjected to conditions that frequently cause DNA damage. The repair of DNA double-stranded breaks (DSBs is thought to rely almost exclusively on homologous recombination (HR, due to a lack of efficient nonhomologous end joining. However, given that the parasite is haploid during this stage of its life cycle, the mechanisms involved in maintaining genome stability are poorly understood. Of particular interest are the subtelomeric regions of the chromosomes, which contain the majority of the multicopy variant antigen-encoding genes responsible for virulence and disease severity. Here, we show that parasites utilize a competitive balance between de novo telomere addition, also called “telomere healing,” and HR to stabilize chromosome ends. Products of both repair pathways were observed in response to DSBs that occurred spontaneously during routine in vitro culture or resulted from experimentally induced DSBs, demonstrating that both pathways are active in repairing DSBs within subtelomeric regions and that the pathway utilized was determined by the DNA sequences immediately surrounding the break. In combination, these two repair pathways enable parasites to efficiently maintain chromosome stability while also contributing to the generation of genetic diversity.

  11. Author Details

    African Journals Online (AJOL)

    Sanie-Jahromi, Fatemeh. Vol 18, No 4 (2017) - Articles Extremely low frequency electromagnetic field in combination with β-Lapachone up-regulates the genes of non-homologous end joining. Abstract PDF. ISSN: 1110-8630. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors ...

  12. RNF4 is required for DNA double-strand break repair in vivo

    DEFF Research Database (Denmark)

    Vyas, R; Kumar, R; Clermont, F

    2013-01-01

    for both homologous recombination (HR) and non-homologous end joining repair. To establish a link between Rnf4 and the DNA damage response (DDR) in vivo, we generated an Rnf4 allelic series in mice. We show that Rnf4-deficiency causes persistent ionizing radiation-induced DNA damage and signaling...

  13. Double-Strand DNA Break Repair in Mycobacteria.

    Science.gov (United States)

    Glickman, Michael S

    2014-10-01

    Discontinuity of both strands of the chromosome is a lethal event in all living organisms because it compromises chromosome replication. As such, a diversity of DNA repair systems has evolved to repair double-strand DNA breaks (DSBs). In part, this diversity of DSB repair systems has evolved to repair breaks that arise in diverse physiologic circumstances or sequence contexts, including cellular states of nonreplication or breaks that arise between repeats. Mycobacteria elaborate a set of three genetically distinct DNA repair pathways: homologous recombination, nonhomologous end joining, and single-strand annealing. As such, mycobacterial DSB repair diverges substantially from the standard model of prokaryotic DSB repair and represents an attractive new model system. In addition, the presence in mycobacteria of a DSB repair system that can repair DSBs in nonreplicating cells (nonhomologous end joining) or when DSBs arise between repeats (single-strand annealing) has clear potential relevance to Mycobacterium tuberculosis pathogenesis, although the exact role of these systems in M. tuberculosis pathogenesis is still being elucidated. In this article we will review the genetics of mycobacterial DSB repair systems, focusing on recent insights.

  14. Mycobacterial UvrD1 is a Ku-dependent DNA helicase that plays a role in multiple DNA repair events, including double-strand break repair.

    Science.gov (United States)

    Sinha, Krishna Murari; Stephanou, Nicolas C; Gao, Feng; Glickman, Michael S; Shuman, Stewart

    2007-05-18

    Mycobacterium tuberculosis and other bacterial pathogens have a Ku-dependent nonhomologous end joining pathway of DNA double-strand break repair. Here we identify mycobacterial UvrD1 as a novel interaction partner for Ku in a genome-wide yeast two-hybrid screen. UvrD1 per se is a vigorous DNA-dependent ATPase but a feeble DNA helicase. Ku stimulates UvrD1 to catalyze ATP-dependent unwinding of 3'-tailed DNAs. UvrD1, Ku, and DNA form a stable ternary complex in the absence of ATP. The Ku binding determinants are located in the distinctive C-terminal segment of UvrD1. A second mycobacterial paralog, UvrD2, is a vigorous Ku-independent DNA helicase. Ablation of UvrD1 sensitizes Mycobacterium smegmatis to killing by ultraviolet and ionizing radiation and to a single chromosomal break generated by I-SceI endonuclease. The physical and functional interactions of bacterial Ku and UvrD1 highlight the potential for cross-talk between components of nonhomologous end joining and nucleotide excision repair pathways.

  15. Productive Homologous and Non-homologous Recombination of Hepatitis C Virus in Cell Culture

    Science.gov (United States)

    Li, Yi-Ping; Mikkelsen, Lotte S.; Gottwein, Judith M.; Bukh, Jens

    2013-01-01

    Genetic recombination is an important mechanism for increasing diversity of RNA viruses, and constitutes a viral escape mechanism to host immune responses and to treatment with antiviral compounds. Although rare, epidemiologically important hepatitis C virus (HCV) recombinants have been reported. In addition, recombination is an important regulatory mechanism of cytopathogenicity for the related pestiviruses. Here we describe recombination of HCV RNA in cell culture leading to production of infectious virus. Initially, hepatoma cells were co-transfected with a replicating JFH1ΔE1E2 genome (genotype 2a) lacking functional envelope genes and strain J6 (2a), which has functional envelope genes but does not replicate in culture. After an initial decrease in the number of HCV positive cells, infection spread after 13–36 days. Sequencing of recovered viruses revealed non-homologous recombinants with J6 sequence from the 5′ end to the NS2–NS3 region followed by JFH1 sequence from Core to the 3′ end. These recombinants carried duplicated sequence of up to 2400 nucleotides. HCV replication was not required for recombination, as recombinants were observed in most experiments even when two replication incompetent genomes were co-transfected. Reverse genetic studies verified the viability of representative recombinants. After serial passage, subsequent recombination events reducing or eliminating the duplicated region were observed for some but not all recombinants. Furthermore, we found that inter-genotypic recombination could occur, but at a lower frequency than intra-genotypic recombination. Productive recombination of attenuated HCV genomes depended on expression of all HCV proteins and tolerated duplicated sequence. In general, no strong site specificity was observed. Non-homologous recombination was observed in most cases, while few homologous events were identified. A better understanding of HCV recombination could help identification of natural recombinants

  16. Mutagenic effects of carbon ions near the range end in plants

    Energy Technology Data Exchange (ETDEWEB)

    Hase, Yoshihiro, E-mail: hase.yoshihiro@jaea.go.jp [Ion Beam Mutagenesis Research Group, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki, Takasaki, Gunma 370-1292 (Japan); Yoshihara, Ryouhei; Nozawa, Shigeki; Narumi, Issay [Ion Beam Mutagenesis Research Group, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki, Takasaki, Gunma 370-1292 (Japan)

    2012-03-01

    To gain insight into the mutagenic effects of accelerated heavy ions in plants, the mutagenic effects of carbon ions near the range end (mean linear energy transfer (LET): 425 keV/{mu}m) were compared with the effects of carbon ions penetrating the seeds (mean LET: 113 keV/{mu}m). Mutational analysis by plasmid rescue of Escherichia coli rpsL from irradiated Arabidopsis plants showed a 2.7-fold increase in mutant frequency for 113 keV/{mu}m carbon ions, whereas no enhancement of mutant frequency was observed for carbon ions near the range end. This suggested that carbon ions near the range end induced mutations that were not recovered by plasmid rescue. An Arabidopsis DNA ligase IV mutant, deficient in non-homologous end-joining repair, showed hyper-sensitivity to both types of carbon-ion irradiation. The difference in radiation sensitivity between the wild type and the repair-deficient mutant was greatly diminished for carbon ions near the range end, suggesting that these ions induce irreparable DNA damage. Mutational analysis of the Arabidopsis GL1 locus showed that while the frequency of generation of glabrous mutant sectors was not different between the two types of carbon-ion irradiation, large deletions (>{approx}30 kb) were six times more frequently induced by carbon ions near the range end. When 352 keV/{mu}m neon ions were used, these showed a 6.4 times increase in the frequency of induced large deletions compared with the 113 keV/{mu}m carbon ions. We suggest that the proportion of large deletions increases with LET in plants, as has been reported for mammalian cells. The nature of mutations induced in plants by carbon ions near the range end is discussed in relation to mutation detection by plasmid rescue and transmissibility to progeny.

  17. AUGMENTING 3D CITY MODEL COMPONENTS BY GEODATA JOINS TO FACILITATE AD-HOC GEOMETRIC-TOPOLOGICALLY SOUND INTEGRATION

    Directory of Open Access Journals (Sweden)

    R. Kaden

    2012-07-01

    Full Text Available Virtual 3D city models are integrated complex compositions of spatial data of different themes, origin, quality, scale, and dimensions. Within this paper, we address the problem of spatial compatibility of geodata aiming to provide support for ad-hoc integration of virtual 3D city models including geodata of different sources and themes like buildings, terrain, and city furniture. In contrast to related work which is dealing with the integration of redundant geodata structured according to different data models and ontologies, we focus on the integration of complex 3D models of the same representation (here: CityGML but regarding to the geometric-topological consistent matching of non-homologous objects, e.g. a building is connected to a road, and their geometric homogenisation. Therefore, we present an approach including a data model for a Geodata Join and the general concept of an integration procedure using the join information. The Geodata Join aims to bridge the lack of information between fragmented geodata by describing the relationship between adjacent objects from different datasets. The join information includes the geometrical representation of those parts of an object, which have a specific/known topological or geometrical relationship to another object. This part is referred to as a Connector and is either described by points, lines, or surfaces of the existing object geometry or by additional join geometry. In addition, the join information includes the specification of the connected object in the other dataset and the description of the topological and geometrical relationship between both objects, which is used to aid the matching process. Furthermore, the Geodata Join contains object-related information like accuracy values and restrictions of movement and deformation which are used to optimize the integration process. Based on these parameters, a functional model including a matching algorithm, transformation methods, and

  18. DNA-dependent protein kinase inhibits AID-induced antibody gene conversion.

    Directory of Open Access Journals (Sweden)

    Adam J L Cook

    2007-04-01

    Full Text Available Affinity maturation and class switching of antibodies requires activation-induced cytidine deaminase (AID-dependent hypermutation of Ig V(DJ rearrangements and Ig S regions, respectively, in activated B cells. AID deaminates deoxycytidine bases in Ig genes, converting them into deoxyuridines. In V(DJ regions, subsequent excision of the deaminated bases by uracil-DNA glycosylase, or by mismatch repair, leads to further point mutation or gene conversion, depending on the species. In Ig S regions, nicking at the abasic sites produced by AID and uracil-DNA glycosylases results in staggered double-strand breaks, whose repair by nonhomologous end joining mediates Ig class switching. We have tested whether nonhomologous end joining also plays a role in V(DJ hypermutation using chicken DT40 cells deficient for Ku70 or the DNA-dependent protein kinase catalytic subunit (DNA-PKcs. Inactivation of the Ku70 or DNA-PKcs genes in DT40 cells elevated the rate of AID-induced gene conversion as much as 5-fold. Furthermore, DNA-PKcs-deficiency appeared to reduce point mutation. The data provide strong evidence that double-strand DNA ends capable of recruiting the DNA-dependent protein kinase complex are important intermediates in Ig V gene conversion.

  19. Efficient disruption of endogenous Bombyx gene by TAL effector nucleases

    Czech Academy of Sciences Publication Activity Database

    Sajwan, Suresh; Takasu, Y.; Tamura, T.; Uchino, K.; Sezutsu, H.; Žurovec, Michal

    2013-01-01

    Roč. 43, č. 1 (2013), s. 17-23 ISSN 0965-1748 R&D Projects: GA ČR GAP305/10/2406 Grant - others:Japan Society for the Promotion of Science(JP) 23580083 Institutional support: RVO:60077344 Keywords : gene targeting * ZFN * nonhomologous end joining Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.420, year: 2013 http://www.sciencedirect.com/science/article/pii/S0965174812001622#

  20. Nonhomologous DNA end joining and chromosome aberrations in human embryonic lung fibroblasts treated with environmental pollutants

    Czech Academy of Sciences Publication Activity Database

    Rössner ml., Pavel; Rössnerová, Andrea; Beskid, Olena; Tabashidze, Nana; Líbalová, Helena; Uhlířová, Kateřina; Topinka, Jan; Šrám, Radim

    763-764, MAY-JUN 2014 (2014), s. 28-38 ISSN 0027-5107 R&D Projects: GA ČR GAP503/11/0084 Institutional support: RVO:68378041 Keywords : benzo[a]pyrene * chromosome aberrations * double-strand DNA breaks Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 3.680, year: 2014

  1. Deciphering the Role of Alternative Non-Homologous End Joining (Alt-NHEJ) DNA Repair in Breast Cancer

    Science.gov (United States)

    2017-12-01

    consecutive TTAGGG repeats. To detect rare reads containing fusion junctions, we exploited the novel sequence arrangement created by the ligation of the 39G...journal.pgen.1001005 (2010). 14 van Kregten, M. et al. T-DNA integration in plants results from polymerase-theta-mediated DNA repair. Nat Plants 2

  2. Chrysler Upset Protrusion Joining Techniques for Joining Dissimilar Metals

    Energy Technology Data Exchange (ETDEWEB)

    Logan, Stephen [FCA US LLC, Auburn Hills, MI (United States)

    2017-09-28

    The project goal was to develop and demonstrate a robust, cost effective, and versatile joining technique, known as Upset Protrusion Joining (UPJ), for joining challenging dissimilar metal com-binations, especially those where one of the metals is a die cast magnesium (Mg) component. Since two of the key obstacles preventing more widespread use of light metals (especially in high volume automotive applications) are 1) a lack of robust joining techniques and 2) susceptibility to galvanic corrosion, and since the majority of the joint combinations evaluated in this project include die cast Mg (the lightest structural metal) as one of the two materials being joined, and since die casting is the most common and cost effective process for producing Mg components, then successful project completion provides a key enabler to high volume application of lightweight materials, thus potentially leading to reduced costs, and encouraging implementation of lightweight multi-material vehicles for significant reductions in energy consumption and reduced greenhouse gas emissions. Eco-nomic benefits to end-use consumers are achieved primarily via the reduction in fuel consumption. Unlike currently available commercial processes, the UPJ process relies on a very robust mechanical joint rather than intermetallic bonding, so the more cathodic material can be coated prior to joining, thus creating a robust isolation against galvanic attack on the more anodic material. Additionally, since the UPJ protrusion is going through a hole that can be pre-drilled or pre-punched prior to coating, the UPJ process is less likely to damage the coating when the joint is being made. Further-more, since there is no additional cathodic material (such as a steel fastener) used to create the joint, there is no joining induced galvanic activity beyond that of the two parent materials. In accordance with its originally proposed plan, this project has successfully developed process variants of UPJ to enable

  3. An RNA secondary structure bias for non-homologous reverse transcriptase-mediated deletions in vivo

    DEFF Research Database (Denmark)

    Duch, Mogens; Carrasco, Maria L; Jespersen, Thomas

    2004-01-01

    Murine leukemia viruses harboring an internal ribosome entry site (IRES)-directed translational cassette are able to replicate, but undergo loss of heterologous sequences upon continued passage. While complete loss of heterologous sequences is favored when these are flanked by a direct repeat......, deletion mutants with junction sites within the heterologous cassette may also be retrieved, in particular from vectors without flanking repeats. Such deletion mutants were here used to investigate determinants of reverse transcriptase-mediated non-homologous recombination. Based upon previous structural...... result from template switching during first-strand cDNA synthesis and that the choice of acceptor sites for non-homologous recombination are guided by non-paired regions. Our results may have implications for recombination events taking place within structured regions of retroviral RNA genomes...

  4. Transition piece for joining together tubular pieces

    International Nuclear Information System (INIS)

    Holko, K.H.

    1981-01-01

    A transition piece for joining together tubular pieces formed respectively from a low alloy or carbon steel and a high temperature alloy containing at least 16% chromium includes a plurality of tubular parts welded together and formed from materials of selected composition with a maximum chromium content difference of 5% between adjacent parts when the chromium content of each part is below 10% and a maximum chromium difference of 7% between adjacent parts when the chromium content of either part is above 10%. The transition parts are also graded as to such characteristics as thermal expansion coefficient. The transition parts at opposite ends of the transition joint have chromium percentages similar to the tubular pieces to which they are to be joined. The parts may be joined by fusion and/or friction welding and parts may be formed by fusion weld deposition. (author)

  5. A polycomb group protein, PHF1, is involved in the response to DNA double-strand breaks in human cell

    Science.gov (United States)

    Hong, Zehui; Jiang, Jie; Lan, Li; Nakajima, Satoshi; Kanno, Shin-ichiro; Koseki, Haruhiko; Yasui, Akira

    2008-01-01

    DNA double-strand breaks (DSBs) represent the most toxic DNA damage arisen from endogenous and exogenous genotoxic stresses and are known to be repaired by either homologous recombination or nonhomologous end-joining processes. Although many proteins have been identified to participate in either of the processes, the whole processes still remain elusive. Polycomb group (PcG) proteins are epigenetic chromatin modifiers involved in gene silencing, cancer development and the maintenance of embryonic and adult stem cells. By screening proteins responding to DNA damage using laser micro-irradiation, we found that PHF1, a human homolog of Drosophila polycomb-like, Pcl, protein, was recruited to DSBs immediately after irradiation and dissociated within 10 min. The accumulation at DSBs is Ku70/Ku80-dependent, and knockdown of PHF1 leads to X-ray sensitivity and increases the frequency of homologous recombination in HeLa cell. We found that PHF1 interacts physically with Ku70/Ku80, suggesting that PHF1 promotes nonhomologous end-joining processes. Furthermore, we found that PHF1 interacts with a number of proteins involved in DNA damage responses, RAD50, SMC1, DHX9 and p53, further suggesting that PHF1, besides the function in PcG, is involved in genome maintenance processes. PMID:18385154

  6. A polycomb group protein, PHF1, is involved in the response to DNA double-strand breaks in human cell

    OpenAIRE

    Hong, Zehui; Jiang, Jie; Lan, Li; Nakajima, Satoshi; Kanno, Shin-ichiro; Koseki, Haruhiko; Yasui, Akira

    2008-01-01

    DNA double-strand breaks (DSBs) represent the most toxic DNA damage arisen from endogenous and exogenous genotoxic stresses and are known to be repaired by either homologous recombination or nonhomologous end-joining processes. Although many proteins have been identified to participate in either of the processes, the whole processes still remain elusive. Polycomb group (PcG) proteins are epigenetic chromatin modifiers involved in gene silencing, cancer development and the maintenance of embry...

  7. C-terminal low-complexity sequence repeats of Mycobacterium smegmatis Ku modulate DNA binding.

    Science.gov (United States)

    Kushwaha, Ambuj K; Grove, Anne

    2013-01-24

    Ku protein is an integral component of the NHEJ (non-homologous end-joining) pathway of DSB (double-strand break) repair. Both eukaryotic and prokaryotic Ku homologues have been characterized and shown to bind DNA ends. A unique feature of Mycobacterium smegmatis Ku is its basic C-terminal tail that contains several lysine-rich low-complexity PAKKA repeats that are absent from homologues encoded by obligate parasitic mycobacteria. Such PAKKA repeats are also characteristic of mycobacterial Hlp (histone-like protein) for which they have been shown to confer the ability to appose DNA ends. Unexpectedly, removal of the lysine-rich extension enhances DNA-binding affinity, but an interaction between DNA and the PAKKA repeats is indicated by the observation that only full-length Ku forms multiple complexes with a short stem-loop-containing DNA previously designed to accommodate only one Ku dimer. The C-terminal extension promotes DNA end-joining by T4 DNA ligase, suggesting that the PAKKA repeats also contribute to efficient end-joining. We suggest that low-complexity lysine-rich sequences have evolved repeatedly to modulate the function of unrelated DNA-binding proteins.

  8. Police must join the fast track to end AIDS by 2030.

    Science.gov (United States)

    Crofts, Nick; Patterson, David

    2016-01-01

    World leaders have committed to ending AIDS by 2030, but stigma and discrimination remain significant obstacles. In particular, police are critical, front-line determinants of risk for many people living with HIV (PLHIV) and members of other key affected populations (KAPs). The negative impact of adverse police behaviours and practices on HIV risk is well documented, and these risks undermine global efforts to end AIDS. Far less well documented, and less common, are attempts to ameliorate this impact by working to change police behaviours. This Special Issue seeks to help redress this imbalance by presenting a selection of original, provocative and important interventions from academics, police officers and other stakeholders concerned with documenting the potential for constructive, progressive and evidence-based approaches to the reduction of HIV risk. We recommend urgent boosting of efforts and funding to engage police in the HIV response. Among other strategies, this needs sustainable funding of programmes and their evaluation, and increased and continuing advocacy and education at all levels to match policy and law reform.

  9. The helicase domain of Polθ counteracts RPA to promote alt-NHEJ.

    Science.gov (United States)

    Mateos-Gomez, Pedro A; Kent, Tatiana; Deng, Sarah K; McDevitt, Shane; Kashkina, Ekaterina; Hoang, Trung M; Pomerantz, Richard T; Sfeir, Agnel

    2017-12-01

    Mammalian polymerase theta (Polθ) is a multifunctional enzyme that promotes error-prone DNA repair by alternative nonhomologous end joining (alt-NHEJ). Here we present structure-function analyses that reveal that, in addition to the polymerase domain, Polθ-helicase activity plays a central role during double-strand break (DSB) repair. Our results show that the helicase domain promotes chromosomal translocations by alt-NHEJ in mouse embryonic stem cells and also suppresses CRISPR-Cas9- mediated gene targeting by homologous recombination (HR). In vitro assays demonstrate that Polθ-helicase activity facilitates the removal of RPA from resected DSBs to allow their annealing and subsequent joining by alt-NHEJ. Consistent with an antagonistic role for RPA during alt-NHEJ, inhibition of RPA1 enhances end joining and suppresses recombination. Taken together, our results reveal that the balance between HR and alt-NHEJ is controlled by opposing activities of Polθ and RPA, providing further insight into the regulation of repair-pathway choice in mammalian cells.

  10. Transient disruption of non-homologous end-joining facilitates targeted genome manipulations in the filamentous fungus Aspergillus nidulans

    DEFF Research Database (Denmark)

    Nielsen, Jakob Blæsbjerg; Nielsen, Michael Lynge; Mortensen, Uffe Hasbro

    2008-01-01

    influences subsequent analyses of the manipulated strain. Our system will facilitate construction of large numbers of defined mutations in A. nidulans. Moreover, as the system can likely be adapted to other filamentous fungi, we expect it will be particularly beneficial in species where NHEJ cannot...... be restored by sexual crossing. (c) 2007 Elsevier Inc. All rights reserved....

  11. Precision Joining Center

    Science.gov (United States)

    Powell, J. W.; Westphal, D. A.

    1991-08-01

    A workshop to obtain input from industry on the establishment of the Precision Joining Center (PJC) was held on July 10-12, 1991. The PJC is a center for training Joining Technologists in advanced joining techniques and concepts in order to promote the competitiveness of U.S. industry. The center will be established as part of the DOE Defense Programs Technology Commercialization Initiative, and operated by EG&G Rocky Flats in cooperation with the American Welding Society and the Colorado School of Mines Center for Welding and Joining Research. The overall objectives of the workshop were to validate the need for a Joining Technologists to fill the gap between the welding operator and the welding engineer, and to assure that the PJC will train individuals to satisfy that need. The consensus of the workshop participants was that the Joining Technologist is a necessary position in industry, and is currently used, with some variation, by many companies. It was agreed that the PJC core curriculum, as presented, would produce a Joining Technologist of value to industries that use precision joining techniques. The advantage of the PJC would be to train the Joining Technologist much more quickly and more completely. The proposed emphasis of the PJC curriculum on equipment intensive and hands-on training was judged to be essential.

  12. Chromosome aberration model combining radiation tracks, chromatin structure, DSB repair and chromatin mobility

    International Nuclear Information System (INIS)

    Friedland, W.; Kundrat, P.

    2015-01-01

    The module that simulates the kinetics and yields of radiation-induced chromosome aberrations within the biophysical code PARTRAC is described. Radiation track structures simulated by Monte Carlo methods are overlapped with multi-scale models of DNA and chromatin to assess the resulting DNA damage. Spatial mobility of individual DNA ends from double-strand breaks is modelled simultaneously with their processing by the non-homologous end-joining enzymes. To score diverse types of chromosome aberrations, the joined ends are classified regarding their original chromosomal location, orientation and the involvement of centromeres. A comparison with experimental data on dicentrics induced by gamma and alpha particles shows that their relative dose dependence is predicted correctly, although the absolute yields are overestimated. The critical model assumptions on chromatin mobility and on the initial damage recognition and chromatin remodelling steps and their future refinements to solve this issue are discussed. (authors)

  13. Development of joining techniques for fabrication of fuel rod simulators

    International Nuclear Information System (INIS)

    Moorhead, A.J.; McCulloch, R.W.; Reed, R.W.; Woodhouse, J.J.

    1980-10-01

    Much of the safety-related thermal-hydraulic tests on nuclear reactors are conducted not in the reactor itself, but in mockup segments of a core that uses resistance-heated fuel rod simulators (FRS) in place of the radioactive fuel rods. Laser welding and furnace brazing techniques are described for joining subassemblies for FRS that have survived up to 1000 h steady-state operation at 700 to 1100 0 C cladding temperatures and over 5000 thermal transients, ranging from 10 to 100 0 C/s. A pulsed-laser welding procedure that includes use of small-diameter filler wire is used to join one end of a resistance heating element of Pt-8 W, Fe-22 Cr-5.5 Al-0.5 Co, or 80 Ni-20 Cr (wt %) to a tubular conductor of an appropriate intermediate material. The other end of the heating element is laser welded to an end plug, which in turn is welded to a central conductor rod

  14. A novel missense RAG-1 mutation results in T−B−NK+ SCID in Athabascan-speaking Dine Indians from the Canadian Northwest Territories

    OpenAIRE

    Xiao, Zheng; Yannone, Steven M; Dunn, Elizabeth; Cowan, Morton J

    2008-01-01

    DNA double-strand repair factors in the non-homologous end joining (NHEJ) pathway resolve DNA double-strand breaks introduced by the recombination-activating gene (RAG) proteins during V(D)J recombination of T and B lymphocyte receptor genes. Defective NHEJ and subsequent failure of V(D)J recombination leads to severe combined immunodeficiency disease (SCID). We originally linked T−B−NK+ SCID in Athabascan-speaking Native Americans in the Southwestern US and Northwest Territories of Canada to...

  15. Joining NbTi superconductors by ultrasonic welding

    International Nuclear Information System (INIS)

    Hafstrom, J.W.; Killpatrick, D.H.; Niemann, R.C.; Purcell, J.R.; Thresh, H.R.

    1976-01-01

    An important consideration in the design and construction of large, high-field, superconducting magnets is the capability to fabricate reliable, high-strength, low-resistance joints. A process for joining NbTi, copper stabilized, superconducting composites by ultrasonic welding is described. This process yields a joint strength comparable to that of the superconducting composite and a resistivity significantly lower than achieved by conventional soft soldering. The superconducting properties of the composite are not affected by the joining process. Scarfing the joint to maintain a constant conductor cross section does not degrade its electrical or mechanical properties. The application of the ultrasonic joining process, including process control, scarfing, and NDT procedures, in the construction of the superconducting magnet (U.S. SCMS) for the joint U. S. -- Soviet MHD program is described

  16. RPA accumulation during class switch recombination represents 5'-3' DNA-end resection during the S-G2/M phase of the cell cycle.

    Science.gov (United States)

    Yamane, Arito; Robbiani, Davide F; Resch, Wolfgang; Bothmer, Anne; Nakahashi, Hirotaka; Oliveira, Thiago; Rommel, Philipp C; Brown, Eric J; Nussenzweig, Andre; Nussenzweig, Michel C; Casellas, Rafael

    2013-01-31

    Activation-induced cytidine deaminase (AID) promotes chromosomal translocations by inducing DNA double-strand breaks (DSBs) at immunoglobulin (Ig) genes and oncogenes in the G1 phase. RPA is a single-stranded DNA (ssDNA)-binding protein that associates with resected DSBs in the S phase and facilitates the assembly of factors involved in homologous repair (HR), such as Rad51. Notably, RPA deposition also marks sites of AID-mediated damage, but its role in Ig gene recombination remains unclear. Here, we demonstrate that RPA associates asymmetrically with resected ssDNA in response to lesions created by AID, recombination-activating genes (RAG), or other nucleases. Small amounts of RPA are deposited at AID targets in G1 in an ATM-dependent manner. In contrast, recruitment in the S-G2/M phase is extensive, ATM independent, and associated with Rad51 accumulation. In the S-G2/M phase, RPA increases in nonhomologous-end-joining-deficient lymphocytes, where there is more extensive DNA-end resection. Thus, most RPA recruitment during class switch recombination represents salvage of unrepaired breaks by homology-based pathways during the S-G2/M phase of the cell cycle. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  17. Mouse embryonic stem cells, but not somatic cells, predominantly use homologous recombination to repair double-strand DNA breaks.

    Science.gov (United States)

    Tichy, Elisia D; Pillai, Resmi; Deng, Li; Liang, Li; Tischfield, Jay; Schwemberger, Sandy J; Babcock, George F; Stambrook, Peter J

    2010-11-01

    Embryonic stem (ES) cells give rise to all cell types of an organism. Since mutations at this embryonic stage would affect all cells and be detrimental to the overall health of an organism, robust mechanisms must exist to ensure that genomic integrity is maintained. To test this proposition, we compared the capacity of murine ES cells to repair DNA double-strand breaks with that of differentiated cells. Of the 2 major pathways that repair double-strand breaks, error-prone nonhomologous end joining (NHEJ) predominated in mouse embryonic fibroblasts, whereas the high fidelity homologous recombinational repair (HRR) predominated in ES cells. Microhomology-mediated end joining, an emerging repair pathway, persisted at low levels in all cell types examined. The levels of proteins involved in HRR and microhomology-mediated end joining were highly elevated in ES cells compared with mouse embryonic fibroblasts, whereas those for NHEJ were quite variable, with DNA Ligase IV expression low in ES cells. The half-life of DNA Ligase IV protein was also low in ES cells. Attempts to increase the abundance of DNA Ligase IV protein by overexpression or inhibition of its degradation, and thereby elevate NHEJ in ES cells, were unsuccessful. When ES cells were induced to differentiate, however, the level of DNA Ligase IV protein increased, as did the capacity to repair by NHEJ. The data suggest that preferential use of HRR rather than NHEJ may lend ES cells an additional layer of genomic protection and that the limited levels of DNA Ligase IV may account for the low level of NHEJ activity.

  18. Human longevity and variation in DNA damage response and repair

    DEFF Research Database (Denmark)

    Debrabant, Birgit; Soerensen, Mette; Flachsbart, Friederike

    2014-01-01

    others. Data were applied on 592 SNPs from 77 genes involved in nine sub-processes: DNA-damage response, base excision repair (BER), nucleotide excision repair, mismatch repair, non-homologous end-joining, homologous recombinational repair (HRR), RecQ helicase activities (RECQ), telomere functioning...... in genotyping procedures and investigated SNPs, potentially inducing differences in the coverage of gene regions. Specifically, five genes were not covered at all in the German data. Therefore, investigations in additional study populations are needed before final conclusion can be drawn....

  19. Fast and scalable inequality joins

    KAUST Repository

    Khayyat, Zuhair

    2016-09-07

    Inequality joins, which is to join relations with inequality conditions, are used in various applications. Optimizing joins has been the subject of intensive research ranging from efficient join algorithms such as sort-merge join, to the use of efficient indices such as (Formula presented.)-tree, (Formula presented.)-tree and Bitmap. However, inequality joins have received little attention and queries containing such joins are notably very slow. In this paper, we introduce fast inequality join algorithms based on sorted arrays and space-efficient bit-arrays. We further introduce a simple method to estimate the selectivity of inequality joins which is then used to optimize multiple predicate queries and multi-way joins. Moreover, we study an incremental inequality join algorithm to handle scenarios where data keeps changing. We have implemented a centralized version of these algorithms on top of PostgreSQL, a distributed version on top of Spark SQL, and an existing data cleaning system, Nadeef. By comparing our algorithms against well-known optimization techniques for inequality joins, we show our solution is more scalable and several orders of magnitude faster. © 2016 Springer-Verlag Berlin Heidelberg

  20. Development of reconstitution method for surveillance specimens using surface activated joining

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Terumi; Kaihara, Shoichiro; Yoshida, Kazuo; Sato, Akira [Ishikawajima-Harima Heavy Industries Co. Ltd., Tokyo (Japan); Onizawa, Kunio; Nishiyama, Yutaka; Fukaya, Kiyoshi; Suzuki, Masahide

    1996-03-01

    Evaluation of embrittlement of reactor vessel steel due to irradiation requires surveillance tests. However, many surveillance specimens are necessary for nuclear plants life extension. Therefore, a specimen reconstitution technique has become important to provide the many specimens for continued surveillance. A surface activated joining (SAJ) method has been developed to join various materials together at low temperatures with little deformation, and is useful to bond irradiated specimens. To assess the validity of this method, Charpy impact tests were carried out, and the characteristics caused by heating during joining were measured. The test results showed the Charpy impact values were almost the same as base materials, and surface activated joining reduced heat affected zone to less than 2 mm. (author).

  1. Nonhomologous recombination between defective poliovirus and coxsackievirus genomes suggests a new model of genetic plasticity for picornaviruses.

    Science.gov (United States)

    Holmblat, Barbara; Jégouic, Sophie; Muslin, Claire; Blondel, Bruno; Joffret, Marie-Line; Delpeyroux, Francis

    2014-08-05

    Most of the circulating vaccine-derived polioviruses (cVDPVs) implicated in poliomyelitis outbreaks in Madagascar have been shown to be recombinants between the type 2 poliovirus (PV) strain of the oral polio vaccine (Sabin 2) and another species C human enterovirus (HEV-C), such as type 17 coxsackie A virus (CA17) in particular. We studied intertypic genetic exchanges between PV and non-PV HEV-C by developing a recombination model, making it possible to rescue defective type 2 PV RNA genomes with a short deletion at the 3' end by the cotransfection of cells with defective or infectious CA17 RNAs. We isolated over 200 different PV/CA17 recombinants, using murine cells expressing the human PV receptor (PVR) and selecting viruses with PV capsids. We found some homologous (H) recombinants and, mostly, nonhomologous (NH) recombinants presenting duplications of parental sequences preferentially located in the regions encoding proteins 2A, 2B, and 3A. Short duplications appeared to be stable, whereas longer duplications were excised during passaging in cultured cells or after multiplication in PVR-transgenic mice, generating H recombinants with diverse sites of recombination. This suggests that NH recombination events may be a transient, intermediate step in the generation and selection of the fittest H recombinants. In addition to the classical copy-choice mechanism of recombination thought to generate mostly H recombinants, there may also be a modular mechanism of recombination, involving NH recombinant precursors, shaping the genomes of recombinant enteroviruses and other picornaviruses. Importance: The multiplication of circulating vaccine-derived polioviruses (cVDPVs) in poorly immunized human populations can render these viruses pathogenic, causing poliomyelitis outbreaks. Most cVDPVs are intertypic recombinants between a poliovirus (PV) strain and another human enterovirus, such as type 17 coxsackie A viruses (CA17). For further studies of the genetic exchanges

  2. News Release: USDA Joins Fair Food Network, State and Local Partners to

    Science.gov (United States)

    vitamin C, calcium, and iron. The Food Bank of Eastern Michigan in Flint is distributing those USDA Foods Joins Fair Food Network, State and Local Partners to Promote Nutrition Resources for Lead-Affected Flint Residents News Release: USDA Joins Fair Food Network, State and Local Partners to Promote Nutrition

  3. The DNA-dependent protein kinase: a multifunctional protein kinase with roles in DNA double strand break repair and mitosis

    OpenAIRE

    Jette, Nicholas; Lees-Miller, Susan P.

    2014-01-01

    The DNA-dependent protein kinase (DNA-PK) is a serine/threonine protein kinase composed of a large catalytic subunit (DNA-PKcs) and the Ku70/80 heterodimer. Over the past two decades, significant progress has been made in elucidating the role of DNA-PK in non-homologous end joining (NHEJ), the major pathway for repair of ionizing radiation-induced DNA double strand breaks in human cells and recently, additional roles for DNA-PK have been reported. In this review, we will describe the biochemi...

  4. Molecular dynamics simulation of joining process of Ag-Au nanowires and mechanical properties of the hybrid nanojoint

    Directory of Open Access Journals (Sweden)

    Su Ding

    2015-05-01

    Full Text Available The nanojoining process of Ag-Au hybrid nanowires at 800K was comprehensively studied by virtue of molecular dynamics (MD simulation. Three kinds of configurations including end-to-end, T-like and X-like were built in the simulation aiming to understand the nanojoining mechanism. The detailed dynamic evolution of atoms, crystal structure transformation and defects development during the nanojoining processes were performed. The results indicate that there are two stages in the nanojoining process of Ag-Au nanowires which are atom diffusion and new bonds formation. Temperature is a key parameter affecting both stages ascribed to the energy supply and the optimum temperature for Ag-Au nanojoint with diameter of 4.08 nm has been discussed. The mechanical properties of the nanojoint were examined with simulation of tensile test on the end-to-end joint. It was revealed that the nanojoint was strong enough to resist fracture at the joining area.

  5. Joining of advanced materials

    CERN Document Server

    Messler, Robert W

    1993-01-01

    Provides an unusually complete and readable compilation of the primary and secondary options for joining conventional materials in non-conventional ways. Provides unique coverage of adhesive bonding using both organic and inorganic adhesives, cements and mortars. Focuses on materials issues without ignoring issues related to joint design, production processing, quality assurance, process economics, and joining performance in service.Joining of advanced materials is a unique treatment of joining of both conventional and advanced metals andalloys, intermetallics, ceramics, glasses, polymers, a

  6. Joining by plastic deformation

    DEFF Research Database (Denmark)

    Mori, Ken-ichiro; Bay, Niels; Fratini, Livan

    2013-01-01

    As the scale and complexity of products such as aircraft and cars increase, demand for new functional processes to join mechanical parts grows. The use of plastic deformation for joining parts potentially offers improved accuracy, reliability and environmental safety as well as creating opportuni......As the scale and complexity of products such as aircraft and cars increase, demand for new functional processes to join mechanical parts grows. The use of plastic deformation for joining parts potentially offers improved accuracy, reliability and environmental safety as well as creating...

  7. Ceramic joining

    Energy Technology Data Exchange (ETDEWEB)

    Loehman, R.E. [Sandia National Lab., Albuquerque, NM (United States)

    1996-04-01

    This paper describes the relation between reactions at ceramic-metal interfaces and the development of strong interfacial bonds in ceramic joining. Studies on a number of systems are described, including silicon nitrides, aluminium nitrides, mullite, and aluminium oxides. Joints can be weakened by stresses such as thermal expansion mismatch. Ceramic joining is used in a variety of applications such as solid oxide fuel cells.

  8. Double-strand break repair-adox: Restoration of suppressed double-strand break repair during mitosis induces genomic instability.

    Science.gov (United States)

    Terasawa, Masahiro; Shinohara, Akira; Shinohara, Miki

    2014-12-01

    Double-strand breaks (DSBs) are one of the severest types of DNA damage. Unrepaired DSBs easily induce cell death and chromosome aberrations. To maintain genomic stability, cells have checkpoint and DSB repair systems to respond to DNA damage throughout most of the cell cycle. The failure of this process often results in apoptosis or genomic instability, such as aneuploidy, deletion, or translocation. Therefore, DSB repair is essential for maintenance of genomic stability. During mitosis, however, cells seem to suppress the DNA damage response and proceed to the next G1 phase, even if there are unrepaired DSBs. The biological significance of this suppression is not known. In this review, we summarize recent studies of mitotic DSB repair and discuss the mechanisms of suppression of DSB repair during mitosis. DSB repair, which maintains genomic integrity in other phases of the cell cycle, is rather toxic to cells during mitosis, often resulting in chromosome missegregation and aberration. Cells have multiple safeguards to prevent genomic instability during mitosis: inhibition of 53BP1 or BRCA1 localization to DSB sites, which is important to promote non-homologous end joining or homologous recombination, respectively, and also modulation of the non-homologous end joining core complex to inhibit DSB repair. We discuss how DSBs during mitosis are toxic and the multiple safeguard systems that suppress genomic instability. © 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

  9. Highly efficient generation of knock-in transgenic medaka by CRISPR/Cas9-mediated genome engineering.

    Science.gov (United States)

    Watakabe, Ikuko; Hashimoto, Hisashi; Kimura, Yukiko; Yokoi, Saori; Naruse, Kiyoshi; Higashijima, Shin-Ichi

    2018-01-01

    Medaka ( Oryzias latipes ) is a popular animal model used in vertebrate genetic analysis. Recently, an efficient (~ 30%) knock-in system via non-homologous end joining (NHEJ) was established in zebrafish using the CRISPR/Cas9 system. If the same technique were applicable in medaka, it would greatly expand the usefulness of this model organism. The question of the applicability of CRISPR/Cas9 in medaka, however, has yet to be addressed. We report the highly efficient generation of knock-in transgenic medaka via non-homologous end joining (NHEJ). Donor plasmid containing a heat-shock promoter and a reporter gene was co-injected with a short guide RNA (sgRNA) targeted for genome digestion, an sgRNA targeted for donor plasmid digestion, and Cas9 mRNA. Broad transgene expression in the expression domain of a target gene was observed in approximately 25% of injected embryos. By raising these animals, we established stable knock-in transgenic fish with several different constructs for five genetic loci, obtaining transgenic founders at efficiencies of > 50% for all five loci. Further, we show that the method is useful for obtaining mutant alleles. In the experiments where transgene integrations were targeted between the transcription start site and the initiation methionine, the resultant transgenic fish became mutant alleles. With its simplicity, design flexibility, and high efficiency, we propose that CRISPR/Cas9-mediated knock-in via NHEJ will become a standard method for the generation of transgenic and mutant medaka.

  10. Engineering a Nickase on the Homing Endonuclease I-DmoI Scaffold

    DEFF Research Database (Denmark)

    Molina, Rafael; Marcaida, María José; Redondo, Pilar

    2015-01-01

    strand break could be an approach to reduce the toxicity associated with non-homologous end joining by promoting the use of homologous recombination to repair the cleavage of a single DNA break. Taking advantage of the sequential DNA cleavage mechanism of I-DmoI LAGLIDADG homing endonuclease, we have......Homing endonucleases are useful tools for genome modification because of their capability to recognize and cleave specifically large DNA targets. These endonucleases generate a DNA double strand break that can be repaired by the DNA damage response machinery. The break can be repaired by homologous...

  11. Induction of DNA deletions after UV-light irradiation in yeast Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Stepanova, A.N.; Koltovaya, N.A.

    2008-01-01

    We study mutagenic action of such a damaging agent as UV light, which can lead to DNA double-strand breaks (DSB). DNA deletions and gross rearrangements occur in process of DSB repair. We show that UV light induces deletion and rearrangement very efficiently. Analysis of efficacy of different types of repair shows that cell tries to repair DSBs with a combination of both homologous recombination (HR) and nonhomologous end joining (NHEJ) if available and that DSB repair by HR is more effective than by NHEJ in growing culture of haploid yeast

  12. A new type of radiosensitive T–B–NK+ severe combined immunodeficiency caused by a LIG4 mutation

    OpenAIRE

    van der Burg, Mirjam; van Veelen, Lieneke R.; Verkaik, Nicole S.; Wiegant, Wouter W.; Hartwig, Nico G.; Barendregt, Barbara H.; Brugmans, Linda; Raams, Anja; Jaspers, Nicolaas G.J.; Zdzienicka, Malgorzata Z.; van Dongen, Jacques J.M.; van Gent, Dik C.

    2005-01-01

    textabstractV(D)J recombination of Ig and TCR loci is a stepwise process during which site-specific DNA double-strand breaks (DSBs) are made by RAG1/RAG2, followed by DSB repair by nonhomologous end joining. Defects in V(D)J recombination result in SCID characterized by absence of mature B and T cells. A subset of T-B-NK+ SCID patients is sensitive to ionizing radiation, and the majority of these patients have mutations in Artemis. We present a patient with a new type of radiosensitive T-B-NK...

  13. Fast and sensitive detection of indels induced by precise gene targeting

    DEFF Research Database (Denmark)

    Yang, Zhang; Steentoft, Catharina; Hauge, Camilla

    2015-01-01

    The nuclease-based gene editing tools are rapidly transforming capabilities for altering the genome of cells and organisms with great precision and in high throughput studies. A major limitation in application of precise gene editing lies in lack of sensitive and fast methods to detect...... and characterize the induced DNA changes. Precise gene editing induces double-stranded DNA breaks that are repaired by error-prone non-homologous end joining leading to introduction of insertions and deletions (indels) at the target site. These indels are often small and difficult and laborious to detect...

  14. Human nucleolus organizers on nonhomologous chromosomes can share the same ribosomal gene variants.

    Science.gov (United States)

    Krystal, M; D'Eustachio, P; Ruddle, F H; Arnheim, N

    1981-01-01

    The distributions of three human ribosomal gene polymorphisms among individual chromosomes containing nucleolus organizers were analyzed by using mouse--human hybrid cells. Different nucleolus organizers can contain the same variant, suggesting the occurrence of genetic exchanges among ribosomal gene clusters on nonhomologous chromosomes. Such exchanges appear to occur less frequently in mice. This difference is discussed in terms of the nucleolar organization and chromosomal location of ribosomal gene clusters in humans and mice. Images PMID:6272316

  15. Join Operations in Temporal Databases

    DEFF Research Database (Denmark)

    Gao, D.; Jensen, Christian Søndergaard; Snodgrass, R.T.

    2005-01-01

    Joins are arguably the most important relational operators. Poor implementations are tantamount to computing the Cartesian product of the input relations. In a temporal database, the problem is more acute for two reasons. First, conventional techniques are designed for the evaluation of joins...... with equality predicates rather than the inequality predicates prevalent in valid-time queries. Second, the presence of temporally varying data dramatically increases the size of a database. These factors indicate that specialized techniques are needed to efficiently evaluate temporal joins. We address...... this need for efficient join evaluation in temporal databases. Our purpose is twofold. We first survey all previously proposed temporal join operators. While many temporal join operators have been defined in previous work, this work has been done largely in isolation from competing proposals, with little...

  16. Synthetic lethality between murine DNA repair factors XLF and DNA-PKcs is rescued by inactivation of Ku70

    DEFF Research Database (Denmark)

    Xing, Mengtan; Bjørås, Magnar; Daniel, Jeremy A

    2017-01-01

    DNA double-strand breaks (DSBs) are recognized and repaired by the Classical Non-Homologous End-Joining (C-NHEJ) and Homologous Recombination pathways. C-NHEJ includes the core Ku70 and Ku80 (or Ku86) heterodimer that binds DSBs and thus promotes recruitment of accessory downstream NHEJ factors XLF......, PAXX, DNA-PKcs, Artemis and other core subunits, XRCC4 and DNA Ligase 4 (Lig4). In the absence of core C-NHEJ factors, DNA repair can be performed by Alternative End-Joining, which likely depends on DNA Ligase 1 and DNA Ligase 3. Genetic inactivation of C-NHEJ factors, such as Ku70, Ku80, XLF, PAXX...... with severe apoptosis in the central nervous system. Here, we demonstrate that inactivation of the Ku70 gene rescues the synthetic lethality between XLF and DNA-PKcs, resulting in triple knockout mice that are indistinguishable from Ku70-deficient littermates by size or levels of genomic instability. Moreover...

  17. Cdk1 Restrains NHEJ through Phosphorylation of XRCC4-like Factor Xlf1

    Directory of Open Access Journals (Sweden)

    Pierre Hentges

    2014-12-01

    Full Text Available Eukaryotic cells use two principal mechanisms for repairing DNA double-strand breaks (DSBs: homologous recombination (HR and nonhomologous end-joining (NHEJ. DSB repair pathway choice is strongly regulated during the cell cycle. Cyclin-dependent kinase 1 (Cdk1 activates HR by phosphorylation of key recombination factors. However, a mechanism for regulating the NHEJ pathway has not been established. Here, we report that Xlf1, a fission yeast XLF ortholog, is a key regulator of NHEJ activity in the cell cycle. We show that Cdk1 phosphorylates residues in the C terminus of Xlf1 over the course of the cell cycle. Mutation of these residues leads to the loss of Cdk1 phosphorylation, resulting in elevated levels of NHEJ repair in vivo. Together, these data establish that Xlf1 phosphorylation by Cdc2Cdk1 provides a molecular mechanism for downregulation of NHEJ in fission yeast and indicates that XLF is a key regulator of end-joining processes in eukaryotic organisms.

  18. End of Life Care

    Science.gov (United States)

    ... Related Topics Choosing Wisely Join our e-newsletter! Aging & Health A to Z End of Life Care ... be used. Ice chips, popsicles, moist swabs, or artificial saliva can help prevent the mouth from becoming ...

  19. Interdependence of the rad50 hook and globular domain functions.

    Science.gov (United States)

    Hohl, Marcel; Kochańczyk, Tomasz; Tous, Cristina; Aguilera, Andrés; Krężel, Artur; Petrini, John H J

    2015-02-05

    Rad50 contains a conserved Zn(2+) coordination domain (the Rad50 hook) that functions as a homodimerization interface. Hook ablation phenocopies Rad50 deficiency in all respects. Here, we focused on rad50 mutations flanking the Zn(2+)-coordinating hook cysteines. These mutants impaired hook-mediated dimerization, but recombination between sister chromatids was largely unaffected. This may reflect that cohesin-mediated sister chromatid interactions are sufficient for double-strand break repair. However, Mre11 complex functions specified by the globular domain, including Tel1 (ATM) activation, nonhomologous end joining, and DNA double-strand break end resection were affected, suggesting that dimerization exerts a broad influence on Mre11 complex function. These phenotypes were suppressed by mutations within the coiled-coil and globular ATPase domains, suggesting a model in which conformational changes in the hook and globular domains are transmitted via the extended coils of Rad50. We propose that transmission of spatial information in this manner underlies the regulation of Mre11 complex functions. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Blind-date Conversation Joining

    Directory of Open Access Journals (Sweden)

    Luca Cesari

    2013-07-01

    Full Text Available We focus on a form of joining conversations among multiple parties in service-oriented applications where a client may asynchronously join an existing conversation without need to know in advance any information about it. More specifically, we show how the correlation mechanism provided by orchestration languages enables a form of conversation joining that is completely transparent to clients and that we call 'blind-date joining'. We provide an implementation of this strategy by using the standard orchestration language WS-BPEL. We then present its formal semantics by resorting to COWS, a process calculus specifically designed for modelling service-oriented applications. We illustrate our approach by means of a simple, but realistic, case study from the online games domain.

  1. Lightning Fast and Space Efficient Inequality Joins

    KAUST Repository

    Khayyat, Zuhair

    2015-09-01

    Inequality joins, which join relational tables on inequality conditions, are used in various applications. While there have been a wide range of optimization methods for joins in database systems, from algorithms such as sort-merge join and band join, to various indices such as B+-tree,R*-tree and Bitmap, inequality joins have received little attention and queries containing such joins are usually very slow. In this paper, we introduce fast inequality join algorithms. We put columns to be joined in sorted arrays and we use permutation arrays to encode positions of tuples in one sorted array w.r.t. the other sorted array. In contrast to sort-merge join, we use space effcient bit-arrays that enable optimizations, such as Bloom filter indices, for fast computation of the join results. We have implemented a centralized version of these algorithms on top of PostgreSQL, and a distributed version on top of Spark SQL. We have compared against well known optimization techniques for inequality joins and show that our solution is more scalable and several orders of magnitude faster.

  2. Lightning fast and space efficient inequality joins

    KAUST Repository

    Khayyat, Zuhair

    2015-09-01

    Inequality joins, which join relational tables on inequality conditions, are used in various applications. While there have been a wide range of optimization methods for joins in database systems, from algorithms such as sort-merge join and band join, to various indices such as B+-tree, R*-tree and Bitmap, inequality joins have received little attention and queries containing such joins are usually very slow. In this paper, we introduce fast inequality join algorithms. We put columns to be joined in sorted arrays and we use permutation arrays to encode positions of tuples in one sorted array w.r.t. the other sorted array. In contrast to sort-merge join, we use space efficient bit-arrays that enable optimizations, such as Bloom filter indices, for fast computation of the join results. We have implemented a centralized version of these algorithms on top of PostgreSQL, and a distributed version on top of Spark SQL. We have compared against well known optimization techniques for inequality joins and show that our solution is more scalable and several orders of magnitude faster.

  3. Lightning fast and space efficient inequality joins

    KAUST Repository

    Khayyat, Zuhair; Lucia, William; Singh, Meghna; Ouzzani, Mourad; Papotti, Paolo; Quiané -Ruiz, Jorge-Arnulfo; Tang, Nan; Kalnis, Panos

    2015-01-01

    Inequality joins, which join relational tables on inequality conditions, are used in various applications. While there have been a wide range of optimization methods for joins in database systems, from algorithms such as sort-merge join and band join, to various indices such as B+-tree, R*-tree and Bitmap, inequality joins have received little attention and queries containing such joins are usually very slow. In this paper, we introduce fast inequality join algorithms. We put columns to be joined in sorted arrays and we use permutation arrays to encode positions of tuples in one sorted array w.r.t. the other sorted array. In contrast to sort-merge join, we use space efficient bit-arrays that enable optimizations, such as Bloom filter indices, for fast computation of the join results. We have implemented a centralized version of these algorithms on top of PostgreSQL, and a distributed version on top of Spark SQL. We have compared against well known optimization techniques for inequality joins and show that our solution is more scalable and several orders of magnitude faster.

  4. Summary of Dissimilar Metal Joining Trials Conducted by Edison Welding Institute

    Energy Technology Data Exchange (ETDEWEB)

    MJ Lambert

    2005-11-18

    Under the direction of the NASA-Glenn Research Center, the Edison Welding Institute (EWI) in Columbus, OH performed a series of non-fusion joining experiments to determine the feasibility of joining refractory metals or refractory metal alloys to Ni-based superalloys. Results, as reported by EWI, can be found in the project report for EWI Project 48819GTH (Attachment A, at the end of this document), dated October 10, 2005. The three joining methods used in this investigation were inertia welding, magnetic pulse welding, and electro-spark deposition joining. Five materials were used in these experiments: Mo-47Re, T-111, Hastelloy X, Mar M-247 (coarse-grained, 0.5 mm to several millimeter average grain size), and Mar M-247 (fine-grained, approximately 50 {micro}m average grain size). Several iterative trials of each material combination with each joining method were performed to determine the best practice joining method. Mo-47Re was found to be joined easily to Hastelloy X via inertia welding, but inertia welding of the Mo-alloy to both Mar M-247 alloys resulted in inconsistent joint strength and large reaction layers between the two metals. T-111 was found to join well to Hastelloy X and coarse-grained Mar M-247 via inertia welding, but joining to fine-grained Mar M-247 resulted in low joint strength. Magnetic pulse welding (MPW) was only successful in joining T-111 tubing to Hastelloy X bar stock. The joint integrity and reaction layer between the metals were found to be acceptable. This single joining trial, however, caused damage to the electromagnetic concentrators used in this process. Subsequent design efforts to eliminate the problem resulted in a loss of power imparted to the accelerating work piece, and results could not be reproduced. Welding trials of Mar M-247 to T-111 resulted in catastrophic failure of the bar stock, even at lower power. Electro-spark deposition joining of Mo-47Re, in which the deposited material was Hastelloy X, did not have a

  5. Summary of Dissimilar Metal Joining Trials Conducted by Edison Welding Institute

    International Nuclear Information System (INIS)

    MJ Lambert

    2005-01-01

    Under the direction of the NASA-Glenn Research Center, the Edison Welding Institute (EWI) in Columbus, OH performed a series of non-fusion joining experiments to determine the feasibility of joining refractory metals or refractory metal alloys to Ni-based superalloys. Results, as reported by EWI, can be found in the project report for EWI Project 48819GTH (Attachment A, at the end of this document), dated October 10, 2005. The three joining methods used in this investigation were inertia welding, magnetic pulse welding, and electro-spark deposition joining. Five materials were used in these experiments: Mo-47Re, T-111, Hastelloy X, Mar M-247 (coarse-grained, 0.5 mm to several millimeter average grain size), and Mar M-247 (fine-grained, approximately 50 (micro)m average grain size). Several iterative trials of each material combination with each joining method were performed to determine the best practice joining method. Mo-47Re was found to be joined easily to Hastelloy X via inertia welding, but inertia welding of the Mo-alloy to both Mar M-247 alloys resulted in inconsistent joint strength and large reaction layers between the two metals. T-111 was found to join well to Hastelloy X and coarse-grained Mar M-247 via inertia welding, but joining to fine-grained Mar M-247 resulted in low joint strength. Magnetic pulse welding (MPW) was only successful in joining T-111 tubing to Hastelloy X bar stock. The joint integrity and reaction layer between the metals were found to be acceptable. This single joining trial, however, caused damage to the electromagnetic concentrators used in this process. Subsequent design efforts to eliminate the problem resulted in a loss of power imparted to the accelerating work piece, and results could not be reproduced. Welding trials of Mar M-247 to T-111 resulted in catastrophic failure of the bar stock, even at lower power. Electro-spark deposition joining of Mo-47Re, in which the deposited material was Hastelloy X, did not have a

  6. FANCD2 Maintains Fork Stability in BRCA1/2-Deficient Tumors and Promotes Alternative End-Joining DNA Repair

    Directory of Open Access Journals (Sweden)

    Zeina Kais

    2016-06-01

    Full Text Available BRCA1/2 proteins function in homologous recombination (HR-mediated DNA repair and cooperate with Fanconi anemia (FA proteins to maintain genomic integrity through replication fork stabilization. Loss of BRCA1/2 proteins results in DNA repair deficiency and replicative stress, leading to genomic instability and enhanced sensitivity to DNA-damaging agents. Recent studies have shown that BRCA1/2-deficient tumors upregulate Polθ-mediated alternative end-joining (alt-EJ repair as a survival mechanism. Whether other mechanisms maintain genomic integrity upon loss of BRCA1/2 proteins is currently unknown. Here we show that BRCA1/2-deficient tumors also upregulate FANCD2 activity. FANCD2 is required for fork protection and fork restart in BRCA1/2-deficient tumors. Moreover, FANCD2 promotes Polθ recruitment at sites of damage and alt-EJ repair. Finally, loss of FANCD2 in BRCA1/2-deficient tumors enhances cell death. These results reveal a synthetic lethal relationship between FANCD2 and BRCA1/2, and they identify FANCD2 as a central player orchestrating DNA repair pathway choice at the replication fork.

  7. Joining uranium to steel

    International Nuclear Information System (INIS)

    Perkins, M.A.

    1976-05-01

    A method has been devised which will allow the joining of uranium to steel by fusion welding through the use of an intermediate material. Uranium-0.5 titanium was joined to AISI 304L stainless steel by using a vanadium insert. Also, a method is now available for selecting possible filler metals when two entirely dissimilar metals need to be joined. This method allows a quantitative ranking to be made of the possible filler metals and thus the most likely candidate can be selected

  8. Authenticated join processing in outsourced databases

    KAUST Repository

    Yang, Yin

    2009-01-01

    Database outsourcing requires that a query server constructs a proof of result correctness, which can be verified by the client using the data owner\\'s signature. Previous authentication techniques deal with range queries on a single relation using an authenticated data structure (ADS). On the other hand, authenticated join processing is inherently more complex than ranges since only the base relations (but not their combination) are signed by the owner. In this paper, we present three novel join algorithms depending on the ADS availability: (i) Authenticated Indexed Sort Merge Join (AISM), which utilizes a single ADS on the join attribute, (ii) Authenticated Index Merge Join (AIM) that requires an ADS (on the join attribute) for both relations, and (iii) Authenticated Sort Merge Join (ASM), which does not rely on any ADS. We experimentally demonstrate that the proposed methods outperform two benchmark algorithms, often by several orders of magnitude, on all performance metrics, and effectively shift the workload to the outsourcing service. Finally, we extend our techniques to complex queries that combine multi-way joins with selections and projections. ©2009 ACM.

  9. Authenticated join processing in outsourced databases

    KAUST Repository

    Yang, Yin; Papadias, Dimitris; Papadopoulos, Stavros; Kalnis, Panos

    2009-01-01

    Database outsourcing requires that a query server constructs a proof of result correctness, which can be verified by the client using the data owner's signature. Previous authentication techniques deal with range queries on a single relation using an authenticated data structure (ADS). On the other hand, authenticated join processing is inherently more complex than ranges since only the base relations (but not their combination) are signed by the owner. In this paper, we present three novel join algorithms depending on the ADS availability: (i) Authenticated Indexed Sort Merge Join (AISM), which utilizes a single ADS on the join attribute, (ii) Authenticated Index Merge Join (AIM) that requires an ADS (on the join attribute) for both relations, and (iii) Authenticated Sort Merge Join (ASM), which does not rely on any ADS. We experimentally demonstrate that the proposed methods outperform two benchmark algorithms, often by several orders of magnitude, on all performance metrics, and effectively shift the workload to the outsourcing service. Finally, we extend our techniques to complex queries that combine multi-way joins with selections and projections. ©2009 ACM.

  10. Identification and analysis of Ku70 and Ku80 homologs in the koji molds Aspergillus sojae and Aspergillus oryzae.

    Science.gov (United States)

    Takahashi, Tadashi; Masuda, Tsutomu; Koyama, Yasuji

    2006-01-01

    Ku genes play a key role in the non-homologous end-joining pathway. We have identified Ku70 and Ku80 homologs in the koji molds Aspergillus sojae and Aspergillus oryzae, and have constructed the disruption mutants of Ku70, Ku80, and Ku70-80 to characterize the phenotypic change in these mutants. Neither Ku70- nor Ku80-disrupted strains show hypersensitivity to the DNA damaging agents methylmethane sulfonate (MMS) and phleomycin. Moreover, undesirable phenotypes, such as poor growth or repressed conidiospore formation, were not observed in the Ku-disrupted A. sojae and A. oryzae.

  11. Genome engineering with TALENs and ZFNs: repair pathways and donor design.

    Science.gov (United States)

    Carroll, Dana; Beumer, Kelly J

    2014-09-01

    Genome engineering with targetable nucleases depends on cellular pathways of DNA repair after target cleavage. Knowledge of how those pathways work, their requirements and their active factors, can guide experimental design and improve outcomes. While many aspects of both homologous recombination (HR) and nonhomologous end joining (NHEJ) are shared by a broad range of cells and organisms, some features are specific to individual situations. This article reviews the influence of repair mechanisms on the results of gene targeting experiments, with an emphasis on lessons learned from experiments with Drosophila. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. [CRISPR/CAS9, the King of Genome Editing Tools].

    Science.gov (United States)

    Bannikov, A V; Lavrov, A V

    2017-01-01

    The discovery of CRISPR/Cas9 brought a hope for having an efficient, reliable, and readily available tool for genome editing. CRISPR/Cas9 is certainly easy to use, while its efficiency and reliability remain the focus of studies. The review describes the general principles of the organization and function of Cas nucleases and a number of important issues to be considered while planning genome editing experiments with CRISPR/Cas9. The issues include evaluation of the efficiency and specificity for Cas9, sgRNA selection, Cas9 variants designed artificially, and use of homologous recombination and nonhomologous end joining in DNA editing.

  13. Live cell imaging reveals at novel view of DNA

    International Nuclear Information System (INIS)

    Moritomi-Yano, Keiko; Yano, Ken-ichi

    2010-01-01

    Non-homologous end-joining (NHEJ) is the major repair pathway for DNA double-strand breaks (DSBs) that are the most severe form of DNA damages. Recently, live cell imaging techniques coupled with laser micro-irradiation were used to analyze the spatio-temporal behavior of the NHEJ core factors upon DSB induction in living cells. Based on the live cell imaging studies, we proposed a novel two-phase model for DSB sensing and protein assembly in the NHEJ pathway. This new model provides a novel view of the dynamic protein behavior on DSBs and broad implications for the molecular mechanism of NHEJ. (author)

  14. Efficient Oligo nucleotide mediated CRISPR-Cas9 Gene Editing in Aspergilli

    DEFF Research Database (Denmark)

    Nødvig, Christina Spuur; Hoof, Jakob Blæsbjerg; Kogle, Martin Engelhard

    2018-01-01

    CRISPR-Cas9 technologies are revolutionizing fungal gene editing. Here we show that survival of specific Cas9/sgRNA mediated DNA double strand breaks (DSBs) depends on the non-homologous end-joining, NHEJ, DNA repair pathway and we use this observation to develop a tool to assess protospacer....... niger, and in A. oryzae indicating that this type of repair may be wide spread in filamentous fungi. Importantly, we demonstrate that by using single-stranded oligo nucleotides for CRISPR-Cas9 mediated gene editing it is possible to introduce specific point mutations as well gene deletions...

  15. Turbo Charge CPU Utilization in Fork/Join Using the ManagedBlocker

    CERN Multimedia

    CERN. Geneva

    2017-01-01

    Fork/Join is a framework for parallelizing calculations using recursive decomposition, also called divide and conquer. These algorithms occasionally end up duplicating work, especially at the beginning of the run. We can reduce wasted CPU cycles by implementing a reserved caching scheme. Before a task starts its calculation, it tries to reserve an entry in the shared map. If it is successful, it immediately begins. If not, it blocks until the other thread has finished its calculation. Unfortunately this might result in a significant number of blocked threads, decreasing CPU utilization. In this talk we will demonstrate this issue and offer a solution in the form of the ManagedBlocker. Combined with the Fork/Join, it can keep parallelism at the desired level.

  16. Parallel trajectory similarity joins in spatial networks

    KAUST Repository

    Shang, Shuo

    2018-04-04

    The matching of similar pairs of objects, called similarity join, is fundamental functionality in data management. We consider two cases of trajectory similarity joins (TS-Joins), including a threshold-based join (Tb-TS-Join) and a top-k TS-Join (k-TS-Join), where the objects are trajectories of vehicles moving in road networks. Given two sets of trajectories and a threshold θ, the Tb-TS-Join returns all pairs of trajectories from the two sets with similarity above θ. In contrast, the k-TS-Join does not take a threshold as a parameter, and it returns the top-k most similar trajectory pairs from the two sets. The TS-Joins target diverse applications such as trajectory near-duplicate detection, data cleaning, ridesharing recommendation, and traffic congestion prediction. With these applications in mind, we provide purposeful definitions of similarity. To enable efficient processing of the TS-Joins on large sets of trajectories, we develop search space pruning techniques and enable use of the parallel processing capabilities of modern processors. Specifically, we present a two-phase divide-and-conquer search framework that lays the foundation for the algorithms for the Tb-TS-Join and the k-TS-Join that rely on different pruning techniques to achieve efficiency. For each trajectory, the algorithms first find similar trajectories. Then they merge the results to obtain the final result. The algorithms for the two joins exploit different upper and lower bounds on the spatiotemporal trajectory similarity and different heuristic scheduling strategies for search space pruning. Their per-trajectory searches are independent of each other and can be performed in parallel, and the mergings have constant cost. An empirical study with real data offers insight in the performance of the algorithms and demonstrates that they are capable of outperforming well-designed baseline algorithms by an order of magnitude.

  17. Parallel trajectory similarity joins in spatial networks

    KAUST Repository

    Shang, Shuo; Chen, Lisi; Wei, Zhewei; Jensen, Christian S.; Zheng, Kai; Kalnis, Panos

    2018-01-01

    The matching of similar pairs of objects, called similarity join, is fundamental functionality in data management. We consider two cases of trajectory similarity joins (TS-Joins), including a threshold-based join (Tb-TS-Join) and a top-k TS-Join (k-TS-Join), where the objects are trajectories of vehicles moving in road networks. Given two sets of trajectories and a threshold θ, the Tb-TS-Join returns all pairs of trajectories from the two sets with similarity above θ. In contrast, the k-TS-Join does not take a threshold as a parameter, and it returns the top-k most similar trajectory pairs from the two sets. The TS-Joins target diverse applications such as trajectory near-duplicate detection, data cleaning, ridesharing recommendation, and traffic congestion prediction. With these applications in mind, we provide purposeful definitions of similarity. To enable efficient processing of the TS-Joins on large sets of trajectories, we develop search space pruning techniques and enable use of the parallel processing capabilities of modern processors. Specifically, we present a two-phase divide-and-conquer search framework that lays the foundation for the algorithms for the Tb-TS-Join and the k-TS-Join that rely on different pruning techniques to achieve efficiency. For each trajectory, the algorithms first find similar trajectories. Then they merge the results to obtain the final result. The algorithms for the two joins exploit different upper and lower bounds on the spatiotemporal trajectory similarity and different heuristic scheduling strategies for search space pruning. Their per-trajectory searches are independent of each other and can be performed in parallel, and the mergings have constant cost. An empirical study with real data offers insight in the performance of the algorithms and demonstrates that they are capable of outperforming well-designed baseline algorithms by an order of magnitude.

  18. On the Expressiveness of Joining

    Directory of Open Access Journals (Sweden)

    Thomas Given-Wilson

    2015-08-01

    Full Text Available The expressiveness of communication primitives has been explored in a common framework based on the pi-calculus by considering four features: synchronism (asynchronous vs synchronous, arity (monadic vs polyadic data, communication medium (shared dataspaces vs channel-based, and pattern-matching (binding to a name vs testing name equality vs intensionality. Here another dimension coordination is considered that accounts for the number of processes required for an interaction to occur. Coordination generalises binary languages such as pi-calculus to joining languages that combine inputs such as the Join Calculus and general rendezvous calculus. By means of possibility/impossibility of encodings, this paper shows coordination is unrelated to the other features. That is, joining languages are more expressive than binary languages, and no combination of the other features can encode a joining language into a binary language. Further, joining is not able to encode any of the other features unless they could be encoded otherwise.

  19. Involvement of DNA repair in telomere maintenance and chromosomal instability in human cells

    International Nuclear Information System (INIS)

    Ayouaz, Ali

    2008-01-01

    Telomeres are a major actor of cell immortalization, precursor of a carcinogenesis process. Thus, it appears that the maintenance of telomeres is crucial in the implementation of carcinogenesis process. Due to their structures and under some conditions, telomeres can be assimilated in some respects to chromosomal breakages. Within this perspective, this research thesis aims at determining under which circumstances telomeres can be taken as targets by DNA repair mechanisms. More precisely, the author addressed the respective contributions of two repair mechanisms (the Non-Homologous End-Joining or NHEJ, and Homologous Recombination or HR) in the maintenance of telomere integrity. The author first discusses knowledge related to the interaction between chromosomal extremities and repair mechanisms. Then, he defines the behaviour of these mechanisms with respect to telomeres. He shows that, in absence of recombination mechanisms, the integrity of telomeres is not affected. Finally, he reports the attempt to determine their respective contributions in telomeric homeostasis [fr

  20. Altered Hematopoiesis in Mice Lacking DNA Polymerase μ Is Due to Inefficient Double-Strand Break Repair

    Science.gov (United States)

    Lucas, Daniel; Escudero, Beatriz; Ligos, José Manuel; Segovia, Jose Carlos; Estrada, Juan Camilo; Terrados, Gloria; Blanco, Luis; Samper, Enrique; Bernad, Antonio

    2009-01-01

    Polymerase mu (Polμ) is an error-prone, DNA-directed DNA polymerase that participates in non-homologous end-joining (NHEJ) repair. In vivo, Polμ deficiency results in impaired Vκ-Jκ recombination and altered somatic hypermutation and centroblast development. In Polμ−/− mice, hematopoietic development was defective in several peripheral and bone marrow (BM) cell populations, with about a 40% decrease in BM cell number that affected several hematopoietic lineages. Hematopoietic progenitors were reduced both in number and in expansion potential. The observed phenotype correlates with a reduced efficiency in DNA double-strand break (DSB) repair in hematopoietic tissue. Whole-body γ-irradiation revealed that Polμ also plays a role in DSB repair in non-hematopoietic tissues. Our results show that Polμ function is required for physiological hematopoietic development with an important role in maintaining early progenitor cell homeostasis and genetic stability in hematopoietic and non-hematopoietic tissues. PMID:19229323

  1. Productive homologous and non-homologous recombination of hepatitis C virus in cell culture

    DEFF Research Database (Denmark)

    Scheel, Troels K H; Galli, Andrea; Li, Yi-Ping

    2013-01-01

    . In addition, recombination is an important regulatory mechanism of cytopathogenicity for the related pestiviruses. Here we describe recombination of HCV RNA in cell culture leading to production of infectious virus. Initially, hepatoma cells were co-transfected with a replicating JFH1ΔE1E2 genome (genotype 2a......) lacking functional envelope genes and strain J6 (2a), which has functional envelope genes but does not replicate in culture. After an initial decrease in the number of HCV positive cells, infection spread after 13-36 days. Sequencing of recovered viruses revealed non-homologous recombinants with J6...

  2. Factors affecting occupational therapists' decision to join their regional professional association: Facteurs influençant la décision des ergothérapeutes de joindre les rangs de leur association professionnelle régionale.

    Science.gov (United States)

    Reyes, Angelica N; Brown, Cary A

    2018-01-01

    Voluntary occupational therapy organizations fill an important role. However, recruitment and retention can be problematic. Little is known about factors influencing occupational therapists to join/maintain membership in professional associations. This study investigated factors influencing occupational therapists' decision to join/remain members of their association. An electronic survey was carried out and data were analyzed using SPSS software and manual categorization of open-ended comments. Two hundred and fifty-four therapists responded. Generation of new ideas, opportunities for professional development, self-improvement, maintenance of standards, improvement of the profession, and discounts on equipment/educational opportunities were significant factors in deciding to join the organization. The factors perceived as priorities varied in relation to participants' year of graduation. More-customized strategies, reflecting priorities that vary during a therapist's career path, may need to be employed to best address recruitment and retention across the range of therapists' needs and goals.

  3. Common Influence Join

    DEFF Research Database (Denmark)

    Yiu, Man Lung; Mamoulis, Nikos; Karras, Panagiotis

    2008-01-01

    We identify and formalize a novel join operator for two spatial pointsets P and Q. The common influence join (CIJ) returns the pairs of points (p,q),p isin P,q isin Q, such that there exists a location in space, being closer to p than to any other point in P and at the same time closer to q than ......-demand, is very efficient in practice, incurring only slightly higher I/O cost than the theoretical lower bound cost for the problem....

  4. Speeding Up Neighbour-Joining Tree Construction

    DEFF Research Database (Denmark)

    Brodal, Gerth Stølting; Fagerberg, Rolf; Mailund, Thomas

    A widely used method for constructing phylogenetic trees is the neighbour-joining method of Saitou and Nei. We develope heuristics for speeding up the neighbour-joining method which generate the same phylogenetic trees as the original method. All heuristics are based on using a quad-tree to guide...... the search for the next pair of nodes to join, but di#er in the information stored in quad-tree nodes, the way the search is performed, and in the way the quad-tree is updated after a join. We empirically evaluate the performance of the heuristics on distance matrices obtained from the Pfam collection...... of alignments, and compare the running time with that of the QuickTree tool, a well-known and widely used implementation of the standard neighbour-joining method. The results show that the presented heuristics can give a significant speed-up over the standard neighbour-joining method, already for medium sized...

  5. Oxidative DNA Damage in Neurons: Implication of Ku in Neuronal Homeostasis and Survival

    Directory of Open Access Journals (Sweden)

    Daniela De Zio

    2012-01-01

    Full Text Available Oxidative DNA damage is produced by reactive oxygen species (ROS which are generated by exogenous and endogenous sources and continuously challenge the cell. One of the most severe DNA lesions is the double-strand break (DSB, which is mainly repaired by nonhomologous end joining (NHEJ pathway in mammals. NHEJ directly joins the broken ends, without using the homologous template. Ku70/86 heterodimer, also known as Ku, is the first component of NHEJ as it directly binds DNA and recruits other NHEJ factors to promote the repair of the broken ends. Neurons are particularly metabolically active, displaying high rates of transcription and translation, which are associated with high metabolic and mitochondrial activity as well as oxygen consumption. In such a way, excessive oxygen radicals can be generated and constantly attack DNA, thereby producing several lesions. This condition, together with defective DNA repair systems, can lead to a high accumulation of DNA damage resulting in neurodegenerative processes and defects in neurodevelopment. In light of recent findings, in this paper, we will discuss the possible implication of Ku in neurodevelopment and in mediating the DNA repair dysfunction observed in certain neurodegenerations.

  6. Efficient Joins with Compressed Bitmap Indexes

    Energy Technology Data Exchange (ETDEWEB)

    Computational Research Division; Madduri, Kamesh; Wu, Kesheng

    2009-08-19

    We present a new class of adaptive algorithms that use compressed bitmap indexes to speed up evaluation of the range join query in relational databases. We determine the best strategy to process a join query based on a fast sub-linear time computation of the join selectivity (the ratio of the number of tuples in the result to the total number of possible tuples). In addition, we use compressed bitmaps to represent the join output compactly: the space requirement for storing the tuples representing the join of two relations is asymptotically bounded by min(h; n . cb), where h is the number of tuple pairs in the result relation, n is the number of tuples in the smaller of the two relations, and cb is the cardinality of the larger column being joined. We present a theoretical analysis of our algorithms, as well as experimental results on large-scale synthetic and real data sets. Our implementations are efficient, and consistently outperform well-known approaches for a range of join selectivity factors. For instance, our count-only algorithm is up to three orders of magnitude faster than the sort-merge approach, and our best bitmap index-based algorithm is 1.2x-80x faster than the sort-merge algorithm, for various query instances. We achieve these speedups by exploiting several inherent performance advantages of compressed bitmap indexes for join processing: an implicit partitioning of the attributes, space-efficiency, and tolerance of high-cardinality relations.

  7. Welding and Joining of Titanium Aluminides

    Science.gov (United States)

    Cao, Jian; Qi, Junlei; Song, Xiaoguo; Feng, Jicai

    2014-01-01

    Welding and joining of titanium aluminides is the key to making them more attractive in industrial fields. The purpose of this review is to provide a comprehensive overview of recent progress in welding and joining of titanium aluminides, as well as to introduce current research and application. The possible methods available for titanium aluminides involve brazing, diffusion bonding, fusion welding, friction welding and reactive joining. Of the numerous methods, solid-state diffusion bonding and vacuum brazing have been most heavily investigated for producing reliable joints. The current state of understanding and development of every welding and joining method for titanium aluminides is addressed respectively. The focus is on the fundamental understanding of microstructure characteristics and processing–microstructure–property relationships in the welding and joining of titanium aluminides to themselves and to other materials. PMID:28788113

  8. Welding and Joining of Titanium Aluminides

    Directory of Open Access Journals (Sweden)

    Jian Cao

    2014-06-01

    Full Text Available Welding and joining of titanium aluminides is the key to making them more attractive in industrial fields. The purpose of this review is to provide a comprehensive overview of recent progress in welding and joining of titanium aluminides, as well as to introduce current research and application. The possible methods available for titanium aluminides involve brazing, diffusion bonding, fusion welding, friction welding and reactive joining. Of the numerous methods, solid-state diffusion bonding and vacuum brazing have been most heavily investigated for producing reliable joints. The current state of understanding and development of every welding and joining method for titanium aluminides is addressed respectively. The focus is on the fundamental understanding of microstructure characteristics and processing–microstructure–property relationships in the welding and joining of titanium aluminides to themselves and to other materials.

  9. Joining of beryllium by braze welding technique: preliminary results

    Energy Technology Data Exchange (ETDEWEB)

    Banaim, P.; Abramov, E. [Ben-Gurion Univ. of the Negev, Beersheba (Israel); Zalkind, S.; Eden, S.

    1998-01-01

    Within the framework of some applications, there is a need to join beryllium parts to each other. Gas Tungsten Arc Braze Welds were made in beryllium using 0.3 mm commercially Aluminum (1100) shim preplaced at the joint. The welds exhibited a tendency to form microcracks in the Fusion Zone and Heat Affected Zone. All the microcracks were backfilled with Aluminum. (author)

  10. Ring-constrained Join

    DEFF Research Database (Denmark)

    Yiu, Man Lung; Karras, Panagiotis; Mamoulis, Nikos

    2008-01-01

    . This new operation has important applications in decision support, e.g., placing recycling stations at fair locations between restaurants and residential complexes. Clearly, RCJ is defined based on a geometric constraint but not on distances between points. Thus, our operation is fundamentally different......We introduce a novel spatial join operator, the ring-constrained join (RCJ). Given two sets P and Q of spatial points, the result of RCJ consists of pairs (p, q) (where p ε P, q ε Q) satisfying an intuitive geometric constraint: the smallest circle enclosing p and q contains no other points in P, Q...

  11. Performance Comparison of the Optimized Inverted Joined Wing Airplane Concept and Classical Configuration Airplanes

    Directory of Open Access Journals (Sweden)

    Sieradzki Adam

    2016-09-01

    Full Text Available The joined wing concept is an unconventional airplane configuration, known since the mid-twenties of the last century. It has several possible advantages, like reduction of the induced drag and weight due to the closed wing concept. The inverted joined wing variant is its rarely considered version, with the front wing being situated above the aft wing. The following paper presents a performance prediction of the recently optimized configuration of this airplane. Flight characteristics obtained numerically were compared with the performance of two classical configuration airplanes of similar category. Their computational fluid dynamics (CFD models were created basing on available documentation, photographs and some inverse engineering methods. The analysis included simulations performed for a scale of 3-meter wingspan inverted joined wing demonstrator and also for real-scale manned airplanes. Therefore, the results of CFD calculations allowed us to assess the competitiveness of the presented concept, as compared to the most technologically advanced airplanes designed and manufactured to date. At the end of the paper, the areas where the inverted joined wing is better than conventional airplane were predicted and new research possibilities were described.

  12. Current topics in DNA double-strand break repair

    International Nuclear Information System (INIS)

    Kobayashi, Junya; Takata, Minoru; Iwabuchi, Kuniyoshi; Miyagawa, Kiyoshi; Sonoda, Eiichiro; Suzuki, Keiji; Tauchi, Hiroshi

    2008-01-01

    DNA double strand break (DSB) is one of the most critical types of damage which is induced by ionizing radiation. In this review, we summarize current progress in investigations on the function of DSB repair-related proteins. We focused on recent findings in the analysis of the function of proteins such as 53BP1, histone H2AX, Mus81-Eme1, Fanc complex, and UBC13, which are found to be related to homologous recombination repair or to non-homologous end joining. In addition to the function of these proteins in DSB repair, the biological function of nuclear foci formation following DSB induction is discussed. (author)

  13. The DNA-dependent protein kinase: a multifunctional protein kinase with roles in DNA double strand break repair and mitosis

    Science.gov (United States)

    Jette, Nicholas; Lees-Miller, Susan P.

    2015-01-01

    The DNA-dependent protein kinase (DNA-PK) is a serine/threonine protein kinase composed of a large catalytic subunit (DNA-PKcs) and the Ku70/80 heterodimer. Over the past two decades, significant progress has been made in elucidating the role of DNA-PK in non-homologous end joining (NHEJ), the major pathway for repair of ionizing radiation-induced DNA double strand breaks in human cells and recently, additional roles for DNA-PK have been reported. In this review, we will describe the biochemistry, structure and function of DNA-PK, its roles in DNA double strand break repair and its newly described roles in mitosis and other cellular processes. PMID:25550082

  14. Suppression of DNA-dependent protein kinase sensitize cells to radiation without affecting DSB repair

    International Nuclear Information System (INIS)

    Gustafsson, Ann-Sofie; Abramenkovs, Andris; Stenerlöw, Bo

    2014-01-01

    Highlights: • We reduced the level of DNA-PKcs with siRNA and examined cells after γ-irradiation. • Low DNA-PKcs levels lead to radiosensitivity but did not affect repair of DSB. • Low DNA-PKcs levels may block progression of mitosis. • DNA-PKcs role in mitotic progression is independent of its role in DSB repair. • We suggest different mechanisms by which loss of DNA-PKcs function sensitize cells. - Abstract: Efficient and correct repair of DNA double-strand break (DSB) is critical for cell survival. Defects in the DNA repair may lead to cell death, genomic instability and development of cancer. The catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) is an essential component of the non-homologous end joining (NHEJ) which is the major DSB repair pathway in mammalian cells. In the present study, by using siRNA against DNA-PKcs in four human cell lines, we examined how low levels of DNA-PKcs affected cellular response to ionizing radiation. Decrease of DNA-PKcs levels by 80–95%, induced by siRNA treatment, lead to extreme radiosensitivity, similar to that seen in cells completely lacking DNA-PKcs and low levels of DNA-PKcs promoted cell accumulation in G2/M phase after irradiation and blocked progression of mitosis. Surprisingly, low levels of DNA-PKcs did not affect the repair capacity and the removal of 53BP1 or γ-H2AX foci and rejoining of DSB appeared normal. This was in strong contrast to cells completely lacking DNA-PKcs and cells treated with the DNA-PKcs inhibitor NU7441, in which DSB repair were severely compromised. This suggests that there are different mechanisms by which loss of DNA-PKcs functions can sensitize cells to ionizing radiation. Further, foci of phosphorylated DNA-PKcs (T2609 and S2056) co-localized with DSB and this was independent of the amount of DNA-PKcs but foci of DNA-PKcs was only seen in siRNA-treated cells. Our study emphasizes on the critical role of DNA-PKcs for maintaining survival after radiation exposure

  15. Suppression of DNA-dependent protein kinase sensitize cells to radiation without affecting DSB repair

    Energy Technology Data Exchange (ETDEWEB)

    Gustafsson, Ann-Sofie, E-mail: ann-sofie.gustafsson@bms.uu.se; Abramenkovs, Andris; Stenerlöw, Bo

    2014-11-15

    Highlights: • We reduced the level of DNA-PKcs with siRNA and examined cells after γ-irradiation. • Low DNA-PKcs levels lead to radiosensitivity but did not affect repair of DSB. • Low DNA-PKcs levels may block progression of mitosis. • DNA-PKcs role in mitotic progression is independent of its role in DSB repair. • We suggest different mechanisms by which loss of DNA-PKcs function sensitize cells. - Abstract: Efficient and correct repair of DNA double-strand break (DSB) is critical for cell survival. Defects in the DNA repair may lead to cell death, genomic instability and development of cancer. The catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) is an essential component of the non-homologous end joining (NHEJ) which is the major DSB repair pathway in mammalian cells. In the present study, by using siRNA against DNA-PKcs in four human cell lines, we examined how low levels of DNA-PKcs affected cellular response to ionizing radiation. Decrease of DNA-PKcs levels by 80–95%, induced by siRNA treatment, lead to extreme radiosensitivity, similar to that seen in cells completely lacking DNA-PKcs and low levels of DNA-PKcs promoted cell accumulation in G2/M phase after irradiation and blocked progression of mitosis. Surprisingly, low levels of DNA-PKcs did not affect the repair capacity and the removal of 53BP1 or γ-H2AX foci and rejoining of DSB appeared normal. This was in strong contrast to cells completely lacking DNA-PKcs and cells treated with the DNA-PKcs inhibitor NU7441, in which DSB repair were severely compromised. This suggests that there are different mechanisms by which loss of DNA-PKcs functions can sensitize cells to ionizing radiation. Further, foci of phosphorylated DNA-PKcs (T2609 and S2056) co-localized with DSB and this was independent of the amount of DNA-PKcs but foci of DNA-PKcs was only seen in siRNA-treated cells. Our study emphasizes on the critical role of DNA-PKcs for maintaining survival after radiation exposure

  16. Chromosome End Repair and Genome Stability in Plasmodium falciparum.

    Science.gov (United States)

    Calhoun, Susannah F; Reed, Jake; Alexander, Noah; Mason, Christopher E; Deitsch, Kirk W; Kirkman, Laura A

    2017-08-08

    The human malaria parasite Plasmodium falciparum replicates within circulating red blood cells, where it is subjected to conditions that frequently cause DNA damage. The repair of DNA double-stranded breaks (DSBs) is thought to rely almost exclusively on homologous recombination (HR), due to a lack of efficient nonhomologous end joining. However, given that the parasite is haploid during this stage of its life cycle, the mechanisms involved in maintaining genome stability are poorly understood. Of particular interest are the subtelomeric regions of the chromosomes, which contain the majority of the multicopy variant antigen-encoding genes responsible for virulence and disease severity. Here, we show that parasites utilize a competitive balance between de novo telomere addition, also called "telomere healing," and HR to stabilize chromosome ends. Products of both repair pathways were observed in response to DSBs that occurred spontaneously during routine in vitro culture or resulted from experimentally induced DSBs, demonstrating that both pathways are active in repairing DSBs within subtelomeric regions and that the pathway utilized was determined by the DNA sequences immediately surrounding the break. In combination, these two repair pathways enable parasites to efficiently maintain chromosome stability while also contributing to the generation of genetic diversity. IMPORTANCE Malaria is a major global health threat, causing approximately 430,000 deaths annually. This mosquito-transmitted disease is caused by Plasmodium parasites, with infection with the species Plasmodium falciparum being the most lethal. Mechanisms underlying DNA repair and maintenance of genome integrity in P. falciparum are not well understood and represent a gap in our understanding of how parasites survive the hostile environment of their vertebrate and insect hosts. Our work examines DNA repair in real time by using single-molecule real-time (SMRT) sequencing focused on the subtelomeric

  17. Trajectory similarity join in spatial networks

    KAUST Repository

    Shang, Shuo

    2017-09-07

    The matching of similar pairs of objects, called similarity join, is fundamental functionality in data management. We consider the case of trajectory similarity join (TS-Join), where the objects are trajectories of vehicles moving in road networks. Thus, given two sets of trajectories and a threshold θ, the TS-Join returns all pairs of trajectories from the two sets with similarity above θ. This join targets applications such as trajectory near-duplicate detection, data cleaning, ridesharing recommendation, and traffic congestion prediction. With these applications in mind, we provide a purposeful definition of similarity. To enable efficient TS-Join processing on large sets of trajectories, we develop search space pruning techniques and take into account the parallel processing capabilities of modern processors. Specifically, we present a two-phase divide-and-conquer algorithm. For each trajectory, the algorithm first finds similar trajectories. Then it merges the results to achieve a final result. The algorithm exploits an upper bound on the spatiotemporal similarity and a heuristic scheduling strategy for search space pruning. The algorithm\\'s per-trajectory searches are independent of each other and can be performed in parallel, and the merging has constant cost. An empirical study with real data offers insight in the performance of the algorithm and demonstrates that is capable of outperforming a well-designed baseline algorithm by an order of magnitude.

  18. Recrafting the neighbor-joining method

    Directory of Open Access Journals (Sweden)

    Pedersen Christian NS

    2006-01-01

    Full Text Available Abstract Background The neighbor-joining method by Saitou and Nei is a widely used method for constructing phylogenetic trees. The formulation of the method gives rise to a canonical Θ(n3 algorithm upon which all existing implementations are based. Results In this paper we present techniques for speeding up the canonical neighbor-joining method. Our algorithms construct the same phylogenetic trees as the canonical neighbor-joining method. The best-case running time of our algorithms are O(n2 but the worst-case remains O(n3. We empirically evaluate the performance of our algoritms on distance matrices obtained from the Pfam collection of alignments. The experiments indicate that the running time of our algorithms evolve as Θ(n2 on the examined instance collection. We also compare the running time with that of the QuickTree tool, a widely used efficient implementation of the canonical neighbor-joining method. Conclusion The experiments show that our algorithms also yield a significant speed-up, already for medium sized instances.

  19. Willmore energy for joining of carbon nanostructures

    Science.gov (United States)

    Sripaturad, P.; Alshammari, N. A.; Thamwattana, N.; McCoy, J. A.; Baowan, D.

    2018-06-01

    Numerous types of carbon nanostructure have been found experimentally, including nanotubes, fullerenes and nanocones. These structures have applications in various nanoscale devices and the joining of these structures may lead to further new configurations with more remarkable properties and applications. The join profile between different carbon nanostructures in a symmetric configuration may be modelled using the calculus of variations. In previous studies, carbon nanostructures were assumed to deform according to perfect elasticity, thus the elastic energy, depending only on the axial curvature, was used to determine the join profile consisting of a finite number of discrete bonds. However, one could argue that the relevant energy should also involve the rotational curvature, especially when its size is comparable to the axial curvature. In this paper, we use the Willmore energy, a natural generalisation of the elastic energy that depends on both the axial and rotational curvatures. Catenoids are absolute minimisers of this energy and pieces of these may be used to join various nanostructures. We focus on the cases of joining a fullerene to a nanotube and joining two fullerenes along a common axis. By comparing our results with the earlier work, we find that both energies give similar joining profiles. Further work on other configurations may reveal which energy provides a better model.

  20. Isolation of Chromatin from Dysfunctional Telomeres Reveals an Important Role for Ring1b in NHEJ-Mediated Chromosome Fusions

    Directory of Open Access Journals (Sweden)

    Cristina Bartocci

    2014-05-01

    Full Text Available When telomeres become critically short, DNA damage response factors are recruited at chromosome ends, initiating a cellular response to DNA damage. We performed proteomic isolation of chromatin fragments (PICh in order to define changes in chromatin composition that occur upon onset of acute telomere dysfunction triggered by depletion of the telomere-associated factor TRF2. This unbiased purification of telomere-associated proteins in functional or dysfunctional conditions revealed the dynamic changes in chromatin composition that take place at telomeres upon DNA damage induction. On the basis of our results, we describe a critical role for the polycomb group protein Ring1b in nonhomologous end-joining (NHEJ-mediated end-to-end chromosome fusions. We show that cells with reduced levels of Ring1b have a reduced ability to repair uncapped telomeric chromatin. Our data represent an unbiased isolation of chromatin undergoing DNA damage and are a valuable resource to map the changes in chromatin composition in response to DNA damage activation.

  1. Structure of a preternary complex involving a prokaryotic NHEJ DNA polymerase.

    Science.gov (United States)

    Brissett, Nigel C; Martin, Maria J; Pitcher, Robert S; Bianchi, Julie; Juarez, Raquel; Green, Andrew J; Fox, Gavin C; Blanco, Luis; Doherty, Aidan J

    2011-01-21

    In many prokaryotes, a specific DNA primase/polymerase (PolDom) is required for nonhomologous end joining (NHEJ) repair of DNA double-strand breaks (DSBs). Here, we report the crystal structure of a catalytically active conformation of Mycobacterium tuberculosis PolDom, consisting of a polymerase bound to a DNA end with a 3' overhang, two metal ions, and an incoming nucleotide but, significantly, lacking a primer strand. This structure represents a polymerase:DNA complex in a preternary intermediate state. This polymerase complex occurs in solution, stabilizing the enzyme on DNA ends and promoting nucleotide extension of short incoming termini. We also demonstrate that the invariant Arg(220), contained in a conserved loop (loop 2), plays an essential role in catalysis by regulating binding of a second metal ion in the active site. We propose that this NHEJ intermediate facilitates extension reactions involving critically short or noncomplementary DNA ends, thus promoting break repair and minimizing sequence loss during DSB repair. Copyright © 2011 Elsevier Inc. All rights reserved.

  2. The study on joining carbon/carbon composites using Ti–Ni–Si compound

    International Nuclear Information System (INIS)

    Wang Jie; Li Kezhi; Song Xinrui; Guo Lingjun; Li Wei; Li Zhaoqian

    2012-01-01

    Highlights: ► First study to join C/C composites using Ti–Ni–Si compound. ► The average joint shear strength can reach up to 23.58 MPa. ► Strong joints are apt to fracture through interlayer, interfaces and C/C matrix. ► Phase compositions in joints are varied with increasing joining temperature. - Abstract: 2D carbon/carbon (C/C) composites have been successfully joined by partial transient liquid phase (PTLP) bonding using Ti–Ni–Si compound as interlayer. The shear strengths of C/C joints were found to be affected observably by joining temperature, joining pressure and holding time. The optimum process parameters were determined at 1150 °C, 30 MPa, 45 min, and the maximum joint shear strength was obtained under this optimum process condition with an average value of 23.58 MPa. Microstructures and compositions of joints were analyzed by scanning electronic microscopy (SEM), energy disperse spectroscopy (EDS) and X-ray diffraction (XRD). Results showed that element interdiffusions and chemical reactions took place in the joining zone and various compounds including Ni 4 Si 7 Ti 4 , TiSi 2 , SiC and TiC were produced, by which compact interlayer and strong interface were formed in C/C joints. The shear morphology indicated there were three fracture modes in C/C joints and the fracture was apt to occur through interlayer, interface and C/C matrix in strong joints. Thermal shock resistance experimental results revealed that joint shear strength has a rapid drop after 5 thermal cycles and then reduces gradually with the increase of thermal cycles between room temperature and 1000 °C.

  3. MICROWAVE JOINING OF ALUMINA CERAMIC AND HYDROXYLAPATITE BIOCERAMIC

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    Microwave joining is a rapid developmental new techniqu e in recent years.This paper introduces a new microwave joining equipment which was made by our lab,succeeds in alumina ceramic-hydroxylapatite bioceramic j o in in the equipment, and analyzes the join situation of join boundary by using s canni ng electron microscope(SEM),this paper analyzes the mechanism of microwave joini ng also.

  4. Prospects of joining multi-material structures

    Science.gov (United States)

    Sankaranarayanan, R.; Hynes, N. Rajesh Jesudoss

    2018-05-01

    Spring up trends and necessities make the pipelines for the brand new Technologies. The same way, Multimaterial structures emerging as fruitful alternatives for the conventional structures in the manufacturing sector. Especially manufacturing of transport vehicles is placing a perfect platform for these new structures. Bonding or joining technology plays a crucial role in the field of manufacturing for sustainability. These latest structures are purely depending on such joining technologies so that multi-material structuring can be possible practically. The real challenge lies on joining dissimilar materials of different properties and nature. Escalation of thermoplastic usage in large structural components also faces similar ambiguity for joining multi-material structures. Adhesive bonding, mechanical fastening and are the answering technologies for multi-material structures. This current paper analysis the prospects of these bonding technologies to meet the challenges of tomorrow.

  5. Borated stainless steel joining technology. Final report

    International Nuclear Information System (INIS)

    Smith, R.J.

    1994-12-01

    EPRI had continued investigating the application of borated stainless steel products within the US commercial nuclear power industry through participation in a wide range of activities. This effort provides the documentation of the data obtained in the development of the ASTM-A887 Specification preparation effort conducted by Applied Science and Technology and the most recent efforts for the development of joining technologies conducted under a joint effort by EPRI, Carpenter Technologies and Sandia National Laboratory under a US DOE CRADA program. The data presented in this report provides the basis for the ASTM specification which has been previously unpublished by EPRI and the data generated in support of the Joining Technology research effort conducted at Sandia. The results of the Sandia research, although terminated prior to the completion, confirms earlier data that the degradation of material properties in fusion welded borated stainless steels occurs in the heat affected zone of the weld area and not in the base material. The data obtained also supports the conclusion that the degradation of material properties can be overcome by post weld heat treatment which can result in material properties near the original unwelded metal

  6. A data mining approach for classifying DNA repair genes into ageing-related or non-ageing-related

    Directory of Open Access Journals (Sweden)

    Vasieva Olga

    2011-01-01

    Full Text Available Abstract Background The ageing of the worldwide population means there is a growing need for research on the biology of ageing. DNA damage is likely a key contributor to the ageing process and elucidating the role of different DNA repair systems in ageing is of great interest. In this paper we propose a data mining approach, based on classification methods (decision trees and Naive Bayes, for analysing data about human DNA repair genes. The goal is to build classification models that allow us to discriminate between ageing-related and non-ageing-related DNA repair genes, in order to better understand their different properties. Results The main patterns discovered by the classification methods are as follows: (a the number of protein-protein interactions was a predictor of DNA repair proteins being ageing-related; (b the use of predictor attributes based on protein-protein interactions considerably increased predictive accuracy of attributes based on Gene Ontology (GO annotations; (c GO terms related to "response to stimulus" seem reasonably good predictors of ageing-relatedness for DNA repair genes; (d interaction with the XRCC5 (Ku80 protein is a strong predictor of ageing-relatedness for DNA repair genes; and (e DNA repair genes with a high expression in T lymphocytes are more likely to be ageing-related. Conclusions The above patterns are broadly integrated in an analysis discussing relations between Ku, the non-homologous end joining DNA repair pathway, ageing and lymphocyte development. These patterns and their analysis support non-homologous end joining double strand break repair as central to the ageing-relatedness of DNA repair genes. Our work also showcases the use of protein interaction partners to improve accuracy in data mining methods and our approach could be applied to other ageing-related pathways.

  7. Xrcc1-dependent and Ku-dependent DNA double-strand break repair kinetics in Arabidopsis plants.

    Science.gov (United States)

    Charbonnel, Cyril; Gallego, Maria E; White, Charles I

    2010-10-01

    Double-strand breakage (DSB) of DNA involves loss of information on the two strands of the DNA fibre and thus cannot be repaired by simple copying of the complementary strand which is possible with single-strand DNA damage. Homologous recombination (HR) can precisely repair DSB using another copy of the genome as template and non-homologous recombination (NHR) permits repair of DSB with little or no dependence on DNA sequence homology. In addition to the well-characterised Ku-dependent non-homologous end-joining (NHEJ) pathway, much recent attention has been focused on Ku-independent NHR. The complex interrelationships and regulation of NHR pathways remain poorly understood, even more so in the case of plants, and we present here an analysis of Ku-dependent and Ku-independent repair of DSB in Arabidopsis thaliana. We have characterised an Arabidopsis xrcc1 mutant and developed quantitative analysis of the kinetics of appearance and loss of γ-H2AX foci as a tool to measure DSB repair in dividing root tip cells of γ-irradiated plants in vivo. This approach has permitted determination of DSB repair kinetics in planta following a short pulse of γ-irradiation, establishing the existence of a Ku-independent, Xrcc1-dependent DSB repair pathway. Furthermore, our data show a role for Ku80 during the first minutes post-irradiation and that Xrcc1 also plays such a role, but only in the absence of Ku. The importance of Xrcc1 is, however, clearly visible at later times in the presence of Ku, showing that alternative end-joining plays an important role in DSB repair even in the presence of active NHEJ. © 2010 The Authors. Journal compilation © 2010 Blackwell Publishing Ltd.

  8. Conjoint Forming - Technologies for Simultaneous Forming and Joining

    International Nuclear Information System (INIS)

    Groche, P; Wohletz, S; Mann, A; Krech, M; Monnerjahn, V

    2016-01-01

    The market demand for new products optimized for e. g. lightweight applications or smart components leads to new challenges in production engineering. Hybrid structures represent one promising approach. They aim at higher product performance by using a suitable combination of different materials. The developments of hybrid structures stimulate the research on joining of dissimilar materials. Since they allow for joining dissimilar materials without external heating technologies based on joining by plastic deformation seem to be of special attractiveness. The paper at hand discusses the conjoint forming approach. This approach combines forming and joining in one process. Two or more workpieces are joined while at least one workpiece is plastically deformed. After presenting the fundamental joining mechanisms, the conjoint forming approach is discussed comprehensively. Examples of conjoint processes demonstrate the effectiveness and reveal the underlying phenomena. (paper)

  9. Ku recruits XLF to DNA double-strand breaks.

    Science.gov (United States)

    Yano, Ken-ichi; Morotomi-Yano, Keiko; Wang, Shih-Ya; Uematsu, Naoya; Lee, Kyung-Jong; Asaithamby, Aroumougame; Weterings, Eric; Chen, David J

    2008-01-01

    XRCC4-like factor (XLF)--also known as Cernunnos--has recently been shown to be involved in non-homologous end-joining (NHEJ), which is the main pathway for the repair of DNA double-strand breaks (DSBs) in mammalian cells. XLF is likely to enhance NHEJ by stimulating XRCC4-ligase IV-mediated joining of DSBs. Here, we report mechanistic details of XLF recruitment to DSBs. Live cell imaging combined with laser micro-irradiation showed that XLF is an early responder to DSBs and that Ku is essential for XLF recruitment to DSBs. Biochemical analysis showed that Ku-XLF interaction occurs on DNA and that Ku stimulates XLF binding to DNA. Unexpectedly, XRCC4 is dispensable for XLF recruitment to DSBs, although photobleaching analysis showed that XRCC4 stabilizes the binding of XLF to DSBs. Our observations showed the direct involvement of XLF in the dynamic assembly of the NHEJ machinery and provide mechanistic insights into DSB recognition.

  10. Discovery and characterization of Ku acetylation in Mycobacterium smegmatis.

    Science.gov (United States)

    Zhou, Ying; Chen, Tao; Zhou, Lin; Fleming, Joy; Deng, Jiaoyu; Wang, Xude; Wang, Liwei; Wang, Yingying; Zhang, Xiaoli; Wei, Wenjing; Bi, Lijun

    2015-03-01

    Lysine acetylation is an important post-translational modification and is known to regulate many eukaryotic cellular processes. Little, however, is known about acetylated proteins in prokaryotes. Here, using immunoblotting, mass spectrometry and mutagenesis studies, we investigate the acetylation dynamics of the DNA repair protein Ku and its relationship with the deacetylase protein Sir2 and the non-homologous end joining (NHEJ) pathway in Mycobacterium smegmatis. We report that acetylation of Ku increases with growth, while NHEJ activity decreases, providing support for the hypothesis that acetylation of Ku may be involved in the DNA damage response in bacteria. Ku has multiple lysine sites. Our results indicate that K29 is an important acetylation site and that deficiency of Sir2 or mutation of K29 affects the quantity of Ku and its acetylation dynamics. Our findings expand knowledge of acetylation targets in prokaryotes and indicate a new direction for further research on bacterial DNA repair mechanisms. © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Differentiation of Human Induced Pluripotent or Embryonic Stem Cells Decreases the DNA Damage Repair by Homologous Recombination

    Directory of Open Access Journals (Sweden)

    Kalpana Mujoo

    2017-11-01

    Full Text Available The nitric oxide (NO-cyclic GMP pathway contributes to human stem cell differentiation, but NO free radical production can also damage DNA, necessitating a robust DNA damage response (DDR to ensure cell survival. How the DDR is affected by differentiation is unclear. Differentiation of stem cells, either inducible pluripotent or embryonic derived, increased residual DNA damage as determined by γ-H2AX and 53BP1 foci, with increased S-phase-specific chromosomal aberration after exposure to DNA-damaging agents, suggesting reduced homologous recombination (HR repair as supported by the observation of decreased HR-related repair factor foci formation (RAD51 and BRCA1. Differentiated cells also had relatively increased fork stalling and R-loop formation after DNA replication stress. Treatment with NO donor (NOC-18, which causes stem cell differentiation has no effect on double-strand break (DSB repair by non-homologous end-joining but reduced DSB repair by HR. Present studies suggest that DNA repair by HR is impaired in differentiated cells.

  12. The C-Terminal Domain of Cernunnos/XLF Is Dispensable for DNA Repair In Vivo▿ †

    Science.gov (United States)

    Malivert, Laurent; Callebaut, Isabelle; Rivera-Munoz, Paola; Fischer, Alain; Mornon, Jean-Paul; Revy, Patrick; de Villartay, Jean-Pierre

    2009-01-01

    The core nonhomologous end-joining DNA repair pathway is composed of seven factors: Ku70, Ku80, DNA-PKcs, Artemis, XRCC4 (X4), DNA ligase IV (L4), and Cernunnos/XLF (Cernunnos). Although Cernunnos and X4 are structurally related and participate in the same complex together with L4, they have distinct functions during DNA repair. L4 relies on X4 but not on Cernunnos for its stability, and L4 is required for optimal interaction of Cernunnos with X4. We demonstrate here, using in vitro-generated Cernunnos mutants and a series of functional assays in vivo, that the C-terminal region of Cernunnos is dispensable for its activity during DNA repair. PMID:19103754

  13. Mutations in XRCC4 cause primordial dwarfism without causing immunodeficiency.

    Science.gov (United States)

    Saito, Shinta; Kurosawa, Aya; Adachi, Noritaka

    2016-08-01

    In successive reports from 2014 to 2015, X-ray repair cross-complementing protein 4 (XRCC4) has been identified as a novel causative gene of primordial dwarfism. XRCC4 is indispensable for non-homologous end joining (NHEJ), the major pathway for repairing DNA double-strand breaks. As NHEJ is essential for V(D)J recombination during lymphocyte development, it is generally believed that abnormalities in XRCC4 cause severe combined immunodeficiency. Contrary to expectations, however, no overt immunodeficiency has been observed in patients with primordial dwarfism harboring XRCC4 mutations. Here, we describe the various XRCC4 mutations that lead to disease and discuss their impact on NHEJ and V(D)J recombination.

  14. Targeted Gene Knock Out Using Nuclease-Assisted Vector Integration: Hemi- and Homozygous Deletion of JAG1.

    Science.gov (United States)

    Gapinske, Michael; Tague, Nathan; Winter, Jackson; Underhill, Gregory H; Perez-Pinera, Pablo

    2018-01-01

    Gene editing technologies are revolutionizing fields such as biomedicine and biotechnology by providing a simple means to manipulate the genetic makeup of essentially any organism. Gene editing tools function by introducing double-stranded breaks at targeted sites within the genome, which the host cells repair preferentially by Non-Homologous End Joining. While the technologies to introduce double-stranded breaks have been extensively optimized, this progress has not been matched by the development of methods to integrate heterologous DNA at the target sites or techniques to detect and isolate cells that harbor the desired modification. We present here a technique for rapid introduction of vectors at target sites in the genome that enables efficient isolation of successfully edited cells.

  15. Flavourings significantly affect inhalation toxicity of aerosol generated from electronic nicotine delivery systems (ENDS).

    Science.gov (United States)

    Leigh, Noel J; Lawton, Ralph I; Hershberger, Pamela A; Goniewicz, Maciej L

    2016-11-01

    E-cigarettes or electronic nicotine delivery systems (ENDS) are designed to deliver nicotine-containing aerosol via inhalation. Little is known about the health effects of flavoured ENDS aerosol when inhaled. Aerosol from ENDS was generated using a smoking machine. Various types of ENDS devices or a tank system prefilled with liquids of different flavours, nicotine carrier, variable nicotine concentrations and with modified battery output voltage were tested. A convenience sample of commercial fluids with flavour names of tobacco, piña colada, menthol, coffee and strawberry were used. Flavouring chemicals were identified using gas chromatography/mass spectrometry. H292 human bronchial epithelial cells were directly exposed to 55 puffs of freshly generated ENDS aerosol, tobacco smoke or air (controls) using an air-liquid interface system and the Health Canada intense smoking protocol. The following in vitro toxicological effects were assessed: (1) cell viability, (2) metabolic activity and (3) release of inflammatory mediators (cytokines). Exposure to ENDS aerosol resulted in decreased metabolic activity and cell viability and increased release of interleukin (IL)-1β, IL-6, IL-10, CXCL1, CXCL2 and CXCL10 compared to air controls. Cell viability and metabolic activity were more adversely affected by conventional cigarettes than most tested ENDS products. Product type, battery output voltage and flavours significantly affected toxicity of ENDS aerosol, with a strawberry-flavoured product being the most cytotoxic. Our data suggest that characteristics of ENDS products, including flavours, may induce inhalation toxicity. Therefore, ENDS users should use the products with caution until more comprehensive studies are performed. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  16. DNA double strand break repair in mammalian cells: role of MRE11 and BLM proteins at the initiation of Non Homologous End Joining (NHEJ)

    International Nuclear Information System (INIS)

    Grabarz, Anastazja

    2011-01-01

    DNA double strand breaks (DSBs) are highly cytotoxic lesions, which can lead to genetic rearrangements. Two pathways are responsible for repairing these lesions: homologous recombination (HR) and non homologous end joining (NHEJ). In our laboratory, an intrachromosomal substrate has been established in order to measure the efficiency and the fidelity of NHEJ in living cells (Guirouilh-Barbat 2004). This approach led us to identify a KU-independent alternative pathway, which uses micro homologies in the proximity of the junction to accomplish repair - the alternative NHEJ (Guirouilh-Barbat 2004, Guirouilh-Barbat et Rass 2007). The goal of my thesis consisted in identifying and characterising major actors of this pathway. In the absence of KU, alternative NHEJ would be initiated by ssDNA resection of damaged ends. We showed that the nuclease activity of MRE11 is necessary for this mechanism. MRE11 overexpression leads to a two fold stimulation of NHEJ efficiency, while the extinction of MRE11 by siRNA results in a two fold decrease. Our results demonstrate that the proteins RAD50 and CtIP act in the same pathway as MRE11. Moreover, in cells deficient for XRCC4, MIRIN - an inhibitor of the MRN complex - leads to a decrease in repair efficiency, implicating MRE11 in alternative NHEJ. We also showed that MRE11 can act in an ATM-dependent and independent manner (Rass et Grabarz Nat Struct Mol Biol 2009). The initiation of break resection needs to be pursued by a more extensive degradation of DNA, which is accomplished in yeast by the proteins Exo1 and Sgs1/Dna2. In human cells, in vitro studies have recently proposed a similar model of a two-step break resection. We chose to elucidate the role of one of the human homologs of Sgs1 - the RecQ helicase BLM - in the resection process. Our experiments show, that he absence of BLM decreases the efficiency of end joining by NHEJ, accompanied by an increase in error-prone events, especially long-range deletions (≥200 nt). This

  17. Induction of human immunodeficiency virus (HIV-1 envelope specific cell-mediated immunity by a non-homologous synthetic peptide.

    Directory of Open Access Journals (Sweden)

    Ammar Achour

    2007-11-01

    Full Text Available Cell mediated immunity, including efficient CTL response, is required to prevent HIV-1 from cell-to-cell transmission. In previous investigations, we have shown that B1 peptide derived by Fourier transformation of HIV-1 primary structures and sharing no sequence homology with the parent proteins was able to generate antiserum which recognizes envelope and Tat proteins. Here we have investigated cellular immune response towards a novel non-homologous peptide, referred to as cA1 peptide.The 20 amino acid sequence of cA1 peptide was predicted using the notion of peptide hydropathic properties; the peptide is encoded by the complementary anti-sense DNA strand to the sense strand of previously described non-homologous A1 peptide. In this report we demonstrate that the cA1 peptide can be a target for major histocompatibility complex (MHC class I-restricted cytotoxic T lymphocytes in HIV-1-infected or envelope-immunized individuals. The cA1 peptide is recognized in association with different MHC class I allotypes and could prime in vitro CTLs, derived from gp160-immunized individuals capable to recognize virus variants.For the first time a theoretically designed immunogen involved in broad-based cell-immune memory activation is described. Our findings may thus contribute to the advance in vaccine research by describing a novel strategy to develop a synthetic AIDS vaccine.

  18. Thin layer joining by gas adsorption

    Energy Technology Data Exchange (ETDEWEB)

    Taga, Yasunori, E-mail: y-taga@isc.chubu.ac.jp; Fukumura, Toshio

    2014-10-01

    Highlights: • We report thin layer molecular joining between glass and COP by gas adsorption. Thickness of joining layer is 1–2 nm and joining process was carried out at low temperature at about 100 °C. • Adhesion strength measured by 180 degree peel test revealed to be 1–10 N/25 mm and the joined stack showed high durability for practical use. - Abstract: Attempt has been made to join borosilicate glass and cycloolefin (COP) polymer film by using gas adsorption method. After corona plasma treat, COP was exposed to (3-glycidoxypropyl) trimethoxysilane (GPS) and glass to (3-aminopropyl) triethoxysilane (APS) both in air atmosphere, resulting in co-adsorption of water vapor in the atmosphere and organosilane gases. Surface characterization of plasma treated and gas adsorbed surfaces was carried out by X-ray photoelectron spectroscopy (XPS) using Mg Kα X-ray source. Joining was carried out by a roll laminator after contact of both surfaces at room temperature, followed by annealing at 130 °C for 10 min. Adhesion strength was evaluated by 180 degree peel test based on ASTM D-903 and durability was examined under the conditions of 60 °C and 95% RH. It was found that after plasma treatment, complex functional groups such as C-H, C-O, C=O, O-C=O and CO{sub 3} were found on COP and O-H on glass. Thickness of GPS gas adsorption layer on COP was evaluated by the XPS to be at least 1.1 nm by taking inelastic mean free path of Si{sub 2p} photoelectron into consideration. Joining force was found to be more than 5 N/25 mm corresponding to almost equal to COP bulk tensile strength. In addition, durability of this adhesion strength remained unchanged over 2000 h even after exposure to the durability test conditions of 60 °C and 95% RH. The results can be explained in terms of formation of H-H hydrogen bonding and Si-O covalent bonding via silanols will be made at the interface as a result of lamination and annealing processes. In conclusion, ultrathin joining method

  19. Spatio-temporal joins on symbolic indoor tracking data

    DEFF Research Database (Denmark)

    Lu, Hua; Yang, Bin; Jensen, Christian S.

    2011-01-01

    and studies probabilistic, spatio-temporal joins on historical indoor tracking data. Two meaningful types of join are defined. They return object pairs that satisfy spatial join predicates either at a time point or during a time interval. The predicates considered include “same X,” where X is a semantic...

  20. Boolean Operations, Joins, and the Extended Low Hierarchy

    OpenAIRE

    Hemaspaandra, Lane A.; Jiang, Zhigen; Rothe, Joerg; Watanabe, Osamu

    1999-01-01

    We prove that the join of two sets may actually fall into a lower level of the extended low hierarchy than either of the sets. In particular, there exist sets that are not in the second level of the extended low hierarchy, EL_2, yet their join is in EL_2. That is, in terms of extended lowness, the join operator can lower complexity. Since in a strong intuitive sense the join does not lower complexity, our result suggests that the extended low hierarchy is unnatural as a complexity measure. We...

  1. Joining elements of silicon carbide

    International Nuclear Information System (INIS)

    Olson, B.A.

    1979-01-01

    A method of joining together at least two silicon carbide elements (e.g.in forming a heat exchanger) is described, comprising subjecting to sufficiently non-oxidizing atmosphere and sufficiently high temperature, material placed in space between the elements. The material consists of silicon carbide particles, carbon and/or a precursor of carbon, and silicon, such that it forms a joint joining together at least two silicon carbide elements. At least one of the elements may contain silicon. (author)

  2. Recrafting the Neighbor-Joining Method

    DEFF Research Database (Denmark)

    Mailund; Brodal, Gerth Stølting; Fagerberg, Rolf

    2006-01-01

    Background: The neighbor-joining method by Saitou and Nei is a widely used method for constructing phylogenetic trees. The formulation of the method gives rise to a canonical Θ(n3) algorithm upon which all existing implementations are based. Methods: In this paper we present techniques for speeding...... up the canonical neighbor-joining method. Our algorithms construct the same phylogenetic trees as the canonical neighbor-joining method. The best-case running time of our algorithms are O(n2) but the worst-case remains O(n3). We empirically evaluate the performance of our algoritms on distance...... matrices obtained from the Pfam collection of alignments. Results: The experiments indicate that the running time of our algorithms evolve as Θ(n2) on the examined instance collection. We also compare the running time with that of the QuickTree tool, a widely used efficient implementation of the canonical...

  3. Join or be excluded from biomedicine? JOINS and Post-colonial Korea.

    Science.gov (United States)

    Ma, Eunjeong

    2015-04-01

    This paper discusses re-emergence of Korean medicine(s) in the global context with a focus on a natural drug JOINS, a highly contentious drug regarding its legal status. By following through its life world, the paper contends that the drug is the embodiment of the postcolonial anxiety that crosses the intersections between the aspiring nation and globalizing strategies of the bio-pharmaceutical industry. JOINS is a natural drug prescribed to alleviate the symptoms of degenerative arthritis. SK Chemicals, Ltd., a giant domestic pharmaceutical company developed the drug by utilizing the knowledge of traditional pharmacopeia and put it on the market in 2001. In the domestic market, the drug is treated as a prescription drug, implying that Western medicine-trained doctors (as opposed to Korean medicine doctors) are entitled to prescribe drugs. It also indicates that the drug has undergone a series of lab tests such as toxicity, efficacy, and clinical trials in compliance with regulatory guidelines. However, the domestic standards are not rigorous enough to satisfy international standards, so that it is exported as a nutritional supplement abroad. The government, the pharmaceutical industry, and the Western medicine profession are happy with how the drug stands domestically and internationally. Rather, it is Korean doctors who try to disrupt the status quo and reclaim their rights to traditional knowledge, who have been alienated from the pharmaceuticalization of traditional knowledge. Thus, the JOINS tablet embodies the complex web of modern Korean society, professional interests, the pharmaceutical industry, and globalization.

  4. Effects of different brazing and welding methods on the fracture load of various orthodontic joining configurations.

    Science.gov (United States)

    Bock, Jens J; Bailly, Jacqueline; Fuhrmann, Robert A

    2009-06-01

    The aim of this study was to compare the fracture load of different joints made by conventional brazing, tungston inert gas (TIG) and laser welding. Six standardized joining configurations of spring hard quality orthodontic wire were investigated: end-to-end, round, cross, 3 mm length, 9 mm length and 6.5 mm to orthodontic band. The joints were made by five different methods: brazing with universal silver solder, two TIG and two laser welding devices. The fracture loads were measured with a universal testing machine (Zwick 005). Data were analysed with the Mann-Whitney-Wilcoxon and Kruskal-Wallis tests. The significance level was set at Pwelding (Pwelding (826 N). No differences between the various TIG or laser welding devices were demonstrated, although it was not possible to join an orthodontic wire to an orthodontic band using TIG welding. For orthodontic purposes laser and TIG welding are solder free alternatives. TIG welding and laser welding showed similar results. The laser technique is an expensive, but sophisticated and simple method.

  5. JOINING DISSIMILAR MATERIALS USING FRICTION STIR SCRIBE TECHNIQUE

    Energy Technology Data Exchange (ETDEWEB)

    Upadhyay, Piyush; Hovanski, Yuri; Jana, Saumyadeep; Fifield, Leonard S.

    2016-09-01

    Development of robust and cost effective method of joining dissimilar materials can provide a critical pathway to enable widespread use of multi-material design and components in mainstream industrial applications. The use of multi-material components such as Steel-Aluminum, Aluminum-Polymer allows design engineers to optimize material utilization based on service requirements and often lead weight and cost reductions. However producing an effective joint between materials with vastly different thermal, microstructural and deformation response is highly problematic using conventional joining and /or fastening methods. This is especially challenging in cost sensitive high volume markets that largely rely on low–cost joining solutions. Friction Stir Scribe technology was developed to meet the demands of joining materials with drastically different properties and melting regimes. The process enables joining of light metals like Magnesium and Aluminum to high temperature materials like Steels and Titanium. Additionally viable joints between polymer composites and metal can also be made using this method. This paper will present state of the art, progress made and challenges associated with this innovative derivative of Friction Stir welding in reference to joining dissimilar metals and polymer/metal combinations.

  6. Joining Dissimilar Materials Using Friction Stir Scribe Technique

    Energy Technology Data Exchange (ETDEWEB)

    Upadhyay, Piyush [Pacific Northwest National Laboratory, Richland 99352, WA e-mail: piyush.upadhyay@pnnl.gov; Hovanski, Yuri [Pacific Northwest National Laboratory, Richland 99352, WA; Jana, Saumyadeep [Pacific Northwest National Laboratory, Richland 99352, WA; Fifield, Leonard S. [Pacific Northwest National Laboratory, Richland 99352, WA

    2016-10-03

    Development of a robust and cost-effective method of joining dissimilar materials could provide a critical pathway to enable widespread use of multi-material designs and components in mainstream industrial applications. The use of multi-material components such as steel-aluminum and aluminum-polymer would allow design engineers to optimize material utilization based on service requirements and could often lead to weight and cost reductions. However, producing an effective joint between materials with vastly different thermal, microstructural, and deformation responses is highly problematic using conventional joining and/or fastening methods. This is especially challenging in cost sensitive, high volume markets that largely rely on low cost joining solutions. Friction stir scribe technology was developed to meet the demands of joining materials with drastically different properties and melting regimes. The process enables joining of light metals like magnesium and aluminum to high temperature materials like steel and titanium. Viable joints between polymer composites and metal can also be made using this method. This paper will present the state of the art, progress made, and challenges associated with this innovative derivative of friction stir welding in reference to joining dissimilar metals and polymer/metal combinations.

  7. Joining of porous silicon carbide bodies

    Science.gov (United States)

    Bates, Carl H.; Couhig, John T.; Pelletier, Paul J.

    1990-05-01

    A method of joining two porous bodies of silicon carbide is disclosed. It entails utilizing an aqueous slip of a similar silicon carbide as was used to form the porous bodies, including the sintering aids, and a binder to initially join the porous bodies together. Then the composite structure is subjected to cold isostatic pressing to form a joint having good handling strength. Then the composite structure is subjected to pressureless sintering to form the final strong bond. Optionally, after the sintering the structure is subjected to hot isostatic pressing to further improve the joint and densify the structure. The result is a composite structure in which the joint is almost indistinguishable from the silicon carbide pieces which it joins.

  8. WRN Exonuclease Structure, Molecular Mechanism, and DNA EndProcessing Role

    Energy Technology Data Exchange (ETDEWEB)

    Perry, J. Jefferson P.; Yannone, Steven M.; Holden, Lauren G.; Hitomi, Chiharu; Asaithamby, Aroumougame; Han, Seungil; Cooper, PriscillaK.; Chen, David J.; Tainer, John A.

    2006-02-15

    WRN is unique among the five human RecQ DNA helicases by having a functional exonuclease domain (WRN-exo) and being defective in the premature aging and cancer-related disorder Werner syndrome. Here, we characterize WRN-exo crystal structures, biochemical activity and participation in DNA end-joining. Metal ion complex structures, active site mutations and activity assays reveal a two-metal-ion mediated nuclease mechanism. The DNA end-binding Ku70/80 complex specifically stimulates WRN-exo activity, and structure-based mutational inactivation of WRN-exo alters DNA end-joining in human cells. We furthermore establish structural and biochemical similarities of WRN-exo to DnaQ family replicative proofreading exonucleases, with WRN-specific adaptations consistent with dsDNA specificity and functionally important conformational changes. These results indicate WRN-exo is a human DnaQ family member and support analogous proof-reading activities that are stimulated by Ku70/80 with implications for WRN functions in age related pathologies and maintenance of genomic integrity.

  9. Efficient string similarity join in multi-core and distributed systems.

    Directory of Open Access Journals (Sweden)

    Cairong Yan

    Full Text Available In big data area a significant challenge about string similarity join is to find all similar pairs more efficiently. In this paper, we propose a parallel processing framework for efficient string similarity join. First, the input is split into some disjoint small subsets according to the joint frequency distribution and the interval distribution of strings. Then the filter-verification strategy is adopted in the computation of string similarity for each subset so that the number of candidate pairs is reduced before an effective pruning strategy is used to improve the performance. Finally, the operation of string join is executed in parallel. Para-Join algorithm based on the multi-threading technique is proposed to implement the framework in a multi-core system while Pada-Join algorithm based on Spark platform is proposed to implement the framework in a cluster system. We prove that Para-Join and Pada-Join cannot only avoid reduplicate computation but also ensure the completeness of the result. Experimental results show that Para-Join can achieve high efficiency and significantly outperform than state-of-the-art approaches, meanwhile, Pada-Join can work on large datasets.

  10. Analysis of factors affecting the implementation of back-end nuclear fuel cycle policy in Korea

    International Nuclear Information System (INIS)

    Choi, Yung Myung; Yang, Maeng Ho; Kim, Hyun Joon; Chung, Hwan Sam; Oh, Keun Bae; Lee, Byung OoK; Ko, Han Suk; Song, Ki Dong; Lee, Man Ki; Moon, Ki Hwan; Lee, Han Myung

    1994-01-01

    In this study, the back-end nuclear fuel cycle acceptability is surveyed and analyzed in the following three aspects. To begin with, the future political situation and energy-environmental issues are analyzed as part of the socio-economic aspect. Secondly, the domestic situation of nuclear industries and the fuel cycle policy of foreign countries are surveyed as the technical aspect. Finally, NPT, IAEA safeguards and nuclear export control regimes are analyzed as the institutional aspect. The unification period of South and North Korea also will greatly affect the implementation of back-end fuel cycle policy, and public attitudes will affect the acquisition of site, construction, and operation of nuclear facilities. An effort to release international restrictions on the back-end fuel cycle is also required to accelerate the implementation of the policy. In this regard, the back-end fuel cycle policy should be clear-cut to avoid misunderstanding with respect to nuclear proliferation. Importantly, agreements with foreign countries should be amended at a mutual equivalent level. (Author) 30 refs., 5 figs., 25 tabs

  11. Analysis of factors affecting the implementation of back-end nuclear fuel cycle policy in Korea

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Yung Myung; Yang, Maeng Ho; Kim, Hyun Joon; Chung, Hwan Sam; Oh, Keun Bae; Lee, Byung OoK; Ko, Han Suk; Song, Ki Dong; Lee, Man Ki; Moon, Ki Hwan; Lee, Han Myung [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1994-01-01

    In this study, the back-end nuclear fuel cycle acceptability is surveyed and analyzed in the following three aspects. To begin with, the future political situation and energy-environmental issues are analyzed as part of the socio-economic aspect. Secondly, the domestic situation of nuclear industries and the fuel cycle policy of foreign countries are surveyed as the technical aspect. Finally, NPT, IAEA safeguards and nuclear export control regimes are analyzed as the institutional aspect. The unification period of South and North Korea also will greatly affect the implementation of back-end fuel cycle policy, and public attitudes will affect the acquisition of site, construction, and operation of nuclear facilities. An effort to release international restrictions on the back-end fuel cycle is also required to accelerate the implementation of the policy. In this regard, the back-end fuel cycle policy should be clear-cut to avoid misunderstanding with respect to nuclear proliferation. Importantly, agreements with foreign countries should be amended at a mutual equivalent level. (Author) 30 refs., 5 figs., 25 tabs.

  12. A Modeling Approach for Plastic-Metal Laser Direct Joining

    Science.gov (United States)

    Lutey, Adrian H. A.; Fortunato, Alessandro; Ascari, Alessandro; Romoli, Luca

    2017-09-01

    Laser processing has been identified as a feasible approach to direct joining of metal and plastic components without the need for adhesives or mechanical fasteners. The present work sees development of a modeling approach for conduction and transmission laser direct joining of these materials based on multi-layer optical propagation theory and numerical heat flow simulation. The scope of this methodology is to predict process outcomes based on the calculated joint interface and upper surface temperatures. Three representative cases are considered for model verification, including conduction joining of PBT and aluminum alloy, transmission joining of optically transparent PET and stainless steel, and transmission joining of semi-transparent PA 66 and stainless steel. Conduction direct laser joining experiments are performed on black PBT and 6082 anticorodal aluminum alloy, achieving shear loads of over 2000 N with specimens of 2 mm thickness and 25 mm width. Comparison with simulation results shows that consistently high strength is achieved where the peak interface temperature is above the plastic degradation temperature. Comparison of transmission joining simulations and published experimental results confirms these findings and highlights the influence of plastic layer optical absorption on process feasibility.

  13. QuickJoin—Fast Neighbour-Joining Tree Reconstruction

    DEFF Research Database (Denmark)

    Mailund; Pedersen, Christian N. Storm

    2004-01-01

    We have built a tool for fast construction of very large phylogenetic trees. The tool uses heuristics for speeding up the neighbour-joining algorithm—while still constructing the same tree as the original neighbour-joining algorithm—making it possible to construct trees for ~8000 species in less...

  14. Fermilab Friends for Science Education | Join Us

    Science.gov (United States)

    Fermilab Friends for Science Education FFSE Home About Us Join Us Support Us Contact Us Join Us improving science (science, technology, engineering and mathematics) education. Your donation allows us to membership dues allow us to create new, innovative science education programs, making the best use of unique

  15. Potentials of Industrie 4.0 and Machine Learning for Mechanical Joining

    OpenAIRE

    Jäckel, Mathias

    2017-01-01

    -Sensitivity analysis of the influence of component properties and joining parameters on the joining result for self-pierce riveting -Possibilities to link mechanical joining technologies with the automotive process chain for quality and flexibility improvements -Potential of using machine learning to reduce automotive product development cycles in relation to mechanical joining -Datamining for machine learning at mechanical joining

  16. Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks.

    Science.gov (United States)

    Uematsu, Naoya; Weterings, Eric; Yano, Ken-ichi; Morotomi-Yano, Keiko; Jakob, Burkhard; Taucher-Scholz, Gisela; Mari, Pierre-Olivier; van Gent, Dik C; Chen, Benjamin P C; Chen, David J

    2007-04-23

    The DNA-dependent protein kinase catalytic subunit (DNA-PK(CS)) plays an important role during the repair of DNA double-strand breaks (DSBs). It is recruited to DNA ends in the early stages of the nonhomologous end-joining (NHEJ) process, which mediates DSB repair. To study DNA-PK(CS) recruitment in vivo, we used a laser system to introduce DSBs in a specified region of the cell nucleus. We show that DNA-PK(CS) accumulates at DSB sites in a Ku80-dependent manner, and that neither the kinase activity nor the phosphorylation status of DNA-PK(CS) influences its initial accumulation. However, impairment of both of these functions results in deficient DSB repair and the maintained presence of DNA-PK(CS) at unrepaired DSBs. The use of photobleaching techniques allowed us to determine that the kinase activity and phosphorylation status of DNA-PK(CS) influence the stability of its binding to DNA ends. We suggest a model in which DNA-PK(CS) phosphorylation/autophosphorylation facilitates NHEJ by destabilizing the interaction of DNA-PK(CS) with the DNA ends.

  17. New Joining Technology for Optimized Metal/Composite Assemblies

    Directory of Open Access Journals (Sweden)

    Holger Seidlitz

    2014-01-01

    Full Text Available The development of a new joining technology, which is used to manufacture high strength hybrid constructions with thermoplastic composites (FRP and metals, is introduced. Similar to natural regulation effects at trees, fibers around the FRP joint become aligned along the lines of force and will not be destroyed by the joining process. This is achieved by the local utilization of the specific plastic flow properties of the FRT and metal component. Compared with usual joining methods—such as flow drill screws, blind and self-piercing rivets—noticeably higher tensile properties can be realized through the novel process management. The load-bearing capability increasing effect could be proved on hybrid joints with hot-dip galvanized steel HX420LAD and orthotropic glass—as well as carbon—fiber reinforced plastics. The results, which were determined in tensile-shear and cross-shear tests according to DIN EN ISO 14273 and DIN EN ISO 14272, are compared with holding loads of established joining techniques with similar joining point diameter and material combinations.

  18. Transcription-associated processes cause DNA double-strand breaks and translocations in neural stem/progenitor cells.

    Science.gov (United States)

    Schwer, Bjoern; Wei, Pei-Chi; Chang, Amelia N; Kao, Jennifer; Du, Zhou; Meyers, Robin M; Alt, Frederick W

    2016-02-23

    High-throughput, genome-wide translocation sequencing (HTGTS) studies of activated B cells have revealed that DNA double-strand breaks (DSBs) capable of translocating to defined bait DSBs are enriched around the transcription start sites (TSSs) of active genes. We used the HTGTS approach to investigate whether a similar phenomenon occurs in primary neural stem/progenitor cells (NSPCs). We report that breakpoint junctions indeed are enriched around TSSs that were determined to be active by global run-on sequencing analyses of NSPCs. Comparative analyses of transcription profiles in NSPCs and B cells revealed that the great majority of TSS-proximal junctions occurred in genes commonly expressed in both cell types, possibly because this common set has higher transcription levels on average than genes transcribed in only one or the other cell type. In the latter context, among all actively transcribed genes containing translocation junctions in NSPCs, those with junctions located within 2 kb of the TSS show a significantly higher transcription rate on average than genes with junctions in the gene body located at distances greater than 2 kb from the TSS. Finally, analysis of repair junction signatures of TSS-associated translocations in wild-type versus classical nonhomologous end-joining (C-NHEJ)-deficient NSPCs reveals that both C-NHEJ and alternative end-joining pathways can generate translocations by joining TSS-proximal DSBs to DSBs on other chromosomes. Our studies show that the generation of transcription-associated DSBs is conserved across divergent cell types.

  19. Silencing of end-joining repair for efficient site-specific gene insertion after TALEN/CRISPR mutagenesis in Aedes aegypti.

    Science.gov (United States)

    Basu, Sanjay; Aryan, Azadeh; Overcash, Justin M; Samuel, Glady Hazitha; Anderson, Michelle A E; Dahlem, Timothy J; Myles, Kevin M; Adelman, Zach N

    2015-03-31

    Conventional control strategies for mosquito-borne pathogens such as malaria and dengue are now being complemented by the development of transgenic mosquito strains reprogrammed to generate beneficial phenotypes such as conditional sterility or pathogen resistance. The widespread success of site-specific nucleases such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 in model organisms also suggests that reprogrammable gene drive systems based on these nucleases may be capable of spreading such beneficial phenotypes in wild mosquito populations. Using the mosquito Aedes aegypti, we determined that mutations in the FokI domain used in TALENs to generate obligate heterodimeric complexes substantially and significantly reduce gene editing rates. We found that CRISPR/Cas9-based editing in the mosquito Ae. aegypti is also highly variable, with the majority of guide RNAs unable to generate detectable editing. By first evaluating candidate guide RNAs using a transient embryo assay, we were able to rapidly identify highly effective guide RNAs; focusing germ line-based experiments only on this cohort resulted in consistently high editing rates of 24-90%. Microinjection of double-stranded RNAs targeting ku70 or lig4, both essential components of the end-joining response, increased recombination-based repair in early embryos as determined by plasmid-based reporters. RNAi-based suppression of Ku70 concurrent with embryonic microinjection of site-specific nucleases yielded consistent gene insertion frequencies of 2-3%, similar to traditional transposon- or ΦC31-based integration methods but without the requirement for an initial docking step. These studies should greatly accelerate investigations into mosquito biology, streamline development of transgenic strains for field releases, and simplify the evaluation of novel Cas9-based gene drive systems.

  20. Microwave joining of SiC ceramics and composites

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, I.; Silberglitt, R.; Tian, Y.L. [FM Technologies, Inc., Fairfax, VA (United States); Katz, J.D. [Los Alamos National Lab., NM (United States)

    1997-04-01

    Potential applications of SiC include components for advanced turbine engines, tube assemblies for radiant burners and petrochemical processing and heat exchangers for high efficiency electric power generation systems. Reliable methods for joining SiC are required in order to cost-effectively fabricate components for these applications from commercially available shapes and sizes. This manuscript reports the results of microwave joining experiments performed using two different types of SiC materials. The first were on reaction bonded SiC, and produced joints with fracture toughness equal to or greater than that of the base material over an extended range of joining temperatures. The second were on continuous fiber-reinforced SiC/SiC composite materials, which were successfully joined with a commercial active brazing alloy, as well as by using a polymer precursor.

  1. Enabling Dissimilar Material Joining Using Friction Stir Scribe Technology

    Energy Technology Data Exchange (ETDEWEB)

    Hovanski, Yuri; Upadyay, Piyush; Kleinbaum, Sarah; Carlson, Blair; Boettcher, Eric; Ruokolainen, Robert

    2017-04-05

    One challenge in adapting welding processes to dissimilar material joining is the diversity of melting temperatures of the different materials. Although the use of mechanical fasteners and adhesives have mostly paved the way for near-term implementation of dissimilar material systems, these processes only accentuate the need for low-cost welding processes capable of joining dissimilar material components regardless of alloy, properties, or melting temperature. Friction stir scribe technology was developed to overcome the challenges of joining dissimilar material components where melting temperatures vary greatly, and properties and/or chemistry are not compatible with more traditional welding processes. Although the friction stir scribe process is capable of joining dissimilar metals and metal/polymer systems, a more detailed evaluation of several aluminum/steel joints is presented herein to demonstrate the ability to both chemically and mechanically join dissimilar materials.

  2. Towards mastering CRISPR-induced gene knock-in in plants: Survey of key features and focus on the model Physcomitrella patens.

    Science.gov (United States)

    Collonnier, Cécile; Guyon-Debast, Anouchka; Maclot, François; Mara, Kostlend; Charlot, Florence; Nogué, Fabien

    2017-05-15

    Beyond its predominant role in human and animal therapy, the CRISPR-Cas9 system has also become an essential tool for plant research and plant breeding. Agronomic applications rely on the mastery of gene inactivation and gene modification. However, if the knock-out of genes by non-homologous end-joining (NHEJ)-mediated repair of the targeted double-strand breaks (DSBs) induced by the CRISPR-Cas9 system is rather well mastered, the knock-in of genes by homology-driven repair or end-joining remains difficult to perform efficiently in higher plants. In this review, we describe the different approaches that can be tested to improve the efficiency of CRISPR-induced gene modification in plants, which include the use of optimal transformation and regeneration protocols, the design of appropriate guide RNAs and donor templates and the choice of nucleases and means of delivery. We also present what can be done to orient DNA repair pathways in the target cells, and we show how the moss Physcomitrella patens can be used as a model plant to better understand what DNA repair mechanisms are involved, and how this knowledge could eventually be used to define more performant strategies of CRISPR-induced gene knock-in. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Joining of Aluminium Alloy and Steel by Laser Assisted Reactive Wetting

    Science.gov (United States)

    Liedl, Gerhard; Vázquez, Rodrigo Gómez; Murzin, Serguei P.

    2018-03-01

    Compounds of dissimilar materials, like aluminium and steel offer an interesting opportunity for the automotive industry to reduce the weight of a car body. Thermal joining of aluminium and steel leads to the formation of brittle intermetallic compounds, which negatively affects the properties of the welded joint. Amongst others, growth of such intermetallic compounds depends on maximum temperature and on the time at certain temperatures. Laser welding with its narrow well seam and its fast heating and cooling cycles provides an excellent opportunity to obtain an ultrathin diffusion zone. Joining of sheet metal DC01 with aluminium alloy AW6016 has been chosen for research. The performed experimental studies showed that by a variation of the beam power and scanning speed it is possible to obtain an ultrathin diffusion zone with narrow intermetallic interlayers. With the aim of supporting further investigation of laser welding of the respective and other dissimilar pairings a multi-physical simulation model has been developed.

  4. On the Complexity of Inner Product Similarity Join

    DEFF Research Database (Denmark)

    Ahle, Thomas Dybdahl; Pagh, Rasmus; Razenshteyn, Ilya

    2016-01-01

    A number of tasks in classification, information retrieval, recommendation systems, and record linkage reduce to the core problem of inner product similarity join (IPS join): identifying pairs of vectors in a collection that have a sufficiently large inner product. IPS join is well understood when...... bounds for (A)LSH-based algorithms. In particular, we show that asymmetry can be avoided by relaxing the LSH definition to only consider the collision probability of distinct elements. A new indexing method for IPS based on linear sketches, implying that our hardness results are not far from being tight...

  5. Effect of epoxy resin on bending momentum in L type corner joins

    Directory of Open Access Journals (Sweden)

    Mehmet Nuri Yıldırım

    2017-11-01

    Full Text Available In the furniture industry, the joining points of frame and box construction furniture according to the loads to be affected by the use place is important for the security of the user and service life of the furniture element. In this direction, it is aimed to determine the diagonal compression and diagonal tensile moment values of "L" type corner joints of box framed construction furniture prepared from solid wood materials. The Pinus Nigra, Fagus Orientalis L and Populus Nigra were used as solid wood materials. Wood-based biscuit joining elements were used in corner joints of the test construction and epoxy resin was used as glue for materials. The static loads were applied to construction according to ASTM-D1037. The results show that, the highest tensile and compression values were obtained from Fagus Orientalis L and the lowest values were obtained from Populus Nigra specimens. In the statistical study, the difference between the tensile and compressive bending moment values of the biscuit connection element was found to be statistically significant. This study indicates that, it is suggested to use of L type joints prepared from Fagus Orientalis L by using epoxy resin and wood based biscuit joining element in frame constructions.

  6. Effects of processing parameters on Be/CuCrZr joining

    International Nuclear Information System (INIS)

    Park, Jeong-Yong; Lee, Jung-Suk; Choi, Byung-Kwon; Park, Sang-Yun; Hong, Bong Guen; Jeong, Yong Hwan; Jung, Ki-Jung

    2007-01-01

    A joining of Be/CuCrZr has been considered as the key technology for the fabrication of the ITER first wall. Among the joining methods, Hot isostatic pressing (HIP), which is one of the diffusion bonding methods, is the most feasible method to join the Be and CuCrZr alloy. In the HIP joining of Be and CuCrZr, the interlayer was used to prevent the formation of brittle intermetallic compounds in the interface. Therefore, it is crucial to select a suitable interlayer for a joining of Be and CuCrZr. On the other hand, the diffusion between Be and CuCrZr would be enhanced with an increase of the HIP joining temperature, thereby increasing the joint strength. However, the HIP joining temperature is limited by the mechanical properties of CuCrZr. During the fabrication process of the ITER first wall, CuCrZr is subjected to several thermal cycles including a solution annealing, a cooling and an aging. The HIP joining of Be and CuCrZr corresponds to the aging of CuCrZr. The HIP joining at a higher temperature would cause a degradation of the mechanical properties of CuCrZr by an overaging effect although it is preferable for an improvement of the joint strength. In this study, the effect of the cooling rate on the mechanical properties of aged CuCrZr was investigated to find the maximum HIP temperature without a degradation of the mechanical properties of CuCrZr

  7. Nonhomologous Recombination between Defective Poliovirus and Coxsackievirus Genomes Suggests a New Model of Genetic Plasticity for Picornaviruses

    Science.gov (United States)

    Holmblat, Barbara; Jégouic, Sophie; Muslin, Claire; Blondel, Bruno; Joffret, Marie-Line

    2014-01-01

    ABSTRACT Most of the circulating vaccine-derived polioviruses (cVDPVs) implicated in poliomyelitis outbreaks in Madagascar have been shown to be recombinants between the type 2 poliovirus (PV) strain of the oral polio vaccine (Sabin 2) and another species C human enterovirus (HEV-C), such as type 17 coxsackie A virus (CA17) in particular. We studied intertypic genetic exchanges between PV and non-PV HEV-C by developing a recombination model, making it possible to rescue defective type 2 PV RNA genomes with a short deletion at the 3′ end by the cotransfection of cells with defective or infectious CA17 RNAs. We isolated over 200 different PV/CA17 recombinants, using murine cells expressing the human PV receptor (PVR) and selecting viruses with PV capsids. We found some homologous (H) recombinants and, mostly, nonhomologous (NH) recombinants presenting duplications of parental sequences preferentially located in the regions encoding proteins 2A, 2B, and 3A. Short duplications appeared to be stable, whereas longer duplications were excised during passaging in cultured cells or after multiplication in PVR-transgenic mice, generating H recombinants with diverse sites of recombination. This suggests that NH recombination events may be a transient, intermediate step in the generation and selection of the fittest H recombinants. In addition to the classical copy-choice mechanism of recombination thought to generate mostly H recombinants, there may also be a modular mechanism of recombination, involving NH recombinant precursors, shaping the genomes of recombinant enteroviruses and other picornaviruses. PMID:25096874

  8. Joining of Gamma Titanium Aluminides

    National Research Council Canada - National Science Library

    Baeslack, William

    2002-01-01

    .... Although organized and presented by joining process, many of the observations made and relationships developed, particularly those regarding the weldability and welding metallurgy of gamma titanium...

  9. [Early prenatal diagnosis of diprosopic syncephalic joined twins].

    Science.gov (United States)

    Picaud, A; Nlome-Nze, A R; Engongha-Beka, T; Ogowet-Igumu, N

    1990-06-01

    The authors summarize the case of diprosopic syncephalic joined twins diagnosed at 22 weeks of pregnancy by ultrasonography performed because of hydramnios. The rate of separation anomalies of monozygotic twins is assessed by a review of the literature: from 1 to twenty to fifty thousands for joined twins to 1 per cent fifty thousand to fifteen millions for diprosopus. The etiology is the result of a late division of the egg between D12 and D16. Often an encephalic diprosopic joined twins cause elevated levels of maternal serum alpha protein. Early ultrasonography permits to consider a vaginal therapeutic abortion.

  10. The antimetabolite ara-CTP blocks long-term memory of conditioned taste aversion.

    Science.gov (United States)

    Wang, Jianpeng; Ren, Keqin; Pérez, Javier; Silva, Alcino J; Peña de Ortiz, Sandra

    2003-01-01

    We examined the hypothesis that processes related to DNA recombination and repair are involved in learning and memory. Rats received intracerebroventricular (i.c.v.) infusions of the antimetabolite 1-beta-D-arabinofuranosylcytosine triphosphate (ara-CTP) or its precursor cytosine arabinoside (ara-C) 30 min prior to conditioned taste aversion (CTA) training. Both ara-CTP and ara-C caused significant impairments in long-term memory (LTM) of CTA. Control experiments indicate that the effect of ara-CTP on CTA memory is related to interference with learning. Furthermore, as it was previously demonstrated for the protein synthesis inhibitor anisomycin, ara-CTP had no effect on CTA memory when it was injected 1 h after training. Importantly, although both ara-CTP and anisomycin significantly blocked LTM in the task, short-term memory (STM) measured 1 h after training was not affected by either of the drugs. Finally, ara-CTP had no effect on in vitro transcription, but it did effectively block nonhomologous DNA end joining (NHEJ) activity of brain protein extracts. We suggest that DNA ligase-mediated DNA recombination and repair processes are necessary for the expression of LTM in the brain.

  11. Clinical and molecular studies in four patients with SRY-positive 46,XX testicular disorders of sex development: implications for variable sex development and genomic rearrangements.

    Science.gov (United States)

    Nakashima, Shinichi; Ohishi, Akira; Takada, Fumio; Kawamura, Hideki; Igarashi, Maki; Fukami, Maki; Ogata, Tsutomu

    2014-10-01

    We report four patients with SRY-positive 46,XX testicular disorders of sex development (46,XX-TDSD) (cases 1-4). Case 1 exhibited underdeveloped external genitalia with hypospadias, case 2 manifested micropenis and cases 3 and 4 showed normal external genitalia. The Xp;Yp translocations occurred between the X- and the Y-differential regions in case 1, between PRKX and inverted PRKY in case 2 and between the X-chromosomal short arm pseudoautosomal region and the Y-differential regions in cases 3 and 4. The distance of the Yp breakpoint from SRY was ~0.75 Mb in case 1, ~6.5 Mb in case 2, ~2.3 Mb in case 3 and ~72 kb in case 4. The Xp;Yp translocation occurred within an 87-bp homologous segment of PRKX and PRKY in case 2, and between non-homologous regions with addition of an 18-bp sequence of unknown origin in case 4. X-inactivation analysis revealed random inactivation in cases 1-4. The results argue against the notion that undermasculinization in 46,XX-TDSD is prone to occur when translocated Yp materials are small (<100 kb of the Y-differential region), and imply that the Xp;Yp translocations result from several mechanisms including non-allelic homologous recombination and non-homologous end joining.

  12. Chromosomal Integrity after UV Irradiation Requires FANCD2-Mediated Repair of Double Strand Breaks.

    Science.gov (United States)

    Federico, María Belén; Vallerga, María Belén; Radl, Analía; Paviolo, Natalia Soledad; Bocco, José Luis; Di Giorgio, Marina; Soria, Gastón; Gottifredi, Vanesa

    2016-01-01

    Fanconi Anemia (FA) is a rare autosomal recessive disorder characterized by hypersensitivity to inter-strand crosslinks (ICLs). FANCD2, a central factor of the FA pathway, is essential for the repair of double strand breaks (DSBs) generated during fork collapse at ICLs. While lesions different from ICLs can also trigger fork collapse, the contribution of FANCD2 to the resolution of replication-coupled DSBs generated independently from ICLs is unknown. Intriguingly, FANCD2 is readily activated after UV irradiation, a DNA-damaging agent that generates predominantly intra-strand crosslinks but not ICLs. Hence, UV irradiation is an ideal tool to explore the contribution of FANCD2 to the DNA damage response triggered by DNA lesions other than ICL repair. Here we show that, in contrast to ICL-causing agents, UV radiation compromises cell survival independently from FANCD2. In agreement, FANCD2 depletion does not increase the amount of DSBs generated during the replication of UV-damaged DNA and is dispensable for UV-induced checkpoint activation. Remarkably however, FANCD2 protects UV-dependent, replication-coupled DSBs from aberrant processing by non-homologous end joining, preventing the accumulation of micronuclei and chromatid aberrations including non-homologous chromatid exchanges. Hence, while dispensable for cell survival, FANCD2 selectively safeguards chromosomal stability after UV-triggered replication stress.

  13. Unraveling the Secondary Metabolism of the Biotechnological Important Filamentous Fungus Trichoderma reesei ( Teleomorph Hypocrea jecorina)

    DEFF Research Database (Denmark)

    Jørgensen, Mikael Skaanning

    that would enable pursuance of the primary objective. The developed molecular tools were assembled into an expression system for high-throughput construction of defined integrated T. reesei mutants and combined inactivation of the non-homologous end joining pathway that facilitates ectopic integration...... of exposed DNA fragments, and a color maker so that the mutants, in which the substrate had been integrated correct, could be identified by their phenotype. A new bidirectional selective marker system was developed based on the pyr2 gene, involved in the pyrimidine biosynthesis pathway, and was included...... essential for biosynthesis of the sorbicillinoids. Hence, genes involved in biosynthesis of this group of polyketides were identified for the first time. Comparative genomics was subsequently used to identify a highly similar polyketide synthase gene cluster in another well-known sorbicillinoid producer...

  14. APOBEC3B-Mediated Cytidine Deamination Is Required for Estrogen Receptor Action in Breast Cancer

    Directory of Open Access Journals (Sweden)

    Manikandan Periyasamy

    2015-10-01

    Full Text Available Estrogen receptor α (ERα is the key transcriptional driver in a large proportion of breast cancers. We report that APOBEC3B (A3B is required for regulation of gene expression by ER and acts by causing C-to-U deamination at ER binding regions. We show that these C-to-U changes lead to the generation of DNA strand breaks through activation of base excision repair (BER and to repair by non-homologous end-joining (NHEJ pathways. We provide evidence that transient cytidine deamination by A3B aids chromatin modification and remodelling at the regulatory regions of ER target genes that promotes their expression. A3B expression is associated with poor patient survival in ER+ breast cancer, reinforcing the physiological significance of A3B for ER action.

  15. DNA double-strand break repair: a tale of pathway choices

    Institute of Scientific and Technical Information of China (English)

    Jing Li; Xingzhi Xu

    2016-01-01

    Deoxyribonucleic acid double-strand breaks (DSBs) are cytotoxic lesions that must be repaired either through homologous recombination (HR) or non-homologous end-joining (NHEJ) pathways.DSB repair is critical for genome integrity,cellular homeostasis and also constitutes the biological foundation for radiotherapy and the majority of chemotherapy.The choice between HR and NHEJ is a complex yet not completely understood process that will entail more future efforts.Herein we review our current understandings about how the choice is made over an antagonizing balance between p53-binding protein 1 and breast cancer 1 in the context of cell cycle stages,downstream effects,and distinct chromosomal histone marks.These exciting areas of research will surely bring more mechanistic insights about DSB repair and be utilized in the clinical settings.

  16. Synapsis-defective mutants reveal a correlation between chromosome conformation and the mode of double-strand break repair during Caenorhabditis elegans meiosis.

    Science.gov (United States)

    Smolikov, Sarit; Eizinger, Andreas; Hurlburt, Allison; Rogers, Eric; Villeneuve, Anne M; Colaiácovo, Mónica P

    2007-08-01

    SYP-3 is a new structural component of the synaptonemal complex (SC) required for the regulation of chromosome synapsis. Both chromosome morphogenesis and nuclear organization are altered throughout the germlines of syp-3 mutants. Here, our analysis of syp-3 mutants provides insights into the relationship between chromosome conformation and the repair of meiotic double-strand breaks (DSBs). Although crossover recombination is severely reduced in syp-3 mutants, the production of viable offspring accompanied by the disappearance of RAD-51 foci suggests that DSBs are being repaired in these synapsis-defective mutants. Our studies indicate that once interhomolog recombination is impaired, both intersister recombination and nonhomologous end-joining pathways may contribute to repair during germline meiosis. Moreover, our studies suggest that the conformation of chromosomes may influence the mode of DSB repair employed during meiosis.

  17. Joining and fabrication techniques for high temperature structures including the first wall in fusion reactor

    International Nuclear Information System (INIS)

    Lee, Ho Jin; Lee, B. S.; Kim, K. B.

    2003-09-01

    The materials for PFC's (Plasma Facing Components) in a fusion reactor are severely irradiated with fusion products in facing the high temperature plasma during the operation. The refractory materials can be maintained their excellent properties in severe operating condition by lowering surface temperature by bonding them to the high thermal conducting materials of heat sink. Hence, the joining and bonding techniques between dissimilar materials is considered to be important in case of the fusion reactor or nuclear reactor which is operated at high temperature. The first wall in the fusion reactor is heated to approximately 1000 .deg. C and irradiated severely by the plasma. In ITER, beryllium is expected as the primary armour candidate for the PFC's; other candidates including W, Mo, SiC, B4C, C/C and Si 3 N 4 . Since the heat affected zones in the PFC's processed by conventional welding are reported to have embrittlement and degradation in the sever operation condition, both brazing and diffusion bonding are being considered as prime candidates for the joining technique. In this report, both the materials including ceramics and the fabrication techniques including joining technique between dissimilar materials for PFC's are described. The described joining technique between the refractory materials and the dissimilar materials may be applicable for the fusion reactor and Generation-4 future nuclear reactor which are operated at high temperature and high irradiation

  18. Development of Joining Technologies for the ITER Blanket First Wall

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Byoung Kwon; Jung, Yang Il; Park, Dong Jun; Kim, Hyun Gil; Park, Sang Yoon; Park, Jeong Yong; Jeong, Yong Hwan; Lee, Dong Won; Kim, Suk Kwon [KAERI, Daejeon (Korea, Republic of)

    2011-01-15

    The design of the ITER blanket first wall includes the Beryllium amour tiles joined to CuCrZr heat sink with stainless steel cooling tubes. For the ITER application, the Be/CuCrZr/SS joint was proposed as a first wall material. The joining of Be/CuCrZr as well as CuCrZr/SS was generally carried out by using a hot isostatic pressing (CuC) in many countries. The joining strength for Be/CuCrZr is relatively lower than that for CuCrZr/SS, since we usually forms surface oxides (BeO) and brittle a metallics with Cu. Therefore, the joining technology for the Be/CuCrZr joint has been investigated. Be is apt to adsorb oxygen in an air atmosphere, so we should be etched to eliminate the surface pre-oxide using a chemical solution and Ar ions in a vacuum chamber. Then we is coated with a first was to prevent further oxidation. The kinds of a first we are chosen to be able to enhance the joining strength as inhibiting excessive be diffusion. The performance of the Be/CuCrZr/SS joint used for the ITER first wall is primarily dependent on the joining strength of the Be/CuCrZr interface. The Cr/Cu and Ti/Cr/Cu interlayers enabled the successful joining of be tile to CuCrZr plate. Moreover, ion-beam assisted deposition (IBAD) increased joining strength of the Be/CuCrZr joint mock-ups. IBAD induced the increased packing of depositing atoms, which resulted in denser and more adhesive interlayers. The interlayers formed by IBAD process revealed about 40% improved resistance to the scratch test. It is suggested that the improved adhesion of coating interlayers enabled tight joining of Be and CuCrZr blocks. As compared to without IBAD coating, the shear strength as well as the 4-point bend strength were increased more than 20% depending on interlayer types and coating conditions

  19. Development of Joining Technologies for the ITER Blanket First Wall

    International Nuclear Information System (INIS)

    Choi, Byoung Kwon; Jung, Yang Il; Park, Dong Jun; Kim, Hyun Gil; Park, Sang Yoon; Park, Jeong Yong; Jeong, Yong Hwan; Lee, Dong Won; Kim, Suk Kwon

    2011-01-01

    The design of the ITER blanket first wall includes the Beryllium amour tiles joined to CuCrZr heat sink with stainless steel cooling tubes. For the ITER application, the Be/CuCrZr/SS joint was proposed as a first wall material. The joining of Be/CuCrZr as well as CuCrZr/SS was generally carried out by using a hot isostatic pressing (CuC) in many countries. The joining strength for Be/CuCrZr is relatively lower than that for CuCrZr/SS, since we usually forms surface oxides (BeO) and brittle a metallics with Cu. Therefore, the joining technology for the Be/CuCrZr joint has been investigated. Be is apt to adsorb oxygen in an air atmosphere, so we should be etched to eliminate the surface pre-oxide using a chemical solution and Ar ions in a vacuum chamber. Then we is coated with a first was to prevent further oxidation. The kinds of a first we are chosen to be able to enhance the joining strength as inhibiting excessive be diffusion. The performance of the Be/CuCrZr/SS joint used for the ITER first wall is primarily dependent on the joining strength of the Be/CuCrZr interface. The Cr/Cu and Ti/Cr/Cu interlayers enabled the successful joining of be tile to CuCrZr plate. Moreover, ion-beam assisted deposition (IBAD) increased joining strength of the Be/CuCrZr joint mock-ups. IBAD induced the increased packing of depositing atoms, which resulted in denser and more adhesive interlayers. The interlayers formed by IBAD process revealed about 40% improved resistance to the scratch test. It is suggested that the improved adhesion of coating interlayers enabled tight joining of Be and CuCrZr blocks. As compared to without IBAD coating, the shear strength as well as the 4-point bend strength were increased more than 20% depending on interlayer types and coating conditions

  20. Inference of Large Phylogenies Using Neighbour-Joining

    DEFF Research Database (Denmark)

    Simonsen, Martin; Mailund, Thomas; Pedersen, Christian Nørgaard Storm

    2011-01-01

    The neighbour-joining method is a widely used method for phylogenetic reconstruction which scales to thousands of taxa. However, advances in sequencing technology have made data sets with more than 10,000 related taxa widely available. Inference of such large phylogenies takes hours or days using...... the Neighbour-Joining method on a normal desktop computer because of the O(n^3) running time. RapidNJ is a search heuristic which reduce the running time of the Neighbour-Joining method significantly but at the cost of an increased memory consumption making inference of large phylogenies infeasible. We present...... two extensions for RapidNJ which reduce the memory requirements and \\makebox{allows} phylogenies with more than 50,000 taxa to be inferred efficiently on a desktop computer. Furthermore, an improved version of the search heuristic is presented which reduces the running time of RapidNJ on many data...

  1. Possible role of repetitious DNA in recombinatory joining during chromosome rearrangement in Drosophila melanogaster

    International Nuclear Information System (INIS)

    Lee, C.S.

    1975-01-01

    It is postulated that certain repetitious DNA components play a role in the recombination processes during chromosome rearrangements. When the distribution of silver grain densities after the in situ hybridization of repetitious DNA and the distribution of chromosome breaks due to x-irradiation are compared, a strong correlation is found for the euchromatic portion of the D. melanogaster salivary X chromosome. These observations justify the postulate above that certain repetitious DNA provides homologous regions in the DNA of broken chromosome ends necessary for proper recombinatory joining. (U.S.)

  2. Development of low temperature solid state joining technology of dissimilar for nuclear heat exchanger tube components

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2010-08-15

    By conventional fusion welding process (TIG), a realization of reliable and sound joints for the nuclear heat exchanger components is very difficult, especially for the parts comprising of the dissimilar metal couples (Ti-STS, Ti-Cu alloy etc.). This is mainly attributed to the formation of brittle intermetallics (Ti{sub x}Cu{sub y}, Ti{sub x}Fe{sub y}, Ti{sub x}Ni{sub y} etc.) and wide difference in physical properties. Moreover, it usually employs very high thermal input, so making it difficult to obtain sound joints due to generations of high residual stresses and degradation of the adjacent base metals, even for similar metal combinations. In this project, the low temperature solid-state joining technology was established by developing new alloy fillers, e.g. the multi-component eutectic based alloys or amorphous alloys, and thereby lowering the joining temperature down to {approx}800 .deg. C without affecting the structural properties of base metals. Based on a low temperature joining, the interlayer engineering technology was then developed to be able to eliminate the brittleness of the joints for strong Ti-STS dissimilar joints, and the diffusion brazing technology of Ti-Ti with a superior joining strength and corrosion-resistance comparable to those of base metal were developed. By using those developed technologies, the joining procedures feasible for the heat exchanger components were finally established for the dissimilar metal joints including Ti tube sheet to super STS tube, Ti tube sheet to super STS tube sheet, and the joints of the Ti tube to Ti tube sheet

  3. Researching the Ends of Identity: Birth and Death on Social Media

    OpenAIRE

    Tama Leaver

    2015-01-01

    This paper argues that expanding the scope of social media studies to examine birth and early life at one end, and death and memorialisation at the other, demonstrates that social media is never just about an individual, but also the way individuals are always already joined together as families, groups, communities and more. Mapping these ends of identity also reveals more of the nuances of everyday social media use and its impact.

  4. Compliance between clinical and genetic diagnosis of choroidal hypoplasia in 103 Norwegian Border Collie puppies.

    Science.gov (United States)

    Grosås, Siv; Lingaas, Frode; Prestrud, Kristin Wear; Ropstad, Ernst-Otto

    2017-11-07

    To describe the frequency of the nonhomologous end-joining factor 1 (NHEJ1) mutation and the compliance between clinical and genetic diagnosis of choroidal hypoplasia (CH) in a group of Norwegian Border Collies. Border collie puppies in the age from 5 to 8 weeks. Puppies included in the study had a complete ophthalmological examination. All findings were recorded, and an ECVO scheme form was issued for each puppy. DNA samples were achieved from buccal swabs. Genetic typing was performed for the 7.8-kb deletion in the gene encoding NHEJ1. Dogs with none, one, or two copies of the mutated allele were classified as free, carriers, and affected, respectively. 103 Border Collie puppies from 16 litters, 52 females and 51 males, were included in the study. Ages ranged from 5.1 to 8.9 weeks. One puppy had clinical findings consistent with CH and optic nerve coloboma compatible with the diagnosis Collie Eye Anomaly (CEA). Findings on ophthalmological examination of the remaining puppies were within normal limits. On genetic testing, 85 puppies were clear of the mutation in the NHEJ1 gene, 17 puppies were carriers, and one puppy was genetically affected. A good compliance between the clinical diagnosis and the genetic test results was found in all of the puppies examined. The allele frequency of the mutation was 6.3%. © 2017 American College of Veterinary Ophthalmologists.

  5. Joining of thin thickness SiCf/SiC composites: research of a joining composition and of an associated elaboration process

    International Nuclear Information System (INIS)

    Jacques, E.

    2012-01-01

    The present work is part of the Fourth Generation Fast Reactors program. One of the key issues is the joining of the SiC f /SiC ceramic matrix composites (CMC) to seal the combustible cladding. At the present time, no chemical composition as a joint is refractive enough to face the expected operating temperatures. The aims of this study are the following: (1) the identification of a joining composition and its associated elaboration process answering the specifications, (2) the validation of a local and fast heating process and (3) the definition of mechanical tests to characterise these joinings. We describe the methodology and the results for joining SiC and SiC f /SiC substrates at solid and liquid state using metallic silicides. Joint integrity and joint strength can be improved by adding small SiC particles to the silicides powders. Chemical reactivity, wettability tests and thermomechanical properties analysis have been carried out on the joints. Cross sections of the assembly were prepared to study the joint/substrate chemical bonding, the cracking and the crack deflection in the vicinity of the interface. Also, one of the challenge consists in using a local heating at a high temperature (around 1800 C) for a short time to avoid the degradation of the composite structure. The assemblies have been then performed in an inductive furnace but in order to prepare the joining technology, trials of local heating have also been investigated with a CO 2 laser beam and a microwave generator. Finally, descriptions of the 4-points bending mechanical test used and the associated results are presented. (author) [fr

  6. Microstructure and properties of ceramics and composites joined by plastic deformation.

    Energy Technology Data Exchange (ETDEWEB)

    Goretta, K. C.; Singh, D.; Chen, N.; Gutierrez-Mora, F.; Lorenzo-Martin, M. de la, Cinta; Dominguez-Rodriguez, A.; Routbort, J. L.; Energy Systems; Univ. of Seville

    2008-12-01

    A review is presented of the design of suitable materials systems for joining by high-temperature plastic deformation, details of the joining techniques, microstructures and properties of the resulting composite bodies, and prospects and limitation for this type of joining technology. Joining parameters and resulting forms are discussed for Al{sub 2}O{sub 3}/mullite particulate composites, Y{sub 2}O{sub 3}-stabilized ZrO{sub 2} particulate/Al{sub 2}O{sub 3} particulate and whisker-reinforced composites, hydroxyapatite bioceramics, La{sub 0.85}Sr{sub 0.15}MnO{sub 3} electronic ceramics, MgF{sub 2} optical ceramics, and Ni{sub 3}Al intermetallics. Results are contrasted with those obtained by other methods of joining brittle, high-temperature materials, with special focus on durability and mechanical properties.

  7. Microstructure and properties of ceramics and composites joined by plastic deformation

    Energy Technology Data Exchange (ETDEWEB)

    Goretta, K.C. [Argonne National Laboratory, Argonne, IL 60439-4838 (United States)], E-mail: ken.goretta@aoard.af.mil; Singh, D.; Chen Nan [Argonne National Laboratory, Argonne, IL 60439-4838 (United States); Gutierrez-Mora, F.; Cinta Lorenzo-Martin, M. de la [Argonne National Laboratory, Argonne, IL 60439-4838 (United States); University of Seville, Seville 41080 (Spain); Dominguez-Rodriguez, A. [University of Seville, Seville 41080 (Spain); Routbort, J.L. [Argonne National Laboratory, Argonne, IL 60439-4838 (United States)

    2008-12-20

    A review is presented of the design of suitable materials systems for joining by high-temperature plastic deformation, details of the joining techniques, microstructures and properties of the resulting composite bodies, and prospects and limitation for this type of joining technology. Joining parameters and resulting forms are discussed for Al{sub 2}O{sub 3}/mullite particulate composites, Y{sub 2}O{sub 3}-stabilized ZrO{sub 2} particulate/Al{sub 2}O{sub 3} particulate and whisker-reinforced composites, hydroxyapatite bioceramics, La{sub 0.85}Sr{sub 0.15}MnO{sub 3} electronic ceramics, MgF{sub 2} optical ceramics, and Ni{sub 3}Al intermetallics. Results are contrasted with those obtained by other methods of joining brittle, high-temperature materials, with special focus on durability and mechanical properties.

  8. Joining and fabrication techniques for high temperature structures including the first wall in fusion reactor

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ho Jin; Lee, B. S.; Kim, K. B

    2003-09-01

    The materials for PFC's (Plasma Facing Components) in a fusion reactor are severely irradiated with fusion products in facing the high temperature plasma during the operation. The refractory materials can be maintained their excellent properties in severe operating condition by lowering surface temperature by bonding them to the high thermal conducting materials of heat sink. Hence, the joining and bonding techniques between dissimilar materials is considered to be important in case of the fusion reactor or nuclear reactor which is operated at high temperature. The first wall in the fusion reactor is heated to approximately 1000 .deg. C and irradiated severely by the plasma. In ITER, beryllium is expected as the primary armour candidate for the PFC's; other candidates including W, Mo, SiC, B4C, C/C and Si{sub 3}N{sub 4}. Since the heat affected zones in the PFC's processed by conventional welding are reported to have embrittlement and degradation in the sever operation condition, both brazing and diffusion bonding are being considered as prime candidates for the joining technique. In this report, both the materials including ceramics and the fabrication techniques including joining technique between dissimilar materials for PFC's are described. The described joining technique between the refractory materials and the dissimilar materials may be applicable for the fusion reactor and Generation-4 future nuclear reactor which are operated at high temperature and high irradiation.

  9. Joining of Aluminium Alloy Sheets by Rectangular Mechanical Clinching

    International Nuclear Information System (INIS)

    Abe, Y.; Mori, K.; Kato, T.

    2011-01-01

    A mechanical clinching has the advantage of low running costs. However, the joint strength is not high. To improve the maximum load of the joined sheets by a mechanical clinching, square and rectangular mechanical clinching were introduced. In the mechanical clinching, the two sheets are mechanically joined by forming an interlock between the lower and upper sheets by the punch and die. The joined length with the interlock was increased by the rectangular punch and die. The deforming behaviours of the sheets in the mechanical clinching were investigated, and then the interlock in the sheets had distribution in the circumference of the projection. Although the interlocks were formed in both projection side and diagonal, the interlock in the diagonal was smaller because of the long contact length between the lower sheet and the die cavity surface. The maximum load of the joined sheets by the rectangular mechanical clinching was two times larger than the load by the round mechanical clinching.

  10. Fast and scalable inequality joins

    KAUST Repository

    Khayyat, Zuhair; Lucia, William; Singh, Meghna; Ouzzani, Mourad; Papotti, Paolo; Quiané -Ruiz, Jorge Arnulfo; Tang, Nan; Kalnis, Panos

    2016-01-01

    an incremental inequality join algorithm to handle scenarios where data keeps changing. We have implemented a centralized version of these algorithms on top of PostgreSQL, a distributed version on top of Spark SQL, and an existing data cleaning system, Nadeef

  11. Dual CRISPR-Cas9 Cleavage Mediated Gene Excision and Targeted Integration in Yarrowia lipolytica.

    Science.gov (United States)

    Gao, Difeng; Smith, Spencer; Spagnuolo, Michael; Rodriguez, Gabriel; Blenner, Mark

    2018-05-29

    CRISPR-Cas9 technology has been successfully applied in Yarrowia lipolytica for targeted genomic editing including gene disruption and integration; however, disruptions by existing methods typically result from small frameshift mutations caused by indels within the coding region, which usually resulted in unnatural protein. In this study, a dual cleavage strategy directed by paired sgRNAs is developed for gene knockout. This method allows fast and robust gene excision, demonstrated on six genes of interest. The targeted regions for excision vary in length from 0.3 kb up to 3.5 kb and contain both non-coding and coding regions. The majority of the gene excisions are repaired by perfect nonhomologous end-joining without indel. Based on this dual cleavage system, two targeted markerless integration methods are developed by providing repair templates. While both strategies are effective, homology mediated end joining (HMEJ) based method are twice as efficient as homology recombination (HR) based method. In both cases, dual cleavage leads to similar or improved gene integration efficiencies compared to gene excision without integration. This dual cleavage strategy will be useful for not only generating more predictable and robust gene knockout, but also for efficient targeted markerless integration, and simultaneous knockout and integration in Y. lipolytica. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. 49 CFR 192.283 - Plastic pipe: Qualifying joining procedures.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 3 2010-10-01 2010-10-01 false Plastic pipe: Qualifying joining procedures. 192... Materials Other Than by Welding § 192.283 Plastic pipe: Qualifying joining procedures. (a) Heat fusion... for making plastic pipe joints by a heat fusion, solvent cement, or adhesive method, the procedure...

  13. Superplasticity and joining of zirconia-based ceramics

    International Nuclear Information System (INIS)

    Dominguez-Rodriguez, A.; Gutierrez-Mora, F.; Jimenez-Melendo, M.; Chaim, R.; Routbort, J. L.

    1999-01-01

    Steady-state creep and joining of alumina/zirconia composites containing alumina volume fractions of 20, 60, and 85% have been investigated between 1,250 and 1,350 C. Superplasticity of these compounds is controlled by grain-boundary sliding and the creep rate is a function of alumina volume fraction, not grain size. Using the principles of superplasticity, pieces of the composite have been joined by applying the stress required to achieve 5 to 10% strain to form a strong interface at temperatures as low as 1,200 C

  14. Superplasticity and joining of zirconia-based ceramics

    International Nuclear Information System (INIS)

    Gutierrez-Mora, F.; Dominguez-Rodriguez, A.; Jimenez-Melendo, M.; Chaim, R.; Ravi, G.B.; Routbort, J.L.

    2000-01-01

    Steady-state creep and joining of alumina/zirconia composites containing alumina volume fractions of 20, 60 and 85% have been investigated between 1,250 and 1,350 C. Superplasticity of these compounds is controlled by grain-boundary sliding and the creep rate is a function of alumina volume fraction, not grain size. Using the principles of superplasticity, pieces of the composite have been joined by applying the stress required to achieve 5 to 10% strain to form a strong interface at temperatures as low as 1,200 C

  15. Development of a procedure for forming assisted thermal joining of tubes

    Science.gov (United States)

    Chen, Hui; Löbbe, Christian; Staupendahl, Daniel; Tekkaya, A. Erman

    2018-05-01

    With the demand of lightweight design in the automotive industry, not only the wall-thicknesses of tubular components of the chassis or spaceframe are continuously decreased. Also the thicknesses of exhaust system parts are reduced to save material and mass. However, thinner tubular parts bring about additional challenges in joining. Welding or brazing methods, which are utilized in joining tubes with specific requirements concerning leak tightness, are sensitive to the gap between the joining partners. Furthermore, a large joining area is required to ensure the durability of the joint. The introduction of a forming step in the assembled state prior to thermal joining can define and control the gap for subsequent brazing or welding. The mechanical pre-joint resulting from the previously described calibration step also results in easier handling of the tubes prior to thermal joining. In the presented investigation, a spinning process is utilized to produce force-fit joints of varying lengths and diameter reduction and form-fit joints with varying geometrical attributes. The spinning process facilitates a high formability and geometrical flexibility, while at the achievable precision is high and the process forces are low. The strength of the joints is used to evaluate the joint quality. Finally, a comparison between joints produced by forming with subsequent brazing and original tube is conducted, which presents the high performance of the developed procedure for forming assisted thermal joining.

  16. Model Checking Processes Specified In Join-Calculus Algebra

    Directory of Open Access Journals (Sweden)

    Sławomir Piotr Maludziński

    2014-01-01

    Full Text Available This article presents a model checking tool used to verify concurrent systems specified in join-calculus algebra. The temporal properties of systems under verification are expressed in CTL logic. Join-calculus algebra with its operational semantics defined by the chemical abstract machine serves as the basic method for the specification of concurrent systems and their synchronization mechanisms, and allows the examination of more complex systems.

  17. Targeted mutagenesis in the silkworm Bombyx mori using zinc finger nuclease mRNA injection

    Czech Academy of Sciences Publication Activity Database

    Takasu, Yoko; Kobayashi, I.; Beumer, K.; Uchino, K.; Sezutsu, H.; Sajwan, S.; Carroll, D.; Tamura, T.; Žurovec, Michal

    2010-01-01

    Roč. 40, č. 10 (2010), s. 759-765 ISSN 0965-1748 R&D Projects: GA AV ČR IAA500070601; GA ČR GAP305/10/2406; GA MŠk(CZ) LC06077 Institutional research plan: CEZ:AV0Z50070508 Keywords : gene targeting * nonhomologous and joining * Lepidoptera Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.018, year: 2010

  18. Glass-Metal Joining in Nuclear Environment: the State of the Art

    International Nuclear Information System (INIS)

    Jacobs, M.

    2007-01-01

    Full text of publication follows: In the ITER fusion machine and in material testing fission reactors, it is not possible to avoid the use of non-metallic materials like glass for example. There is therefore a need to apply metal to glass joints. This problem arose already at the beginning of the 19. century when the electric light bulb was invented. Nowadays this type of glass-metal joint is very successful and widely used in the electronic industry. In the case of ITER and material testing reactors, glass-metal joints are necessary for the fixation of the optical windows and optical fibres to a metal structure to perform diagnostics. These types of joints are still difficult to make and their behaviour is not fully understood. A joint between glass and metal for a nuclear or fusion application has indeed to resist high temperatures and high neutron fluences, while keeping a good mechanical strength and remaining leak tight. These characteristics are difficult to obtain under these severe conditions. This paper presents an overview of the different joining technologies that can be used to join glass to metal in a severe nuclear environment. The working mechanism of the technologies are explained, together with their respective advantages and drawbacks. Three different types of joining are discussed: fastening, liquid phase joining and solid phase joining. Fastening is a mechanical attachment technique, not achieving easily hermetic seals. Liquid and solid phase joining on the other hand form a real bond, what makes the joint much stronger. The most important technologies using liquid phase joining are adhesive bonding, fusion welding and brazing. In the case of the solid phase joining the choices are ultrasonic torsion welding, diffusion bonding and electrostatic bonding. If it is usually not possible to join the glass directly to the metal, an interlayer must be used. One speaks then of indirect joining. The paper will conclude with a discussion on the best

  19. Taiwan university joins search for antimatter

    CERN Multimedia

    Chiu, Y

    2002-01-01

    National Cheng Kung University announced yesterday that it will be joining the Alpha Magnetic Spectrometer, an international scientific research project aimed at looking for antimatter in space (1 page).

  20. High-energy electron beams for ceramic joining

    Science.gov (United States)

    Turman, Bob N.; Glass, S. J.; Halbleib, J. A.; Helmich, D. R.; Loehman, Ron E.; Clifford, Jerome R.

    1995-03-01

    Joining of structural ceramics is possible using high melting point metals such as Mo and Pt that are heated with a high energy electron beam, with the potential for high temperature joining. A 10 MeV electron beam can penetrate through 1 cm of ceramic, offering the possibility of buried interface joining. Because of transient heating and the lower heat capacity of the metal relative to the ceramic, a pulsed high power beam has the potential for melting the metal without decomposing or melting the ceramic. We have demonstrated the feasibility of the process with a series of 10 MeV, 1 kW electron beam experiments. Shear strengths up to 28 MPa have been measured. This strength is comparable to that reported in the literature for bonding silicon nitride (Si3N4) to molybdenum with copper-silver-titanium braze, but weaker than that reported for Si3N4 - Si3N4 with gold-nickel braze. The bonding mechanism appears to be formation of a thin silicide layer. Beam damage to the Si3N4 was also assessed.

  1. Top scientists join Stephen Hawking at Perimeter Institute

    Science.gov (United States)

    Banks, Michael

    2009-03-01

    Nine leading researchers are to join Stephen Hawking as visiting fellows at the Perimeter Institute for Theoretical Physics in Ontario, Canada. The researchers, who include string theorists Leonard Susskind from Stanford University and Asoka Sen from the Harisch-Chandra Research Institute in India, will each spend a few months of the year at the institute as "distinguished research chairs". They will be joined by another 30 scientists to be announced at a later date.

  2. Creation of Novel Protein Variants with CRISPR/Cas9-Mediated Mutagenesis: Turning a Screening By-Product into a Discovery Tool.

    Directory of Open Access Journals (Sweden)

    Katherine F Donovan

    Full Text Available CRISPR/Cas9 screening has proven to be a versatile tool for genomics research. Based on unexpected results from a genome-wide screen, we developed a CRISPR/Cas9-mediated approach to mutagenesis, exploiting the allelic diversity generated by error-prone non-homologous end-joining (NHEJ to identify novel gain-of-function and drug resistant alleles of the MAPK signaling pathway genes MEK1 and BRAF. We define the parameters of a scalable technique to easily generate cell populations containing thousands of endogenous allelic variants to map gene functions. Further, these results highlight an unexpected but important phenomenon, that Cas9-induced gain-of-function alleles are an inherent by-product of normal Cas9 loss-of-function screens and should be investigated during analysis of data from large-scale positive selection screens.

  3. Biological defense mechanisms against DNA double-strand break and their possible medical applications

    International Nuclear Information System (INIS)

    Matsumoto, Yoshihisa

    2011-01-01

    Radiation is now widely used for clinical diagnosis and therapeutics. On the other hand, radiation influences various tissues represented by immunological and reproductive systems, and is also recognized as one of the cause of carcinogenesis. Such pleiotropic effects of radiation are mediated through generation of damages on DNA molecule, vitally important genetic macromolecule. Among various types of DNA damages, double-strand break (DSB) is considered most critical and, therefore, responsible for biological effects. DSB is repaired mainly through two pathways: non-homologous end joining (NHEJ) and homologous recombination (HR). Understanding of these mechanisms has been greatly deepened in past 20 years and is now providing a promising approach toward cancer therapy. We have studied the mechanisms of NHEJ, focusing especially on the role of phosphorylation and the assembly of machinery therein, which will be introduced below. (author)

  4. In normal human fibroblasts variation in DSB repair capacity cannot be ascribed to radiation-induced changes in the localisation, expression or activity of major NHEJ proteins

    DEFF Research Database (Denmark)

    Kasten-Pisula, Ulla; Vronskaja, Svetlana; Overgaard, Jens

    2008-01-01

    in the activity of the DNA-PK complex induced upon irradiation. CONCLUSIONS: For normal human fibroblasts, the level or activity of NHEJ proteins measured prior to or after irradiation cannot be used to predict the DSB repair capacity or cellular radiosensitivity. Udgivelsesdato: 2008-Mar......BACKGROUND AND PURPOSE: The aim of the present study was to test whether for normal human fibroblasts the variation in double-strand break (DSB) repair capacity results from radiation-induced differences in localisation, expression or activity of major non-homologous end-joining (NHEJ) proteins....... MATERIALS AND METHODS: Experiments were performed with 11 normal human fibroblast strains AF01-11. NHEJ proteins were determined by Western blot and DNA-PK activity by pulldown-assay. RESULTS: The four NHEJ proteins tested (Ku70, Ku80, XRCC4 and DNA-PKcs) were found to be localised almost exclusively...

  5. Repair of endogenous and ionizing radiation-induced DNA damages: mechanisms and biological functions

    International Nuclear Information System (INIS)

    Boiteux, S.

    2002-01-01

    The cellular DNA is continuously exposed to endogenous and exogenous stress. Oxidative stress due to cellular metabolism is the major cause of endogenous DNA damage. On the other hand, ionizing radiation (IR) is an important exogenous stress. Both induce similar DNA damages: damaged bases, abasic sites and strand breakage. Most of these lesions are lethal and/or mutagenic. The survival of the cell is managed by efficient and accurate DNA repair mechanisms that remove lesions before their replication or transcription. DNA repair pathways involved in the removal of IR-induced lesions are briefly described. Base excision repair (BER) is mostly involved in the removal of base damage, abasic sites and single strand breaks. In contrast, DNA double strand breaks are mostly repaired by non-homologous end joining (NHEJ) or homologous recombination (HR). How DNA repair pathways prevent cancer process is also discussed. (author)

  6. Regulation of homologous recombination repair protein Rad51 by Ku70

    International Nuclear Information System (INIS)

    Du Liqing; Liu Qiang; Wang Yan; Xu Chang; Cao Jia; Fu Yue; Chen Fenghua; Fan Feiyue

    2013-01-01

    Objective: To explore the regulative effect of non-homologous end joining (NHEJ)protein Ku70 on homologous recombination repair protein Rad51, and to investigate the synergistic mechanism of homologous recombination repair in combination with NHEJ. Methods: Observed Rad51 protein expression after transfect Ku70 small interfering RNA or Ku70 plasmid DNA into tumor cells using Western blot. Results: Expression of Rad51 was obviously reduced after pretreated with Ku70 small interfering RNA. And with the increasing expression of Ku70 protein after transfection of Ku70 plasmid DNA PGCsi3.0-hKu70 into tumor cell lines, the Rad51 protein expression was increased. Conclusion: Ku70 protein has regulating effect on gene expression of Rad51, and it might participate in the collaboration between homologous recombination repair and NHEJ. (authors)

  7. Recombinant protein production from stable mammalian cell lines and pools.

    Science.gov (United States)

    Hacker, David L; Balasubramanian, Sowmya

    2016-06-01

    We highlight recent developments for the production of recombinant proteins from suspension-adapted mammalian cell lines. We discuss the generation of stable cell lines using transposons and lentivirus vectors (non-targeted transgene integration) and site-specific recombinases (targeted transgene integration). Each of these methods results in the generation of cell lines with protein yields that are generally superior to those achievable through classical plasmid transfection that depends on the integration of the transfected DNA by non-homologous DNA end-joining. This is the main reason why these techniques can also be used for the generation of stable cell pools, heterogenous populations of recombinant cells generated by gene delivery and genetic selection without resorting to single cell cloning. This allows the time line from gene transfer to protein production to be reduced. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Pulsed Joining Of Body-In-White Components

    Energy Technology Data Exchange (ETDEWEB)

    Bonnen, John [Ford Motor Company, Dearborn, MI (United States)

    2014-09-30

    The objective of this project was to develop cost efficient high quality pulsed welding (PW) technology for joining 6xxx Aluminum and High Strength Steel (with tensile strength above 580MPa) components enabling broad usage of hydroformed parts and leading to substantial weight reduction of cars and trucks to reduce US demand on petroleum, lower carbon emissions and energy expenditures. In general, pulsed welding is a form of impact welding where two dissimilar metal pieces are joined by accelerating one to velocities exceeding 300m/s at which point the first piece strikes the second and forms a weld. In this work, two methods were used to accelerate the flyer material: Electro-Magnetic (EM) pulse and Electro-Hydraulic (EH) pulse launching. The advantage of pulsed welding techniques is that welds can be formed between two materials that cannot otherwise be welded: high strength aluminum and high strength steel. The technical objectives of the project included: 1) developing cost affordable production feasible tooling design for PW of 6xxx aluminum to High Strength steel with strengths above 580MPa; 2) demonstrating that fabricated joints can exceed the required service load strength initially at the coupon level and then at the component level; 3) developing fundamental understanding of the mechanisms of joint formation and conditions leading to formation of high quality PW joint; and 4) creating a numerical model predicting the tooling and electric discharge parameters necessary for the joint formation and that satisfy the targeted strength parameters. The project successfully developed: 1) EM and EH pulsed welds between high strength aluminum with tensile strengths exceeding 240MPa and steels exceeding 580MPa; 2) pulsed welds of extrusions with strengths exceeding project requirements; 3) EM and EH flyer launch models and 4) weld interface formation models. However, the grant holder, Ford Motor Company, could see no path to commercialization and the work was

  9. ROMANIA'S MACROECONOMIC ACHIEVEMENTS FOR JOINING THE UNIQUE EUROPEAN CURENCY

    Directory of Open Access Journals (Sweden)

    SILVIA POPESCU

    2011-04-01

    Full Text Available The Romanian government has announced plans to join the eurozone by 2015. Currently, the leu is not yet part of ERM II but plans to join in 2010-2012. The economic advantages of the monetary union grow with expansion of the Euro zone. There is also a high level of skepticism; the main fear about the Euro is the inflation –that is considerable promoted by the Euro currency’s exchange rate in comparison with 2002; another restraint is due to member states inability to establish their own interest rates. The IMF arose the option of joining the Euro zone criteria relaxing. A one-sided Euro’s joining was suggested by International Monetary Fund on March-April 2009, in a confidential report mentioned by The Financial Times as the emergent states in Central and Eastern Europe to be able to pass to the unique currency, but not being represented in the Central European Bank Board. By its side, CEB considers that emergent states of the European Union must not pass to the unique currency unilaterally, because such a fact could under-mine the trust in Euro currency worldwide. This option would hardly deepen the macroeconomic controversies inside the Euro zone and would contradict the previous conditions already imposed. An acceptable solution could be the fastening of emergent countries joining the Exchange Rate Mechanism 2, after they are aware of risks arisen by such a step. The European Commission endorses in the Convergence Report on 2010 that Romania doesn’t meet any criteria needed by passing to the unique European currency, respectively: prices stability; budget position of the government; stability of exchange rate; interest convergence on long run and there are also law impediments. Our paper discusses arguments for a faster passing to the Euro currency versus arguments for a late joining the Euro currency in Romania.

  10. Mutations that affect meiosis in male mice influence the dynamics of the mid-preleptotene and bouquet stages

    International Nuclear Information System (INIS)

    Liebe, B.; Petukhova, G.; Barchi, M.; Bellani, M.; Braselmann, H.; Nakano, T.; Pandita, T.K.; Jasin, M.; Fornace, A.; Meistrich, M.L.; Baarends, W.M.; Schimenti, J.; Lange, T. de; Keeney, S.; Camerini-Otero, R.D.; Scherthan, H.

    2006-01-01

    Meiosis pairs and segregates homologous chromosomes and thereby forms haploid germ cells to compensate the genome doubling at fertilization. Homologue pairing in many eukaryotic species depends on formation of DNA double strand breaks (DSBs) during early prophase I when telomeres begin to cluster at the nuclear periphery (bouquet stage). By fluorescence in situ hybridization criteria, we observe that mid-preleptotene and bouquet stage frequencies are altered in male mice deficient for proteins required for recombination, ubiquitin conjugation and telomere length control. The generally low frequencies of mid-preleptotene spermatocytes were significantly increased in male mice lacking recombination proteins SPO11, MEI1, MLH1, KU80, ubiquitin conjugating enzyme HR6B, and in mice with only one copy of the telomere length regulator Terf1. The bouquet stage was significantly enriched in Atm -/- , Spo11 -/- , Mei1 m1Jcs/m1Jcs , Mlh1 -/- , Terf1 +/- and Hr6b -/- spermatogenesis, but not in mice lacking recombination proteins DMC1 and HOP2, the non-homologous end-joining DNA repair factor KU80 and the ATM downstream effector GADD45a. Mice defective in spermiogenesis (Tnp1 -/- , Gmcl1 -/- , Asm -/- ) showed wild-type mid-preleptotene and bouquet frequencies. A low frequency of bouquet spermatocytes in Spo11 -/- Atm -/- spermatogenesis suggests that DSBs contribute to the Atm -/- -correlated bouquet stage exit defect. Insignificant changes of bouquet frequencies in mice with defects in early stages of DSB repair (Dmc1 -/- , Hop2 -/- ) suggest that there is an ATM-specific influence on bouquet stage duration. Altogether, it appears that several pathways influence telomere dynamics in mammalian meiosis

  11. Microwave joining of SiC

    Energy Technology Data Exchange (ETDEWEB)

    Silberglitt, R.; Ahmad, I. [FM Technologies, Inc., Fairfax, VA (United States); Black, W.M. [George Mason Univ., Fairfax, VA (United States)] [and others

    1995-05-01

    The purpose of this work is to optimize the properties of SiC-SiC joints made using microwave energy. The current focus is on optimization of time-temperature profiles, production of SiC from chemical precursors, and design of new applicators for joining of long tubes.

  12. Joining of SiCf/SiC composites for thermonuclear fusion reactors

    International Nuclear Information System (INIS)

    Ferraris, M.; Badini, C.; Montorsi, M.; Appendino, P.; Scholz, H.W.

    1994-01-01

    Due to their favourable radiological behaviour, SiC f /SiC composites are promising structural materials for future use in fusion reactors. A problem to cope with is the joining of the ceramic composite material (CMC) to itself for more complex structures. Maintenance concepts for a reactor made of SiC f /SiC will demand a method of joining. The joining agents should comply with the low-activation approach of the base material. With the acceptable elements Si and Mg, sandwich structures of composite/metal/composite were prepared in Ar atmosphere at temperatures just above the melting points of the metals. Another promising route is the use of joining agents of boro-silicate glasses: their composition can be tailored to obtain softening temperatures of interest for fusion applications. The glassy joint can be easily ceramised to improve thermomechanical properties. The joining interfaces were investigated by SEM-EDS, XRD and mechanical tests. ((orig.))

  13. Why join the Staff Association

    CERN Multimedia

    Association du personnel

    2011-01-01

    Becoming a member of the Staff Association (SA) is above all a personal choice, showing that the joining person’s commitment and adherence to values such as solidarity, social cohesion, etc.In September, the SA launches a membership campaign to convince a maximum number to join, to inform, arouse interest and support. Posters, emails and individual contacts are part of the campaign programme, just like this editorial. As far as individual contacts are concerned, we ask you to give time and lend an ear to the delegates of your department in the Staff Council, who will approach you, in order to make an open and constructive discussion possible. Do not hesitate to ask questions and let them know your thoughts about the SA, as (constructive) criticism enables us to progress. The Staff Association and its role of collective representation The Staff Association, via its delegates, represents collectively all staff of the Organization before the Director-General and Member States. To do this, staff rep...

  14. Flexible Friction Stir Joining Technology

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Zhili [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Lim, Yong Chae [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Mahoney, Murray [MegaStir Technologies LLC, Orem, UT (United States); Sanderson, Samuel [MegaStir Technologies LLC, Orem, UT (United States); Larsen, Steve [MegaStir Technologies LLC, Orem, UT (United States); Steel, Russel [MegaStir Technologies LLC, Orem, UT (United States); Fleck, Dale [MegaStir Technologies LLC, Orem, UT (United States); Fairchild, Doug P [ExxonMobil, Upstream Research Company (URC), Houston, TX (United States); Wasson, Andrew J [ExxonMobil, Upstream Research Company (URC), Houston, TX (United States); Babb, Jon [MegaStir Technologies LLC, Orem, UT (United States); Higgins, Paul [MegaStir Technologies LLC, Orem, UT (United States)

    2015-07-23

    Reported herein is the final report on a U.S. Department of Energy (DOE) Advanced Manufacturing Office (AMO) project with industry cost-share that was jointly carried out by Oak Ridge National Laboratory (ORNL), ExxonMobil Upstream Research Company (ExxonMobil), and MegaStir Technologies (MegaStir). The project was aimed to advance the state of the art of friction stir welding (FSW) technology, a highly energy-efficient solid-state joining process, for field deployable, on-site fabrications of large, complex and thick-sectioned structures of high-performance and high-temperature materials. The technology innovations developed herein attempted to address two fundamental shortcomings of FSW: 1) the inability for on-site welding and 2) the inability to weld thick section steels, both of which have impeded widespread use of FSW in manufacturing. Through this work, major advance has been made toward transforming FSW technology from a “specialty” process to a mainstream materials joining technology to realize its pervasive energy, environmental, and economic benefits across industry.

  15. Chromosomal Integrity after UV Irradiation Requires FANCD2-Mediated Repair of Double Strand Breaks.

    Directory of Open Access Journals (Sweden)

    María Belén Federico

    2016-01-01

    Full Text Available Fanconi Anemia (FA is a rare autosomal recessive disorder characterized by hypersensitivity to inter-strand crosslinks (ICLs. FANCD2, a central factor of the FA pathway, is essential for the repair of double strand breaks (DSBs generated during fork collapse at ICLs. While lesions different from ICLs can also trigger fork collapse, the contribution of FANCD2 to the resolution of replication-coupled DSBs generated independently from ICLs is unknown. Intriguingly, FANCD2 is readily activated after UV irradiation, a DNA-damaging agent that generates predominantly intra-strand crosslinks but not ICLs. Hence, UV irradiation is an ideal tool to explore the contribution of FANCD2 to the DNA damage response triggered by DNA lesions other than ICL repair. Here we show that, in contrast to ICL-causing agents, UV radiation compromises cell survival independently from FANCD2. In agreement, FANCD2 depletion does not increase the amount of DSBs generated during the replication of UV-damaged DNA and is dispensable for UV-induced checkpoint activation. Remarkably however, FANCD2 protects UV-dependent, replication-coupled DSBs from aberrant processing by non-homologous end joining, preventing the accumulation of micronuclei and chromatid aberrations including non-homologous chromatid exchanges. Hence, while dispensable for cell survival, FANCD2 selectively safeguards chromosomal stability after UV-triggered replication stress.

  16. Social Pharmacy and Clinical Pharmacy-Joining Forces.

    Science.gov (United States)

    Almarsdottir, Anna Birna; Granas, Anne Gerd

    2015-12-22

    This commentary seeks to define the areas of social pharmacy and clinical pharmacy to uncover what they have in common and what still sets them apart. Common threats and challenges of the two areas are reviewed in order to understand the forces in play. Forces that still keep clinical and social pharmacy apart are university structures, research traditions, and the management of pharmacy services. There are key (but shrinking) differences between clinical and social pharmacy which entail the levels of study within pharmaceutical sciences, the location in which the research is carried out, the choice of research designs and methods, and the theoretical foundations. Common strengths and opportunities are important to know in order to join forces. Finding common ground can be developed in two areas: participating together in multi-disciplinary research, and uniting in a dialogue with internal and external key players in putting forth what is needed for the profession of pharmacy. At the end the question is posed, "What's in a name?" and we argue that it is important to emphasize what unifies the families of clinical pharmacy and social pharmacy for the benefit of both fields, pharmacy in general, and society at large.

  17. Comparative analysis of the practice of China and Russia joining the WTO

    Directory of Open Access Journals (Sweden)

    Anastasia Sergeyevna Loginova

    2015-09-01

    Full Text Available Objective to examine the positive and negative experience of China39s joining the World Trade Organization WTO that joined it on terms similar to the Russian ones for the best understanding of the advantages and disadvantages of joining and building the appropriate policy. Methods the objective of the work has been achieved through the application of both general and specific scientific methods. In particular the analysis and synthesis allowed to study the social and economic context of the state in the framework of joining the WTO the consequences of such membership and to identify the key areas for improvement of this step for Russia. Statistical techniques were used in the process of gathering information about the key economic indicators. The fundamental method was comparativelegal which made it possible to draw parallels in joining the WTO by various states. Results basing on the measures taken by China both before and after joining the WTO as well as evaluation of their implication the conclusion was made about its successful experience. The actions before joining were very effective. In particular they are applicable to the fields of agriculture automotive industry investment policy etc. The measures of nonaddress subsidizing maintaining the socioeconomic stability in the country the export orientation of the economy along with functioning of special economic zones with joint ventures are in the authors39 opinion constructive actions for Russiarsquos adaptation to the WTO. Scientific novelty for the first time the thesis is put forward about the similarity of the Russia and China conditions when joining the WTO. The analysis is made of the complex measures preceding and following the joining. The necessity is grounded of reflecting the China39s positive and negative experiences in the Russian policy. Practical significance the main provisions and conclusions can be used in the research activity in addressing the issues of Russiarsquos

  18. Joining of aluminum sheet and glass fiber reinforced polymer using extruded pins

    Science.gov (United States)

    Conte, Romina; Buhl, Johannes; Ambrogio, Giuseppina; Bambach, Markus

    2018-05-01

    The present contribution proposes a new approach for joining sheet metal and fiber reinforced composites. The joining process draws upon a Friction Stir Forming (FSF) process, which is performed on the metal sheet to produce slender pins. These pins are used to pierce through the composite. Joining is complete by forming a locking head out of the part if the pin sticks out of the composite. Pins of different diameters and lengths were produced from EN AW-1050 material, which were joined to glass fiber reinforced polyamide-6. The strength of the joint has been experimentally tested in order to understand the effect of the process temperature on the pins strength and therefore on the joining. The results demonstrate the feasibility of this new technique, which uses no excess material.

  19. Joining of parts via magnetic heating of metal aluminum powders

    Science.gov (United States)

    Baker, Ian

    2013-05-21

    A method of joining at least two parts includes steps of dispersing a joining material comprising a multi-phase magnetic metal-aluminum powder at an interface between the at least two parts to be joined and applying an alternating magnetic field (AMF). The AMF has a magnetic field strength and frequency suitable for inducing magnetic hysteresis losses in the metal-aluminum powder and is applied for a period that raises temperature of the metal-aluminum powder to an exothermic transformation temperature. At the exothermic transformation temperature, the metal-aluminum powder melts and resolidifies as a metal aluminide solid having a non-magnetic configuration.

  20. Recent progress in micro and nano-joining

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Y; Hu, A; Khan, M I; Wu, W; Tam, B; Yavuz, M [Centre for Advanced Materials Joining Department of Mechanical and Mechatronics Engineering, University of Waterloo 200 University Avenue West, Waterloo, N2L 3G1 (Canada)], E-mail: nzhou@uwaterloo.ca

    2009-05-01

    Micro and nano-joining has been identified as a key enabling technology in the construction of micromechanical and microelectronic devices. The current article reviews recent progress in micro and nano-joining. In particular, laser micro-welding (LMW) of crossed 316 LVM stainless steel (SS) wire was compared to conventional resistance micro-welding (RMW) and was successfully employed in welding a Pt-Ir /SS dissimilar combination. Welding of Au nanoparticles was realized using femtosecond laser irradiation and its application in the surface enhanced Raman spectroscopy was investigated. Brazing between carbon nanotube (CNT) bundles and Ni electrodes was attained in vacuum, resulting in the development of a novel CNT filament of incandescent lamps.

  1. In Vitro Expansion of Bone Marrow Derived Mesenchymal Stem Cells Alters DNA Double Strand Break Repair of Etoposide Induced DNA Damage

    Directory of Open Access Journals (Sweden)

    Ian Hare

    2016-01-01

    Full Text Available Mesenchymal stem cells (MSCs are of interest for use in diverse cellular therapies. Ex vivo expansion of MSCs intended for transplantation must result in generation of cells that maintain fidelity of critical functions. Previous investigations have identified genetic and phenotypic alterations of MSCs with in vitro passage, but little is known regarding how culturing influences the ability of MSCs to repair double strand DNA breaks (DSBs, the most severe of DNA lesions. To investigate the response to DSB stress with passage in vitro, primary human MSCs were exposed to etoposide (VP16 at various passages with subsequent evaluation of cellular damage responses and DNA repair. Passage number did not affect susceptibility to VP16 or the incidence and repair kinetics of DSBs. Nonhomologous end joining (NHEJ transcripts showed little alteration with VP16 exposure or passage; however, homologous recombination (HR transcripts were reduced following VP16 exposure with this decrease amplified as MSCs were passaged in vitro. Functional evaluations of NHEJ and HR showed that MSCs were unable to activate NHEJ repair following VP16 stress in cells after successive passage. These results indicate that ex vivo expansion of MSCs alters their ability to perform DSB repair, a necessary function for cells intended for transplantation.

  2. In Vitro Expansion of Bone Marrow Derived Mesenchymal Stem Cells Alters DNA Double Strand Break Repair of Etoposide Induced DNA Damage.

    Science.gov (United States)

    Hare, Ian; Gencheva, Marieta; Evans, Rebecca; Fortney, James; Piktel, Debbie; Vos, Jeffrey A; Howell, David; Gibson, Laura F

    2016-01-01

    Mesenchymal stem cells (MSCs) are of interest for use in diverse cellular therapies. Ex vivo expansion of MSCs intended for transplantation must result in generation of cells that maintain fidelity of critical functions. Previous investigations have identified genetic and phenotypic alterations of MSCs with in vitro passage, but little is known regarding how culturing influences the ability of MSCs to repair double strand DNA breaks (DSBs), the most severe of DNA lesions. To investigate the response to DSB stress with passage in vitro, primary human MSCs were exposed to etoposide (VP16) at various passages with subsequent evaluation of cellular damage responses and DNA repair. Passage number did not affect susceptibility to VP16 or the incidence and repair kinetics of DSBs. Nonhomologous end joining (NHEJ) transcripts showed little alteration with VP16 exposure or passage; however, homologous recombination (HR) transcripts were reduced following VP16 exposure with this decrease amplified as MSCs were passaged in vitro. Functional evaluations of NHEJ and HR showed that MSCs were unable to activate NHEJ repair following VP16 stress in cells after successive passage. These results indicate that ex vivo expansion of MSCs alters their ability to perform DSB repair, a necessary function for cells intended for transplantation.

  3. A cell cycle-dependent regulatory circuit composed of 53BP1-RIF1 and BRCA1-CtIP controls DNA repair pathway choice.

    Science.gov (United States)

    Escribano-Díaz, Cristina; Orthwein, Alexandre; Fradet-Turcotte, Amélie; Xing, Mengtan; Young, Jordan T F; Tkáč, Ján; Cook, Michael A; Rosebrock, Adam P; Munro, Meagan; Canny, Marella D; Xu, Dongyi; Durocher, Daniel

    2013-03-07

    DNA double-strand break (DSB) repair pathway choice is governed by the opposing activities of 53BP1 and BRCA1. 53BP1 stimulates nonhomologous end joining (NHEJ), whereas BRCA1 promotes end resection and homologous recombination (HR). Here we show that 53BP1 is an inhibitor of BRCA1 accumulation at DSB sites, specifically in the G1 phase of the cell cycle. ATM-dependent phosphorylation of 53BP1 physically recruits RIF1 to DSB sites, and we identify RIF1 as the critical effector of 53BP1 during DSB repair. Remarkably, RIF1 accumulation at DSB sites is strongly antagonized by BRCA1 and its interacting partner CtIP. Lastly, we show that depletion of RIF1 is able to restore end resection and RAD51 loading in BRCA1-depleted cells. This work therefore identifies a cell cycle-regulated circuit, underpinned by RIF1 and BRCA1, that governs DSB repair pathway choice to ensure that NHEJ dominates in G1 and HR is favored from S phase onward. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Joining of AZ31 and AZ91 Mg alloys by friction stir welding

    Directory of Open Access Journals (Sweden)

    B. Ratna Sunil

    2015-12-01

    Full Text Available Two dissimilar magnesium (Mg alloy sheets, one with low aluminium (AZ31 and another with high aluminium (AZ91 content, were successfully joined by friction stir welding (FSW. The effect of process parameters on the formation of hot cracks was investigated. A sound metallurgical joint was obtained at optimized process parameters (1400 rpm with 25 mm/min feed which contained fine grains and distributed β (Mg17Al12 phase within the nugget zone. An increasing trend in the hardness measurements has also confirmed more amount of dissolution of aluminium within the nugget zone. A sharp interface between nugget zone and thermo mechanical affected zone (TMAZ was clearly noticed at the AZ31 Mg alloy side (advancing but not on the AZ91 Mg alloy side (retreating. From the results it can be concluded that FSW can be effectively used to join dissimilar metals, particularly difficult to process metals such as Mg alloys, and hot cracking can be completely eliminated by choosing appropriate process parameters to achieve sound joint.

  5. Molecular Basis for DNA Double-Strand Break Annealing and Primer Extension by an NHEJ DNA Polymerase

    Directory of Open Access Journals (Sweden)

    Nigel C. Brissett

    2013-11-01

    Full Text Available Nonhomologous end-joining (NHEJ is one of the major DNA double-strand break (DSB repair pathways. The mechanisms by which breaks are competently brought together and extended during NHEJ is poorly understood. As polymerases extend DNA in a 5′-3′ direction by nucleotide addition to a primer, it is unclear how NHEJ polymerases fill in break termini containing 3′ overhangs that lack a primer strand. Here, we describe, at the molecular level, how prokaryotic NHEJ polymerases configure a primer-template substrate by annealing the 3′ overhanging strands from opposing breaks, forming a gapped intermediate that can be extended in trans. We identify structural elements that facilitate docking of the 3′ ends in the active sites of adjacent polymerases and reveal how the termini act as primers for extension of the annealed break, thus explaining how such DSBs are extended in trans. This study clarifies how polymerases couple break-synapsis to catalysis, providing a molecular mechanism to explain how primer extension is achieved on DNA breaks.

  6. Structure and function of DNA polymerase μ

    International Nuclear Information System (INIS)

    Matsumoto, Takuro; Maezawa, So

    2013-01-01

    DNA polymerases are enzymes playing the central role in DNA metabolism, including DNA replication, DNA repair and recombination. DNA polymerase μ (pol μ DNA polymerase λ (pol λ) and terminal deoxynucleotidyltransferase (TdT) in X family DNA polymerases function in non-homologous end-joining (NHEJ), which is the predonmiant repair pathway for DNA double-strand breaks (DSBs). NHEJ involves enzymes that capture both ends of the broken DNA strand, bring them together in a synaptic DNA-protein complex, and repair the DSB. Pol μ and pol λ fill in the gaps at the junction to maintain the genomic integrity. TdT synthesizes N region at the junction during V(D)J recombination and promotes diversity of immunoglobulin or T-cell receptor gene. Among these three polymerases, the regulatory mechanisms of pol μ remain rather unclear. We have approached the mechanism of pol μ from both sides of structure and cellular dynamics. Here, we propose some new insights into pol μ and the probable NHEJ model including our findings. (author)

  7. Performance of electro-plated and joined components for divertor application

    International Nuclear Information System (INIS)

    Krauss, Wolfgang; Lorenz, Julia; Konys, Jürgen

    2013-01-01

    Highlights: • Active interlayers of Ni and Pd were electroplated on W to assist joining. • Demonstrator types of W-steel and W–W joints were successfully fabricated. • Diffusion processes increase operation temperature above brazing temperature. • Ni electro-plating is less sensitive to variation of deposition parameters than Pd. • Shear tests showed values in resistance comparable to those of commercial fillers. -- Abstract: A general challenge in divertor development, independently of design type and cooling medium water or helium, is the reliable and adapted joining of components. Depending on the design variants, the characteristics of the joints will be focused on functional or structural behavior to guarantee e.g. good thermal conductivity and sufficient mechanical strength. All variants will have in common that tungsten is the plasma facing material. Thus, material combinations to be joined will range from Cu base over steel to tungsten. Especially tungsten shows lacks in adapted joining due to its metallurgical behavior ranging from immiscibility over bad wetting up to brittle intermetallic phase formation. Joining assisted by electro-chemical deposition of functional and filler layers showed that encouraging progress was achieved in wetting applying nickel interlayers. Nickel proved to be a good reference material but alternative elements (e.g. Pd, Fe) may be more attractive in fusion to manufacture suitable joints. Replacing of Ni as activator element by Pd for W/W or W/steel joints was achieved and joining with Cu-filler was successfully performed. Manufactured joints were characterized applying metallurgical testing and SEM/EDX analyses demonstrating the applicability of Pd activator. First shear tests showed that the joints exhibit mechanical stability sufficient for technical application

  8. How Students' Perceptions of the School Climate Influence Their Choice to Upstand, Bystand, or Join Perpetrators of Bullying

    Science.gov (United States)

    Ferráns, Silvia Diazgranados; Selman, Robert L.

    2014-01-01

    The authors of this article, Silvia Diazgranados Ferráns and Robert Selman, use an emergent framework to explore how the rules of the school culture at different perceived school climates affect early adolescents' decisions to upstand, bystand, or join the perpetrators when they witness peer aggression and bullying. Through a grounded theory…

  9. Cohesive delamination and frictional contact on joining surface via XFEM

    Directory of Open Access Journals (Sweden)

    Francesco Parrinello

    2018-02-01

    Full Text Available In the present paper, the complex mechanical behaviour of the surfaces joining two differentbodies is analysed by a cohesive-frictional interface constitutive model. The kinematical behaviouris characterized by the presence of discontinuous displacement fields, that take place at the internalconnecting surfaces, both in the fully cohesive phase and in the delamination one. Generally, in order tocatch discontinuous displacement fields, internal connecting surfaces (adhesive layers are modelled bymeans of interface elements, which connect, node by node, the meshes of the joined bodies, requiringthe mesh to be conforming to the geometry of the single bodies and to the relevant connecting surface.In the present paper, the Extended Finite Element Method (XFEM is employed to model, both fromthe geometrical and from the kinematical point of view, the whole domain, including the connectedbodies and the joining surface. The joining surface is not discretized by specific finite elements, butit is defined as an internal discontinuity surface, whose spatial position inside the mesh is analyticallydefined. The proposed approach is developed for two-dimensional composite domains, formed by twoor more material portions joined together by means of a zero thickness adhesive layer. The numericalresults obtained with the proposed approach are compared with the results of the classical interfacefinite element approach. Some examples of delamination and frictional contact are proposed withlinear, circular and curvilinear adhesive layer.

  10. Fastening, coupling and joining technique between diaspora and irredenta

    Science.gov (United States)

    Bauer, C.-O.

    1980-06-01

    The problem of eliminating the present divergence and shattering (diaspora) in the treatment of problems of the fastening, coupling, and joining technique on different technical branches is examined. It is shown that by an appropriate independence the fastening, coupling and joining techniques can recognize and consequently utilize the numerous performance reserves which are concealed by the present organization and action due to the lack of systematically tended works.

  11. POSSIBLE RELATED FUNCTIONS OF THE NON-HOMOLOGOUS CO-REGULATED GENE PAIR PDCD10 AND SERPINI1

    Directory of Open Access Journals (Sweden)

    Concetta Scimone

    2017-04-01

    Full Text Available Gene expression in mammalians is a very finely controlled mechanism, and bidirectional promoters can be considered one of the most compelling examples of the accuracy of genic expression coordination. As recently reported, a bidirectional promoter regulates the expression of the PDCD10(whose mutations cause familial Cerebral Cavernous Malformations (CCMs and SERPINI1 gene pair, even though they are non-homologous genes. The aim of this study was to identify any potential common roles of these two coregulated genes. An in-silico approach was used to identify functional correlations, using the BioGraph, IPA® and Cytoscape tools and the KEGG pathway database. The results obtained show that PDCD10 and SERPINI1 may co-regulate some cellular processes, particularly those related to focal adhesion maintenance. All common pathways identified for PDCD10 and SERPINI1 are closely associated with the pathogenic characteristics of CCMs; we thus hypothesize that genes involved in these networks may contribute to the development of CCMs.

  12. An Adaptive Genetic Algorithm with Dynamic Population Size for Optimizing Join Queries

    OpenAIRE

    Vellev, Stoyan

    2008-01-01

    The problem of finding the optimal join ordering executing a query to a relational database management system is a combinatorial optimization problem, which makes deterministic exhaustive solution search unacceptable for queries with a great number of joined relations. In this work an adaptive genetic algorithm with dynamic population size is proposed for optimizing large join queries. The performance of the algorithm is compared with that of several classical non-determinis...

  13. Five Performance Enhancements for Hybrid Hash Join

    National Research Council Canada - National Science Library

    Graefe, Goetz

    1992-01-01

    .... We discuss five performance enhancements for hybrid hash join algorithms, namely data compression, large cluster sizes and multi-level recursion, role reversal of build and probe inputs, histogram...

  14. Rhenium Mechanical Properties and Joining Technology

    Science.gov (United States)

    Reed, Brian D.; Biaglow, James A.

    1996-01-01

    Iridium-coated rhenium (Ir/Re) provides thermal margin for high performance and long life radiation cooled rockets. Two issues that have arisen in the development of flight Ir/Re engines are the sparsity of rhenium (Re) mechanical property data (particularly at high temperatures) required for engineering design, and the inability to directly electron beam weld Re chambers to C103 nozzle skirts. To address these issues, a Re mechanical property database is being established and techniques for creating Re/C103 transition joints are being investigated. This paper discusses the tensile testing results of powder metallurgy Re samples at temperatures from 1370 to 2090 C. Also discussed is the evaluation of Re/C103 transition pieces joined by both, explosive and diffusion bonding. Finally, the evaluation of full size Re transition pieces, joined by inertia welding, as well as explosive and diffusion bonding, is detailed.

  15. Exploring the deleterious SNPs in XRCC4 gene using computational approach and studying their association with breast cancer in the population of West India.

    Science.gov (United States)

    Singh, Preety K; Mistry, Kinnari N; Chiramana, Haritha; Rank, Dharamshi N; Joshi, Chaitanya G

    2018-05-20

    Non-homologous end joining (NHEJ) pathway has pivotal role in repair of double-strand DNA breaks that may lead to carcinogenesis. XRCC4 is one of the essential proteins of this pathway and single-nucleotide polymorphisms (SNPs) of this gene are reported to be associated with cancer risks. In our study, we first used computational approaches to predict the damaging variants of XRCC4 gene. Tools predicted rs79561451 (S110P) nsSNP as the most deleterious SNP. Along with this SNP, we analysed other two SNPs (rs3734091 and rs6869366) to study their association with breast cancer in population of West India. Variant rs3734091 was found to be significantly associated with breast cancer while rs6869366 variant did not show any association. These SNPs may influence the susceptibility of individuals to breast cancer in this population. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Infrared transient-liquid-phase joining of SCS-6/{beta}21S titanium matrix composite

    Energy Technology Data Exchange (ETDEWEB)

    Blue, C.A.; Sikka, V.K. [Oak Ridge National Lab., TN (United States). Materials Processing Group; Blue, R.A.; Lin, R.Y. [Univ. of Cincinnati, OH (United States). Dept. of Materials Science and Engineering

    1996-02-01

    Fiber-reinforced titanium matrix composites (TMCs) are among the advanced materials being considered for use in the aerospace industry due to their light weight, high strength, and high modulus. A rapid infrared joining process has been developed for the joining of composites and advanced materials. Rapid infrared joining has been shown not to have many of the problems associated with conventional joining methods. Two models were utilized to predict the joint evolution and fiber reaction zone growth. TMC, 16-ply SCS-6/{beta}21S, has been successfully joined with total processing times of under 2 min utilizing the rapid infrared joining technique. The process utilizes a 50 C/sec ramping rate, 17-{micro}m Ti-15Cu-15Ni wt % filler material between the faying surfaces; a joining temperature of 1,100 C; and 120 sec of time to join the composite material. Joint shear strength testing of the rapid infrared joints at temperatures as high as 800 C has revealed no joint failures. Also, due to the rapid cooling of the process, no poststabilization of the matrix material is necessary to prevent the formation of a brittle omega phase during subsequent use of the TMC at intermediate temperatures, 270 to 430 C, for up to 20 h.

  17. Social Pharmacy and Clinical Pharmacy—Joining Forces

    Directory of Open Access Journals (Sweden)

    Anna Birna Almarsdottir

    2015-12-01

    Full Text Available This commentary seeks to define the areas of social pharmacy and clinical pharmacy to uncover what they have in common and what still sets them apart. Common threats and challenges of the two areas are reviewed in order to understand the forces in play. Forces that still keep clinical and social pharmacy apart are university structures, research traditions, and the management of pharmacy services. There are key (but shrinking differences between clinical and social pharmacy which entail the levels of study within pharmaceutical sciences, the location in which the research is carried out, the choice of research designs and methods, and the theoretical foundations. Common strengths and opportunities are important to know in order to join forces. Finding common ground can be developed in two areas: participating together in multi-disciplinary research, and uniting in a dialogue with internal and external key players in putting forth what is needed for the profession of pharmacy. At the end the question is posed, “What’s in a name?” and we argue that it is important to emphasize what unifies the families of clinical pharmacy and social pharmacy for the benefit of both fields, pharmacy in general, and society at large.

  18. Joining of fuel pin end plugs. Pulsed magnetic welding (PMW), pressurized resistance welding (PRW) and their inspection

    International Nuclear Information System (INIS)

    Kamimura, Katsuichiro; Seki, Masayuki

    1996-01-01

    In Power Reactor and Nuclear Fuel Development Corporation, in order to attain the high burnup of FBR fuel, the development of new cladding tube materials mainly aiming at the improvement of swelling resistance has been advanced. Oxide dispersion-strengthened ferritic steel has excellent swelling characteristics and high temperature creep strength, but the strength of its welded parts lowers remarkably. As the result of the investigation of solid phase joining, the conclusion that PMW and PRW are promising was obtained. So far, the manufacture of a welder was started first, the welding test was advanced, and the ultrasonic flaw detection technology of high accuracy was developed for the inspection of welding defects. The features, the principle of welding, the welders and the examples of application of the PMW and the PRW are reported. The features of the ultrasonic inspection apparatus are explained. The inspection apparatus comprises 5 pulse motors for driving probes and one pulse motor for turning a sample. The example of flaw detection test results is shown. (K.I.)

  19. Evolutionary dynamics and sites of illegitimate recombination revealed in the interspersion and sequence junctions of two nonhomologous satellite DNAs in cactophilic Drosophila species.

    Science.gov (United States)

    Kuhn, G C S; Teo, C H; Schwarzacher, T; Heslop-Harrison, J S

    2009-05-01

    Satellite DNA (satDNA) is a major component of genomes but relatively little is known about the fine-scale organization of unrelated satDNAs residing at the same chromosome location, and the sequence structure and dynamics of satDNA junctions. We studied the organization and sequence junctions of two nonhomologous satDNAs, pBuM and DBC-150, in three species from the neotropical Drosophila buzzatii cluster (repleta group). In situ hybridization to microchromosomes, interphase nuclei and extended DNA fibers showed frequent interspersion of the two satellites in D. gouveai, D. antonietae and, to a lesser extent, D. seriema. We isolated by PCR six pBuM x DBC-150 junctions: four are exclusive to D. gouveai and two are exclusive to D. antonietae. The six junction breakpoints occur at different positions within monomers, suggesting independent origin. Four junctions showed abrupt transitions between the two satellites, whereas two junctions showed a distinct 10 bp tandem duplication before the junction. Unlike pBuM, DBC-150 junction repeats are more variable than randomly cloned monomers and showed diagnostic features in common to a 3-monomer higher-order repeat seen in the sister species D. serido. The high levels of interspersion between pBuM and DBC-150 repeats suggest extensive rearrangements between the two satellites, maybe favored by specific features of the microchromosomes. Our interpretation is that the junctions evolved by multiples events of illegitimate recombination between nonhomologous satDNA repeats, with subsequent rounds of unequal crossing-over expanding the copy number of some of the junctions.

  20. In cellulo phosphorylation of XRCC4 Ser320 by DNA-PK induced by DNA damage

    International Nuclear Information System (INIS)

    Sharma, Mukesh Kumar; Imamichi, Shoji; Fukuchi, Mikoto; Samarth, Ravindra Mahadeo; Tomita, Masanori; Matsumoto, Yoshihisa

    2016-01-01

    XRCC4 is a protein associated with DNA Ligase IV, which is thought to join two DNA ends at the final step of DNA double-strand break repair through non-homologous end joining. In response to treatment with ionizing radiation or DNA damaging agents, XRCC4 undergoes DNA-PK-dependent phosphorylation. Furthermore, Ser260 and Ser320 (or Ser318 in alternatively spliced form) of XRCC4 were identified as the major phosphorylation sites by purified DNA-PK in vitro through mass spectrometry. However, it has not been clear whether these sites are phosphorylated in vivo in response to DNA damage. In the present study, we generated an antibody that reacts with XRCC4 phosphorylated at Ser320 and examined in cellulo phosphorylation status of XRCC4 Ser320. The phosphorylation of XRCC4 Ser320 was induced by γ-ray irradiation and treatment with Zeocin. The phosphorylation of XRCC4 Ser320 was detected even after 1 Gy irradiation and increased in a manner dependent on radiation dose. The phosphorylation was observed immediately after irradiation and remained mostly unchanged for up to 4 h. The phosphorylation was inhibited by DNA-PK inhibitor NU7441 and was undetectable in DNA-PKcs-deficient cells, indicating that the phosphorylation was mainly mediated by DNA-PK. These results suggested potential usefulness of the phosphorylation status of XRCC4 Ser320 as an indicator of DNA-PK functionality in living cells

  1. Distinct kinetics of human DNA ligases I, IIIalpha, IIIbeta, and IV reveal direct DNA sensing ability and differential physiological functions in DNA repair

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xi; Ballin, Jeff D.; Della-Maria, Julie; Tsai, Miaw-Sheue; White, Elizabeth J.; Tomkinson, Alan E.; Wilson, Gerald M.

    2009-05-11

    The three human LIG genes encode polypeptides that catalyze phosphodiester bond formation during DNA replication, recombination and repair. While numerous studies have identified protein partners of the human DNA ligases (hLigs), there has been little characterization of the catalytic properties of these enzymes. In this study, we developed and optimized a fluorescence-based DNA ligation assay to characterize the activities of purified hLigs. Although hLigI joins DNA nicks, it has no detectable activity on linear duplex DNA substrates with short, cohesive single-strand ends. By contrast, hLigIII{beta} and the hLigIII{alpha}/XRCC1 and hLigIV/XRCC4 complexes are active on both nicked and linear duplex DNA substrates. Surprisingly, hLigIV/XRCC4, which is a key component of the major non-homologous end joining (NHEJ) pathway, is significantly less active than hLigIII on a linear duplex DNA substrate. Notably, hLigIV/XRCC4 molecules only catalyze a single ligation event in the absence or presence of ATP. The failure to catalyze subsequent ligation events reflects a defect in the enzyme-adenylation step of the next ligation reaction and suggests that, unless there is an in vivo mechanism to reactivate DNA ligase IV/XRCC4 following phosphodiester bond formation, the cellular NHEJ capacity will be determined by the number of adenylated DNA ligaseIV/XRCC4 molecules.

  2. Why do people join trade unions?

    DEFF Research Database (Denmark)

    Toubøl, Jonas; Jensen, Carsten Strøby

    level on union recruitment, which is not done before. Workplace union density is taken to measure the strength of the workplace’s custom of being union member creating an instrumental incentive to join the union. Self-placement on a political left-right scale measures political attitude taken...

  3. Tungsten foil laminate for structural divertor applications - Joining of tungsten foils

    Science.gov (United States)

    Reiser, Jens; Rieth, Michael; Möslang, Anton; Dafferner, Bernhard; Hoffmann, Jan; Mrotzek, Tobias; Hoffmann, Andreas; Armstrong, D. E. J.; Yi, Xiaoou

    2013-05-01

    This paper is the fourth in our series on tungsten laminates. The aim of this paper is to discuss laminate synthesis, meaning the joining of tungsten foils. It is obvious that the properties of the tungsten laminate strongly depend on the combination of (i) interlayer and (ii) joining technology, as this combination defines (i) the condition of the tungsten foil after joining (as-received or recrystallised) as well as (ii) the characteristics of the interface between the tungsten foil and the interlayer (wettability or diffusion leading to a solid solution or the formation of intermetallics). From the example of tungsten laminates joined by brazing with (i) an eutectic silver copper brazing filler, (ii) copper, (iii) titanium, and (iv) zirconium, the microstructure will be discussed, with special focus on the interface. Based on our assumptions of the mechanism of the extraordinary ductility of tungsten foil we present three syntheses strategies and make recommendations for the synthesis of high temperature tungsten laminates.

  4. Aluminum and stainless steel tubes joined by simple ring and welding process

    Science.gov (United States)

    Townhill, A.

    1967-01-01

    Duranel ring is used to join aluminum and stainless steel tubing. Duranel is a bimetal made up of roll-bonded aluminum and stainless steel. This method of joining the tubing requires only two welding operations.

  5. p53 regulates the repair of DNA double-strand breaks by both homologous and non-homologous recombination

    International Nuclear Information System (INIS)

    Willers, H.; Powell, S.N.; Dahm-Daphi, J.

    2003-01-01

    Full text: p53 is known to suppress spontaneous homologous recombination (HR), while its role in non-homologous recombination (NHR) remains to be clarified. Here, we sought to determine the influence of p53 on the repair of chromosomal double-strand breaks (DSBs) by HR or NHR using specially designed recombination substrates that integrate into the genome. Isogenic mouse fibroblast pairs with or without expression of exogenous p53 protein were utilized. A reporter plasmid carrying a mutated XGPRT gene was chromosomally integrated and DSBs were generated within the plasmid by the I-SceI endonuclease. Subsequent homology-mediated repair from an episomal donor resulted in XGPRT reconstitution and cellular resistance to a selection antibiotic. Analogously, the repair of chromosomal I-SceI breaks by NHR using another novel reporter plasmid restored XGPRT translation. For p53-null cells, the mean frequency of I-SceI break repair via HR was 5.5 x 10 -4 . The p53-Val135 mutant, which previously has been shown to suppress spontaneous HR by 14-fold employing the same cell system and reporter gene, only caused a 2- to 3-fold suppression of break-induced HR. In contrast, a dramatic effect of p53 on repair via NHR was found. Preliminary sequence analysis indicated that there was at least a 1000-fold reduction of illegitimate repair events resulting in loss of sequence at the break sites. The observed effects were mediated by p53 mutants defective in regulation of the cell-cycle and apoptosis. The main findings were: (1) p53 virtually blocked illegitimate rejoining of chromosomal ends. (2) The suppression of homologous DSB repair was less pronounced than the inhibition of spontaneous HR. We hypothesize that p53 allows to a certain extent error-free homology-dependent repair to proceed, while blocking error-prone NHR. The data support and extent a previous model, in which p53 maintains genomic stability by regulating recombination independently of its transactivation function

  6. Factors affecting the diffusion of online end user literature searching.

    Science.gov (United States)

    Ash, J S

    1999-01-01

    The aim of this study was to identify factors that affect diffusion of usage of online end user literature searching. Fifteen factors clustered into three attribute sets (innovation attributes, organizational attributes, and marketing attributes) were measured to study their effect on the diffusion of online searching within institutions. A random sample of sixty-seven academic health sciences centers was selected and then 1,335 library and informatics staff members at those institutions were surveyed by mail with electronic mail follow-up. Multiple regression analysis was performed. The survey yielded a 41% response rate with electronic mail follow-up being particularly effective. Two dependent variables, internal diffusion (spread of diffusion) and infusion (depth of diffusion), were measured. There was little correlation between them, indicating they measured different things. Fifteen independent variables clustered into three attribute sets were measured. The innovation attributes set was significant for both internal diffusion and infusion. Significant individual variables were visibility for internal diffusion and image enhancement effects (negative relation) as well as visibility for infusion (depth of diffusion). Organizational attributes were also significant predictors for both dependent variables. No individual variables were significant for internal diffusion. Communication, management support (negative relation), rewards, and existence of champions were significant for infusion. Marketing attributes were not significant predictors. Successful diffusion of online end user literature searching is dependent on the visibility of the systems, communication among, rewards to, and peers of possible users who promote use (champions). Personal image enhancement effects have a negative relation to infusion, possibly because the use of intermediaries is still seen as the more luxurious way to have searches done. Management support also has a negative relation to

  7. Atom probe tomography of intermetallic phases and interfaces formed in dissimilar joining between Al alloys and steel

    International Nuclear Information System (INIS)

    Lemmens, B.; Springer, H.; Duarte, M.J.; De Graeve, I.; De Strycker, J.; Raabe, D.; Verbeken, K.

    2016-01-01

    While Si additions to Al are widely used to reduce the thickness of the brittle intermetallic seam formed at the interface during joining of Al alloys to steel, the underlying mechanisms are not clarified yet. The developed approach for the site specific atom probe tomography analysis revealed Si enrichments at grain and phase boundaries between the θ (Fe 4 Al 13 ) and η (Fe 2 Al 5 ) phase, up to about ten times that of the concentration in Al. The increase in Si concentration could play an important role for the growth kinetics of the intermetallic phases formed for example in hot-dip aluminizing of steel. - Highlights: •Si additions to Al reduce thickness of intermetallic seam in joining with steel. •Approach developed for the site specific APT analysis of the intermetallic seam •Si enrichment at grain and phase boundaries possibly affects growth of intermetallics.

  8. Teens join the MoEDAL collaboration

    CERN Multimedia

    Stephanie Hills

    2013-01-01

    The principal investigator for any institute joining an experimental collaboration is generally a self-assured researcher with evident leadership skills and an in-depth knowledge of their subject gained over many years. Katherine Evans fits the brief in every respect, except that she is 17 years old and her research institute is the Langton Star Centre, based at the Simon Langton Grammar School for Boys. The school has just joined the MoEDAL experiment.   Teacher Becky Parker (left) with two students from the Simon Langton Grammar School for Boys in the MoEDAL experimental area. MoEDAL, the latest LHC experiment has detectors located close to the interaction point of the LHCb experiment. This new experiment is designed to search for the highly ionizing avatars of new physics at the LHC, specifically the magnetic monopole or dyon and other highly ionizing stable massive particles from a number of beyond-the-Standard-Model scenarios. MoEDAL was approved in 2010 and is due to start taking data i...

  9. Joining of thermoplastic substrates by microwaves

    Science.gov (United States)

    Paulauskas, Felix L.; Meek, Thomas T.

    1997-01-01

    A method for joining two or more items having surfaces of thermoplastic material includes the steps of depositing an electrically-conductive material upon the thermoplastic surface of at least one of the items, and then placing the other of the two items adjacent the one item so that the deposited material is in intimate contact with the surfaces of both the one and the other items. The deposited material and the thermoplastic surfaces contacted thereby are then exposed to microwave radiation so that the thermoplastic surfaces in contact with the deposited material melt, and then pressure is applied to the two items so that the melted thermoplastic surfaces fuse to one another. Upon discontinuance of the exposure to the microwave energy, and after permitting the thermoplastic surfaces to cool from the melted condition, the two items are joined together by the fused thermoplastic surfaces. The deposited material has a thickness which is preferably no greater than a skin depth, .delta..sub.s, which is related to the frequency of the microwave radiation and characteristics of the deposited material in accordance with an equation.

  10. Joining of metals to structural ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Sistiaga, J M; Salvador, J M

    1988-01-01

    A wide review is made on metal-ceramics joining by brazing, mainly by active metal containing brazing filler alloys and solid state welding that is diffusion welding and hot isostatic pressure (HIP). Both the basic aspects of the processes and the mechanisms involved are considsered. At last, different joint testing ands evaluation procedures are presented. (Author)

  11. Joining of metals to structural ceramics

    International Nuclear Information System (INIS)

    Sistiaga, J.M.; Salvador, J.M.

    1988-01-01

    A wide review is made on metal-ceramics joining by brazing, mainly by active metal containing brazing filler alloys and solid state welding that is diffusion welding and hot isostatic pressure (HIP). Both the basic aspects of the processes and the mechanisms involved are considered. At last, different joint testing and evaluation procedures are presented. (Author)

  12. Alternative end-joining of DNA breaks

    NARCIS (Netherlands)

    Schendel, Robin van

    2016-01-01

    DNA is arguably the most important molecule found in any organism, as it contains all information to perform cellular functions and enables continuity of species. It is continuously exposed to DNA-damaging agents both from endogenous and exogenous sources. To protect DNA against these sources of DNA

  13. History-based joins : semantics, soundness and implementation

    NARCIS (Netherlands)

    Hee, van K.M.; Oanea, O.I.; Serebrenik, A.; Sidorova, N.; Voorhoeve, M.; Dustdar, S.; Fiadeiro, J.L.; Sheth, A.

    2006-01-01

    In this paper we study the use of case history for control structures in workflow processes. In particular we introduce a historydependent join. History dependent control offers much more modeling power than classical control structures and it solves several semantical problems of industrial

  14. Tungsten foil laminate for structural divertor applications – Joining of tungsten foils

    Energy Technology Data Exchange (ETDEWEB)

    Reiser, Jens, E-mail: jens.reiser@kit.edu [Karlsruhe Institute of Technology (KIT), Institute for Applied Materials (IAM-AWP) (Germany); Rieth, Michael; Möslang, Anton; Dafferner, Bernhard; Hoffmann, Jan [Karlsruhe Institute of Technology (KIT), Institute for Applied Materials (IAM-AWP) (Germany); Mrotzek, Tobias; Hoffmann, Andreas [PLANSEE SE, Reutte (Austria); Armstrong, D.E.J.; Yi, Xiaoou [University of Oxford, Department of Materials (United Kingdom)

    2013-05-15

    This paper is the fourth in our series on tungsten laminates. The aim of this paper is to discuss laminate synthesis, meaning the joining of tungsten foils. It is obvious that the properties of the tungsten laminate strongly depend on the combination of (i) interlayer and (ii) joining technology, as this combination defines (i) the condition of the tungsten foil after joining (as-received or recrystallised) as well as (ii) the characteristics of the interface between the tungsten foil and the interlayer (wettability or diffusion leading to a solid solution or the formation of intermetallics). From the example of tungsten laminates joined by brazing with (i) an eutectic silver copper brazing filler, (ii) copper, (iii) titanium, and (iv) zirconium, the microstructure will be discussed, with special focus on the interface. Based on our assumptions of the mechanism of the extraordinary ductility of tungsten foil we present three syntheses strategies and make recommendations for the synthesis of high temperature tungsten laminates.

  15. Machining, joining and modifications of advanced materials

    CERN Document Server

    Altenbach, Holm

    2016-01-01

    This book presents the latest advances in mechanical and materials engineering applied to the machining, joining and modification of modern engineering materials. The contributions cover the classical fields of casting, forming and injection moulding as representative manufacturing methods, whereas additive manufacturing methods (rapid prototyping and laser sintering) are treated as more innovative and recent technologies that are paving the way for the manufacturing of shapes and features that traditional methods are unable to deliver. The book also explores water jet cutting as an innovative cutting technology that avoids the heat build-up typical of classical mechanical cutting. It introduces readers to laser cutting as an alternative technology for the separation of materials, and to classical bonding and friction stir welding approaches in the context of joining technologies. In many cases, forming and machining technologies require additional post-treatment to achieve the required level of surface quali...

  16. Regulation of the DNA Damage Response by DNA-PKcs Inhibitory Phosphorylation of ATM.

    Science.gov (United States)

    Zhou, Yi; Lee, Ji-Hoon; Jiang, Wenxia; Crowe, Jennie L; Zha, Shan; Paull, Tanya T

    2017-01-05

    Ataxia-telangiectasia mutated (ATM) regulates the DNA damage response as well as DNA double-strand break repair through homologous recombination. Here we show that ATM is hyperactive when the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) is chemically inhibited or when the DNA-PKcs gene is deleted in human cells. Pre-incubation of ATM protein with active DNA-PKcs also significantly reduces ATM activity in vitro. We characterize several phosphorylation sites in ATM that are targets of DNA-PKcs and show that phospho-mimetic mutations at these residues significantly inhibit ATM activity and impair ATM signaling upon DNA damage. In contrast, phospho-blocking mutations at one cluster of sites increase the frequency of apoptosis during normal cell growth. DNA-PKcs, which is integral to the non-homologous end joining pathway, thus negatively regulates ATM activity through phosphorylation of ATM. These observations illuminate an important regulatory mechanism for ATM that also controls DNA repair pathway choice. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. FACS-Assisted CRISPR-Cas9 Genome Editing Facilitates Parkinson's Disease Modeling

    Directory of Open Access Journals (Sweden)

    Jonathan Arias-Fuenzalida

    2017-11-01

    Full Text Available Genome editing and human induced pluripotent stem cells hold great promise for the development of isogenic disease models and the correction of disease-associated mutations for isogenic tissue therapy. CRISPR-Cas9 has emerged as a versatile and simple tool for engineering human cells for such purposes. However, the current protocols to derive genome-edited lines require the screening of a great number of clones to obtain one free of random integration or on-locus non-homologous end joining (NHEJ-containing alleles. Here, we describe an efficient method to derive biallelic genome-edited populations by the use of fluorescent markers. We call this technique FACS-assisted CRISPR-Cas9 editing (FACE. FACE allows the derivation of correctly edited polyclones carrying a positive selection fluorescent module and the exclusion of non-edited, random integrations and on-target allele NHEJ-containing cells. We derived a set of isogenic lines containing Parkinson's-disease-associated mutations in α-synuclein and present their comparative phenotypes.

  18. TALE nucleases and next generation GM crops.

    KAUST Repository

    Mahfouz, Magdy M.

    2011-04-01

    Site-specific and adaptable DNA binding domains are essential modules to develop genome engineering technologies for crop improvement. Transcription activator-like effectors (TALEs) proteins are used to provide a highly specific and adaptable DNA binding modules. TALE chimeric nucleases (TALENs) were used to generate site-specific double strand breaks (DSBs) in vitro and in yeast, Caenorhabditis elegans, mammalian and plant cells. The genomic DSBs can be generated at predefined and user-selected loci and repaired by either the non-homologous end joining (NHEJ) or homology dependent repair (HDR). Thus, TALENs can be used to achieve site-specific gene addition, stacking, deletion or inactivation. TALE-based genome engineering tools should be powerful to develop new agricultural biotechnology approaches for crop improvement. Here, we discuss the recent research and the potential applications of TALENs to accelerate the generation of genomic variants through targeted mutagenesis and to produce a non-transgenic GM crops with the desired phenotype.

  19. Distinct roles of FANCO/RAD51C in DNA damage signaling and repair: implications for fanconi anemia and breast cancer susceptibility

    International Nuclear Information System (INIS)

    Nagaraju, G.; Somyajit, K.; Subramanya, S.

    2012-01-01

    Unrepaired or misrepaired chromosomal double-strand breaks (DSBs) can cause gross chromosomal rearrangements which eventually can lead to tumorigenesis through inactivation of tumor suppressor genes or activation of oncogenes. There are two major mechanisms of DSB repair: non-homologous end joining (NHEJ) and homologous recombination (HR). DSBs that are generated during S and G2 phase of the cell are preferentially repaired by sister chromatid recombination (SCR), an HR pathway that utilizes neighboring sister chromatid as a template. Since the copied information is accurate, SCR is potentially an error-free pathway. HR also plays a critical role in the repair of daughter strand gaps (DSGs) that arise as a result of replication fork stalling and facilitates replication fork recovery. Furthermore, in collaboration with nucleotide excision repair and translesion synthesis, HR is involved in the repair of DNA interstrand cross-links (ICLs). Thus, HR is important for the maintenance of genome integrity and its dysfunction can lead to various genetic disorders and cancer

  20. Small molecules enhance CRISPR genome editing in pluripotent stem cells.

    Science.gov (United States)

    Yu, Chen; Liu, Yanxia; Ma, Tianhua; Liu, Kai; Xu, Shaohua; Zhang, Yu; Liu, Honglei; La Russa, Marie; Xie, Min; Ding, Sheng; Qi, Lei S

    2015-02-05

    The bacterial CRISPR-Cas9 system has emerged as an effective tool for sequence-specific gene knockout through non-homologous end joining (NHEJ), but it remains inefficient for precise editing of genome sequences. Here we develop a reporter-based screening approach for high-throughput identification of chemical compounds that can modulate precise genome editing through homology-directed repair (HDR). Using our screening method, we have identified small molecules that can enhance CRISPR-mediated HDR efficiency, 3-fold for large fragment insertions and 9-fold for point mutations. Interestingly, we have also observed that a small molecule that inhibits HDR can enhance frame shift insertion and deletion (indel) mutations mediated by NHEJ. The identified small molecules function robustly in diverse cell types with minimal toxicity. The use of small molecules provides a simple and effective strategy to enhance precise genome engineering applications and facilitates the study of DNA repair mechanisms in mammalian cells. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Escape from Telomere-Driven Crisis Is DNA Ligase III Dependent

    Directory of Open Access Journals (Sweden)

    Rhiannon E. Jones

    2014-08-01

    Full Text Available Short dysfunctional telomeres are capable of fusion, generating dicentric chromosomes and initiating breakage-fusion-bridge cycles. Cells that escape the ensuing cellular crisis exhibit large-scale genomic rearrangements that drive clonal evolution and malignant progression. We demonstrate that there is an absolute requirement for fully functional DNA ligase III (LIG3, but not ligase IV (LIG4, to facilitate the escape from a telomere-driven crisis. LIG3- and LIG4-dependent alternative (A and classical (C nonhomologous end-joining (NHEJ pathways were capable of mediating the fusion of short dysfunctional telomeres, both displaying characteristic patterns of microhomology and deletion. Cells that failed to escape crisis exhibited increased proportions of C-NHEJ-mediated interchromosomal fusions, whereas those that escaped displayed increased proportions of intrachromosomal fusions. We propose that the balance between inter- and intrachromosomal telomere fusions dictates the ability of human cells to escape crisis and is influenced by the relative activities of A- and C-NHEJ at short dysfunctional telomeres.

  2. IgD class switching is initiated by microbiota and limited to mucosa-associated lymphoid tissue in mice

    Science.gov (United States)

    Choi, Jin Huk; Wang, Kuan-wen; Zhang, Duanwu; Zhan, Xiaowei; Wang, Tao; Bu, Chun-Hui; Behrendt, Cassie L.; Zeng, Ming; Wang, Ying; Misawa, Takuma; Li, Xiaohong; Tang, Miao; Zhan, Xiaoming; Scott, Lindsay; Hildebrand, Sara; Murray, Anne R.; Moresco, Eva Marie Y.; Hooper, Lora V.; Beutler, Bruce

    2017-01-01

    Class-switch recombination (CSR) alters the Ig isotype to diversify antibody effector functions. IgD CSR is a rare event, and its regulation is poorly understood. We report that deficiency of 53BP1, a DNA damage-response protein, caused age-dependent overproduction of secreted IgD resulting from increased IgD CSR exclusively within B cells of mucosa-associated lymphoid tissues. IgD overproduction was dependent on activation-induced cytidine deaminase, hematopoietic MyD88 expression, and an intact microbiome, against which circulating IgD, but not IgM, was reactive. IgD CSR occurred via both alternative nonhomologous end-joining and homologous recombination pathways. Microbiota-dependent IgD CSR also was detected in nasal-associated lymphoid tissue of WT mice. These results identify a pathway, present in WT mice and hyperactivated in 53BP1-deficient mice, by which microbiota signal via Toll-like receptors to elicit IgD CSR. PMID:28137874

  3. Modification of ethylene sensitivity in ornamental plants using CRISPR/Cas9

    DEFF Research Database (Denmark)

    Kemp, Oliver; Favero, Bruno Trevenzoli; Hegelund, Josefine Nymark

    2017-01-01

    ; the Clustered Regularly Interspaced Palindromic Repeats (CRISPR) RNA guided Cas9 DNA nuclease (CRISPR/Cas9). CRISPR/Cas9 may be employed to introduce targeted double-stranded breaks (DSBs) at desired sites in the host genome. The DSBs will be repaired by the non-homologous end-joining (NHEJ) repair mechanism...... which often results in small indels and consequently gene knockout. The CRISPR/Cas9 system consists of a protein DNA nuclease (Cas9) which is guided to the target sequence by a small RNA molecule (sgRNA) that recognizes a 20 bp target sequence in the genome situated immediately downstream of a 3 bp...... protospacer adjacent motif (PAM). The sgRNA confers the sequence specificity of the CRISPR/Cas9 complex and may thus be designed to target virtually any sequence, a feature that has made it the method of choice within precise genetic engineering. Although most research with CRISPR/Cas9 has been conducted...

  4. Genomic analysis of murine DNA-dependent protein kinase

    International Nuclear Information System (INIS)

    Fujimori, A.; Abe, M.

    2003-01-01

    Full text: The gene of catalytic subunit of DNA dependent protein kinase is responsible gene for SCID mice. The molecules play a critical role in non-homologous end joining including the V(D)J recombination. Contribution of the molecules to the difference of radiosensitivity and the susceptibility to cancer has been suggested. Here we show the entire nucleotide sequence of approximately 193 kbp and 84 kbp genomic regions encoding the entire DNA-PKcs gene in the mouse and chicken respectively. Retroposon was found in the intron 51 of mouse genomic DNA-PKcs gene but in human and chicken. Comparative analysis of these two species strongly suggested that only two genes, DNA-PKcs and MCM4, exist in the region of both species. Several conserved sequences and cis elements, however, were predicted. Recently, the orthologous region for the human DNA-PKcs locus was completed. The results of further comparative study will be discussed

  5. IgD class switching is initiated by microbiota and limited to mucosa-associated lymphoid tissue in mice.

    Science.gov (United States)

    Choi, Jin Huk; Wang, Kuan-Wen; Zhang, Duanwu; Zhan, Xiaowei; Wang, Tao; Bu, Chun-Hui; Behrendt, Cassie L; Zeng, Ming; Wang, Ying; Misawa, Takuma; Li, Xiaohong; Tang, Miao; Zhan, Xiaoming; Scott, Lindsay; Hildebrand, Sara; Murray, Anne R; Moresco, Eva Marie Y; Hooper, Lora V; Beutler, Bruce

    2017-02-14

    Class-switch recombination (CSR) alters the Ig isotype to diversify antibody effector functions. IgD CSR is a rare event, and its regulation is poorly understood. We report that deficiency of 53BP1, a DNA damage-response protein, caused age-dependent overproduction of secreted IgD resulting from increased IgD CSR exclusively within B cells of mucosa-associated lymphoid tissues. IgD overproduction was dependent on activation-induced cytidine deaminase, hematopoietic MyD88 expression, and an intact microbiome, against which circulating IgD, but not IgM, was reactive. IgD CSR occurred via both alternative nonhomologous end-joining and homologous recombination pathways. Microbiota-dependent IgD CSR also was detected in nasal-associated lymphoid tissue of WT mice. These results identify a pathway, present in WT mice and hyperactivated in 53BP1-deficient mice, by which microbiota signal via Toll-like receptors to elicit IgD CSR.

  6. RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency.

    Science.gov (United States)

    Song, Jun; Yang, Dongshan; Xu, Jie; Zhu, Tianqing; Chen, Y Eugene; Zhang, Jifeng

    2016-01-28

    Zinc-finger nuclease, transcription activator-like effector nuclease and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) are becoming major tools for genome editing. Importantly, knock-in in several non-rodent species has been finally achieved thanks to these customizable nucleases; yet the rates remain to be further improved. We hypothesize that inhibiting non-homologous end joining (NHEJ) or enhancing homology-directed repair (HDR) will improve the nuclease-mediated knock-in efficiency. Here we show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. Our work presents tools to nuclease-mediated knock-in animal production, and sheds light on improving gene-targeting efficiencies on pluripotent stem cells.

  7. RS-1 enhances CRISPR/Cas9- and TALEN-mediated knock-in efficiency

    Science.gov (United States)

    Song, Jun; Yang, Dongshan; Xu, Jie; Zhu, Tianqing; Chen, Y. Eugene; Zhang, Jifeng

    2016-01-01

    Zinc-finger nuclease, transcription activator-like effector nuclease and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) are becoming major tools for genome editing. Importantly, knock-in in several non-rodent species has been finally achieved thanks to these customizable nucleases; yet the rates remain to be further improved. We hypothesize that inhibiting non-homologous end joining (NHEJ) or enhancing homology-directed repair (HDR) will improve the nuclease-mediated knock-in efficiency. Here we show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. Our work presents tools to nuclease-mediated knock-in animal production, and sheds light on improving gene-targeting efficiencies on pluripotent stem cells. PMID:26817820

  8. Carcinoma esophagus with xeroderma pigmentosa: Case report on a rare association

    Directory of Open Access Journals (Sweden)

    P Guru Sai Ratna Priya

    2018-01-01

    Full Text Available Radiation in patients with diseases such as xeroderma pigmentosa (XP, systemic lupus erythematosus, and other connective diseases is a matter of concern because of higher incidence of toxicities. Here with, we are reporting a case of carcinoma esophagus with XP, who tolerated the treatment well with sufficiently prolonged palliation of symptoms, after treatment with external beam radiotherapy. This might be attributed to the different mechanisms of DNA damage and repair mechanisms for ultraviolet (UV rays and X-rays. UV rays cause DNA damage by dimer formation whereas X-rays will cause single- or double-stranded breaks in DNA. The repair mechanisms for UV rays are nucleotide excision repair and translesion synthesis while for X-rays, they are base excision repair, homologous recombination, and nonhomologous end joining, and these repair mechanisms for X-rays are intact in a XP patient. Hence, they can be been treated with high dose of radiation, and they do tolerate the treatment well.

  9. Effect of Ti and Si interlayer materials on the joining of SiC ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Yang Il; Park, Jung Hwan; Kim, Hyun Gil; Park, Dong Jun; Park, Jeong Yong; Kim, Weon Ju [LWR Fuel Technology Division, Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2016-08-15

    SiC-based ceramic composites are currently being considered for use in fuel cladding tubes in light-water reactors. The joining of SiC ceramics in a hermetic seal is required for the development of ceramic-based fuel cladding tubes. In this study, SiC monoliths were diffusion bonded using a Ti foil interlayer and additional Si powder. In the joining process, a very low uniaxial pressure of ∼0.1 MPa was applied, so the process is applicable for joining thin-walled long tubes. The joining strength depended strongly on the type of SiC material. Reaction-bonded SiC (RB-SiC) showed a higher joining strength than sintered SiC because the diffusion reaction of Si was promoted in the former. The joining strength of sintered SiC was increased by the addition of Si at the Ti interlayer to play the role of the free Si in RB-SiC. The maximum joint strength obtained under torsional stress was ∼100 MPa. The joint interface consisted of TiSi{sub 2}, Ti{sub 3}SiC{sub 2}, and SiC phases formed by a diffusion reaction of Ti and Si.

  10. Joining of HHF materials applying electroplating technology

    Energy Technology Data Exchange (ETDEWEB)

    Krauss, Wolfgang, E-mail: wolfgang.krauss@kit.edu; Lorenz, Julia; Konys, Jürgen

    2014-10-15

    Highlights: • Electroplating of fillers is industrially relevant for brazing in fusion. • Interlayers (Ni or Pd) improve adherence and reduce failure risks. • Tungsten and Eurofer joints successfully fabricated by electroplating. • Mechanical and non-destructive testing integrated into qualification. • Shear strength of W joints comparable with conventionally brazed steel. - Abstract: Tungsten will be used as armor material for blanket shielding and is designated as high heat flux material for divertors, beyond application of improved W composite alloys as structural material. Independent from design (water- or helium-cooled), a successful development is inherently correlated with joining of tungsten with functional components. Depending on the design variants, the fabricated joints have to guarantee specific functional or structural properties, e.g., good thermal conductivity or mechanical load transmission. Tungsten shows lacks in adapted joining due to its metallurgical behavior ranging from immiscibility over bad wetting to brittle intermetallic phase formation. Electroplating has shown to overcome such drawbacks and that homogeneous functional (e.g., Ni or Pd) and filler (e.g., Cu) layers can be deposited. In this paper the progress achieved in development of electroplating processes for joining W to W or steel to steel will be shown. The main focus will be the characterization of the processed joints applying metallurgical investigations including SEM/EDX analyses and non-destructive testing. The mechanical stability of the produced joints is demonstrated by presenting recent shear test data. The W–W joints failed due to cracking in W, whereas the steel–steel joints cracked in the brazing zone at about 200 N/mm{sup 2} load.

  11. Loss of Ubr2, an E3 ubiquitin ligase, leads to chromosome fragility and impaired homologous recombinational repair

    International Nuclear Information System (INIS)

    Ouyang, Yan; Kwon, Yong Tae; An, Jee Young; Eller, Danny; Tsai, S.-C.; Diaz-Perez, Silvia; Troke, Joshua J.; Teitell, Michael A.; Marahrens, York

    2006-01-01

    The N-end rule pathway of protein degradation targets proteins with destabilizing N-terminal residues. Ubr2 is one of the E3 ubiquitin ligases of the mouse N-end rule pathway. We have previously shown that Ubr2 -/- male mice are infertile, owing to the arrest of spermatocytes between the leptotene/zygotene and pachytene of meiosis I, the failure of chromosome pairing, and subsequent apoptosis. Here, we report that mouse fibroblast cells derived from Ubr2 -/- embryos display genome instability. The frequency of chromosomal bridges and micronuclei were much higher in Ubr2 -/- fibroblasts than in +/+ controls. Metaphase chromosome spreads from Ubr2 -/- cells revealed a high incidence of spontaneous chromosomal gaps, indicating chromosomal fragility. These fragile sites were generally replicated late in S phase. Ubr2 -/- cells were hypersensitive to mitomycin C, a DNA cross-linking agent, but displayed normal sensitivity to gamma-irradiation. A reporter assay showed that Ubr2 -/- cells are significantly impaired in the homologous recombination repair of a double strand break. In contrast, Ubr2 -/- cells appeared normal in an assay for non-homologous end joining. Our results therefore unveil the role of the ubiquitin ligase Ubr2 in maintaining genome integrity and in homologous recombination repair

  12. Loss of Ubr2, an E3 ubiquitin ligase, leads to chromosome fragility and impaired homologous recombinational repair

    Energy Technology Data Exchange (ETDEWEB)

    Ouyang, Yan [Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Kwon, Yong Tae [Center for Pharmacogenetics and Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA 15261 (United States); An, Jee Young [Center for Pharmacogenetics and Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA 15261 (United States); Eller, Danny [Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Tsai, S.-C. [Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Diaz-Perez, Silvia [Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Troke, Joshua J. [Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Teitell, Michael A. [Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States); Marahrens, York [Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095 (United States)]. E-mail: ymarahrens@mednet.ucla.edu

    2006-04-11

    The N-end rule pathway of protein degradation targets proteins with destabilizing N-terminal residues. Ubr2 is one of the E3 ubiquitin ligases of the mouse N-end rule pathway. We have previously shown that Ubr2{sup -/-} male mice are infertile, owing to the arrest of spermatocytes between the leptotene/zygotene and pachytene of meiosis I, the failure of chromosome pairing, and subsequent apoptosis. Here, we report that mouse fibroblast cells derived from Ubr2{sup -/-} embryos display genome instability. The frequency of chromosomal bridges and micronuclei were much higher in Ubr2{sup -/-} fibroblasts than in +/+ controls. Metaphase chromosome spreads from Ubr2{sup -/-} cells revealed a high incidence of spontaneous chromosomal gaps, indicating chromosomal fragility. These fragile sites were generally replicated late in S phase. Ubr2{sup -/-} cells were hypersensitive to mitomycin C, a DNA cross-linking agent, but displayed normal sensitivity to gamma-irradiation. A reporter assay showed that Ubr2{sup -/-} cells are significantly impaired in the homologous recombination repair of a double strand break. In contrast, Ubr2{sup -/-} cells appeared normal in an assay for non-homologous end joining. Our results therefore unveil the role of the ubiquitin ligase Ubr2 in maintaining genome integrity and in homologous recombination repair.

  13. Differential radiosensitivity phenotypes of DNA-PKcs mutations affecting NHEJ and HRR systems following irradiation with gamma-rays or very low fluences of alpha particles.

    Science.gov (United States)

    Lin, Yu-Fen; Nagasawa, Hatsumi; Little, John B; Kato, Takamitsu A; Shih, Hung-Ying; Xie, Xian-Jin; Wilson, Paul F; Brogan, John R; Kurimasa, Akihiro; Chen, David J; Bedford, Joel S; Chen, Benjamin P C

    2014-01-01

    We have examined cell-cycle dependence of chromosomal aberration induction and cell killing after high or low dose-rate γ irradiation in cells bearing DNA-PKcs mutations in the S2056 cluster, the T2609 cluster, or the kinase domain. We also compared sister chromatid exchanges (SCE) production by very low fluences of α-particles in DNA-PKcs mutant cells, and in homologous recombination repair (HRR) mutant cells including Rad51C, Rad51D, and Fancg/xrcc9. Generally, chromosomal aberrations and cell killing by γ-rays were similarly affected by mutations in DNA-PKcs, and these mutant cells were more sensitive in G1 than in S/G2 phase. In G1-irradiated DNA-PKcs mutant cells, both chromosome- and chromatid-type breaks and exchanges were in excess than wild-type cells. For cells irradiated in late S/G2 phase, mutant cells showed very high yields of chromatid breaks compared to wild-type cells. Few exchanges were seen in DNA-PKcs-null, Ku80-null, or DNA-PKcs kinase dead mutants, but exchanges in excess were detected in the S2506 or T2609 cluster mutants. SCE induction by very low doses of α-particles is resulted from bystander effects in cells not traversed by α-particles. SCE seen in wild-type cells was completely abolished in Rad51C- or Rad51D-deficient cells, but near normal in Fancg/xrcc9 cells. In marked contrast, very high levels of SCEs were observed in DNA-PKcs-null, DNA-PKcs kinase-dead and Ku80-null mutants. SCE induction was also abolished in T2609 cluster mutant cells, but was only slightly reduced in the S2056 cluster mutant cells. Since both non-homologous end-joining (NHEJ) and HRR systems utilize initial DNA lesions as a substrate, these results suggest the possibility of a competitive interference phenomenon operating between NHEJ and at least the Rad51C/D components of HRR; the level of interaction between damaged DNA and a particular DNA-PK component may determine the level of interaction of such DNA with a relevant HRR component.

  14. Five reasons to join local Chamber of Commerce and Industry

    OpenAIRE

    Verbovskii, Vladislav; Kosov, Vladimir; Chaika (Chayka), Yuliya Aleksandrovna

    2016-01-01

    The article describes five useful things that Chamber of Commerce and Industry membership may give to business owners after joining it. These things are the reasons to become a part of business community formed by Chambers of Commerce that may be considered by those business owner who are deciding whether to join Chamber of Commerce or not. Mentioned reasons are given with examples related to Tomsk Chamber of Commerce and Industry and are relevant to any Chamber of Commerce located in Russian...

  15. Evaluation of the feasibility of joining titanium alloy to heavymet tungsten alloy

    Energy Technology Data Exchange (ETDEWEB)

    1978-06-07

    Information is presented on a program to select and evaluate methods of brazing and/or explosively welding Ti-6Al-4V titanium alloy to Heavymet, a tungsten-base metal containing up to about 20% alloying elements (nickel, copper, etc.) to improve its ductility and other mechanical properties. Designs permitting the reliable production of joints between these base metals were of interest too. While this investigation was primarily concerned with an engineering study of the problems associated with joining these base metals in the required configuration, limited experimental studies were conducted also. The joining methods are reviewed individually. Recommendations for developing a viable titanium-tungsten joining procedure are discussed.

  16. Evaluation of the feasibility of joining titanium alloy to heavymet tungsten alloy

    International Nuclear Information System (INIS)

    1978-01-01

    Information is presented on a program to select and evaluate methods of brazing and/or explosively welding Ti-6Al-4V titanium alloy to Heavymet, a tungsten-base metal containing up to about 20% alloying elements (nickel, copper, etc.) to improve its ductility and other mechanical properties. Designs permitting the reliable production of joints between these base metals were of interest too. While this investigation was primarily concerned with an engineering study of the problems associated with joining these base metals in the required configuration, limited experimental studies were conducted also. The joining methods are reviewed individually. Recommendations for developing a viable titanium-tungsten joining procedure are discussed

  17. Inhibitors of the proteasome suppress homologous DNA recombination in mammalian cells.

    Science.gov (United States)

    Murakawa, Yasuhiro; Sonoda, Eiichiro; Barber, Louise J; Zeng, Weihua; Yokomori, Kyoko; Kimura, Hiroshi; Niimi, Atsuko; Lehmann, Alan; Zhao, Guang Yu; Hochegger, Helfrid; Boulton, Simon J; Takeda, Shunichi

    2007-09-15

    Proteasome inhibitors are novel antitumor agents against multiple myeloma and other malignancies. Despite the increasing clinical application, the molecular basis of their antitumor effect has been poorly understood due to the involvement of the ubiquitin-proteasome pathway in multiple cellular metabolisms. Here, we show that treatment of cells with proteasome inhibitors has no significant effect on nonhomologous end joining but suppresses homologous recombination (HR), which plays a key role in DNA double-strand break (DSB) repair. In this study, we treat human cells with proteasome inhibitors and show that the inhibition of the proteasome reduces the efficiency of HR-dependent repair of an artificial HR substrate. We further show that inhibition of the proteasome interferes with the activation of Rad51, a key factor for HR, although it does not affect the activation of ATM, gammaH2AX, or Mre11. These data show that the proteasome-mediated destruction is required for the promotion of HR at an early step. We suggest that the defect in HR-mediated DNA repair caused by proteasome inhibitors contributes to antitumor effect, as HR plays an essential role in cellular proliferation. Moreover, because HR plays key roles in the repair of DSBs caused by chemotherapeutic agents such as cisplatin and by radiotherapy, proteasome inhibitors may enhance the efficacy of these treatments through the suppression of HR-mediated DNA repair pathways.

  18. Low concentrations of antimony impair DNA damage signaling and the repair of radiation-induced DSB in HeLa S3 cells.

    Science.gov (United States)

    Koch, Barbara; Maser, Elena; Hartwig, Andrea

    2017-12-01

    Antimony is utilized in a large variety of industrial applications, leading to significant environmental and occupational exposure. Mainly based on animal experiments, the IARC and MAK Commission have classified antimony and its inorganic compounds as Group 2B or 2 carcinogens, respectively. However, the underlying mode(s) of action are still largely unknown. In the present study, we investigated the impact of non-cytotoxic up to cytotoxic concentrations of SbCl 3 on DNA DSB repair and cell cycle control in HeLa S3 cells. We induced DSB by γ-irradiation and analyzed inhibitory actions of antimony on potential molecular targets of the DSB repair machinery. Antimony disturbed cell cycle control, affecting phosphorylation of Chk1. Furthermore, the repair of DSB was impaired in the presence of antimony, as monitored by pulsed-field gel electrophoresis and γH2AX foci formation of cells in G1 and G2 phase. Specifically, BRCA1 and RAD51 were identified as molecular targets. Our results point towards an interference with both non-homologous end-joining (NHEJ) and homologous recombination (HR), and inhibitory effects may be explained by interactions with critical cysteine groups; this needs to be further investigated. Altogether, the results provide further evidence for the impairment of DNA repair processes as one underlying mechanism in antimony-induced carcinogenicity.

  19. Reduction of spontaneous somatic mutation frequency by a low-dose X irradiation of Drosophila larvae and possible involvement of DNA single-strand damage repair.

    Science.gov (United States)

    Koana, Takao; Takahashi, Takashi; Tsujimura, Hidenobu

    2012-03-01

    The third instar larvae of Drosophila were irradiated with X rays, and the somatic mutation frequency in their wings was measured after their eclosion. In the flies with normal DNA repair and apoptosis functions, 0.2 Gy irradiation at 0.05 Gy/min reduced the frequency of the so-called small spot (mutant cell clone with reduced reproductive activity) compared with that in the sham-irradiated flies. When apoptosis was suppressed using the baculovirus p35 gene, the small spot frequency increased four times in the sham-irradiated control group, but the reduction by the 0.2-Gy irradiation was still evident. In a non-homologous end joining-deficient mutant, the small spot frequency was also reduced by 0.2 Gy radiation. In a mutant deficient in single-strand break repair, no reduction in the small spot frequency by 0.2 Gy radiation was observed, and the small spot frequency increased with the radiation dose. Large spot (mutant cell clone with normal reproductive activity) frequency was not affected by suppression of apoptosis and increased monotonically with radiation dose in wild-type larvae and in mutants for single- or double-strand break repair. It is hypothesized that some of the small spots resulted from single-strand damage and, in wild-type larvae, 0.2 Gy radiation activated the normal single-strand break repair gene, which reduced the background somatic mutation frequency.

  20. Similarities between long interspersed element-1 (LINE-1) reverse transcriptase and telomerase.

    Science.gov (United States)

    Kopera, Huira C; Moldovan, John B; Morrish, Tammy A; Garcia-Perez, Jose Luis; Moran, John V

    2011-12-20

    Long interspersed element-1 (LINE-1 or L1) retrotransposons encode two proteins (ORF1p and ORF2p) that contain activities required for conventional retrotransposition by a mechanism termed target-site primed reverse transcription. Previous experiments in XRCC4 or DNA protein kinase catalytic subunit-deficient CHO cell lines, which are defective for the nonhomologous end-joining DNA repair pathway, revealed an alternative endonuclease-independent (ENi) pathway for L1 retrotransposition. Interestingly, some ENi retrotransposition events in DNA protein kinase catalytic subunit-deficient cells are targeted to dysfunctional telomeres. Here we used an in vitro assay to detect L1 reverse transcriptase activity to demonstrate that wild-type or endonuclease-defective L1 ribonucleoprotein particles can use oligonucleotide adapters that mimic telomeric ends as primers to initiate the reverse transcription of L1 mRNA. Importantly, these ribonucleoprotein particles also contain a nuclease activity that can process the oligonucleotide adapters before the initiation of reverse transcription. Finally, we demonstrate that ORF1p is not strictly required for ENi retrotransposition at dysfunctional telomeres. Thus, these data further highlight similarities between the mechanism of ENi L1 retrotransposition and telomerase.

  1. Numerical modelling in friction lap joining of aluminium alloy and carbon-fiber-reinforced-plastic sheets

    Science.gov (United States)

    Das, A.; Bang, H. S.; Bang, H. S.

    2018-05-01

    Multi-material combinations of aluminium alloy and carbon-fiber-reinforced-plastics (CFRP) have gained attention in automotive and aerospace industries to enhance fuel efficiency and strength-to-weight ratio of components. Various limitations of laser beam welding, adhesive bonding and mechanical fasteners make these processes inefficient to join metal and CFRP sheets. Friction lap joining is an alternative choice for the same. Comprehensive studies in friction lap joining of aluminium to CFRP sheets are essential and scare in the literature. The present work reports a combined theoretical and experimental study in joining of AA5052 and CFRP sheets using friction lap joining process. A three-dimensional finite element based heat transfer model is developed to compute the temperature fields and thermal cycles. The computed results are validated extensively with the corresponding experimentally measured results.

  2. Thermally joining and/or coating or thermally separating the workpieces having heat-sensitive coating, comprises restoring coating by thermally coating the coating material after thermally joining and/or coating or thermally separating

    OpenAIRE

    Riedel, Frank; Winkelmann, Ralf; Puschmann, Markus

    2011-01-01

    The method for thermally joining and/or coating or thermally separating the workpieces (1), which have a heat-sensitive coating (2), comprises restoring the coating by thermally coating a coating material (3) after thermally joining and/or coating or thermally separating the workpieces. A part of the thermal energy introduced in the workpiece for joining and/or coating or separating or in the workpieces is used for thermally coating the coating material. Two workpieces are welded or soldered ...

  3. Welding and joining of single crystals of BCC refractory metals

    International Nuclear Information System (INIS)

    Hiraoka, Yutaka; Fujii, Tadayuki

    1989-01-01

    Welding and joining is one of key technologies for the wider utilizations of a material. In the present work, the applicability of welding and joining for a single crystal of BCC refractory metal was investigated. Electron-beam welding and tungsten-inert-gas welding by a melt-run technique, and high-temperature brazing by using brazing metals such as Mo-40%Ru alloy, vanadium or platinum were conducted for molybdenum single crystal which had been prepared by means of secondary recrystallization. 12 refs.,12 figs., 2 tabs. (Author)

  4. Glass-ceramic joint and method of joining

    Science.gov (United States)

    Meinhardt, Kerry D [Richland, WA; Vienna, John D [West Richland, WA; Armstrong, Timothy R [Clinton, TN; Pederson, Larry R [Kennewick, WA

    2003-03-18

    The present invention is a glass-ceramic material and method of making useful for joining a solid ceramic component and at least one other solid component. The material is a blend of M1-M2-M3, wherein M1 is BaO, SrO, CaO, MgO, or combinations thereof, M2 is Al.sub.2 O.sub.3, present in the blend in an amount from 2 to 15 mol %, M3 is SiO.sub.2 with up to 50 mol % B.sub.2 O.sub.3 that substantially matches a coefficient of thermal expansion of the solid electrolyte. According to the present invention, a series of glass ceramics in the M1-Al.sub.2 O.sub.3 -M3 system can be used to join or seal both tubular and planar solid oxide fuel cells, oxygen electrolyzers, and membrane reactors for the production of syngas, commodity chemicals and other products.

  5. Joining Dental Ceramic Layers With Glass

    Science.gov (United States)

    Saied, MA; Lloyd, IK; Haller, WK; Lawn, BR

    2011-01-01

    Objective Test the hypothesis that glass-bonding of free-form veneer and core ceramic layers can produce robust interfaces, chemically durable and aesthetic in appearance and, above all, resistant to delamination. Methods Layers of independently produced porcelains (NobelRondo™ Press porcelain, Nobel BioCare AB and Sagkura Interaction porcelain, Elephant Dental) and matching alumina or zirconia core ceramics (Procera alumina, Nobel BioCare AB, BioZyram yttria stabilized tetragonal zirconia polycrystal, Cyrtina Dental) were joined with designed glasses, tailored to match thermal expansion coefficients of the components and free of toxic elements. Scanning electron microprobe analysis was used to characterize the chemistry of the joined interfaces, specifically to confirm interdiffusion of ions. Vickers indentations were used to drive controlled corner cracks into the glass interlayers to evaluate the toughness of the interfaces. Results The glass-bonded interfaces were found to have robust integrity relative to interfaces fused without glass, or those fused with a resin-based adhesive. Significance The structural integrity of the interfaces between porcelain veneers and alumina or zirconia cores is a critical factor in the longevity of all-ceramic dental crowns and fixed dental prostheses. PMID:21802131

  6. Study of international published experiences in joining copper and copper-alloys

    International Nuclear Information System (INIS)

    Dahlgren, Aa.

    1997-04-01

    This study has revealed a number of joining processes to be used when manufacturing copper-canisters for the final storage of high level nuclear waste. However, the decision on which material and which joining process to be used has to be based on the design criterions. The welding procedure has to be qualified, i.e. it shall be demonstrated whether the procedure is capable of fulfilling specified requirements. 32 refs

  7. What happens during a Join? - Dissecting CPU and Memory Optimization Effects

    OpenAIRE

    Manegold, Stefan; Boncz, Peter; Kersten, Martin

    2000-01-01

    textabstractPerformance of modern hardware increasingly depends on proper utilization of both the memory cache hierarchy and parallel execution possibilities in todays super-scalar CPUs. Recent database research has demonstrated that database system performance severely suffers from poor utilization of these resources. In previous work, we presented join algorithms that strongly accelerate large equi-join by tuning the memory access pattern to match the characteristics of the memory cache sub...

  8. Functional Interplay of the Mre11 Nuclease and Ku in the Response to Replication-Associated DNA Damage ▿

    Science.gov (United States)

    Foster, Steven S.; Balestrini, Alessia; Petrini, John H. J.

    2011-01-01

    The Mre11 complex is a central component of the DNA damage response, with roles in damage sensing, molecular bridging, and end resection. We have previously shown that in Saccharomyces cerevisiae, Ku70 (yKu70) deficiency reduces the ionizing radiation sensitivity of mre11Δ mutants. In this study, we show that yKu70 deficiency suppressed the camptothecin (CPT) and methyl methanesulfonate (MMS) sensitivity of nuclease-deficient mre11-3 and sae2Δ mutants in an Exo1-dependent manner. CPT-induced G2/M arrest, γ-H2AX persistence, and chromosome breaks were elevated in mre11-3 mutants. These outcomes were reduced by yKu70 deficiency. Given that the genotoxic effects of CPT are manifest during DNA replication, these data suggest that Ku limits Exo1-dependent double-strand break (DSB) resection during DNA replication, inhibiting the initial processing steps required for homology-directed repair. We propose that Mre11 nuclease- and Sae2-dependent DNA end processing, which initiates DSB resection prevents Ku from engaging DSBs, thus promoting Exo1-dependent resection. In agreement with this idea, we show that Ku affinity for binding to short single-stranded overhangs is much lower than for blunt DNA ends. Collectively, the data define a nonhomologous end joining (NHEJ)-independent, S-phase-specific function of the Ku heterodimer. PMID:21876003

  9. DNA-binding determinants promoting NHEJ by human Polμ.

    Science.gov (United States)

    Martin, Maria Jose; Juarez, Raquel; Blanco, Luis

    2012-12-01

    Non-homologous end-joining (NHEJ), the preferred pathway to repair double-strand breaks (DSBs) in higher eukaryotes, relies on a collection of molecular tools to process the broken ends, including specific DNA polymerases. Among them, Polµ is unique as it can catalyze DNA synthesis upon connection of two non-complementary ends. Here, we demonstrate that this capacity is intrinsic to Polµ, not conferred by other NHEJ factors. To understand the molecular determinants of its specific function in NHEJ, the interaction of human Polµ with DNA has been directly visualized by electromobility shift assay and footprinting assays. Stable interaction with a DNA gap requires the presence of a recessive 5'-P, thus orienting the catalytic domain for primer and nucleotide binding. Accordingly, recognition of the 5'-P is crucial to align the two DNA substrates of the NHEJ reaction. Site-directed mutagenesis demonstrates the relevance of three specific residues (Lys(249), Arg(253) and Arg(416)) in stabilizing the primer strand during end synapsis, allowing a range of microhomology-induced distortions beneficial for NHEJ. Moreover, our results suggest that the Polµ BRCT domain, thought to be exclusively involved in interaction with NHEJ core factors, has a direct role in binding the DNA region neighbor to the 5'-P, thus boosting Polµ-mediated NHEJ reactions.

  10. Stresses imposed by coolant channel end shield interaction in 200 MWe PHWR

    International Nuclear Information System (INIS)

    Mehra, V.K.; Singh, R.K.; Soni, R.S.; Kushwaha, H.S.; Kakodkar, A.

    1983-01-01

    End shield of 200 MWe Pressurised Heavy Water Reactor (PHWR) is a composite tube sheet structure consisting of two circular tube sheets joined together by lattice tubes. Each lattice tube houses a coolant channel assembly which is connected to the end shield through shock absorber device. End shield assembly is suspended in the vault by hanger rods and its horizontal position is controlled by a set of pre-compressed springs. Coolant channel assemblies elongate due to their exposure to fast neutron flux in the reactor. This permanent elongation is monitored periodically. When growth of the channel exceeds a present value, it is prevented from further elongation by the shock absorbing device. Resultant force exerted on the end shield makes it move. This paper describes a numerical method used for evaluating these forces and movement of the end shield. Stresses produced by these forces are calculated by using finite element method. Typical stress values are verified by strain gauge measurements. (orig.)

  11. Joining of silicon carbide using interlayer with matching coefficient of thermal expansion

    International Nuclear Information System (INIS)

    Perham, T.

    1996-11-01

    The primary objective of this study is to develop a technique for joining a commercially available Silicon Carbide that gives good room temperature strength and the potential for good high temperature strength. One secondary objective is that the joining technique be adaptable to SiC f /SiC composites and/or Nickel based superalloys, and another secondary objective is that the materials provide good neutron irradiation resistance and low activation for potential application inside nuclear fusion reactors. The joining techniques studied here are: (1) reaction bonding with Al-Si/Si/SiC/C; (2) reaction/infiltration with calcium aluminum silicate; (3) ion exchange mechanism to form calcium hexaluminate (a refractory cement); and (4) oxide frit brazing with cordierite

  12. Calculation of force and time of contact formation at diffusion metal joining

    Energy Technology Data Exchange (ETDEWEB)

    Sukhanov, V E [Tsentral' nyj Nauchno-Issledovatel' skij Inst. Chernoj Metallurgii, Moscow (USSR); Grushevskij, A V [Moskovskij Stankoinstrumental' nyj Inst., Moscow (USSR); Surovtsev, A P

    1989-03-01

    An analytical model of contact for mation at diffusion joining is suggested. It is based on the introduction of a rough surface with roughnesses in the form of absolutely rigid spherical segnunts into a smooth laminar body. Mathematical expressions, permitting to calculate maximum welding force (pressure) providing close contact of the surfaces welded and time for contact formation between rough surfaces joined, are obtained. Divergence of calculational and experimental data does not exceed 20%. It is confirmed that the most intensive formation of joining occurs in the initial period of welding -the stage of formation of a physical contact, when deformation processes proceed in tensively. Finite formation of a strength joint occurs at the stage of diffusion interaction.

  13. Effect of workplace incivility on end-of-work negative affect: examining individual and organizational moderators in a daily diary study.

    Science.gov (United States)

    Zhou, Zhiqing E; Yan, Yu; Che, Xin Xuan; Meier, Laurenz L

    2015-01-01

    Although previous studies have linked workplace incivility with various negative outcomes, they mainly focused on the long-term effects of chronic exposure to workplace incivility, whereas targets' short-term reactions to incivility episodes have been largely neglected. Using a daily diary design, the current study examined effects of daily workplace incivility on end-of-work negative affect and explored potential individual and organizational moderators. Data collected from 76 full-time employees across 10 consecutive working days revealed that daily workplace incivility positively predicted end-of-work negative affect while controlling for before-work negative affect. Further, the relationship was stronger for people with low emotional stability, high hostile attribution bias, external locus of control, and people experiencing low chronic workload and more chronic organizational constraints, as compared with people with high emotional stability, low hostile attribution bias, internal locus of control, and people experiencing high chronic workload and fewer chronic organizational constraints, respectively. (PsycINFO Database Record (c) 2014 APA, all rights reserved). PsycINFO Database Record (c) 2014 APA, all rights reserved.

  14. Combustion Joining for Composite Fabrication

    Science.gov (United States)

    2009-10-25

    Inert preheating Process beginning T e m p e r a t u r e , o C Time, s I = 600 Amps D = 10 mm Joule preheating only up to Tig UNCLASSIFIED • C...Honeywell Corp (South Bend, IN) • Currently build aircraft brake disks from carbon fibers • use a long (~ 100 day) CVD process to densify • Brake wear...oxidation with every landing A380 -rejected take off test C-C brakes UNCLASSIFIED Joining C-Based Materials • Difficult task – Carbon cannot be welded

  15. High-temperature stability of laser-joined silicon carbide components

    Energy Technology Data Exchange (ETDEWEB)

    Herrmann, Marion, E-mail: marion.herrmann@tu-dresden.de; Lippmann, Wolfgang; Hurtado, Antonio

    2013-11-15

    Silicon carbide is recommended for applications in energy technology due to its good high-temperature corrosion resistance, mechanical durability, and abrasion resistance. The prerequisite for use is often the availability of suitable technologies for joining or sealing the components. A laser-induced process using fillers and local heating of the components represents a possible low-cost option. Investigations in which yttrium aluminosilicate glass was used for laser-induced brazing of SiC components of varying geometry are presented. A four-point bending strength of 112 MPa was found for these joints. In burst tests, laser-joined components were found to withstand internal pressures of up to 54 MPa. Helium leak tests yielded leak rates of less than 10{sup –8} mbar l s{sup −1}, even after 300 h at 900 °C. In contrast, the assemblies showed an increased leak rate after annealing at 1050 °C. The short process time of the laser technique – in the range of a few seconds to a few minutes – results in high temperature gradients and transients. SEM analysis showed that the filler in the seam predominantly solidifies in a glassy state. Crystallization occurred during later thermal loading of the joined components, with chemical equilibrium being established. Differences in seam structures yielded from different cooling rates in the laser process could not be equalized by annealing. The results demonstrated the long-term stability of laser-brazed SiC assemblies to temperatures in the range of glass transformation (900 °C) of the yttrium aluminosilicate filler. In technological investigations, the suitability of the laser joining technique for sealing of SiC components with a geometry approximating that of a fuel element sleeve pin (pin) in a gas-cooled fast reactor was proven.

  16. Biosynthesis of amidated joining peptide from pro-adrenocorticotropin-endorphin

    Energy Technology Data Exchange (ETDEWEB)

    Cullen, E.I.; Mains, R.E. (Johns Hopkins Univ. School of Medicine, Baltimore, MD (USA))

    1987-09-01

    Joining peptide is the major alpha-amidated product of pro-ACTH/endorphin (PAE) in AtT-20 corticotropic tumor cells. To study intracellular joining peptide synthesis, affinity purified antibodies directed against gamma-MSH, joining peptide, and ACTH were used to immunoprecipitate extracts from biosynthetically labeled AtT-20 cells. Immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by tryptic peptide mapping on HPLC. In steady labeling experiments, radioactivity in amidated joining peptide (JP) increased roughly linearly with time, in the manner of a final product, whereas radioactivity associated with PAE (1-94)NH2 reached a constant value after 2-4 h, indicating that PAE(1-94)NH2 is an intermediate in the biosynthesis of JP. Radioactivity appeared in ACTH(1-39) well before JP, consistent with a cleavage order in which ACTH is cleaved from PAE(1-95) before JP sequences are cleaved from PAE(1-74). This conclusion was supported by tryptic peptide analyses of immunoprecipitates, which indicated that less than 5% of JP-related material is cleaved from PAE(1-74) before being cleaved from ACTH-related sequences. After a pulse label, radioactivity in PAE(1-94)NH2 reached a peak value after 1 h of chase and declined with a half-life of less than 1 h. Amidated JP increased to a constant level after 2 h of chase. Enough radiolabeled PAE(1-94)NH2 was detected to account for about half of the radioactivity found in amidated JP, indicating that about half of JP-related material is first cleaved from PAE(1-95) before being amidated. This result was corroborated using HPLC purification to determine both amidated and glycine-extended forms of JP.

  17. Biosynthesis of amidated joining peptide from pro-adrenocorticotropin-endorphin

    International Nuclear Information System (INIS)

    Cullen, E.I.; Mains, R.E.

    1987-01-01

    Joining peptide is the major alpha-amidated product of pro-ACTH/endorphin (PAE) in AtT-20 corticotropic tumor cells. To study intracellular joining peptide synthesis, affinity purified antibodies directed against gamma-MSH, joining peptide, and ACTH were used to immunoprecipitate extracts from biosynthetically labeled AtT-20 cells. Immunoprecipitates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by tryptic peptide mapping on HPLC. In steady labeling experiments, radioactivity in amidated joining peptide (JP) increased roughly linearly with time, in the manner of a final product, whereas radioactivity associated with PAE (1-94)NH2 reached a constant value after 2-4 h, indicating that PAE(1-94)NH2 is an intermediate in the biosynthesis of JP. Radioactivity appeared in ACTH(1-39) well before JP, consistent with a cleavage order in which ACTH is cleaved from PAE(1-95) before JP sequences are cleaved from PAE(1-74). This conclusion was supported by tryptic peptide analyses of immunoprecipitates, which indicated that less than 5% of JP-related material is cleaved from PAE(1-74) before being cleaved from ACTH-related sequences. After a pulse label, radioactivity in PAE(1-94)NH2 reached a peak value after 1 h of chase and declined with a half-life of less than 1 h. Amidated JP increased to a constant level after 2 h of chase. Enough radiolabeled PAE(1-94)NH2 was detected to account for about half of the radioactivity found in amidated JP, indicating that about half of JP-related material is first cleaved from PAE(1-95) before being amidated. This result was corroborated using HPLC purification to determine both amidated and glycine-extended forms of JP

  18. In-Network Processing of an Iceberg Join Query in Wireless Sensor Networks Based on 2-Way Fragment Semijoins

    Directory of Open Access Journals (Sweden)

    Hyunchul Kang

    2015-03-01

    Full Text Available We investigate the in-network processing of an iceberg join query in wireless sensor networks (WSNs. An iceberg join is a special type of join where only those joined tuples whose cardinality exceeds a certain threshold (called iceberg threshold are qualified for the result. Processing such a join involves the value matching for the join predicate as well as the checking of the cardinality constraint for the iceberg threshold. In the previous scheme, the value matching is carried out as the main task for filtering non-joinable tuples while the iceberg threshold is treated as an additional constraint. We take an alternative approach, meeting the cardinality constraint first and matching values next. In this approach, with a logical fragmentation of the join operand relations on the aggregate counts of the joining attribute values, the optimal sequence of 2-way fragment semijoins is generated, where each fragment semijoin employs a Bloom filter as a synopsis of the joining attribute values. This sequence filters non-joinable tuples in an energy-efficient way in WSNs. Through implementation and a set of detailed experiments, we show that our alternative approach considerably outperforms the previous one.

  19. In-Network Processing of an Iceberg Join Query in Wireless Sensor Networks Based on 2-Way Fragment Semijoins

    Science.gov (United States)

    Kang, Hyunchul

    2015-01-01

    We investigate the in-network processing of an iceberg join query in wireless sensor networks (WSNs). An iceberg join is a special type of join where only those joined tuples whose cardinality exceeds a certain threshold (called iceberg threshold) are qualified for the result. Processing such a join involves the value matching for the join predicate as well as the checking of the cardinality constraint for the iceberg threshold. In the previous scheme, the value matching is carried out as the main task for filtering non-joinable tuples while the iceberg threshold is treated as an additional constraint. We take an alternative approach, meeting the cardinality constraint first and matching values next. In this approach, with a logical fragmentation of the join operand relations on the aggregate counts of the joining attribute values, the optimal sequence of 2-way fragment semijoins is generated, where each fragment semijoin employs a Bloom filter as a synopsis of the joining attribute values. This sequence filters non-joinable tuples in an energy-efficient way in WSNs. Through implementation and a set of detailed experiments, we show that our alternative approach considerably outperforms the previous one. PMID:25774710

  20. Xeroderma pigmentosum, complementation group D expression in H1299 lung cancer cells following benzo[a]pyrene exposure as well as in head and neck cancer patients.

    Science.gov (United States)

    Lin, Chang-Shen; Chiou, Wen-Yen; Lee, Ka-Wo; Chen, Tzu-Fen; Lin, Yuan-Jen; Huang, Jau-Ling

    2016-01-01

    DNA repair genes play critical roles in response to carcinogen-induced and anticancer therapy-induced DNA damage. Benzo[a]pyrene (BaP), the most carcinogenic polycyclic aromatic hydrocarbon (PAH), is classified as a group 1 carcinogen by International Agency for Research on Cancer. The aims of this study were to (1) evaluate the effects of BaP on DNA repair activity and expression of DNA repair genes in vitro and (2) examine the role of xeroderma pigmentosum, complementation group D (XPD) mRNA expression in human head and neck cancers. Host cell reactivation assay showed that BaP inhibited nucleotide excision repair in H1299 lung cancer cells. DNA repair through the non-homologous end-joining pathway was not affected by BaP. Real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) and Western blot demonstrated that XPD was downregulated by BaP treatment. BaP exposure did not apparently affect expression of another 11 DNA repair genes. BaP treatment increased the DNA damage marker γ-H2AX and ultraviolet (UV) sensitivity, supporting an impairment of DNA repair in BaP-treated cells. XPD expression was also examined by quantitative RT-PCR in 68 head and neck cancers, and a lower XPD mRNA level was found in smokers' cancer specimens. Importantly, reduced XPD expression was correlated with patient 5-year overall survival rate (35 vs. 56%) and was an independent prognostic factor (hazard ratio: 2.27). Data demonstrated that XPD downregulation was correlated with BaP exposure and human head and neck cancer survival.

  1. Trend and Development of Semisolid Metal Joining Processing

    Directory of Open Access Journals (Sweden)

    M. N. Mohammed

    2015-01-01

    Full Text Available The semisolid metal joining (SSMJ process or thixojoining process has recently been developed based on the principles of SSM processing, which is a technology that involves the formation of metal alloys between solidus and liquidus temperatures. Thixojoining has many potential benefits, which has encouraged researchers to carry out feasibility studies on various materials that could be utilized in this process and which could transform the production of metal components. This paper reviews the findings in the literature to date in this evolving field, specifically, the experimental details, technology considerations for industrialization, and advantages and disadvantages of the various types of SSMJ methods that have been proposed. It also presents details of the range of materials that have been joined by using the SSMJ process. Furthermore, it highlights the huge potential of this process and future directions for further research.

  2. Joining dissimilar materials using Friction Stir scribe technique

    Energy Technology Data Exchange (ETDEWEB)

    Upadhyay, Piyush; Hovanski, Yuri; Jana, Saumyadeep; Fifield, Leonard S.

    2016-10-03

    The ability to effectively join materials with vastly different melting points like Aluminum-Steel, Polymer composites - metals has been one of the road blocks in realizing multi-material components for light weighting efforts. Friction stir scribe (FSS) technique is a promising method that produces continuous overlap joint between materials with vastly different melting regimes and high temperature flow characteristics. FSS uses an offset cutting tool at the tip of the FSW pin to create an insitu mechanical interlock between material interfaces. With investments from Vehicle Technology office, US DOE and several automotive manufacturers and suppliers PNNL is developing the FSS process and has demonstrated viability of joining several material combinations. Details of welding trails, unique challenges and mitigation strategies in different material combinations will be discussed. Joint characterization including mechanical tests and joint performances will also be presented.

  3. Bad Luck When Joining the Shortest Queue

    NARCIS (Netherlands)

    Blanc, J.P.C.

    2008-01-01

    A frequent observation in service systems with queues in parallel is that customers in other queues tend to be served faster than those in one’s own queue. This paper quantifies the probability that one’s service would have started earlier if one had joined another queue than the queue that was

  4. On a possible empirical meaning of meets and joins for quantum propositions

    Science.gov (United States)

    Herbut, Fedor

    1984-09-01

    Following up an idea of Jauch, an empirical basis is worked out for the meets (and joins) of quantum propositions through approximate measurement to an arbitrary degree of accuracy of special yes-no effects constructed from chains of yes-no measurements. This is done with a view to interpreting the meets (and joins) as Dedekind's cuts in analogy with irrationals.

  5. On a possible empirical meaning of meets and joins for quantum propositions

    International Nuclear Information System (INIS)

    Herbut, F.

    1984-01-01

    Following up an idea of Jauch, an empirical basis is worked out for the meets (and joins) of quantum propositions through approximate measurement to an arbitrary degree of accuracy of special yes-no effects constructed from chains of yes-no measurements. This is done with a view to interpreting the meets (and joins) as Dedekind's cuts in analogy with irrationals. (orig.)

  6. A New Approach to Joining Dissimilar Ceramic Oxides for Chemical Sensors

    International Nuclear Information System (INIS)

    Zhuiykov, Serge

    2009-01-01

    Conventional joining of dissimilar oxides for sensing electrodes (SE) of chemical sensors has been pivotal to the development of various sensors and is vital to their further development. However, it is shown that the uncertainty (of a fundamental nature) in the properties of dissimilar oxides in SE causes the determination of their sensing characteristics to be ambiguous. Characteristics are different for such controlled parameters as pyrolysis temperature, crystal structure, particle's morphology and size, chemical and phase composition, the coefficient of thermal expansion (CTE), surface architecture, the bulk and surface stoichiometry and type and conductivity of additives. Here, we provide an alternative approach for joining dissimilar metal-oxides for chemical sensors SE. The approach relies on the development of at least one transient liquid oxide phase on the ceramic-SE interface. These results constitute key points relevant to selection oxides for joining, sintering temperatures and heating/cooling temperature rates.

  7. References and arrow notation instead of join operation in query languages

    Directory of Open Access Journals (Sweden)

    Alexandr Savinov

    2012-10-01

    Full Text Available We study properties of the join operation in query languages and describe some of its major drawbacks. We provide strong arguments against using joins as a main construct for retrieving related data elements in general purpose query languages and argue for using references instead. Since conventional references are quite restrictive when applied to data modeling and query languages, we propose to use generalized references as they are defined in the concept-oriented model (COM. These references are used by two new operations, called projection and de-projection, which are denoted by right and left arrows and therefore this access method is referred to as arrow notation. We demonstrate advantages of the arrow notation in comparison to joins and argue that it makes queries simpler, more natural, easier to understand, and the whole query writing process more productive and less error-prone.

  8. The Swedish Automotive Cluster : How will it be affected by foreign acquisitions at the end of the value chain?

    OpenAIRE

    Sundqvist, Helena

    2011-01-01

    In this master thesis it is analyzed how the Swedish automotive cluster will be affected by foreign acquisitions at the end of the value chain. This is a relevant topic as the Swedish automotive industry is a very important industry in Sweden. It is of interest to evaluate how the recent shifts of ownership for the Swedish automotive manufacturers will affect the Swedish automotive cluster as a whole. It is also relevant from a theoretical point of view and this thesis draws on the extensive ...

  9. Efficient algorithms for factorization and join of blades

    NARCIS (Netherlands)

    Fontijne, D.; Dorst, L.; Bayro-Corrochano, E.; Scheuermann, G.

    2010-01-01

    Subspaces are powerful tools for modeling geometry. In geometric algebra, they are represented using blades and constructed using the outer product. Producing the actual geometrical intersection (meet) and union (join) of subspaces, rather than the simplified linearizations often used in

  10. Efficient algorithms for factorization and join of blades

    NARCIS (Netherlands)

    Fontijne, D.

    2008-01-01

    Subspaces are powerful tools for modeling geometry. In geometric algebra, they are represented using blades and constructed using the outer product. To produce the actual geometrical intersection (Meet) and union (Join) of subspaces, rather than the simplified linearizations often used in

  11. Women's Heart Disease: Join the Heart Truth Community

    Science.gov (United States)

    ... this page please turn JavaScript on. Feature: Women's Heart Disease Join The Heart Truth Community Past Issues / Winter 2014 Table of Contents National Symbol The centerpiece of The Heart Truth ® is The Red Dress ® which was introduced ...

  12. Functional intersection of ATM and DNA-dependent protein kinase catalytic subunit in coding end joining during V(D)J recombination

    DEFF Research Database (Denmark)

    Lee, Baeck-Seung; Gapud, Eric J; Zhang, Shichuan

    2013-01-01

    V(D)J recombination is initiated by the RAG endonuclease, which introduces DNA double-strand breaks (DSBs) at the border between two recombining gene segments, generating two hairpin-sealed coding ends and two blunt signal ends. ATM and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) ar......V(D)J recombination is initiated by the RAG endonuclease, which introduces DNA double-strand breaks (DSBs) at the border between two recombining gene segments, generating two hairpin-sealed coding ends and two blunt signal ends. ATM and DNA-dependent protein kinase catalytic subunit (DNA......-PKcs) are serine-threonine kinases that orchestrate the cellular responses to DNA DSBs. During V(D)J recombination, ATM and DNA-PKcs have unique functions in the repair of coding DNA ends. ATM deficiency leads to instability of postcleavage complexes and the loss of coding ends from these complexes. DNA...... when ATM is present and its kinase activity is intact. The ability of ATM to compensate for DNA-PKcs kinase activity depends on the integrity of three threonines in DNA-PKcs that are phosphorylation targets of ATM, suggesting that ATM can modulate DNA-PKcs activity through direct phosphorylation of DNA...

  13. Joining of Tungsten Armor Using Functional Gradients

    International Nuclear Information System (INIS)

    John Scott O'Dell

    2006-01-01

    The joining of low thermal expansion armor materials such as tungsten to high thermal expansion heat sink materials has been a major problem in plasma facing component (PFC) development. Conventional planar bonding techniques have been unable to withstand the high thermal induced stresses resulting from fabrication and high heat flux testing. During this investigation, innovative functional gradient joints produced using vacuum plasma spray forming techniques have been developed for joining tungsten armor to copper alloy heat sinks. A model was developed to select the optimum gradient architecture. Based on the modeling effort, a 2mm copper rich gradient was selected. Vacuum plasma pray parameters and procedures were then developed to produce the functional gradient joint. Using these techniques, dual cooling channel, medium scale mockups (32mm wide x 400mm length) were produced with vacuum plasma spray formed tungsten armor. The thickness of the tungsten armor was up to 5mm thick. No evidence of debonding at the interface between the heat sink and the vacuum plasma sprayed material was observed.

  14. [Joining WHO of Republic of Korea and the projects in the 1950s].

    Science.gov (United States)

    Lee, Sun Ho

    2014-04-01

    The Republic of Korea(ROK) and the World Health Organization(WHO) have done many projects successfully from 1949, in which the government of First Republic joined the WHO. However the relation between the ROK and the WHO have not been studied very much so far. The main purpose of this research, which could be done by the support of WHO, is connected with three questions. First research point would be "how could the ROK joined WHO in 1949 and what's the meaning of it? And the what's the difference in the process for the WHO between the ROK of 1949 and the DPRK(Democratic People's Republic of Korea) of 1973?" The first president of the ROK, Rhee Syngman, who had received his Ph. D.(about international politics) from Princeton University in 1910, was strongly interested in joining international institutes like UN, WHO. The ROK that could join WHO on 17 August 1949, with the approval of Assembly on 25 May 1949, was one of the founder members of the Western Pacific Region. By joining WHO, the ROK could get chance to increase the level of public health and its administration in 1950's. But the DPRK manage to became a member of WHO on 19 May 1973 and joined the South-East Asia Region. The joining of DPRK was influenced by the easing of the cold war after the Nixon Doctrine and the joining of the China(People's Republic of China). Second research point would be "What kind of roll did the WHO take in the First Republic?" Yet the public health administration of the First Republic that had been made in the period of US army military government was been strongly influenced by USA, the roll of WHO was also important in the 1950's. Last research point would be "What kind of the projects did the ROK and the WHO take part in during the period of he First Republic? How could evaluate the results?" The ROK and the WHO handled the projects including health services, communicable disease prevention and control, control of noncommunicable diseases, and protection of health. Specially

  15. Characterization of Mycobacterium smegmatis PolD2 and PolD1 as RNA/DNA polymerases homologous to the POL domain of bacterial DNA ligase D.

    Science.gov (United States)

    Zhu, Hui; Bhattarai, Hitesh; Yan, Han-Guang; Shuman, Stewart; Glickman, Michael S

    2012-12-21

    Mycobacteria exploit nonhomologous end-joining (NHEJ) to repair DNA double-strand breaks. The core NHEJ machinery comprises the homodimeric DNA end-binding protein Ku and DNA ligase D (LigD), a modular enzyme composed of a C-terminal ATP-dependent ligase domain (LIG), a central 3'-phosphoesterase domain (PE), and an N-terminal polymerase domain (POL). LigD POL is proficient at adding templated and nontemplated deoxynucleotides and ribonucleotides to DNA ends in vitro and is the catalyst in vivo of unfaithful NHEJ events involving nontemplated single-nucleotide additions to blunt DSB ends. Here, we identify two mycobacterial proteins, PolD1 and PolD2, as stand-alone homologues of the LigD POL domain. Biochemical characterization of PolD1 and PolD2 shows that they resemble LigD POL in their monomeric quaternary structures, their ability to add templated and nontemplated nucleotides to primer-templates and blunt ends, and their preference for rNTPs versus dNTPs. Deletion of polD1, polD2, or both from a Mycobacterium smegmatis strain carrying an inactivating mutation in LigD POL failed to reveal a role for PolD1 or PolD2 in templated nucleotide additions during NHEJ of 5'-overhang DSBs or in clastogen resistance. Whereas our results document the existence and characteristics of new stand-alone members of the LigD POL family of RNA/DNA polymerases, they imply that other polymerases can perform fill-in synthesis during mycobacterial NHEJ.

  16. Joining Silicon Carbide Components for Space Propulsion, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — This SBIR Phase I program will identify the joining materials and demonstrate the processes that are suited for construction of advanced ceramic matrix composite...

  17. Contribution to the joining technique of SiC-ceramic using metallic interlayers

    International Nuclear Information System (INIS)

    Gottselig, B.; Gyarmati, E.; Naoumidis, A.; Nickel, H.

    1989-07-01

    For ceramics to be feasible for technical uses suitable joining techniques must be developed that allow reliable ceramic-ceramic and ceramic-metal connections. As yet such procedures exist, based only on empirical studies omitting the reaction behaviour of the joining materials and the specific properties of the reaction products. For this reason the reaction behaviour of silicon carbide with selected metals and subsequently the compatibility of these reaction layers with the ceramic substrate were investigated. The results gained were then applied to silicon carbide joints using intermediate metallic layers. With the reaction phase Ti 3 SiC 2 , found to be the most suitable in basic experiments, joining strengths could be obtained relative tc the mean strength and Weibull modulus comparable to those of the ceramic starting material. (orig.) [de

  18. Joining technology development summary, 189a OH024

    International Nuclear Information System (INIS)

    Edmonds, D.P.

    1978-01-01

    Joining technology is discussed for Breeder Reactor technology including welding development and mechanical properties of weldments for austenitic stainless steels, high-temperature structural design data for austenitic stainless steel weldments and advanced transition joints is discussed. Technology developed in this program is briefly reviewed

  19. Nonhomologous Synapsis and Reduced Crossing over in a Heterozygous Paracentric Inversion in Saccharomyces Cerevisiae

    Science.gov (United States)

    Dresser, M. E.; Ewing, D. J.; Harwell, S. N.; Coody, D.; Conrad, M. N.

    1994-01-01

    Homologous chromosome synapsis (``homosynapsis'') and crossing over are well-conserved aspects of meiotic chromosome behavior. The long-standing assumption that these two processes are causally related has been challenged recently by observations in Saccharomyces cerevisiae of significant levels of crossing over (1) between small sequences at nonhomologous locations and (2) in mutants where synapsis is abnormal or absent. In order to avoid problems of local sequence effects and of mutation pleiotropy, we have perturbed synapsis by making a set of isogenic strains that are heterozygous and homozygous for a large chromosomal paracentric inversion covering a well marked genetic interval and then measured recombination. We find that reciprocal recombination in the marked interval in heterozygotes is reduced variably across the interval, on average to ~55% of that in the homozygotes, and that positive interference still modulates crossing over. Cytologically, stable synapsis across the interval is apparently heterologous rather than homologous, consistent with the interpretation that stable homosynapsis is required to initiate or consummate a large fraction of the crossing over observed in wild-type strains. When crossing over does occur in heterozygotes, dicentric and acentric chromosomes are formed and can be visualized and quantitated on blots though not demonstrated in viable spores. We find that there is no loss of dicentric chromosomes during the two meiotic divisions and that the acentric chromosome is recovered at only 1/3 to 1/2 of the expected level. PMID:7851761

  20. COMPETITIVENESS ОF CHINESE ECONOMY AFTER JOINING WORLD TRADE ORGANIZATION

    Directory of Open Access Journals (Sweden)

    V. S. Shlik

    2008-01-01

    Full Text Available The paper contains analysis of the China’s membership in the World Trade Organization (WTO. The advantages and disadvantages of WTO joining and also prospects and unforeseen difficulties are given in the paper. The People’s Republic of China has already been a full-member of WTO for 6 years. The Chinese Government has mainly fulfilled all the conditions which were imposed for WTO joining. They concerned customs legislation that presupposed a considerable reduction of custom duties, an access to the service market for foreign investors, an increase of responsibility for violation of intellectual property rights. Many of these conditions have been fulfilled in advance.

  1. Joining method for pressure tube and martensitic stainless steel tube

    International Nuclear Information System (INIS)

    Kimoto, Hiroshi; Koike, Hiromitsu.

    1993-01-01

    In a joining portion of zirconium alloy and a stainless steel, the surface of martensitic stainless steel being in contact with Zr and Zr alloy is applied with a laser quenching solidification treatment before expanding joining of them to improve the surface. This can provide the surface with refined coagulated cell tissues and make deposits and impurities homogeneous and solubilized. As a result, the surface of the martensitic stainless steel has highly corrosion resistance, to suppress contact corrosion with Zr and Zr alloy. Accordingly, even if it is exposed to high temperature water of 200 to 350degC, failures of Zr and Zr alloy can be suppressed. (T.M.)

  2. Research and Development Opportunities for Joining Technologies in HVAC&R

    Energy Technology Data Exchange (ETDEWEB)

    Goetzler, William [Navigant Consulting, Burlington, MA (United States); Guernsey, Matt [Navigant Consulting, Burlington, MA (United States); Young, Jim [Navigant Consulting, Burlington, MA (United States)

    2015-10-01

    The Building Technologies Office (BTO) works with researchers and industry partners to develop and deploy technologies that can substantially reduce energy consumption and greenhouse gas (GHG) emissions in residential and commercial buildings. This opportunity assessment aims to advance BTO’s energy savings, GHG reduction, and other program goals by identifying research and development (R&D) initiatives for joining technologies in heating, ventilation, air-conditioning, and refrigeration (HVAC&R) systems. Improving joining technologies for HVAC&R equipment has the potential to increase lifetime equipment operating efficiency, decrease equipment and project cost, and most importantly reduce hydroflourocarbon (HFC) refrigerant leakage to support HFC phasedown and GHG reduction goals.

  3. Status of Joining Thin Sheet and Thin Wall Tubes of 14YWT

    Energy Technology Data Exchange (ETDEWEB)

    Hoelzer, David T. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Unocic, Kinga A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Tang, Wei [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Feng, Zhili [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2015-09-18

    Beginning this fiscal year, the FCRD research project initiated an investigation on joining thin sections of the advanced ODS 14YWT ferritic alloy. Friction stir welding (FSW) was investigated as a method to join thin plate and tubing of 14YWT since it is a solid state joining method that has been shown in past studies to be a promising method for joining plates of ODS alloys, such as 14YWT. However, this study will attempt to be the first to demonstrate if FSW can successfully join thin plates and thin wall tubing of 14YWT. In the first FSW attempt, a 1.06 cm thick plate of 14YWT (SM13 heat) was successfully rolled at 1000ºC to the target thickness of 0.1 cm with no edge cracking. This achievement is a highlight since previous attempts to roll 14YWT plates have resulted in extensive cracking. For the FSW run, a pin tool being developed by the ORNL FSW Process Development effort was used. The first FSW run successfully produced a bead-on-plate weld in the 0.1 cm thick plate. The quality of the weld zone appears very good with no evidence of large defects such as cavities. The microstructural characterization study of the bead-on-plate weld zone has been initiated to compare the results of the microstructure analysis with those obtained in the reference microstructural analysis of the 14YWT (SM13 heat) that showed ultra-fine grain size of 0.43 μm and a high number density of ~2-5 nm sizes oxygen-enriched nanoclusters.

  4. Alternative electro-chemically based processing routes for joining of plasma facing components

    Energy Technology Data Exchange (ETDEWEB)

    Krauss, Wolfgang, E-mail: wolfgang.krauss@kit.edu [Karlsruhe Institute of Technology (KIT), Institute for Materials Research III, Hermann-von-Helmholtz-Platz 1 76344 Eggenstein-Leopoldshafen (Germany); Lorenz, Julia; Holstein, Nils; Konys, Juergen [Karlsruhe Institute of Technology (KIT), Institute for Materials Research III, Hermann-von-Helmholtz-Platz 1 76344 Eggenstein-Leopoldshafen (Germany)

    2011-10-15

    Tungsten is considered in fusion technology as functional and structural material in the area of blanket and divertor for future application in DEMO. The KIT design of a He-cooled divertor includes joints between W and W-alloys as well as of W with Eurofer-steel. The main challenges range from expansion mismatch problem for tungsten/steel joints over metallurgical reactions with brittle phase formation to crack stopping ability and excellent surface wetting. These requirements were only met partly and insufficiently in the past e.g. by direct Cu-casting of tungsten onto steel. Both, the joining needs and the observed failure scenarios of conventionally joined components initiated the development of improved joining technologies based on electro-chemical processing routes. As electrolytes aqueous and aprotic, water free, system are integrated into this development line. In the first step principle requirements are presented to guarantee a reproducible and adherent deposition of scales based on Ni and Cu acting as inter layers and filler, respectively, to generate a real metallurgical bonding as demonstrate by 1100 deg. C joining tests. The development field aprotic systems based on ionic liquids is discussed with respect to enable development of refractory metal based fillers with focus high temperature W-W brazing.

  5. A review of the US joining technologies for plasma facing components in the ITER fusion reactor

    International Nuclear Information System (INIS)

    Odegard, B.C. Jr.; Cadden, C.H.; Watson, R.D.; Slattery, K.T.

    1998-02-01

    This paper is a review of the current joining technologies for plasma facing components in the US for the International Thermonuclear Experimental Reactor (ITER) project. Many facilities are involved in this project. Many unique and innovative joining techniques are being considered in the quest to join two candidate armor plate materials (beryllium and tungsten) to a copper base alloy heat sink (CuNiBe, OD copper, CuCrZr). These techniques include brazing and diffusion bonding, compliant layers at the bond interface, and the use of diffusion barrier coatings and diffusion enhancing coatings at the bond interfaces. The development and status of these joining techniques will be detailed in this report

  6. Load Balancing Issues with Constructing Phylogenetic Trees using Neighbour-Joining Algorithm

    International Nuclear Information System (INIS)

    Al Mamun, S M

    2012-01-01

    Phylogenetic tree construction is one of the most important and interesting problems in bioinformatics. Constructing an efficient phylogenetic tree has always been a research issue. It needs to consider both the correctness and the speed of the tree construction. In this paper, we implemented the neighbour-joining algorithm, using Message Passing Interface (MPI) for constructing the phylogenetic tree. Performance is efficacious, comparing to the best sequential algorithm. From this paper, it would be clear to the researchers that how load balance can make a great effect for constructing phylogenetic trees using neighbour-joining algorithm.

  7. Coordination of Rad1-Rad10 interactions with Msh2-Msh3, Saw1 and RPA is essential for functional 3' non-homologous tail removal.

    Science.gov (United States)

    Eichmiller, Robin; Medina-Rivera, Melisa; DeSanto, Rachel; Minca, Eugen; Kim, Christopher; Holland, Cory; Seol, Ja-Hwan; Schmit, Megan; Oramus, Diane; Smith, Jessica; Gallardo, Ignacio F; Finkelstein, Ilya J; Lee, Sang Eun; Surtees, Jennifer A

    2018-04-06

    Double strand DNA break repair (DSBR) comprises multiple pathways. A subset of DSBR pathways, including single strand annealing, involve intermediates with 3' non-homologous tails that must be removed to complete repair. In Saccharomyces cerevisiae, Rad1-Rad10 is the structure-specific endonuclease that cleaves the tails in 3' non-homologous tail removal (3' NHTR). Rad1-Rad10 is also an essential component of the nucleotide excision repair (NER) pathway. In both cases, Rad1-Rad10 requires protein partners for recruitment to the relevant DNA intermediate. Msh2-Msh3 and Saw1 recruit Rad1-Rad10 in 3' NHTR; Rad14 recruits Rad1-Rad10 in NER. We created two rad1 separation-of-function alleles, rad1R203A,K205A and rad1R218A; both are defective in 3' NHTR but functional in NER. In vitro, rad1R203A,K205A was impaired at multiple steps in 3' NHTR. The rad1R218A in vivo phenotype resembles that of msh2- or msh3-deleted cells; recruitment of rad1R218A-Rad10 to recombination intermediates is defective. Interactions among rad1R218A-Rad10 and Msh2-Msh3 and Saw1 are altered and rad1R218A-Rad10 interactions with RPA are compromised. We propose a model in which Rad1-Rad10 is recruited and positioned at the recombination intermediate through interactions, between Saw1 and DNA, Rad1-Rad10 and Msh2-Msh3, Saw1 and Msh2-Msh3 and Rad1-Rad10 and RPA. When any of these interactions is altered, 3' NHTR is impaired.

  8. People's Republic of China joins ITER

    International Nuclear Information System (INIS)

    Huo Yuping

    2003-01-01

    The People's Republic of China is the largest developing country with a projected population of 1.6 - 2 billion people and an energy consumption growing from the current 1.3 Billion Tons Coal Equivalent (TCE) to more than 4 Billion TCE by 2050. This large demand needs to be accommodated in a sustainable way, requiring energy generation in an environmentally friendly way. Fusion is one of the most promising candidates to solve this important issue. This explains why in the second half of 2002, the ITER Participants' delegations to the ITER Negotiations received expression of interest from the People's Republic of China in the possibility of Chinese participation in ITER, including joining the ongoing Negotiations. The speed with which the Chinese authorities had made their decision to participate in the ITER Negotiations was impressive. The Prime Minister and the State Council had already confirmed their decision to apply to join ITER as soon as possible, and Mr. Xu Guanhua, Chinese Minister of Science and Technology, wrote on behalf of his government, on 10 January 2003, to the four heads of delegation in the ITER Negotiations, requesting that China participate in the present ITER Negotiations, pointing out that China intends to provide a substantial contribution to the Project, comparable to what is currently envisaged by some of the participants in the present Negotiations

  9. Method of and apparatus for use in joining tubular components and tube assemblies made thereby

    International Nuclear Information System (INIS)

    Percival, S.R.

    1979-01-01

    A method of joining difficult to weld materials involves the forming of a rolled joint. A particular application is the joining of zirconium alloy calandria tubes to stainless steel tube-plates in a SGHWR. (UK)

  10. Advanced fusion welding processes, solid state joining and a successful marriage. [production of aerospace structures

    Science.gov (United States)

    Miller, F. R.

    1972-01-01

    Joining processes for aerospace systems combine fusion welding and solid state joining during production of metal structures. Detailed characteristics of electron beam welding, plasma arc welding, diffusion welding, inertia welding and weldbond processes are discussed.

  11. Governing Bodies That Consider Joining or Creating Federations

    Science.gov (United States)

    Howarth, Eleanor

    2015-01-01

    This article reports the findings of a research study exploring what motivates governing bodies to consider joining or creating a federation, their experiences of the process and the barriers faced. The research methodology involved semi-structured telephone interviews, face-to-face interviews and analysis of relevant documentation. The varied…

  12. Joining Forces: The Case of Alliant International University

    Science.gov (United States)

    Leslie, Heather

    2013-01-01

    Mergers and acquisitions are a prevalent force in higher education as more colleges and universities are joining forces to expand resources, enhance missions, or prevent closures. This study examines the merger of Alliant University (formerly California School of Professional Psychology) with United States International University to create what…

  13. 46 CFR 56.75-30 - Pipe joining details.

    Science.gov (United States)

    2010-10-01

    ... COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE ENGINEERING PIPING SYSTEMS AND... cut or machined square and the edges shall be held closely together to insure a satisfactory joint. (b... satisfactory for the brazing alloy to be employed, method of heating, and material to be joined. The annular...

  14. The development of joining doped graphite to copper for first wall application in HT-7 tokamak

    Energy Technology Data Exchange (ETDEWEB)

    Zhou Zhangjian, E-mail: zhouzhj@mater.ustb.edu.cn [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Zhong Zhihong [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China); Chen Junling [Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, Anhui 230031 (China); Ge Changchun [School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083 (China)

    2010-12-15

    Two joining methods have been developed for joining carbon based plasma facing material to copper based heat sink material for the potential application in HT-7 and EAST tokamak. The first joining method is based on brazing technique by using a rapidly solidified foil-type Ti-Zr based amorphous filler with a melting temperature of 850 deg. C. The other joining method is direct active metal casting-casting the premixed powders of copper and active transition metals on the mechanical machined carbon surface directly. SEM observations demonstrate high quality of joining surface for both joints. The brazing technique is more promising for fabrication joint with larger size compared with the direct active alloy casting method. High heat flux test using an e-beam device was performed on the actively cooled C/Cu joint fabricated by brazing method. There has no damage occurred on the joint after heat loading at 6 MW/m{sup 2}.

  15. International Symposium on Interfacial Joining and Surface Technology (IJST2013)

    International Nuclear Information System (INIS)

    Takahashi, Yasuo

    2014-01-01

    Interfacial joining (bonding) is a widely accepted welding process and one of the environmentally benign technologies used in industrial production. As the bonding temperature is lower than the melting point of the parent materials, melting of the latter is kept to a minimum. The process can be based on diffusion bonding, pressure welding, friction welding, ultrasonic bonding, or brazing-soldering, all of which offer many advantages over fusion welding. In addition, surface technologies such as surface modification, spraying, coating, plating, and thin-film formation are necessary for advanced manufacturing, fabrication, and electronics packaging. Together, interfacial joining and surface technology (IJST) will continue to be used in various industrial fields because IJST is a very significant form of environmentally conscious materials processing. The international symposium of IJST 2013 was held at Icho Kaikan, Osaka University, Japan from 27–29 November, 2013. A total of 138 participants came from around the world to attend 56 oral presentations and 36 posters presented at the symposium, and to discuss the latest research and developments on interfacial joining and surface technologies. This symposium was also held to commemorate the 30th anniversary of the Technical Commission on Interfacial Joining of the Japan Welding Society. On behalf of the chair of the symposium, it is my great pleasure to present this volume of IOP Conference Series: Materials Science and Engineering (MSE). Among the presentations, 43 papers are published here, and I believe all of the papers have provided the welding community with much useful information. I would like to thank the authors for their enthusiastic and excellent contributions. Finally, I would like to thank all members of the committees, secretariats, participants, and everyone who contributed to this symposium through their support and invaluable effort for the success of IJST 2013. Yasuo Takahashi Chair of IJST 2013

  16. International Symposium on Interfacial Joining and Surface Technology (IJST2013)

    Science.gov (United States)

    Takahashi, Yasuo

    2014-08-01

    Interfacial joining (bonding) is a widely accepted welding process and one of the environmentally benign technologies used in industrial production. As the bonding temperature is lower than the melting point of the parent materials, melting of the latter is kept to a minimum. The process can be based on diffusion bonding, pressure welding, friction welding, ultrasonic bonding, or brazing-soldering, all of which offer many advantages over fusion welding. In addition, surface technologies such as surface modification, spraying, coating, plating, and thin-film formation are necessary for advanced manufacturing, fabrication, and electronics packaging. Together, interfacial joining and surface technology (IJST) will continue to be used in various industrial fields because IJST is a very significant form of environmentally conscious materials processing. The international symposium of IJST 2013 was held at Icho Kaikan, Osaka University, Japan from 27-29 November, 2013. A total of 138 participants came from around the world to attend 56 oral presentations and 36 posters presented at the symposium, and to discuss the latest research and developments on interfacial joining and surface technologies. This symposium was also held to commemorate the 30th anniversary of the Technical Commission on Interfacial Joining of the Japan Welding Society. On behalf of the chair of the symposium, it is my great pleasure to present this volume of IOP Conference Series: Materials Science and Engineering (MSE). Among the presentations, 43 papers are published here, and I believe all of the papers have provided the welding community with much useful information. I would like to thank the authors for their enthusiastic and excellent contributions. Finally, I would like to thank all members of the committees, secretariats, participants, and everyone who contributed to this symposium through their support and invaluable effort for the success of IJST 2013. Yasuo Takahashi Chair of IJST 2013

  17. Studying DNA Looping by Single-Molecule FRET

    OpenAIRE

    Le, Tung T.; Kim, Harold D.

    2014-01-01

    Bending of double-stranded DNA (dsDNA) is associated with many important biological processes such as DNA-protein recognition and DNA packaging into nucleosomes. Thermodynamics of dsDNA bending has been studied by a method called cyclization which relies on DNA ligase to covalently join short sticky ends of a dsDNA. However, ligation efficiency can be affected by many factors that are not related to dsDNA looping such as the DNA structure surrounding the joined sticky ends, and ligase can als...

  18. The Mechanism of Solid State Joining THA with AlMg3Mn Alloy

    Directory of Open Access Journals (Sweden)

    Kaczorowski M.

    2014-06-01

    Full Text Available The results of experimental study of solid state joining of tungsten heavy alloy (THA with AlMg3Mn alloy are presented. The aim of these investigations was to study the mechanism of joining two extremely different materials used for military applications. The continuous rotary friction welding method was used in the experiment. The parameters of friction welding process i.e. friction load and friction time in whole studies were changed in the range 10 to 30kN and 0,5 to 10s respectively while forging load and time were constant and equals 50kN and 5s. The results presented here concerns only a small part whole studies which were described elsewhere. These are focused on the mechanism of joining which can be adhesive or diffusion controlled. The experiment included macro- and microstructure observations which were supplemented with SEM investigations. The goal of the last one was to reveal the character of fracture surface after tensile test and to looking for anticipated diffusion of aluminum into THA matrix. The results showed that joining of THA with AlMg2Mn alloy has mainly adhesive character, although the diffusion cannot be excluded.

  19. Accelerating foreign-key joins using asymmetric memory channels

    NARCIS (Netherlands)

    Pirk, H.; Manegold, S.; Kersten, M.

    2011-01-01

    Indexed Foreign-Key Joins expose a very asymmetric access pattern: the Foreign-Key Index is sequentially scanned whilst the Primary-Key table is target of many quasi-random lookups which is the dominant cost factor. To reduce the costs of the random lookups the fact-table can be (re-) partitioned at

  20. DNA-dependent protein kinase (DAN-PK), a key enzyme in the re-ligation of DNA double-strand breaks

    International Nuclear Information System (INIS)

    Hennequin, C.; Averbeck, D.

    1999-01-01

    Repair pathways of DNA are now defined and some important findings have been discovered in the last few years. DNA non-homologous end-joining (NEH) is a crucial process in the repair of radiation-induced double-strand breaks (DSBs). NHEj implies at least three steps: the DNA free-ends must get closer, preparation of the free-ends by exonucleases and then a transient hybridization in a region of DNA with weak homology. DNA-dependent protein kinase (DNA-PK) is the key enzyme in this process. DNA-PK is a nuclear serine/threonine kinase that comprises three components: a catalytic subunit (DNA-PK cs ) and two regulatory subunits, DNA-binding proteins, Ku80 and Ku70. The severe combined immuno-deficient (scid) mice are deficient in DNA-PK cs : this protein is involved both in DNA repair and in the V(D)J recombination of immunoglobulin and T-cell receptor genes. It is a protein-kinase of the P13-kinase family and which can phosphorylate Ku proteins, p53 and probably some other proteins still unknown. DNA-PK is an important actor of DSBs repair (induced by ionising radiations or by drugs like etoposide), but obviously it is not the only mechanism existing in the cell for this function. Some others, like homologous recombination, seem also to have a great importance for cell survival. (authors)

  1. Development of a double beam process for joining aluminum and steel

    Science.gov (United States)

    Frank, Sascha

    2014-02-01

    Multi-material structures pose an attractive option for overcoming some of the central challenges in lightweight design. An exceptionally high potential for creating cost-effective lightweight solutions is attributed to the combination of steel and aluminum. However, these materials are also particularly difficult to join due to their tendency to form intermetallic compounds (IMCs). The growth of these compounds is facilitated by high temperatures and long process times. Due to their high brittleness, IMCs can severely weaken a joint. Thus, it is only possible to create durable steel-aluminum joints when the formation of IMCs can be limited to a non-critical level. To meet this goal, a new joining method has been designed. The method is based on the combination of a continuous wave (pw) and a pulsed laser (pw) source. Laser beams from both sources are superimposed in a common process zone. This makes it possible to apply the advantages of laser brazing to mixed-metal joints without requiring the use of chemical fluxes. The double beam technology was first tested in bead-on-plate experiments using different filler wire materials. Based on the results of these tests, a process for joining steel and aluminum in a double-flanged configuration is now being developed. The double flanged seams are joined using zinc- or aluminum-based filler wires. Microsections of selected seams show that it is possible to achieve good base material wetting while limiting the growth of IMCs to acceptable measures. In addition, the results of tensile tests show that high joint strengths can be achieved.

  2. Understanding the spark plasma sintering from the view of materials joining

    International Nuclear Information System (INIS)

    Dong, Peng; Wang, Zhe; Wang, Wenxian; Chen, Shaoping; Zhou, Jun

    2016-01-01

    Spark plasma sintering (SPS) is an attractive consolidation process. However, the mechanism behind this process is still an open topic for debate. This paper presents the first attempt to understand the SPS mechanism from perspective of materials joining. For this, TiNi_f/Al composites were fabricated by SPS, and the interfacial microstructures were investigated using field emission scanning electron microscopy and transmission electron microscopy. According to the experimental results, several joining processes were reflected well during SPS, involving micro-arc welding, electric resistance welding and diffusion welding. The proposed understanding of SPS will be helpful to the control of sintering quality.

  3. Loosening and damage mechanism of thread-joined structures in nuclear power equipment

    International Nuclear Information System (INIS)

    Tang Hui

    1999-01-01

    The author proposes a loosening mechanism of thread-joined structures under vibrate environments in the nuclear power equipment and structures, which is on the base of the macro and imperceptible-mechanics analysis. It has answered the problems on the seizing, the adhesive wearing, the generation of cracks, the thread-tooth fracture. So it has a conclusion that the loosening of thread-joined structures is essential trend, in other words, the locking property of thread-pair is failure under vibrate environments

  4. Larval traits carry over to affect post-settlement behaviour in a common coral reef fish.

    Science.gov (United States)

    Dingeldein, Andrea L; White, J Wilson

    2016-07-01

    Most reef fishes begin life as planktonic larvae before settling to the reef, metamorphosing and entering the benthic adult population. Different selective forces determine survival in the planktonic and benthic life stages, but traits established in the larval stage may carry over to affect post-settlement performance. We tested the hypothesis that larval traits affect two key post-settlement fish behaviours: social group-joining and foraging. Certain larval traits of reef fishes are permanently recorded in the rings in their otoliths. In the bluehead wrasse (Thalassoma bifasciatum), prior work has shown that key larval traits recorded in otoliths (growth rate, energetic condition at settlement) carry over to affect post-settlement survival on the reef, with higher-larval-condition fish experiencing less post-settlement mortality. We hypothesized that this selective mortality is mediated by carry-over effects on post-settlement antipredator behaviours. We predicted that better-condition fish would forage less and be more likely to join groups, both behaviours that would reduce predation risk. We collected 550 recently settled bluehead wrasse (Thalassoma bifasciatum) from three reef sites off St. Croix (USVI) and performed two analyses. First, we compared each settler's larval traits to the size of its social group to determine whether larval traits influenced group-joining behaviour. Secondly, we observed foraging behaviour in a subset of grouped and solitary fish (n = 14) for 1-4 days post-settlement. We then collected the fish and tested whether larval traits influenced the proportion of time spent foraging. Body length at settlement, but not condition, affected group-joining behaviour; smaller fish were more likely to remain solitary or in smaller groups. However, both greater length and better condition were associated with greater proportions of time spent foraging over four consecutive days post-settlement. Larval traits carry over to affect post

  5. Intranuclear Delivery of a Novel Antibody-Derived Radiosensitizer Targeting the DNA-Dependent Protein Kinase Catalytic Subunit

    Energy Technology Data Exchange (ETDEWEB)

    Xiong Hairong [Institute of Molecular Medicine and Genetics, Georgia Health Sciences University, Augusta, GA (Georgia); State Key Laboratory of Virology, Institute of Medical Virology, Wuhan University School of Medicine, Wuhan (China); Lee, Robert J. [Division of Pharmaceutics, College of Pharmacy, Ohio State University, Columbus, OH (United States); Haura, Eric B. [Thoracic Oncology and Experimental Therapeutics Programs, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL (United States); Edwards, John G. [Apeliotus Technologies, Inc., Atlanta, GA (United States); Dynan, William S. [Institute of Molecular Medicine and Genetics, Georgia Health Sciences University, Augusta, GA (Georgia); Li Shuyi, E-mail: sli@georgiahealth.edu [Institute of Molecular Medicine and Genetics, Georgia Health Sciences University, Augusta, GA (Georgia); Apeliotus Technologies, Inc., Atlanta, GA (United States)

    2012-07-01

    Purpose: To inhibit DNA double-strand break repair in tumor cells by delivery of a single-chain antibody variable region fragment (ScFv 18-2) to the cell nucleus. ScFv 18-2 binds to a regulatory region of the DNA-dependent protein kinase (DNA-PK), an essential enzyme in the nonhomologous end-joining pathway, and inhibits DNA end-joining in a cell-free system and when microinjected into single cells. Development as a radiosensitizer has been limited by the lack of a method for intranuclear delivery to target cells. We investigated a delivery method based on folate receptor-mediated endocytosis. Methods and Materials: A recombinant ScFv 18-2 derivative was conjugated to folate via a scissile disulfide linker. Folate-ScFv 18-2 was characterized for its ability to be internalized by tumor cells and to influence the behavior of ionizing radiation-induced repair foci. Radiosensitization was measured in a clonogenic survival assay. Survival curves were fitted to a linear-quadratic model, and between-group differences were evaluated by an F test. Sensitization ratios were determined based on mean inhibitory dose. Results: Human KB and NCI-H292 lung cancer cells treated with folate-conjugated ScFv 18-2 showed significant radiosensitization (p < 0.001). Sensitization enhancement ratios were 1.92 {+-} 0.42 for KB cells and 1.63 {+-} 0.13 for NCI-H292 cells. Studies suggest that treatment inhibits repair of radiation-induced DSBs, as evidenced by the persistence of {gamma}-H2AX-stained foci and by inhibition of staining with anti-DNA-PKcs phosphoserine 2056. Conclusions: Folate-mediated endocytosis is an effective method for intranuclear delivery of an antibody-derived DNA repair inhibitor.

  6. Characterization of joining sites of a viral histone H4 on host insect chromosomes.

    Directory of Open Access Journals (Sweden)

    Sunil Kumar

    Full Text Available A viral histone H4 (CpBV-H4 is encoded in a polydnavirus, Cotesia plutellae bracovirus (CpBV. It plays a crucial role in parasitism of an endoparasitoid wasp, C. plutellae, against diamondback moth, Plutella xylostella, by altering host gene expression in an epigenetic mode by its N-terminal tail after joining host nucleosomes. Comparative transcriptomic analysis between parasitized and nonparasitized P. xylostella by RNA-Seq indicated that 1,858 genes were altered at more than two folds in expression levels at late parasitic stage, including 877 up-regulated genes and 981 down-regulated genes. Among parasitic factors altering host gene expression, CpBV-H4 alone explained 16.3% of these expressional changes. To characterize the joining sites of CpBV-H4 on host chromosomes, ChIP-Seq (chromatin immunoprecipitation followed by deep sequencing was applied to chromatins extracted from parasitized larvae. It identified specific 538 ChIP targets. Joining sites were rich (60.2% in AT sequence. Almost 40% of ChIP targets included short nucleotide repeat sequences presumably recognizable by transcriptional factors and chromatin remodeling factors. To further validate these CpBV-H4 targets, CpBV-H4 was transiently expressed in nonparasitized host at late larval stage and subjected to ChIP-Seq. Two kinds of ChIP-Seqs shared 51 core joining sites. Common targets were close (within 1 kb to genes regulated at expression levels by CpBV-H4. However, other host genes not close to CpBV-H4 joining sites were also regulated by CpBV-H4. These results indicate that CpBV-H4 joins specific chromatin regions of P. xylostella and controls about one sixth of the total host genes that were regulated by C. plutellae parasitism in an epigenetic mode.

  7. Targeting Phosphatidylinositol 4-Kinase IIIα for Radiosensitization: A Potential Model of Drug Repositioning Using an Anti-Hepatitis C Viral Agent

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Jeanny [Department of Radiation Oncology, Graduate School of Medicine, Seoul National University, Seoul (Korea, Republic of); Kim, Dan Hyo; Park, Ji Min [Medical Science Research Institute, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of); Park, Young Hee [Department of Radiation Oncology, Graduate School of Medicine, Seoul National University, Seoul (Korea, Republic of); Hwang, Yeo Hyun [Medical Science Research Institute, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of); Wu, Hong-Gyun [Department of Radiation Oncology, Graduate School of Medicine, Seoul National University, Seoul (Korea, Republic of); Institute of Radiation Medicine, Seoul National University, Seoul (Korea, Republic of); Shin, Kyung Hwan [Department of Radiation Oncology, Graduate School of Medicine, Seoul National University, Seoul (Korea, Republic of); Kim, In Ah, E-mail: inah228@snu.ac.kr [Department of Radiation Oncology, Graduate School of Medicine, Seoul National University, Seoul (Korea, Republic of); Medical Science Research Institute, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of); Institute of Radiation Medicine, Seoul National University, Seoul (Korea, Republic of); Cancer Research Institute, Seoul National University, Seoul (Korea, Republic of)

    2016-11-15

    Purpose: To investigate which isotype of phosphatidylinositol 4-kinase (PI4K) may affect radiosensitivity and examine whether anti–hepatitis C viral (HCV) agents, some of which have been shown to inhibit PI4K IIIα activity, could be repositioned as a radiosensitizer in human cancer cells. Methods and Materials: U251, BT474, and HepG2 cell lines and normal human astrocyte were used. Ribonucleic acid interference, clonogenic assays, Western blotting, immunofluorescence, annexin V assay, lysotracker staining, and β-galactosidase assay were performed. Results: Of the 4 PI4K isotypes, specific inhibition of IIIα increased radiosensitivity. For pharmacologic inhibition of PI4K IIIα, we screened 9 anti-HCV agents by half-maximal inhibitory concentration assay. Simeprevir was selected, and its inhibition of PI4K IIIα activity was confirmed. Combination of simeprevir treatment and radiation significantly attenuated expression of phospho-phospho-PKC and phospho-Akt and increased radiation-induced cell death in tested cell lines. Pretreatment with simeprevir prolonged γH2AX foci formation and down-regulation of phospho-DNA-PKcs, indicating impairment of nonhomologous end-joining repair. Cells pretreated with simeprevir exhibited mixed modes of cell death, including apoptosis and autophagy. Conclusion: These data demonstrate that targeting PI4K IIIα using an anti-HCV agent is a viable approach to enhance the therapeutic efficacy of radiation therapy in various human cancers, such as glioma, breast, and hepatocellular carcinoma.

  8. Friction riveting as an alternative mechanical fastening to join engineering plastics

    Science.gov (United States)

    Gagliardi, Francesco; Conte, Romina; Bentrovato, Renato; Simeoli, Giorgio; Russo, Pietro; Ambrogio, Giuseppina

    2018-05-01

    Friction Rivecting is a quite new joining process to connect multi-material structures. In brief, a metallic rivet is dipped rotating inside matrixes, usually made of plastics, increasing its original diameter. The use of high-performance plastics is more suitable being their higher mechanical and thermal properties important to avoid material degradation and to allow strong part connections. High-speed friction welding system has been usually used to perform the process. In the work here proposed, the joints have been achieved by means of a traditional milling machine and the attention has been focused on a widely used engineering plastic, i.e. polyamide 6 (PA6) with and without glass fiber reinforcement. A specific speed multiplier has been attached into the mandrel of the used machine to increase the reachable rotational speed. Moreover, rivets made of Titanium Grade 2 and of an Aluminum Alloy, the AA-6060, are utilized. The influence that the heating and the forging length can have on the quality of the obtained junctions, considering a fixed joining depth, has been tested and investigated. The performed connections have been judged by tensile tests, which were set to quantify the maximum strength of the joints for a transverse speed of 1,0 mm/min. Barreling effect can be observed close to the tip, which loses the initial shape of a cylinder characterized by straight vertical walls. Finally, the possible degradation of the polymer, due to temperature increment, has been also evaluated close to the working zone. According to that, it has to be highlighted that the process needs a heating balance, which is necessary to get sound joints. The compromise has, on one side, to allow the rivet penetration and deformation, and on the other side, to avoid the degradation of the polymer, which would affect its properties and a proper rivet deformation.

  9. Influence of Hub Parameters on Joining Forces and Torque Transmission Output of Plastically-Joined Shaft-Hub-Connections with a Knurled Contact Surface

    Directory of Open Access Journals (Sweden)

    Lukáš Suchý

    2018-04-01

    Full Text Available A knurled interference fit is a machine part connection made by a plastic joining, which includes the advantages of commonly-used shaft-hub-connections. The combination of the friction and form fit, which are responsible for torque transmission, results in a higher power density than conventional connections. In this paper, parameter gaps are bridged with the aim of enhance the design calculation of the knurled interference fit. Experimental investigations on the shaft chamfer angle (100Cr6 and hub-diameter-ratio (AlSi1MgMn were performed. The analytical approaches are developed for calculating the joining force and maximal torque capacity by accounting for experimentally investigated loss of load transmission at high hub-diameter-ratios and high shaft chamfer angles. The presented calculation approach is an accurate tool for the assessment of early machine designs of the knurled interference fit and helps to save from having to perform time-extensive tests.

  10. Joining-Splitting Interaction of Noncritical String

    Science.gov (United States)

    Hadasz, Leszek; Jaskólski, Zbigniew

    The joining-splitting interaction of noncritical bosonic string is analyzed in the light-cone formulation. The Mandelstam method of constructing tree string amplitudes is extended to the bosonic massive string models of the discrete series. The general properties of the Liouville longitudinal excitations which are necessary and sufficient for the Lorentz covariance of the light-cone amplitudes are derived. The results suggest that the covariant and the light-cone approach are equivalent also in the noncritical dimensions. Some aspects of unitarity of interacting noncritical massive string theory are discussed.

  11. ATM loss leads to synthetic lethality in BRCA1 BRCT mutant mice associated with exacerbated defects in homology-directed repair.

    Science.gov (United States)

    Chen, Chun-Chin; Kass, Elizabeth M; Yen, Wei-Feng; Ludwig, Thomas; Moynahan, Mary Ellen; Chaudhuri, Jayanta; Jasin, Maria

    2017-07-18

    BRCA1 is essential for homology-directed repair (HDR) of DNA double-strand breaks in part through antagonism of the nonhomologous end-joining factor 53BP1. The ATM kinase is involved in various aspects of DNA damage signaling and repair, but how ATM participates in HDR and genetically interacts with BRCA1 in this process is unclear. To investigate this question, we used the Brca1 S1598F mouse model carrying a mutation in the BRCA1 C-terminal domain of BRCA1. Whereas ATM loss leads to a mild HDR defect in adult somatic cells, we find that ATM inhibition leads to severely reduced HDR in Brca1 S1598F cells. Consistent with a critical role for ATM in HDR in this background, loss of ATM leads to synthetic lethality of Brca1 S1598F mice. Whereas both ATM and BRCA1 promote end resection, which can be regulated by 53BP1, 53bp1 deletion does not rescue the HDR defects of Atm mutant cells, in contrast to Brca1 mutant cells. These results demonstrate that ATM has a role in HDR independent of the BRCA1-53BP1 antagonism and that its HDR function can become critical in certain contexts.

  12. CCTop: An Intuitive, Flexible and Reliable CRISPR/Cas9 Target Prediction Tool.

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    Manuel Stemmer

    Full Text Available Engineering of the CRISPR/Cas9 system has opened a plethora of new opportunities for site-directed mutagenesis and targeted genome modification. Fundamental to this is a stretch of twenty nucleotides at the 5' end of a guide RNA that provides specificity to the bound Cas9 endonuclease. Since a sequence of twenty nucleotides can occur multiple times in a given genome and some mismatches seem to be accepted by the CRISPR/Cas9 complex, an efficient and reliable in silico selection and evaluation of the targeting site is key prerequisite for the experimental success. Here we present the CRISPR/Cas9 target online predictor (CCTop, http://crispr.cos.uni-heidelberg.de to overcome limitations of already available tools. CCTop provides an intuitive user interface with reasonable default parameters that can easily be tuned by the user. From a given query sequence, CCTop identifies and ranks all candidate sgRNA target sites according to their off-target quality and displays full documentation. CCTop was experimentally validated for gene inactivation, non-homologous end-joining as well as homology directed repair. Thus, CCTop provides the bench biologist with a tool for the rapid and efficient identification of high quality target sites.

  13. Telomeres and genomic damage repair. Their implication in human pathology

    International Nuclear Information System (INIS)

    Perez, Maria del R.; Dubner, Diana; Michelin, Severino; Gisone, Pablo; Carosella, Edgardo D.

    2002-01-01

    Telomeres, functional complexed that protect eukaryotic chromosome ends, participate in the regulation of cell proliferation and could play a role in the stabilization of genomic regions in response to genotoxic stress. Their significance in human pathology becomes evident in several diseases sharing genomic instability as a common trait, in which alterations of the telomere metabolism have been demonstrated. Many of them are also associated with hypersensitivity to ionizing radiation and cancer susceptibility. Besides the specific proteins belonging to the telomeric complex, other proteins involved in the DNA repair machinery, such as ATM, BRCA1, BRCA2, PARP/tankyrase system, DNA-PK and RAD50-MRE11-NBS1 complexes, are closely related with the telomere. This suggests that the telomere sequesters DNA repair proteins for its own structure maintenance, with could also be released toward damaged sites in the genomic DNA. This communication describes essential aspects of telomere structure and function and their links with homologous recombination, non-homologous end-joining (NHEJ), V(D)J system and mismatch-repair (MMR). Several pathological conditions exhibiting alterations in some of these mechanisms are also considered. The cell response to ionizing radiation and its relationship with the telomeric metabolism is particularly taken into account as a model for studying genotoxicity. (author)

  14. Cores, Joins and the Fano-Flow Conjectures

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    Jin Ligang

    2018-02-01

    Full Text Available The Fan-Raspaud Conjecture states that every bridgeless cubic graph has three 1-factors with empty intersection. A weaker one than this conjecture is that every bridgeless cubic graph has two 1-factors and one join with empty intersection. Both of these two conjectures can be related to conjectures on Fano-flows. In this paper, we show that these two conjectures are equivalent to some statements on cores and weak cores of a bridgeless cubic graph. In particular, we prove that the Fan-Raspaud Conjecture is equivalent to a conjecture proposed in [E. Steffen, 1-factor and cycle covers of cubic graphs, J. Graph Theory 78 (2015 195–206]. Furthermore, we disprove a conjecture proposed in [G. Mazzuoccolo, New conjectures on perfect matchings in cubic graphs, Electron. Notes Discrete Math. 40 (2013 235–238] and we propose a new version of it under a stronger connectivity assumption. The weak oddness of a cubic graph G is the minimum number of odd components (i.e., with an odd number of vertices in the complement of a join of G. We obtain an upper bound of weak oddness in terms of weak cores, and thus an upper bound of oddness in terms of cores as a by-product.

  15. Ubiquitin-specific protease 5 is required for the efficient repair of DNA double-strand breaks.

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    Satoshi Nakajima

    Full Text Available During the DNA damage response (DDR, ubiquitination plays an important role in the recruitment and regulation of repair proteins. However, little is known about elimination of the ubiquitination signal after repair is completed. Here we show that the ubiquitin-specific protease 5 (USP5, a deubiquitinating enzyme, is involved in the elimination of the ubiquitin signal from damaged sites and is required for efficient DNA double-strand break (DSB repair. Depletion of USP5 sensitizes cells to DNA damaging agents, produces DSBs, causes delayed disappearance of γH2AX foci after Bleocin treatment, and influences DSB repair efficiency in the homologous recombination pathway but not in the non-homologous end joining pathway. USP5 co-localizes to DSBs induced by laser micro-irradiation in a RAD18-dependent manner. Importantly, polyubiquitin chains at sites of DNA damage remained for longer periods in USP5-depleted cells. Our results show that disassembly of polyubiquitin chains by USP5 at sites of damage is important for efficient DSB repair.

  16. The Impact of Hedgehog Signaling Pathway on DNA Repair Mechanisms in Human Cancer

    International Nuclear Information System (INIS)

    Meng, Erhong; Hanna, Ann; Samant, Rajeev S.; Shevde, Lalita A.

    2015-01-01

    Defined cellular mechanisms have evolved that recognize and repair DNA to protect the integrity of its structure and sequence when encountering assaults from endogenous and exogenous sources. There are five major DNA repair pathways: mismatch repair, nucleotide excision repair, direct repair, base excision repair and DNA double strand break repair (including non-homologous end joining and homologous recombination repair). Aberrant activation of the Hedgehog (Hh) signaling pathway is a feature of many cancer types. The Hh pathway has been documented to be indispensable for epithelial-mesenchymal transition, invasion and metastasis, cancer stemness, and chemoresistance. The functional transcription activators of the Hh pathway include the GLI proteins. Inhibition of the activity of GLI can interfere with almost all DNA repair types in human cancer, indicating that Hh/GLI functions may play an important role in enabling tumor cells to survive lethal types of DNA damage induced by chemotherapy and radiotherapy. Thus, Hh signaling presents an important therapeutic target to overcome DNA repair-enabled multi-drug resistance and consequently increase chemotherapeutic response in the treatment of cancer

  17. Xenopus egg extract: A powerful tool to study genome maintenance mechanisms.

    Science.gov (United States)

    Hoogenboom, Wouter S; Klein Douwel, Daisy; Knipscheer, Puck

    2017-08-15

    DNA repair pathways are crucial to maintain the integrity of our genome and prevent genetic diseases such as cancer. There are many different types of DNA damage and specific DNA repair mechanisms have evolved to deal with these lesions. In addition to these repair pathways there is an extensive signaling network that regulates processes important for repair, such as cell cycle control and transcription. Despite extensive research, DNA damage repair and signaling are not fully understood. In vitro systems such as the Xenopus egg extract system, have played, and still play, an important role in deciphering the molecular details of these processes. Xenopus laevis egg extracts contain all factors required to efficiently perform DNA repair outside a cell, using mechanisms conserved in humans. These extracts have been used to study several genome maintenance pathways, including mismatch repair, non-homologous end joining, ICL repair, DNA damage checkpoint activation, and replication fork stability. Here we describe how the Xenopus egg extract system, in combination with specifically designed DNA templates, contributed to our detailed understanding of these pathways. Copyright © 2017. Published by Elsevier Inc.

  18. Characterization of Genomic Deletion Efficiency Mediated by Clustered Regularly Interspaced Palindromic Repeats (CRISPR)/Cas9 Nuclease System in Mammalian Cells*♦

    Science.gov (United States)

    Canver, Matthew C.; Bauer, Daniel E.; Dass, Abhishek; Yien, Yvette Y.; Chung, Jacky; Masuda, Takeshi; Maeda, Takahiro; Paw, Barry H.; Orkin, Stuart H.

    2014-01-01

    The clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 nuclease system has provided a powerful tool for genome engineering. Double strand breaks may trigger nonhomologous end joining repair, leading to frameshift mutations, or homology-directed repair using an extrachromosomal template. Alternatively, genomic deletions may be produced by a pair of double strand breaks. The efficiency of CRISPR/Cas9-mediated genomic deletions has not been systematically explored. Here, we present a methodology for the production of deletions in mammalian cells, ranging from 1.3 kb to greater than 1 Mb. We observed a high frequency of intended genomic deletions. Nondeleted alleles are nonetheless often edited with inversions or small insertion/deletions produced at CRISPR recognition sites. Deleted alleles also typically include small insertion/deletions at predicted deletion junctions. We retrieved cells with biallelic deletion at a frequency exceeding that of probabilistic expectation. We demonstrate an inverse relationship between deletion frequency and deletion size. This work suggests that CRISPR/Cas9 is a robust system to produce a spectrum of genomic deletions to allow investigation of genes and genetic elements. PMID:24907273

  19. DNA-PK, ATM and ATR collaboratively regulate p53-RPA interaction to facilitate homologous recombination DNA repair.

    Science.gov (United States)

    Serrano, M A; Li, Z; Dangeti, M; Musich, P R; Patrick, S; Roginskaya, M; Cartwright, B; Zou, Y

    2013-05-09

    Homologous recombination (HR) and nonhomologous end joining (NHEJ) are two distinct DNA double-stranded break (DSB) repair pathways. Here, we report that DNA-dependent protein kinase (DNA-PK), the core component of NHEJ, partnering with DNA-damage checkpoint kinases ataxia telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR), regulates HR repair of DSBs. The regulation was accomplished through modulation of the p53 and replication protein A (RPA) interaction. We show that upon DNA damage, p53 and RPA were freed from a p53-RPA complex by simultaneous phosphorylations of RPA at the N-terminus of RPA32 subunit by DNA-PK and of p53 at Ser37 and Ser46 in a Chk1/Chk2-independent manner by ATR and ATM, respectively. Neither the phosphorylation of RPA nor of p53 alone could dissociate p53 and RPA. Furthermore, disruption of the release significantly compromised HR repair of DSBs. Our results reveal a mechanism for the crosstalk between HR repair and NHEJ through the co-regulation of p53-RPA interaction by DNA-PK, ATM and ATR.

  20. More efficient repair of DNA double-strand breaks in skeletal muscle stem cells compared to their committed progeny

    Directory of Open Access Journals (Sweden)

    Leyla Vahidi Ferdousi

    2014-11-01

    Full Text Available The loss of genome integrity in adult stem cells results in accelerated tissue aging and is possibly cancerogenic. Adult stem cells in different tissues appear to react robustly to DNA damage. We report that adult skeletal stem (satellite cells do not primarily respond to radiation-induced DNA double-strand breaks (DSBs via differentiation and exhibit less apoptosis compared to other myogenic cells. Satellite cells repair these DNA lesions more efficiently than their committed progeny. Importantly, non-proliferating satellite cells and post-mitotic nuclei in the fiber exhibit dramatically distinct repair efficiencies. Altogether, reduction of the repair capacity appears to be more a function of differentiation than of the proliferation status of the muscle cell. Notably, satellite cells retain a high efficiency of DSB repair also when isolated from the natural niche. Finally, we show that repair of DSB substrates is not only very efficient but, surprisingly, also very accurate in satellite cells and that accurate repair depends on the key non-homologous end-joining factor DNA-PKcs.

  1. More efficient repair of DNA double-strand breaks in skeletal muscle stem cells compared to their committed progeny.

    Science.gov (United States)

    Vahidi Ferdousi, Leyla; Rocheteau, Pierre; Chayot, Romain; Montagne, Benjamin; Chaker, Zayna; Flamant, Patricia; Tajbakhsh, Shahragim; Ricchetti, Miria

    2014-11-01

    The loss of genome integrity in adult stem cells results in accelerated tissue aging and is possibly cancerogenic. Adult stem cells in different tissues appear to react robustly to DNA damage. We report that adult skeletal stem (satellite) cells do not primarily respond to radiation-induced DNA double-strand breaks (DSBs) via differentiation and exhibit less apoptosis compared to other myogenic cells. Satellite cells repair these DNA lesions more efficiently than their committed progeny. Importantly, non-proliferating satellite cells and post-mitotic nuclei in the fiber exhibit dramatically distinct repair efficiencies. Altogether, reduction of the repair capacity appears to be more a function of differentiation than of the proliferation status of the muscle cell. Notably, satellite cells retain a high efficiency of DSB repair also when isolated from the natural niche. Finally, we show that repair of DSB substrates is not only very efficient but, surprisingly, also very accurate in satellite cells and that accurate repair depends on the key non-homologous end-joining factor DNA-PKcs. Copyright © 2014. Published by Elsevier B.V.

  2. Coexposure to benzo[a]pyrene plus UVA induced DNA double strand breaks: visualization of Ku assembly in the nucleus having DNA lesions

    International Nuclear Information System (INIS)

    Toyooka, Tatsushi; Ibuki, Yuko; Koike, Manabu; Ohashi, Norio; Takahashi, Sentaro; Goto, Rensuke

    2004-01-01

    Benzo[a]pyrene (BaP) is a ubiquitous environmental pollutant with potential carcinogenicity. It has been shown that BaP, upon UVA irradiation, synergistically induced oxidative DNA damage, but other DNA damage was not confirmed. In this study, we examined whether coexposure to BaP plus UVA induces double strand breaks (DSBs) using xrs-5 cells, deficient in the repair of DSBs (Ku80 mutant), and whether Ku translocates involving the formation of DSBs. BaP plus UVA had a significant cytotoxic effect on CHO-K1 cells and an even more drastic effect on Ku80-deficient, xrs-5 cells, suggesting that the DSBs were generated by coexposure to BaP plus UVA. The DSBs were repaired in CHO-K1 cells within 30 min, but not in xrs-5 cells, indicating the involvement of a non-homologous end joining, which needs Ku proteins. Furthermore, we succeeded in visualizing that Ku80 rapidly assembled to the exposed region, in which DSBs might be generated, and clarified that the presence of both Ku70 and Ku80 was important for their accumulation

  3. A quantitative model of the major pathways for radiation-induced DNA double-strand break repair

    International Nuclear Information System (INIS)

    Belov, O.V.; Krasavin, E.A.; Lyashko, M.S.; Batmunkh, M.; Sweilam, N.H.

    2014-01-01

    We have developed a model approach to simulate the major pathways of DNA double-strand break (DSB) repair in mammalian and human cells. The proposed model shows a possible mechanistic explanation of the basic regularities of DSB processing through the nonhomologous end-joining (NHEJ), homologous recombination (HR), and single-strand annealing (SSA). It reconstructs the time-courses of radiation-induced foci specific to particular repair processes including the major intermediate stages. The model is validated for ionizing radiations of a wide range of linear energy transfer (0.2-236 keV/μm) including a relatively broad spectrum of heavy ions. The appropriate set of reaction rate constants was suggested to satisfy the kinetics of DSB rejoining for the considered types of exposure. The simultaneous assessment of three repair pathways allows one to describe their possible biological relations in response to radiation. With the help of the proposed approach, we reproduce several experimental data sets on γ-H2AX foci remaining in different types of cells including those defective in NHEJ, HR, or SSA functions.

  4. Genome-Wide Mutational Signature of the Chemotherapeutic Agent Mitomycin C in Caenorhabditis elegans.

    Science.gov (United States)

    Tam, Annie S; Chu, Jeffrey S C; Rose, Ann M

    2015-11-12

    Cancer therapy largely depends on chemotherapeutic agents that generate DNA lesions. However, our understanding of the nature of the resulting lesions as well as the mutational profiles of these chemotherapeutic agents is limited. Among these lesions, DNA interstrand crosslinks are among the more toxic types of DNA damage. Here, we have characterized the mutational spectrum of the commonly used DNA interstrand crosslinking agent mitomycin C (MMC). Using a combination of genetic mapping, whole genome sequencing, and genomic analysis, we have identified and confirmed several genomic lesions linked to MMC-induced DNA damage in Caenorhabditis elegans. Our data indicate that MMC predominantly causes deletions, with a 5'-CpG-3' sequence context prevalent in the deleted regions of DNA. Furthermore, we identified microhomology flanking the deletion junctions, indicative of DNA repair via nonhomologous end joining. Based on these results, we propose a general repair mechanism that is likely to be involved in the biological response to this highly toxic agent. In conclusion, the systematic study we have described provides insight into potential sequence specificity of MMC with DNA. Copyright © 2016 Tam et al.

  5. Genome-Wide Mutational Signature of the Chemotherapeutic Agent Mitomycin C in Caenorhabditis elegans

    Directory of Open Access Journals (Sweden)

    Annie S. Tam

    2016-01-01

    Full Text Available Cancer therapy largely depends on chemotherapeutic agents that generate DNA lesions. However, our understanding of the nature of the resulting lesions as well as the mutational profiles of these chemotherapeutic agents is limited. Among these lesions, DNA interstrand crosslinks are among the more toxic types of DNA damage. Here, we have characterized the mutational spectrum of the commonly used DNA interstrand crosslinking agent mitomycin C (MMC. Using a combination of genetic mapping, whole genome sequencing, and genomic analysis, we have identified and confirmed several genomic lesions linked to MMC-induced DNA damage in Caenorhabditis elegans. Our data indicate that MMC predominantly causes deletions, with a 5′-CpG-3′ sequence context prevalent in the deleted regions of DNA. Furthermore, we identified microhomology flanking the deletion junctions, indicative of DNA repair via nonhomologous end joining. Based on these results, we propose a general repair mechanism that is likely to be involved in the biological response to this highly toxic agent. In conclusion, the systematic study we have described provides insight into potential sequence specificity of MMC with DNA.

  6. The Impact of Hedgehog Signaling Pathway on DNA Repair Mechanisms in Human Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Meng, Erhong; Hanna, Ann; Samant, Rajeev S.; Shevde, Lalita A., E-mail: lsamant@uab.edu [Department of Pathology, Comprehensive Cancer Center, University of Alabama at Birmingham, WTI320D, 1824 6th Avenue South, Birmingham, AL 35233 (United States)

    2015-07-21

    Defined cellular mechanisms have evolved that recognize and repair DNA to protect the integrity of its structure and sequence when encountering assaults from endogenous and exogenous sources. There are five major DNA repair pathways: mismatch repair, nucleotide excision repair, direct repair, base excision repair and DNA double strand break repair (including non-homologous end joining and homologous recombination repair). Aberrant activation of the Hedgehog (Hh) signaling pathway is a feature of many cancer types. The Hh pathway has been documented to be indispensable for epithelial-mesenchymal transition, invasion and metastasis, cancer stemness, and chemoresistance. The functional transcription activators of the Hh pathway include the GLI proteins. Inhibition of the activity of GLI can interfere with almost all DNA repair types in human cancer, indicating that Hh/GLI functions may play an important role in enabling tumor cells to survive lethal types of DNA damage induced by chemotherapy and radiotherapy. Thus, Hh signaling presents an important therapeutic target to overcome DNA repair-enabled multi-drug resistance and consequently increase chemotherapeutic response in the treatment of cancer.

  7. High-efficiency targeted editing of large viral genomes by RNA-guided nucleases.

    Science.gov (United States)

    Bi, Yanwei; Sun, Le; Gao, Dandan; Ding, Chen; Li, Zhihua; Li, Yadong; Cun, Wei; Li, Qihan

    2014-05-01

    A facile and efficient method for the precise editing of large viral genomes is required for the selection of attenuated vaccine strains and the construction of gene therapy vectors. The type II prokaryotic CRISPR-Cas (clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas)) RNA-guided nuclease system can be introduced into host cells during viral replication. The CRISPR-Cas9 system robustly stimulates targeted double-stranded breaks in the genomes of DNA viruses, where the non-homologous end joining (NHEJ) and homology-directed repair (HDR) pathways can be exploited to introduce site-specific indels or insert heterologous genes with high frequency. Furthermore, CRISPR-Cas9 can specifically inhibit the replication of the original virus, thereby significantly increasing the abundance of the recombinant virus among progeny virus. As a result, purified recombinant virus can be obtained with only a single round of selection. In this study, we used recombinant adenovirus and type I herpes simplex virus as examples to demonstrate that the CRISPR-Cas9 system is a valuable tool for editing the genomes of large DNA viruses.

  8. High-efficiency targeted editing of large viral genomes by RNA-guided nucleases.

    Directory of Open Access Journals (Sweden)

    Yanwei Bi

    2014-05-01

    Full Text Available A facile and efficient method for the precise editing of large viral genomes is required for the selection of attenuated vaccine strains and the construction of gene therapy vectors. The type II prokaryotic CRISPR-Cas (clustered regularly interspaced short palindromic repeats (CRISPR-associated (Cas RNA-guided nuclease system can be introduced into host cells during viral replication. The CRISPR-Cas9 system robustly stimulates targeted double-stranded breaks in the genomes of DNA viruses, where the non-homologous end joining (NHEJ and homology-directed repair (HDR pathways can be exploited to introduce site-specific indels or insert heterologous genes with high frequency. Furthermore, CRISPR-Cas9 can specifically inhibit the replication of the original virus, thereby significantly increasing the abundance of the recombinant virus among progeny virus. As a result, purified recombinant virus can be obtained with only a single round of selection. In this study, we used recombinant adenovirus and type I herpes simplex virus as examples to demonstrate that the CRISPR-Cas9 system is a valuable tool for editing the genomes of large DNA viruses.

  9. PAXX Is an Accessory c-NHEJ Factor that Associates with Ku70 and Has Overlapping Functions with XLF

    Directory of Open Access Journals (Sweden)

    Satish K. Tadi

    2016-10-01

    Full Text Available In mammalian cells, classical non-homologous end joining (c-NHEJ is critical for DNA double-strand break repair induced by ionizing radiation and during V(DJ recombination in developing B and T lymphocytes. Recently, PAXX was identified as a c-NHEJ core component. We report here that PAXX-deficient cells exhibit a cellular phenotype uncharacteristic of a deficiency in c-NHEJ core components. PAXX-deficient cells display normal sensitivity to radiomimetic drugs, are proficient in transient V(DJ recombination assays, and do not shift toward higher micro-homology usage in plasmid repair assays. Although PAXX-deficient cells lack c-NHEJ phenotypes, PAXX forms a stable ternary complex with Ku bound to DNA. Formation of this complex involves an interaction with Ku70 and requires a bare DNA extension for stability. Moreover, the relatively weak Ku-dependent stimulation of LIG4/XRCC4 activity by PAXX is unmasked by XLF ablation. Thus, PAXX plays an accessory role during c-NHEJ that is largely overlapped by XLF’s function.

  10. Germline excision of transgenes in Aedes aegypti by homing endonucleases.

    Science.gov (United States)

    Aryan, Azadeh; Anderson, Michelle A E; Myles, Kevin M; Adelman, Zach N

    2013-01-01

    Aedes (Ae.) aegypti is the primary vector for dengue viruses (serotypes1-4) and chikungunya virus. Homing endonucleases (HEs) are ancient selfish elements that catalyze double-stranded DNA breaks (DSB) in a highly specific manner. In this report, we show that the HEs Y2-I-AniI, I-CreI and I-SceI are all capable of catalyzing the excision of genomic segments from the Ae. aegypti genome in a heritable manner. Y2-I-AniI demonstrated the highest efficiency at two independent genomic targets, with 20-40% of Y2-I-AniI-treated individuals producing offspring that had lost the target transgene. HE-induced DSBs were found to be repaired via the single-strand annealing (SSA) and non-homologous end-joining (NHEJ) pathways in a manner dependent on the availability of direct repeat sequences in the transgene. These results support the development of HE-based gene editing and gene drive strategies in Ae. aegypti, and confirm the utility of HEs in the manipulation and modification of transgenes in this important vector.

  11. Reciprocal Regulation between DNA-PKcs and Snail1 Conferring Genomic Instability

    International Nuclear Information System (INIS)

    Seo, Haeng Ran; Lee, Hae June; Jin, Yeung Bae; Bae, Sang Woo; Lee, Yun Sil; Kim, Nam Hee; Kim, Hyun Sil; Nam, Hyung Wook; Yook, Jong In

    2010-01-01

    Although the roles of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) involving non-homologous end joining (NHEJ) of DNA repair are well recognized, the biological mechanisms and regulators by which DNA-PKcs regulate genomic instability are not clearly defined. We show herein that DNA-PKcs activity resulting from DNA damage caused by ionizing radiation (IR) phosphorylates Snail1 at serine 100, which results in increased Snail1 expression and its function by inhibition of GSK-3-mediated phosphorylation. Furthermore, Snail1 phosphorylated at serine 100 can reciprocally inhibit kinase activity of DNA-PKcs, resulting in an inhibition to recruit DNA-PKcs or Ku70/80 to a DNA double-strand break site, and ultimately inhibition of DNA repair activity. The impairment of repair activity by a direct interaction between Snail1 and DNA-PKcs increases the resistance to DNA damaging agents, such as IR, and genomic instability. Our findings provide a novel cellular mechanism for induction of genomic instability by reciprocal regulation of DNA-PKcs and Snail1

  12. Gel-based proteomic approach to unravel the extreme radiation resistance of deinococcus radiodurans

    International Nuclear Information System (INIS)

    Basu, Bhakti; Apte, Shree Kumar

    2013-01-01

    The extremophile, Deinococcus radiodurans, is endowed with an extraordinary DNA repair ability and oxidative stress alleviation mechanisms that render it virtually resistant to all types of DNA damaging stressors such as ionizing radiations, UV or years of desiccation. Following DNA damage, the microbe reassembles its complete genome from multiple DNA fragments with impeccable fidelity. The deinococcal genome encodes functional homologues of both prokaryotic and eukaryotic DNA repair pathways, such as RecFOR mediated homologous recombination (HR), nucleotide/base excision repair (NER/BER), strand annealing (SA) and non-homologous end joining (NHEJ), but lacks homologues for universal prokaryotic DNA repair pathways such as RecBCD mediated HR, photo-reactivation and SOS response. It also harbors multiple enzymatic and non-enzymatic oxidative stress defense mechanisms. Proteomic approaches were employed to study the response of D. radiodurans to LD50 dose of gamma irradiation during the post-irradiation growth arrest phase by two dimensional protein electrophoresis coupled with mass spectrometry to reveal kinetics and dynamics of DNA repair, oxidative stress alleviation and resynthesis of damaged proteins, preceding growth recovery

  13. Laser beam joining of non-oxidic ceramics for ultra high temperature resistant joints

    International Nuclear Information System (INIS)

    Lippmann, W.; Knorr, J.; Wolf, R.; Reinecke, A.M.; Rasper, R.

    2004-01-01

    The excellent technical properties of silicon carbide (SiC) and silicon nitride (Si 3 N 4 ) ceramics, such as resistance to extreme temperatures, oxidation, mechanical wear, aggressive chemical substances and radioactive radiation and also its high thermal conductivity and good temperature-shock resistance, make these ceramics ideally suited for use in the field of nuclear technology. However, their practical use has been limited so far because of the unavailability of effective joining techniques for these ceramics, especially for high temperature applications. A new joining technology (CERALINK registered ) has been developed in a network project which allowed high temperature resistant and vacuum-tight joining of SiC or Si 3 N 4 ceramics. A power laser is used as heat source, which makes it possible to join ceramic components in free atmosphere in combination with a pure oxidic braze filler. As no furnace is necessary, there are no limitations on the component dimensions by the furnace-geometry. During the joining process, the heated area can be limited to the seam area so that this technology can also be used to encapsulate materials with a low melting point. The seam has a high mechanical strength, it is resistant to a wide range of chemicals and radiation and it is also vacuum-tight. The temperature resistance can be varied by variation of the braze filler composition - usually between 1,400 C and >1,600 C. Beside the optimum filler it is also important to select the suitable laser wavelength. The paper will demonstrate the influence of different wave lengths, i. e. various laser types, on the seam quality. Examples are chosen to illustrate the strengths and limitations of the new technology

  14. Individual capacity for DNA repair and maintenance of genomic integrity: a fertile ground for studies in the field of assisted reproduction

    Directory of Open Access Journals (Sweden)

    Radoslava Vazharova

    2016-05-01

    Full Text Available Many factors may affect the chances for successful pregnancy, especially at a later age. Fertility evaluations including genetic analysis are recommended to couples that have not achieved pregnancy within 6–12 months of unprotected intercourse. This review discusses some of the common polymorphisms in genes coding for proteins functioning in DNA damage identification and repair and maintenance of genomic integrity that may affect the chances of success in natural conception as well as in assisted reproduction (AR. Common polymorphisms in genes coding for proteins functioning in DNA damage identification and repair and maintenance of genomic integrity may affect the chances of success in assisted reproduction as well as in natural conception. The effects of carriership of different alleles of key genes of DNA repair may have differential effects in men and women and at different ages, suggesting complex interactions with the mechanisms controlling cell and tissue aging and programmed cell death. Future studies in the field are needed in order to elucidate the genotype–phenotype relationships and to translate the knowledge about individual repair capacity and maintenance of genomic integrity to potential clinical applications. Abbreviations: aCGH: microarray-based comparative genomic hybridization; AR: assisted reproduction; ATM: ataxia-telangiectasia mutated; ATP: adenosine triphosphate; BER: base excision repair; BFE: basic fertility evaluation; DMSO: dimethyl sulfoxide; FSH: follicle-stimulating hormone; GNRHR: gonadotropin-releasing hormone receptor; HMG: high-mobility group; ICSI: intracytoplasmic sperm injection; IUI: intrauterine insemination; IVF: in vitro fertilization; LH: luteinizing hormone; LIF: leukaemia inhibitory factor; MTR: methionine synthase; MTRR: methionine synthase reductase; NGS: next-generation sequencing; NER: nucleotide excision repair; NHEJ: non-homologous end joining; PAH: polycyclic aromatic hydrocarbons; PCOS

  15. A PHF8 homolog in C. elegans promotes DNA repair via homologous recombination.

    Directory of Open Access Journals (Sweden)

    Changrim Lee

    Full Text Available PHF8 is a JmjC domain-containing histone demethylase, defects in which are associated with X-linked mental retardation. In this study, we examined the roles of two PHF8 homologs, JMJD-1.1 and JMJD-1.2, in the model organism C. elegans in response to DNA damage. A deletion mutation in either of the genes led to hypersensitivity to interstrand DNA crosslinks (ICLs, while only mutation of jmjd-1.1 resulted in hypersensitivity to double-strand DNA breaks (DSBs. In response to ICLs, JMJD-1.1 did not affect the focus formation of FCD-2, a homolog of FANCD2, a key protein in the Fanconi anemia pathway. However, the dynamic behavior of RPA-1 and RAD-51 was affected by the mutation: the accumulations of both proteins at ICLs appeared normal, but their subsequent disappearance was retarded, suggesting that later steps of homologous recombination were defective. Similar changes in the dynamic behavior of RPA-1 and RAD-51 were seen in response to DSBs, supporting a role of JMJD-1.1 in homologous recombination. Such a role was also supported by our finding that the hypersensitivity of jmjd-1.1 worms to ICLs was rescued by knockdown of lig-4, a homolog of Ligase 4 active in nonhomologous end-joining. The hypersensitivity of jmjd-1.1 worms to ICLs was increased by rad-54 knockdown, suggesting that JMJD-1.1 acts in parallel with RAD-54 in modulating chromatin structure. Indeed, the level of histone H3 Lys9 tri-methylation, a marker of heterochromatin, was higher in jmjd-1.1 cells than in wild-type cells. We conclude that the histone demethylase JMJD-1.1 influences homologous recombination either by relaxing heterochromatin structure or by indirectly regulating the expression of multiple genes affecting DNA repair.

  16. Frequency of joined disabilities of children with cerebral palsy in Tuzla canton

    Directory of Open Access Journals (Sweden)

    Mirela Babajić

    2013-12-01

    Full Text Available Introduction: Cerebral palsy (CP connotes a group of non-progressive, but often variable symptoms of motor impairment of movement and posture, as well as other impairments which are a consequenceof anomalies or brain impairment in different phases of its development. CP is a pathological condition characterised in the fi rst place by motor function impairment to which other disorders such as: visual andhearing impairment, intellectual defi cit, emotional problems, behaviour disorder, speech disorder, epileptic seizure and similar can join. The aim of this study is to determine frequency of joined disabilities ofchildren with cerebral palsy in Tuzla Canton.Methods: The research covers a total sample of 48 examinees, chronological age from 2-19 years, in Tuzla Canton. Research instrument was a Structural Questionnaire for the parents of children and adolescentswith cerebral palsy. Research data were processed by nonparametric statistics method. Basic statistical parameters of frequency and percentages were calculated, and tabular presentation was made.Results: After classification of examinees as per frequency of joined disabilities was done, work results have shown that speech impairment occurred with 35.4 % of children, visual impairment 33.3 %, epilepsy29.3 %, whereas hearing impairment occurred with 2 % of children.Conclusion: In research of frequency of joined disabilities of children with cerebral palsy in Tuzla Canton, most expressed are speech and visual disorders with children, then epilepsy, whereas a small percentageof children are with hearing disorder.

  17. Joining SiC/SiC composites for fusion applications

    International Nuclear Information System (INIS)

    Henager, C.J.

    2007-01-01

    Full text of publication follows: The use of SiC-reinforced composites for fusion reactors or other nuclear applications will likely require some form of joining in order to form large structural or functional sections. Joints must be able to allow load transfer to the composite material so that the fiber reinforcements are able to carry their designed loads and the joint should not fail in a brittle manner. In addition, joints must be tolerant of radiation damage similar to the composite material to maintain the structural integrity of the joined section. Other requirements of interest are low activation joints and ease of joining application and processing, which are a difficult set of criteria to meet. Research at PNNL has been directed at high-strength joints using solid-state displacement reactions and pre-ceramic polymer joints that are easy to apply and that could be used in field repair situations. This research will report on the further development and testing of such joints using a double notch shear (DNS) specimen that is suitable for in-reactor testing. The results reveal that the solid-state joints are stronger than the polymer-based joints but require high-temperature, high-pressure processing. The polymer-based joints can be applied with reduced pressure and temperature processing compared to the solid-state joints. The polymer-based joints may have adequate strength for some applications where ease of application and in situ processing are required. Although irradiation testing is not reported in this study the use of the DNS specimen is discussed with regard to future neutron irradiations. (authors)

  18. Study on the Joining Strength of Spot Welding using POMISPOT Device

    International Nuclear Information System (INIS)

    Mohd Azhar Ahmad; Siti Aiasah Hashim; Mohd Rizal Chulan

    2015-01-01

    Welding is a process to join metals. Spot welding is commonly used for specific purposes such joining in small areas or making temporary joints. POMISPOT is a spot welder that was designed and built by the ADC group, using capacitive resistance method. This study was made to obtain the welding strength that can be made by this spot welder. The study used stainless steel pieces of different thickness and by varying the applied voltage. The strength of welded pieces is tested by applying loads. The relationship between the thickness, voltage and welding strength will be used as the basis of specifications of this tool. (author)

  19. Activities of HIP joining of plasma-facing armors in the blanket first-wall in Korea

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Yang-Il, E-mail: yijung@kaeri.re.kr [Korea Atomic Energy Research Institute, Daedeok-daero, Daejeon 34057 (Korea, Republic of); Park, Jeong-Yong; Choi, Byoung-Kwon; Lee, Jung-Suk; Kim, Hyun-Gil; Park, Dong-Jun; Park, Jung-Hwan; Kim, Suk-Kwon; Lee, Dong-Won [Korea Atomic Energy Research Institute, Daedeok-daero, Daejeon 34057 (Korea, Republic of); Cho, Seungyon [National Fusion Research Institute, Gwahak-ro, Yuseong, Daejeon 34133 (Korea, Republic of)

    2016-11-01

    Highlights: • HIP joints of Be/CuCrZr, Be/FMS, W/FMS were demonstrated. • The process conditions for HIP joining were developed. • For the joining of Be, coating interlayers as well as thick diffusion barrier was developed. • For the joining of W, double-staged HIP was applied for the joint integrity. • No significant defects nor a brittle failure were observed along the joint interface. - Abstract: Joining technology for dissimilar materials was developed for the fabrication of an ITER blanket first-wall, which consisted of Be, CuCrZr, and stainless steel (SS). The Be/CuCrZr/SS joint was fabricated using a hot isostatic pressing (HIP) method. Beryllium armor was joined to the CuCrZr/SS block at 580 °C under 100 MPa. The optimal interlayer coatings of Cr/Cu and Ti/Cr/Cu were developed using an ion-beam assisted physical vapor deposition. Beryllium is also a candidate armor material for the TBM first-wall. Successful joining of Be to ferritic-martensitic steel (FMS) was accomplished using an HIP method by introducing the thick diffusion barrier. A thick diffusion barrier of a Cu foil(10 μm) limited the excessive diffusion and prevented the formation of brittle phases at the Be/FMS interface. Be and FMS were bonded at 650–850 °C; however, a temperature of lower than 750 °C was recommended to avoid material degradation of FMS. In addition, the joining of W to FMS has been developed. Tungsten is another armor material applicable to more severe plasma conditions. The large difference in the thermal expansion between W and FMS was resolved by introducing the Ti interlayer and Mo separator. Moreover, the double-staged HIP (the first stage at 900 °C and 100 MPa and the second stage at 750 °C and 70 MPa) was applied to suppress the edge delamination of W/FMS joints during thermal history.

  20. Trex2 enables spontaneous sister chromatid exchanges without facilitating DNA double-strand break repair.

    Science.gov (United States)

    Dumitrache, Lavinia C; Hu, Lingchuan; Son, Mi Young; Li, Han; Wesevich, Austin; Scully, Ralph; Stark, Jeremy; Hasty, Paul

    2011-08-01

    Trex2 is a 3' → 5' exonuclease that removes 3'-mismatched sequences in a biochemical assay; however, its biological function remains unclear. To address biology we previously generated trex2(null) mouse embryonic stem (ES) cells and expressed in these cells wild-type human TREX2 cDNA (Trex2(hTX2)) or cDNA with a single-amino-acid change in the catalytic domain (Trex2(H188A)) or in the DNA-binding domain (Trex2(R167A)). We found the trex2(null) and Trex2(H188A) cells exhibited spontaneous broken chromosomes and trex2(null) cells exhibited spontaneous chromosomal rearrangements. We also found ectopically expressed human TREX2 was active at the 3' ends of I-SceI-induced chromosomal double-strand breaks (DSBs). Therefore, we hypothesized Trex2 participates in DNA DSB repair by modifying 3' ends. This may be especially important for ends with damaged nucleotides. Here we present data that are unexpected and prompt a new model. We found Trex2-altered cells (null, H188A, and R167A) were not hypersensitive to camptothecin, a type-1 topoisomerase inhibitor that induces DSBs at replication forks. In addition, Trex2-altered cells were not hypersensitive to γ-radiation, an agent that causes DSBs throughout the cell cycle. This observation held true even in cells compromised for one of the two major DSB repair pathways: homology-directed repair (HDR) or nonhomologous end joining (NHEJ). Trex2 deletion also enhanced repair of an I-SceI-induced DSB by both HDR and NHEJ without affecting pathway choice. Interestingly, however, trex2(null) cells exhibited reduced spontaneous sister chromatid exchanges (SCEs) but this was not due to a defect in HDR-mediated crossing over. Therefore, reduced spontaneous SCE could be a manifestation of the same defect that caused spontaneous broken chromosomes and spontaneous chromosomal rearrangements. These unexpected data suggest Trex2 does not enable DSB repair and prompt a new model that posits Trex2 suppresses the formation of broken

  1. Biological durability of wood in relation to end-use - Part 1. Towards a European standard for laboratory testing of the biological durability of wood

    NARCIS (Netherlands)

    Acker, Van J.; Stevens, M.; Carey, J.; Sierra-Alvarez, R.; Militz, H.; Bayon, Le I.; Kleist, G.; Peek, R.D.

    2003-01-01

    The determination of biological durability of wood is an issue requiring sufficient reliability regarding end-use related prediction of performance. Five test institutes joined efforts to check standard test methods and to improve methodology and data interpretation for assessment of natural

  2. Factors affecting dermatological manifestations in patients with end stage renal disease

    International Nuclear Information System (INIS)

    Anees, M.; Gull, S.; Nazeer, A.

    2018-01-01

    To determine skin changes in patients of End Stage Renal Disease (ESRD) on maintenance hemodialysis (MHD) and factors affecting these changes. Study Design:Cross-sectional observational study. Place and Duration of Study:Nephrology Department, Mayo Hospital, Lahore in collaboration with Dermatology Department, King Edward Medical University, Lahore, from October 2015 to January 2016. Methodology:Two hundred patients who were undergoing MHD for more than three months were included in the study. Patients' demographic data, laboratory reports and dialysis records were noted in a predesigned questionnaire. Skin examination was carried out by consultant dermatologist after patient's permission. Results:Among 200 patients included in study, 105 were males and rest of them were females. Major causes of ESRD were Diabetes Mellitus (n=83, 41.5%, followed by Hypertension (n=80, 40%), Nephrolithiasis (n=15, 7.5%) and Chronic glomerulonephritis (n=5, 2.5%). At least one cutaneous finding was present in every patient. Common skin findings observed were pigmentation (86%), xerosis (83%), pallor (79%), pruritus (69%), acquired ichthyosis (50.5%), and bacterial skin infections (18.5%). Among them, nail manifestations were half-and-half nails (52%), nychomycosis (30.5%), onycholysis (20.5%), subungual hyperkeratosis (23.5%), and Mee's lines (7.5). Among hair changes were sparse scalp hair (38.5%), brittle and lustreless hair (28%). The factors contributing to skin changes were patient's age, cause of ESRD, anti HCV positivity, high urea and creatinine levels, duration and frequency of hemodialysis, hemoglobin levels, calcium phosphate product and socioeconomic status. Some skin manifestations were interrelated with each other like xerosis with pruritus (p<0.001), pruritus with bacterial infection (p<0.022), acquired Ichthyosis (p=0.008) and hair changes (p=0.035). Conclusion:ESRD patients on hemodialysis develop various skin changes during the course of disease

  3. DNA requirements for interaction of the C-terminal region of Ku80 with the DNA-dependent protein kinase catalytic subunit (DNA-PKcs).

    Science.gov (United States)

    Radhakrishnan, Sarvan Kumar; Lees-Miller, Susan P

    2017-09-01

    Non-homologous end joining (NHEJ) is the major pathway for the repair of ionizing radiation induced DNA double strand breaks (DSBs) in human cells. Critical to NHEJ is the DNA-dependent interaction of the Ku70/80 heterodimer with the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) to form the DNA-PK holoenzyme. However, precisely how Ku recruits DNA-PKcs to DSBs ends to enhance its kinase activity has remained enigmatic, with contradictory findings reported in the literature. Here we address the role of the Ku80 C-terminal region (CTR) in the DNA-dependent interaction of Ku70/80 with DNA-PKcs using purified components and defined DNA structures. Our results show that the Ku80 CTR is required for interaction with DNA-PKcs on short segments of blunt ended 25bp dsDNA or 25bp dsDNA with a 15-base poly dA single stranded (ss) DNA extension, but this requirement is less stringent on longer dsDNA molecules (35bp blunt ended dsDNA) or 25bp duplex DNA with either a 15-base poly dT or poly dC ssDNA extension. Moreover, the DNA-PKcs-Ku complex preferentially forms on 25 bp DNA with a poly-pyrimidine ssDNA extension.Our work clarifies the role of the Ku80 CTR and dsDNA ends on the interaction of DNA-PKcs with Ku and provides key information to guide assembly and biology of NHEJ complexes. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Joining technology—A challenge for the use of SiC components in HTRs

    Energy Technology Data Exchange (ETDEWEB)

    Herrmann, M., E-mail: marion.herrmann@tu-dresden.de; Meisel, P.; Lippmann, W.; Hurtado, A.

    2016-09-15

    The availability of suitable joining technologies is paramount to the further advancement of ceramic components and their use in HTRs. Among other joining technologies, a modified brazing technology using a laser beam for heating the components to be joined has been developed at TU Dresden. The laser-induced heating behavior of the ceramic material is determined by the interactions between the material and the laser beam. This was shown in two different silicon carbide materials (SSiC and SiC{sub f}-reinforced ceramic material) using a diode laser with wavelengths 808 nm and 940 nm. The laser-based technique was illustrated by three different examples: sealing of monolithic SiC with a pin configuration for fuel claddings, sealing of SiC heat pipes with a length of 1 m, and demonstration of the transferability of the laser technique to fiber-reinforced components by means of a SiC{sub f}/SiCN material. Because the covalent bonding of SiC does not allow conventional welding, much research has been devoted to developing alternative filler systems. Glass or glass–ceramic fillers enable the tailoring of properties such as CTE and viscosity. Glasses are thermally stable up to their glass transition temperatures. It was shown that the crystallization of the yttrium aluminosilicate glass composition of the present work allows it to be used at 1050 °C without any significant changes occurring in braze tightness. For the SiC heat pipes with sodium as the working fluid, a sodium-resistant metal braze consisting of Ni–Ti–Si was formed. The long-term resistance of this filler to sodium at 800 °C was proven. The results demonstrate the possibility of using the laser-based joining technique for the joining of different SiC materials as well as for different brazing materials.

  5. Laser direct joining of metal and plastic

    International Nuclear Information System (INIS)

    Katayama, Seiji; Kawahito, Yousuke

    2008-01-01

    We have developed an innovative rapid laser direct joining process of metal and plastic lap plates without adhesives or glues. The joints made between a Type 304 stainless steel plate and a polyethylene terephthalate (PET) plastic sheet of 30 mm width possessed tensile shear loads of about 3000 N. Transmission electron microscope photographs of the joint demonstrated that Type 304 and the PET were bonded on the atomic, molecular or nanostructural level through a Cr oxide film

  6. Summary of Prior Work on Joining of Oxide Dispersion-Strengthened Alloys

    Energy Technology Data Exchange (ETDEWEB)

    Wright, Ian G [ORNL; Tatlock, Gordon J [ORNL; Badairy, H. [University of Liverpool; Chen, C-L. [University of Liverpool

    2009-08-01

    There is a range of joining techniques available for use with ODS alloys, but care should be exercised in matching the technique to the final duty requirements of the joint. The goal for joining ODS alloys is a joint with no local disruption of the distribution of the oxide dispersion, and no significant change in the size and orientation of the alloy microstructure. Not surprisingly, the fusion welding processes typically employed with wrought alloys produce the least satisfactory results with ODS alloys, but some versions, such as fusion spot welding, and the laser and electron-beam welding technologies, have demonstrated potential for producing sound joints. Welds made using solid-state spot welding reportedly have exhibited parent metal properties. Thus, it is possible to employ processes that result in significant disruption of the alloy microstructure, as long as the processing parameters are adjustment to minimize the extent of or influence of the changes in the alloy microstructure. Selection among these joining approaches largely depends on the particular application and component configuration, and an understanding of the relationships among processing, alloy microstructure, and final properties is key. Recent developments have resulted in friction welding evolving to be a prime method for joining ODS sheet products, and variants of brazing/diffusion bonding have shown excellent promise for use with tubes and pipes. The techniques that come closest to the goal defined above involve solid-state diffusion bonding and, in particular, it has been found that secondary recrystallization of joints made by pulsed plasma-assisted diffusion can produce the desired, continuous, large alloy grain structure through the joint. Such joints have exhibited creep rupture failure at >82% of the load needed to fail the monolithic parent alloy at 1000 C.

  7. Use of Friction Stir Welding and Friction Stir Processing for Advanced Nuclear Fuels and Materials Joining Applications

    International Nuclear Information System (INIS)

    J. I. Cole; J. F. Jue

    2006-01-01

    Application of the latest developments in materials technology may greatly aid in the successful pursuit of next generation reactor and transmutation technologies. One such area where significant progress is needed is joining of advanced fuels and materials. Rotary friction welding, also referred to as friction stir welding (FSW), has shown great promise as a method for joining traditionally difficult to join materials such as aluminum alloys. This relatively new technology, first developed in 1991, has more recently been applied to higher melting temperature alloys such as steels, nickel-based and titanium alloys. An overview of the FSW technology is provided and two specific nuclear fuels and materials applications where the technique may be used to overcome limitations of conventional joining technologies are highlighted

  8. Retrieval of Spatial Join Pattern Instances from Sensor Networks

    DEFF Research Database (Denmark)

    Yiu, Man Lung; Mamoulis, Nikos; Bakiras, Spiridon

    2009-01-01

    We study the continuous evaluation of spatial join queries and extensions thereof, defined by interesting combinations of sensor readings (events) that co-occur in a spatial neighborhood. An example of such a pattern is "a high temperature reading in the vicinity of at least four high-pressure re...

  9. Joining of SiC ceramics and SiC/SiC composites

    Energy Technology Data Exchange (ETDEWEB)

    Rabin, B.H. [Idaho National Engineering Lab., Idaho Falls, ID (United States)

    1996-08-01

    This project has successfully developed a practical and reliable method for fabricating SiC ceramic-ceramic joints. This joining method will permit the use of SiC-based ceramics in a variety of elevated temperature fossil energy applications. The technique is based on a reaction bonding approach that provides joint interlayers compatible with SiC, and excellent joint mechanical properties at temperatures exceeding 1000{degrees}C. Recent emphasis has been given to technology transfer activities, and several collaborative research efforts are in progress. Investigations are focusing on applying the joining method to sintered {alpha}-SiC and fiber-reinforced SiC/SiC composites for use in applications such as heat exchangers, radiant burners and gas turbine components.

  10. Georneys joins AGU's blog network

    Science.gov (United States)

    Viñas, Maria-José

    2011-07-01

    A blog on geological musings, wanderings, and adventures, called Georneys, has joined AGU's network of Earth and space science blogs. With the addition of Georneys, on 11 July, the AGU Blogosphere (http://blogs.agu.org), as the network is known, has grown to showcase eight independent blogs since its launch last fall. “One reason I write this blog is to maintain my sanity as I finish up my Ph.D. In the midst of much stress, long days in lab, and long nights writing thesis chapters, I write to remind myself of why I love geology,” says Georneys blogger Evelyn Mervine. “I also write to document some of my geological adventures and to share my love of geology with others.”

  11. Using and joining a franchised private sector provider network in Myanmar.

    Science.gov (United States)

    O'Connell, Kathryn; Hom, Mo; Aung, Tin; Theuss, Marc; Huntington, Dale

    2011-01-01

    Quality is central to understanding provider motivations to join and remain within a social franchising network. Quality also appears as a key issue from the client's perspective, and may influence why a client chooses to use a franchised provider over another type of provider. The dynamic relationships between providers of social franchising clinics and clients who use these services have not been thoroughly investigated in the context of Myanmar, which has an established social franchising network. This study examines client motivations to use a Sun Quality Health network provider and provider motivations to join and remain in the Sun Quality Health network. Taken together, these two aims provide an opportunity to explore the symbiotic relationship between client satisfaction and provider incentives to increase the utilization of reproductive health care services. Results from a series of focus group discussions with clients of reproductive health services and franchised providers shows that women chose health services provided by franchised private sector general practitioners because of its perceived higher quality, associated with the availability of effective, affordable, drugs. A key finding of the study is associated with providers. Provider focus group discussions indicate that a principle determinate for joining and remaining in the Sun Quality Health Network was serving the poor.

  12. Joining and Performance of Alkali Metal Thermal-to-electric Converter (AMTEC)

    Energy Technology Data Exchange (ETDEWEB)

    Suh, Min-Soo; Lee, Wook-Hyun; Woo, Sang-Kuk [Korea Institute of Energy Research, Daejeon (Korea, Republic of)

    2017-07-15

    The alkali-Metal Thermal-to-electric Converter (AMTEC) is one of the promising static energy conversion technologies for the direct conversion of thermal energy to electrical energy. The advantages over a conventional energy converter are its high theoretical conversion efficiency of 40% and power density of 500 W/kg. The working principle of an AMTEC battery is the electrochemical reaction of the sodium through an ion conducting electrolyte. Sodium ion pass through the hot side of the beta”-alumina solid electrolyte (BASE) primarily as a result of the pressure difference. This pressure difference across the BASE has a significant effect on the overall performance of the AMTEC system. In order to build the high pressure difference across the BASE, hermeticity is required for each joined components for high temperature range of 900°C. The AMTEC battery was manufactured by utilizing robust joining technology of BASE/insulator/metal flange interfaces of the system for both structural and electrical stability. The electrical potential difference between the anode and cathode sides, where the electrons emitted from sodium ionization and recombined into sodium, was characterized as the open-circuit voltage. The efforts of technological improvement were concentrated on a high-power output and conversion efficiency. This paper discusses about the joining and performance of the AMTEC systems.

  13. Using and joining a franchised private sector provider network in Myanmar.

    Directory of Open Access Journals (Sweden)

    Kathryn O'Connell

    Full Text Available BACKGROUND: Quality is central to understanding provider motivations to join and remain within a social franchising network. Quality also appears as a key issue from the client's perspective, and may influence why a client chooses to use a franchised provider over another type of provider. The dynamic relationships between providers of social franchising clinics and clients who use these services have not been thoroughly investigated in the context of Myanmar, which has an established social franchising network. This study examines client motivations to use a Sun Quality Health network provider and provider motivations to join and remain in the Sun Quality Health network. Taken together, these two aims provide an opportunity to explore the symbiotic relationship between client satisfaction and provider incentives to increase the utilization of reproductive health care services. METHODS AND FINDINGS: Results from a series of focus group discussions with clients of reproductive health services and franchised providers shows that women chose health services provided by franchised private sector general practitioners because of its perceived higher quality, associated with the availability of effective, affordable, drugs. A key finding of the study is associated with providers. Provider focus group discussions indicate that a principle determinate for joining and remaining in the Sun Quality Health Network was serving the poor.

  14. Using and Joining a Franchised Private Sector Provider Network in Myanmar

    Science.gov (United States)

    O'Connell, Kathryn; Hom, Mo; Aung, Tin; Theuss, Marc; Huntington, Dale

    2011-01-01

    Background Quality is central to understanding provider motivations to join and remain within a social franchising network. Quality also appears as a key issue from the client's perspective, and may influence why a client chooses to use a franchised provider over another type of provider. The dynamic relationships between providers of social franchising clinics and clients who use these services have not been thoroughly investigated in the context of Myanmar, which has an established social franchising network. This study examines client motivations to use a Sun Quality Health network provider and provider motivations to join and remain in the Sun Quality Health network. Taken together, these two aims provide an opportunity to explore the symbiotic relationship between client satisfaction and provider incentives to increase the utilization of reproductive health care services. Methods and Findings Results from a series of focus group discussions with clients of reproductive health services and franchised providers shows that women chose health services provided by franchised private sector general practitioners because of its perceived higher quality, associated with the availability of effective, affordable, drugs. A key finding of the study is associated with providers. Provider focus group discussions indicate that a principle determinate for joining and remaining in the Sun Quality Health Network was serving the poor. PMID:22180781

  15. Role of DNA-PK in cellular responses to DNA double-strand breaks

    International Nuclear Information System (INIS)

    Chen, D.J.

    2003-01-01

    DNA double-strand breaks (DSBs) are probably the most dangerous of the many different types of DNA damage that occur within the cell. DSBs are generated by exogenous agents such as ionizing radiation (IR) or by endogenously generated reactive oxygen species and occur as intermediates during meiotic and V(D)J recombination. The repair of DSBs is of paramount importance to the cell as misrepair of DSBs can lead to cell death or promote tumorigenesis. In eukaryotes there exists two distinct mechanisms for DNA DSB repair: homologous recombination (HR) and non-homologous end joining (NHEJ). In mammalian cells, however, it is clear that nonhomologous repair of DSBs is highly active and plays a major role in conferring radiation resistance to the cell. The NHEJ machinery minimally consists of the DNA-dependent Protein Kinase (DNA-PK) and a complex of XRCC4 and DNA Ligase IV. The DNA-PK complex is composed of a 470 kDa catalytic subunit (DNA-PKcs), and the heterodimeric Ku70 and Ku80 DNA end-binding complex. DNA-PKcs is a PI-3 kinase with homology to ATM and ATR in its C-terminal kinase domain. The DNA-PK complex protects and tethers the ends, and directs assembly and, perhaps, the activation of other NHEJ proteins. We have previously demonstrated that the kinase activity of DNA-PK is essential for DNA DSB repair and V(D)J recombination. It is, therefore, of immense interest to determine the in vivo targets of DNA-PKcs and the mechanisms by which phosphorylation of these targets modulates NHEJ. Recent studies have resulted in the identification of a number of protein targets that are phosphorylated by and/or interact with DNA-PKcs. Our laboratory has recently identified autophosphorylation site(s) on DNA-PKcs. We find that phosphorylation at these sites in vivo is an early and essential response to DSBs and demonstrate, for the first time, the localization of DNA-PKcs to the sites of DNA damage in vivo. Furthermore, mutation of these phosphorylation sites in mammalian

  16. Smells familiar: group-joining decisions of predatory mites are mediated by olfactory cues of social familiarity.

    Science.gov (United States)

    Muleta, Muluken G; Schausberger, Peter

    2013-09-01

    Group-living animals frequently have to trade off the costs and benefits of leaving an established group and joining another group. Owing to their high fitness relevance, group-joining decisions are commonly nonrandom and may be based on traits of both individual members and the group such as life stage, body size, social status and group density or size, respectively. Many group-living animals are able to recognize and to associate preferentially with familiar individuals, i.e. those encountered before. Hence, after dispersing from established groups, animals commonly have to decide whether to join a new familiar or unfamiliar group. Using binary choice situations we assessed the effects of social familiarity on group-joining behaviour of the plant-inhabiting predatory mite Phytoseiulus persimilis . Group living in P. persimilis is brought about by the patchy distribution of its spider mite prey and mutual conspecific attraction. In the first experiment, gravid predator females given a choice between spider mite patches occupied by unfamiliar and familiar groups of females strongly preferred to join familiar groups and to deposit their eggs in these patches. Preference for socially familiar groups was robust across biases of spider mite prey densities between choice options. The second experiment revealed that the predatory mite females can smell social familiarity from a distance. Females subjected to odour choice situations in artificial cages were more strongly attracted to the odour of familiar than unfamiliar groups. We argue that P. persimilis females preferentially join socially familiar groups because a familiar social environment relaxes competition and optimizes foraging and reproduction.

  17. Smells familiar: group-joining decisions of predatory mites are mediated by olfactory cues of social familiarity☆

    Science.gov (United States)

    Muleta, Muluken G.; Schausberger, Peter

    2013-01-01

    Group-living animals frequently have to trade off the costs and benefits of leaving an established group and joining another group. Owing to their high fitness relevance, group-joining decisions are commonly nonrandom and may be based on traits of both individual members and the group such as life stage, body size, social status and group density or size, respectively. Many group-living animals are able to recognize and to associate preferentially with familiar individuals, i.e. those encountered before. Hence, after dispersing from established groups, animals commonly have to decide whether to join a new familiar or unfamiliar group. Using binary choice situations we assessed the effects of social familiarity on group-joining behaviour of the plant-inhabiting predatory mite Phytoseiulus persimilis. Group living in P. persimilis is brought about by the patchy distribution of its spider mite prey and mutual conspecific attraction. In the first experiment, gravid predator females given a choice between spider mite patches occupied by unfamiliar and familiar groups of females strongly preferred to join familiar groups and to deposit their eggs in these patches. Preference for socially familiar groups was robust across biases of spider mite prey densities between choice options. The second experiment revealed that the predatory mite females can smell social familiarity from a distance. Females subjected to odour choice situations in artificial cages were more strongly attracted to the odour of familiar than unfamiliar groups. We argue that P. persimilis females preferentially join socially familiar groups because a familiar social environment relaxes competition and optimizes foraging and reproduction. PMID:24027341

  18. Influence of friction stir welding parameters on titanium-aluminum heterogeneous lap joining configuration

    Science.gov (United States)

    Picot, Florent; Gueydan, Antoine; Hug, Éric

    2017-10-01

    Lap joining configuration for Friction Stir Welding process is a methodology mostly dedicated to heterogeneous bonding. This welding technology was applied to join pure titanium with pure aluminum by varying the rotation speed and the movement speed of the tool. Regardless of the process parameters, it was found that the maximum strength of the junction remains almost constant. Microstructural observations by means of Scanning Electron Microscopy and Energy Dispersive Spectrometry analysis enable to describe the interfacial join and reveal asymmetric Cold Lap Defects on the sides of the junction. Chemical analysis shows the presence of one exclusive intermetallic compound through the interface identified as TiAl3. This compound is responsible of the crack spreading of the junction during the mechanical loading. The original version of this article supplied to AIP Publishing contained an accidental inversion of the authors, names. An updated version of this article, with the authors names formatted correctly was published on 20 October 2017.

  19. Development of a Dynamic, End-to-End Free Piston Stirling Convertor Model

    Science.gov (United States)

    Regan, Timothy F.; Gerber, Scott S.; Roth, Mary Ellen

    2003-01-01

    A dynamic model for a free-piston Stirling convertor is being developed at the NASA Glenn Research Center. The model is an end-to-end system model that includes the cycle thermodynamics, the dynamics, and electrical aspects of the system. The subsystems of interest are the heat source, the springs, the moving masses, the linear alternator, the controller and the end-user load. The envisioned use of the model will be in evaluating how changes in a subsystem could affect the operation of the convertor. The model under development will speed the evaluation of improvements to a subsystem and aid in determining areas in which most significant improvements may be found. One of the first uses of the end-to-end model will be in the development of controller architectures. Another related area is in evaluating changes to details in the linear alternator.

  20. Optimizing main-memory join on modern hardware

    OpenAIRE

    Boncz, Peter; Manegold, Stefan; Kersten, Martin

    2002-01-01

    textabstractIn the past decade, the exponential growth in commodity CPUs speed has far outpaced advances in memory latency. A second trend is that CPU performance advances are not only brought by increased clock rate, but also by increasing parallelism inside the CPU. Current database systems have not yet adapted to these trends, and show poor utilization of both CPU and memory resources on current hardware. In this article, we show how these resources can be optimized for large joins and tra...

  1. Boldness affects foraging decisions in barnacle geese: an experimental approach

    NARCIS (Netherlands)

    Kurvers, R.H.J.M.; Nolet, B.A.; Prins, H.H.T.; Ydenberg, R.C.; Oers, van K.

    2012-01-01

    Individuals foraging in groups constantly need to make decisions, such as when to leave a group, when to join a group, and when to move collectively to another feeding site. In recent years, it has become evident that personality may affect these foraging decisions, but studies where individuals are

  2. Comparative analysis of different joining techniques to improve the passive fit of cobalt-chromium superstructures.

    Science.gov (United States)

    Barbi, Francisco C L; Camarini, Edevaldo T; Silva, Rafael S; Endo, Eliana H; Pereira, Jefferson R

    2012-12-01

    The influence of different joining techniques on passive fit at the interface structure/abutment of cobalt-chromium (Co-Cr) superstructures has not yet been clearly established. The purpose of this study was to compare 3 different techniques of joining Co-Cr superstructures by measuring the resulting marginal misfit in a simulated prosthetic assembly. A specially designed metal model was used for casting, sectioning, joining, and measuring marginal misfit. Forty-five cast bar-type superstructures were fabricated in a Co-Cr alloy and randomly assigned by drawing lots to 3 groups (n=15) according to the joining method used: conventional gas-torch brazing (G-TB), laser welding (LW), and tungsten inert gas welding (TIG). Joined specimens were assembled onto abutment analogs in the metal model with the 1-screw method. The resulting marginal misfit was measured with scanning electron microscopy (SEM) at 3 different points: distal (D), central (C), and mesial (M) along the buccal aspect of both abutments: A (tightened) and B (without screw). The Levene test was used to evaluate variance homogeneity and then the Welsch ANOVA for heteroscedastic data (α=.05). Significant differences were found on abutment A between groups G-TB and LW (P=.013) measured mesially and between groups G-TB and TIG (P=.037) measured centrally. On abutment B, significant differences were found between groups G-TB and LW (Plaser method. Copyright © 2012 The Editorial Council of the Journal of Prosthetic Dentistry. Published by Mosby, Inc. All rights reserved.

  3. Open-ended and Open-door Treatment Groups for Young People with Mental Illness.

    Science.gov (United States)

    Miller, Rachel; Mason, Susan E

    2012-01-01

    The concept of open-ended groups is expanded to include an open-door model (OEOD) wherein members with severe mental illnesses, including schizophrenia disorders and bi-polar, can join, leave, and re-enter groups as their life circumstances dictate their availability and willingness for treatment. This model is grounded on the work of Schopler and Galinsky's (1984/2006) and Galinsky and Schopler's (1989) theses on the value and processes of open-ended groups and includes perspectives on mutual aid and group development. Groupwork with the OEOD format is illustrated with examples taken from a group of 79 participants diagnosed with first-episode schizophrenia/schizoaffective disorders, 40 of who had co-occurring substance abuse. Of the 79 participants in the OEOD group program, 70 (89%) remained in treatment for the maximum of 3 years. The over-all value of group treatment for this population is reviewed along with the small number of available publications on open-ended and open-door-type groups.

  4. Performance Comparison of the Optimized Inverted Joined Wing Airplane Concept and Classical Configuration Airplanes

    OpenAIRE

    Sieradzki Adam; Dziubiński Adam; Galiński Cezary

    2016-01-01

    The joined wing concept is an unconventional airplane configuration, known since the mid-twenties of the last century. It has several possible advantages, like reduction of the induced drag and weight due to the closed wing concept. The inverted joined wing variant is its rarely considered version, with the front wing being situated above the aft wing. The following paper presents a performance prediction of the recently optimized configuration of this airplane. Flight characteristics obtaine...

  5. Affective science and avoidant end-of-life communication: Can the science of emotion help physicians talk with their patients about the end of life?

    Science.gov (United States)

    Soodalter, Jesse A; Siegle, Greg J; Klein-Fedyshin, Michele; Arnold, Robert; Schenker, Yael

    2018-05-01

    Despite believing end-of-life (EOL) discussions with patients are important, doctors often do not have them. Multiple factors contribute to this shortfall, which interventions including reimbursement changes and communication skills training have not significantly improved to date. One commonly cited but under-researched reason for physician avoidance of EOL discussion is emotional difficulty. High occupational demand for frequent difficult discussions may overload physicians' normal emotional functioning, leading to avoidance or failure. We propose that cognitive, behavioral, and neuroscience evidence from affective science may offer helpful insights into this problem. Data from other populations show that strong emotion impairs cognitive performance and multiple demands can overload cognitive resources. We discuss several affective processes that may apply to physicians attempting EOL discussions. We then discuss selected interventions that have been shown to modify some of these processes and associated behavioral outcomes. Evidence for change in behavioral outcomes of interest includes performance and mood enhancement in healthy populations. We suggest that such mechanistically-targeted interventions may be hypothesized to help decrease physician avoidance of EOL discussion. Physicians may be motivated to adopt such interventions in order to enhance normal emotional functioning to meet supra-normal occupational demand. We propose this as a promising area of future study. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Design of Helical Self-Piercing Rivet for Joining Aluminum Alloy and High-Strength Steel Sheets

    International Nuclear Information System (INIS)

    Kim, W. Y.; Kim, D. B.; Park, J. G; Kim, D. H.; Kim, K. H.; Lee, I. H.; Cho, H. Y.

    2014-01-01

    A self-piercing rivet (SPR) is a mechanical component for joining dissimilar material sheets such as those of aluminum alloy and steel. Unlike conventional rivets, the SPR directly pierces sheets without the need for drilling them beforehand. However, the regular SPR can undergo buckling when it pierces a high-strength steel sheet, warranting the design of a helical SPR. In this study, the joining and forging processes using the helical SPR were simulated using the commercial FEM code, DEFORM-3D. High-tensile-strength steel sheets of different strengths were joined with aluminum alloy sheets using the designed helical SPR. The simulation results were found to agree with the experimental results, validating the optimal design of a helical SPR that can pierce high-strength steel sheets

  7. Design of Helical Self-Piercing Rivet for Joining Aluminum Alloy and High-Strength Steel Sheets

    Energy Technology Data Exchange (ETDEWEB)

    Kim, W. Y.; Kim, D. B.; Park, J. G; Kim, D. H.; Kim, K. H.; Lee, I. H.; Cho, H. Y. [Chungbuk National University, Cheongju (Korea, Republic of)

    2014-07-15

    A self-piercing rivet (SPR) is a mechanical component for joining dissimilar material sheets such as those of aluminum alloy and steel. Unlike conventional rivets, the SPR directly pierces sheets without the need for drilling them beforehand. However, the regular SPR can undergo buckling when it pierces a high-strength steel sheet, warranting the design of a helical SPR. In this study, the joining and forging processes using the helical SPR were simulated using the commercial FEM code, DEFORM-3D. High-tensile-strength steel sheets of different strengths were joined with aluminum alloy sheets using the designed helical SPR. The simulation results were found to agree with the experimental results, validating the optimal design of a helical SPR that can pierce high-strength steel sheets.

  8. POSSIBILITY OF PHARMACOMODULATION OF THE HYPERSENSITIVE RHINITIS JOINED WITH THE NASAL POLYPOSIS

    Directory of Open Access Journals (Sweden)

    Dejan Ursulović

    2001-11-01

    Full Text Available The research goai is to examine the effects of the local corticosteroidapplication to the number of eosinophils in the nasal secretion of the patients withhypersensitive rhinitis joined with the nasal polyposis. The study comprises 13patients with hypersensitive rhinitis joined with the nasal polyposis; 9 of them madeup the experimental group. The local corticosteroid (bechomethasone dipropionatein water spray was given at 12 hours in individual doses of 200 micrograms to theexperimental group patients in six weeks. During the treatment it was confirmed thatthere was a highly important reduction of the number of eosinophils of the nasalsecretion in the experimental group patients.

  9. Feasibility of ceramic joining with high energy electron beams

    International Nuclear Information System (INIS)

    Turman, B.N.; Glass, S.J.; Halbleib, J.A.; Helmich, D.R.; Loehman, R.E.; Clifford, J.R.

    1995-01-01

    Joining structural ceramics is possible using high melting point metals such as Mo and Pt that are heated with a high energy electron beam, with the potential for producing joints with high temperature capability. A 10 MeV electron beam can penetrate through 1 cm of ceramic, offering the possibility of buried interface joining. Because of transient heating and the lower heat capacity of the metal relative to the ceramic, a pulsed high power beam has the potential for melting the metal without decomposing or melting the adjacent ceramic. The authors have demonstrated the feasibility of the process with a series of 10 MeV, 1 kW electron beam experiments. Shear strengths up to 28 NTa have been measured for Si 3 N 4 -Mo-Si 3 N 4 . These modest strengths are due to beam non-uniformity and the limited area of bonding. The bonding mechanism appears to be a thin silicide reaction layer. Si 3 N 4 -Si 3 N 4 joints with no metal layer were also produced, apparently bonded an yttrium apatite grain boundary phase

  10. Surface plasmon resonance assisted rapid laser joining of glass

    Energy Technology Data Exchange (ETDEWEB)

    Zolotovskaya, Svetlana A.; Tang, Guang; Abdolvand, Amin, E-mail: a.abdolvand@dundee.ac.uk [School of Engineering, Physics and Mathematics, University of Dundee, Dundee DD1 4HN (United Kingdom); Wang, Zengbo [School of Electronic Engineering, Bangor University, Bangor LL57 1UT (United Kingdom)

    2014-08-25

    Rapid and strong joining of clear glass to glass containing randomly distributed embedded spherical silver nanoparticles upon nanosecond pulsed laser irradiation (∼40 ns and repetition rate of 100 kHz) at 532 nm is demonstrated. The embedded silver nanoparticles were ∼30–40 nm in diameter, contained in a thin surface layer of ∼10 μm. A joint strength of 12.5 MPa was achieved for a laser fluence of only ∼0.13 J/cm{sup 2} and scanning speed of 10 mm/s. The bonding mechanism is discussed in terms of absorption of the laser energy by nanoparticles and the transfer of the accumulated localised heat to the surrounding glass leading to the local melting and formation of a strong bond. The presented technique is scalable and overcomes a number of serious challenges for a widespread adoption of laser-assisted rapid joining of glass substrates, enabling applications in the manufacture of microelectronic devices, sensors, micro-fluidic, and medical devices.

  11. Advanced Glycation End-Products affect transcription factors regulating insulin gene expression

    International Nuclear Information System (INIS)

    Puddu, A.; Storace, D.; Odetti, P.; Viviani, G.L.

    2010-01-01

    Advanced Glycation End-Products (AGEs) are generated by the covalent interaction of reducing sugars with proteins, lipids or nucleic acids. AGEs are implicated in diabetic complications and pancreatic β-cell dysfunction. We previously demonstrated that exposure of the pancreatic islet cell line HIT-T15 to high concentrations of AGEs leads to a significant decrease of insulin secretion and content. Insulin gene transcription is positively regulated by the beta cell specific transcription factor PDX-1 (Pancreatic and Duodenal Homeobox-1). On the contrary, the forkhead transcription factor FoxO1 inhibits PDX-1 gene transcription. Activity of FoxO1 is regulated by post-translational modifications: phosphorylation deactivates FoxO1, and acetylation prevents FoxO1 ubiquitination. In this work we investigated whether AGEs affect expression and subcellular localization of PDX-1 and FoxO1. HIT-T15 cells were cultured for 5 days in presence of AGEs. Cells were then lysed and processed for subcellular fractionation. We determined intracellular insulin content, then we assessed the expression and subcellular localization of PDX-1, FoxO1, phosphoFoxO1 and acetylFoxO1. As expected intracellular insulin content was lower in HIT-T15 cells cultured with AGEs. The results showed that AGEs decreased expression and nuclear localization of PDX-1, reduced phosphorylation of FoxO1, and increased expression and acetylation of FoxO1. These results suggest that AGEs decrease insulin content unbalancing transcription factors regulating insulin gene expression.

  12. The Oklahoma Attorney General's Task Force report on the State of End-of-Life Health Care, 2005.

    Science.gov (United States)

    Edmondson, W A Drew

    2005-05-01

    This article includes the recommendations submitted by the 15 members of the Oklahoma Attorney General's Task Force in their Report on the State of End-of-Life Health Care. The task force was created on April 21, 2004, and their report was accepted by Attorney General W.A. Drew Edmondson at a press conference April 11, 2005. It has been forwarded to members of the Oklahoma Legislature, relevant state agencies and organizations with an invitation to join with members of the task force to continue efforts to improve end-of-life care for Oklahomans. Copies of the report are available upon request to the Office of Attorney General.

  13. Advanced manufacturing technologies modern machining, advanced joining, sustainable manufacturing

    CERN Document Server

    2017-01-01

    This book provides details and collective information on working principle, process mechanism, salient features, and unique applications of various advanced manufacturing techniques and processes belong. The book is divided in three sessions covering modern machining methods, advanced repair and joining techniques and, finally, sustainable manufacturing. The latest trends and research aspects of those fields are highlighted.

  14. Cancer and non-cancer risk at low doses of radiation: biological basis of radiation-environment interplay

    International Nuclear Information System (INIS)

    Sasaki, Masao S.

    2013-01-01

    Cancer and non-cancer risk at low doses of ionizing radiation remains poorly defined due to ambiguity at low doses caused by limitations in statistical power and information available on interplay with environment. To deal with these problems, a novel non-parametric statistics was developed based on artificial neural networks theorem and applied to cancer and non-cancer risk in A-bomb survivors. The analysis revealed several unique features at low doses that could not be accounted for by nominal radiation dose alone. They include (1) threshold that varies with organ, gender and age, including cardiovascular diseases, (2) prevalence of infectious diseases, and (3) suppression of pathogenesis of HTLV1. The threshold is unique as it is manifested as negative excess relative risk, a reduction of spontaneous rate at low doses. The response is consistent with currently emerging laboratory data on DNA double-strand break (DSB) repair pathway choice and its sustainability as epigenetic memory in accordance with histone code theory. In response to DSB, of radiation or DNA replication arrest origin, distinct and competitively operating repair pathways are instigated. Activation by low doses of restitution-directed canonical non-homologous end-joining (C-NHEJ) suppresses both error-prone alternative end-joining (Alt-NHEJ) and homologous recombination (HR). The latter two present major pathways to mutagenesis at stalled replication folk associated with endogenous and exogenous genotoxin such as tobacco smoke metabolites and AID-associated somatic hypermutation and class switch recombination in Ig gene. Suppression of these error-prone pathways by low doses of low LET radiation is consistent with the reduction of cancer occurrence by environmental genotoxin, immunodiversity and stable integration of retrovirus DNA, providing a significant modulator of dose linearity at low doses. Whole picture may bring about a new landscape of cancer and non-cancer molecular epidemiology which

  15. HIP joining of Be/CuCrZr for fabrication of ITER first wall

    International Nuclear Information System (INIS)

    Park, Jeong-Yong; Choi, Byung-Kwon; Park, Sang-Yoon; Kim, Hyun-Gil; Kim, Jun-Hwan; Hong, Bong Guen; Jeong, Yong Hwan

    2007-01-01

    The effects of different interlayer types and the temperature have been investigated in order to optimize the HIP joining conditions of Be to CuCrZr. Ti/Cu and Cr/Cu interlayers were coated onto a Be surface before a HIP joining. HIP temperature was changed from 580 to 620 deg. C. No intermetallic compounds or defects were formed at the interface of the joint specimens. The joining strength measured by the four-point bend test and the shear test was higher in the joint specimen with Ti/Cu interlayers when compared to that with Cr/Cu interalyers and it was increased with an increase of the HIP temperature from 580 to 620 deg. C. The yield strength of the aged CuCrZr was higher than 150 MPa at 250 deg. C up to an aging temperature of 620 deg. C after a water quenching. It is therefore suggested that the HIP temperature of the Be/CuCrZr joint could be increased to 620 deg. C based on the results obtained from this study

  16. Mechanical and microstructural integrity of nickel-titanium and stainless steel laser joined wires

    International Nuclear Information System (INIS)

    Vannod, J.; Bornert, M.; Bidaux, J.-E.; Bataillard, L.; Karimi, A.; Drezet, J.-M.; Rappaz, M.; Hessler-Wyser, A.

    2011-01-01

    The biomedical industry shows increasing interest in the joining of dissimilar metals, especially with the aim of developing devices that combine different mechanical and corrosive properties. As an example, nickel-titanium shape memory alloys joined to stainless steel are very promising for new invasive surgery devices, such as guidewires. A fracture mechanics study of such joined wires was carried out using in situ tensile testing and scanning electron microscopy imaging combined with chemical analysis, and revealed an unusual fracture behaviour at superelastic stress. Nanoindentation was performed to determine the mechanical properties of the welded area, which were used as an input for mechanical computation in order to understand this unexpected behaviour. Automated image correlation allowed verification of the mechanical modelling and a reduced stress-strain model is proposed to explain the special fracture mechanism. This study reveals the fact that tremendous property changes at the interface between the NiTi base wire and the weld area have more impact on the ultimate tensile strength than the chemical composition variation across the welded area.

  17. JOINING OF ECONOMIC ENTITIES AS A POSSIBILITY OF SURVIVAL ON THE MODERN MARKET

    Directory of Open Access Journals (Sweden)

    Nikolina Pavičić Rešetar

    2016-12-01

    Full Text Available In today's global market, a major challenge for every economic entity is to be competitive and achieve its objectives. The method of survival shows that those companies with the lowest costs will survive and remain on the market. G. J. Stigler explained this technique on the market share of different sized companies in the steel industry in the United States. In his method he groups enterprises by size (small, medium and large. Historically, it is known that economic entities have been joining together since the Middle Ages. At that time, traders who formed associations were not engaged only in trade, but also in transportation and monetary activity. Economic entities are still joining together today, mostly in the form of a cooperative, cluster, cooperation, public private partnership or holding. The reason for that lies in the reduction of costs, risks and competition and the possibility of increasing the capacity of economic entities. Also, it should be noted that there is a synergy effect when economic entities join together. We are witnesses of many failed businesses, which did not respond to the economic crisis and the needs of the market. The aim of this paper is to try to connect the technique of survival and joining of economic entities and show how a merger is one of the survival models that in conditions of economic crisis provides options to business entities. Only companies with the lowest costs and high-quality technology can survive and compete in today's market.

  18. Increasing FSW join strength by optimizing feed rate, rotating speed and pin angle

    Science.gov (United States)

    Darmadi, Djarot B.; Purnowidodo, Anindito; Siswanto, Eko

    2017-10-01

    Principally the join in Friction Stir Welding (FSW) is formed due to mechanical bonding. At least there are two factors determines the quality of this join, first is the temperature in the area around the interface and secondly the intense of mixing forces in nugget zone to create the mechanical bonding. The adequate temperature creates good flowability of the nugget zone and an intensive mixing force produces homogeneous strong bonding. Based on those two factors in this research the effects of feed rate, rotating speed and pin angle of the FSW process to the tensile strength of resulted join are studied. The true experimental method was used. Feed rate was varied at 24, 42, 55 and 74 mm/minutes and from the experimental results, it can be concluded that the higher feed rate decreases the tensile strength of weld join and it is believed due to the lower heat embedded in the material. Inversely, the higher rotating speed increases the join’s tensile strength as a result of higher heat embedded in base metal and higher mixing force in the nugget zone. The rotating speed were 1842, 2257 and 2904 RPMs. The pin angle determines the direction of mixing force. With variation of pin angle: 0°, 4°, 8° and 12° the higher pin angle generally increases the tensile strength because of more intensive mixing force. For 12° pin angle the lower tensile strength is found since the force tends to push out the nugget area from the joint gap.

  19. CuPb rheocast alloy as joining material for CFC composites

    Science.gov (United States)

    Salvo, M.; Lemoine, P.; Ferraris, M.; Appendino Montorsi, M.; Matera, R.

    1995-10-01

    High heat flux components for future use in thermonuclear fusion reactors are designed as layered structures. The assembling of the different parts (armour, heat sink and external structure) requires a joint which could withstand large heat loads and thermal stresses. In this paper we examined a 50 wt% PbCu rheocast alloy (RCA) as joining material for the armour/heat sink joint. The alloy was prepared in vacuum in a rotational furnace and was characterized by SEM-EDS analysis and heating microscopy. The obtained microstructure was globular as foreseen and it remained after prolonged heating at 650°C. The alloy showed very good ductility: sheets of about 200 μm were rolled starting from about 1 × 1 × 1 cm 3 cubes. The alloy was successful in joining both the armour and the heat sink materials, respectively, carbon fibre reinforced composites and copper. Initial mechanical testing shows that the technique is viable for the foreseen applications in the field of thermonuclear fusion reactors.

  20. Cu-Pb rheocast alloy as joining material for CFC composites

    International Nuclear Information System (INIS)

    Salvo, M.

    1995-01-01

    High heat flux components for future use in thermonuclear fusion reactors are designed as layered structures. The assembling of the different parts (armour, heat sink and external structure) requires a joint which could withstand large heat loads and thermal stresses. In this paper we examined a 50 wt% Pb-Cu rheocast alloy (RCA) as joining material for the armour/heat sink joint. The alloy was prepared in vacuum in a rotational furnace and was characterized by SEM-EDS analysis and heating microscopy. The obtained microstructure was globular as foreseen and it remained after prolonged heating at 650 C. The alloy showed very good ductility: sheets of about 200 μm were rolled starting from about 1x1x1 cm 3 cubes. The alloy was successful in joining both the armour and the heat sink materials, respectively, carbon fibre reinforced composites and copper. Initial mechanical testing shows that the technique is viable for the foreseen applications in the field of thermonuclear fusion reactors. (orig.)