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Sample records for t-dna insertion line

  1. Brachypodium distachyon T-DNA insertion lines: a model pathosystem to study nonhost resistance to wheat stripe rust

    PubMed Central

    An, Tianyue; Cai, Yanli; Zhao, Suzhen; Zhou, Jianghong; Song, Bo; Bux, Hadi; Qi, Xiaoquan

    2016-01-01

    Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most destructive diseases and can cause severe yield losses in many regions of the world. Because of the large size and complexity of wheat genome, it is difficult to study the molecular mechanism of interaction between wheat and PST. Brachypodium distachyon has become a model system for temperate grasses’ functional genomics research. The phenotypic evaluation showed that the response of Brachypodium distachyon to PST was nonhost resistance (NHR), which allowed us to present this plant-pathogen system as a model to explore the immune response and the molecular mechanism underlying wheat and PST. Here we reported the generation of about 7,000 T-DNA insertion lines based on a highly efficient Agrobacterium-mediated transformation system. Hundreds of mutants either more susceptible or more resistant to PST than that of the wild type Bd21 were obtained. The three putative target genes, Bradi5g17540, BdMYB102 and Bradi5g11590, of three T-DNA insertion mutants could be involved in NHR of Brachypodium distachyon to wheat stripe rust. The systemic pathologic study of this T-DNA mutants would broaden our knowledge of NHR, and assist in breeding wheat cultivars with durable resistance. PMID:27138687

  2. Implementing reverse genetics in Rosaceae: analysis of T-DNA flanking sequences of insertional mutant lines in the diploid strawberry, Fragaria vesca.

    PubMed

    Oosumi, Teruko; Ruiz-Rojas, Juan Jairo; Veilleux, Richard E; Dickerman, Allan; Shulaev, Vladimir

    2010-09-01

    Reverse genetics is used for functional genomics research in model plants. To establish a model system for the systematic reverse genetics research in the Rosaceae family, we analyzed genomic DNA flanking the T-DNA insertions in 191 transgenic plants of the diploid strawberry, Fragaria vesca. One hundred and seventy-six T-DNA flanking sequences were amplified from the right border (RB) and 37 from the left border (LB) by thermal asymmetric interlaced PCR. Analysis of the T-DNA nick positions revealed that T-DNA was most frequently nicked at the cleavage sites. Analysis of 11 T-DNA integration sites indicated that T-DNA was integrated into the F. vesca genome by illegitimate recombination, as reported in other model plants: Arabidopsis, rice and tobacco. First, deletion of DNA was found at T-DNA integration target sites in all transgenic plants tested. Second, microsimilarities of a few base pairs between the left and/or right ends of the T-DNA and genomic sites were found in all transgenic plants tested. Finally, filler DNA was identified in four break-points. Out of 191 transgenic plants, T-DNA flanking sequences of 79 plants (41%) showed significant similarity to genes, elements or proteins of other plant species and 67 (35%) of the sequences are still unknown strawberry gene fragments. T-DNA flanking sequences of 126 plants (66%) showed homology to plant ESTs. This is the first report of T-DNA integration in a sizeable population of a rosaceous species. We have shown in this paper that T-DNA integration in strawberry is not random but directed by sequence microsimilarities in the host genome.

  3. Embryonic Lethals and T-DNA Insertional Mutagenesis in Arabidopsis.

    PubMed Central

    Errampalli, D; Patton, D; Castle, L; Mickelson, L; Hansen, K; Schnall, J; Feldmann, K; Meinke, D

    1991-01-01

    T-DNA insertional mutagenesis represents a promising approach to the molecular isolation of genes with essential functions during plant embryo development. We describe in this report the isolation and characterization of 18 mutants of Arabidopsis thaliana defective in embryo development following seed transformation with Agrobacterium tumefaciens. Random T-DNA insertion was expected to result in a high frequency of recessive embryonic lethals because many target genes are required for embryogenesis. The cointegrate Ti plasmid used in these experiments contained the nopaline synthase and neomycin phosphotransferase gene markers. Nopaline assays and resistance to kanamycin were used to estimate the number of functional inserts present in segregating families. Nine families appeared to contain a T-DNA insert either within or adjacent to the mutant gene. Eight families were clearly not tagged with a functional insert and appeared instead to contain mutations induced during the transformation process. DNA gel blot hybridization with internal and right border probes revealed a variety of rearrangements associated with T-DNA insertion. A general strategy is presented to simplify the identification of tagged embryonic mutants and facilitate the molecular isolation of genes required for plant embryogenesis. PMID:12324593

  4. Intronic T-DNA insertion in Arabidopsis NBR1 conditionally affects wild-type transcript level

    PubMed Central

    Rodríguez, Milagros Collados; Wawrzyńska, Anna; Sirko, Agnieszka

    2014-01-01

    Abstract The SALK_135513 line of Arabidopsis thaliana is annotated by GenBank to have the T-DNA insertion in the fourth exon of NBR1 (At4g24690). Careful molecular analyses of the homozygous plants of SALK_135513 line indicated the place of T-DNA insertion in the fourth intron. Unexpectedly, 2 kinds of NBR1 transcripts, the wild-type and the mutated, resulting from alternative splicing events, were detected in those plants. Our findings explain the problems encountered by us with phenotypic evaluation of this line and emphasize the necessity for independent verification of the exact insertion site followed by careful expression studies when working with Arabidopsis T-DNA insertional mutants. PMID:25482782

  5. Intronic T-DNA insertion in Arabidopsis NBR1 conditionally affects wild-type transcript level.

    PubMed

    Rodríguez, Milagros Collados; Wawrzyńska, Anna; Sirko, Agnieszka

    2014-01-01

    The SALK_135513 line of Arabidopsis thaliana is annotated by GenBank to have the T-DNA insertion in the fourth exon of NBR1 (At4g24690). Careful molecular analyses of the homozygous plants of SALK_135513 line indicated the place of T-DNA insertion in the fourth intron. Unexpectedly, 2 kinds of NBR1 transcripts, the wild-type and the mutated, resulting from alternative splicing events, were detected in those plants. Our findings explain the problems encountered by us with phenotypic evaluation of this line and emphasize the necessity for independent verification of the exact insertion site followed by careful expression studies when working with Arabidopsis T-DNA insertional mutants.

  6. A User’s Guide to the Arabidopsis T-DNA Insertional Mutant Collections

    PubMed Central

    O'Malley, Ronan C.; Barragan, Cesar C.; Ecker, Joseph R.

    2016-01-01

    Summary The T-DNA sequence-indexed mutant collections contain insertional mutants for most Arabidopsis thaliana genes and have played an important role in plant biology research for almost two decades. By providing a large source of mutant alleles for in vivo characterization of gene function, this resource has been leveraged thousands of times to study a wide-range of problems in plant biology. Our primary goal in this chapter is to provide a general guide to strategies for the effective use of the data and materials in these collections. To do this, we provide a general introduction to the T-DNA insertional sequence-indexed mutant collections with a focus on how best to use the available data sources for good line selection. As isolation of a homozygous line is a common next step once a potential disruption line has been identified, the second half of the chapter will provide a step-by-step guide for the design and implementation of a T-DNA genotyping pipeline. Finally, we describe interpretation of genotyping results and include a troubleshooting section for common types of segregation distortions that we have observed. In this chapter we introduce both basic concepts and specific applications to new and more experienced users of the collections for the design and implementation of small- to large-scale genotyping pipelines. PMID:25757780

  7. A user's guide to the Arabidopsis T-DNA insertion mutant collections.

    PubMed

    O'Malley, Ronan C; Barragan, Cesar C; Ecker, Joseph R

    2015-01-01

    The T-DNA sequence-indexed mutant collections contain insertional mutants for most Arabidopsis thaliana genes and have played an important role in plant biology research for almost two decades. By providing a large source of mutant alleles for in vivo characterization of gene function, this resource has been leveraged thousands of times to study a wide range of problems in plant biology. Our primary goal in this chapter is to provide a general guide to strategies for the effective use of the data and materials in these collections. To do this, we provide a general introduction to the T-DNA insertional sequence-indexed mutant collections with a focus on how best to use the available data sources for good line selection. As isolation of a homozygous line is a common next step once a potential disruption line has been identified, the second half of the chapter provides a step-by-step guide for the design and implementation of a T-DNA genotyping pipeline. Finally, we describe interpretation of genotyping results and include a troubleshooting section for common types of segregation distortions that we have observed. In this chapter we introduce both basic concepts and specific applications to both new and more experienced users of the collections for the design and implementation of small- to large-scale genotyping pipelines.

  8. T-DNA integration into the Arabidopsis genome depends on sequences of pre-insertion sites

    PubMed Central

    Brunaud, Véronique; Balzergue, Sandrine; Dubreucq, Bertrand; Aubourg, Sébastien; Samson, Franck; Chauvin, Stéphanie; Bechtold, Nicole; Cruaud, Corinne; DeRose, Richard; Pelletier, Georges; Lepiniec, Loïc; Caboche, Michel; Lecharny, Alain

    2002-01-01

    A statistical analysis of 9000 flanking sequence tags characterizing transferred DNA (T-DNA) transformants in Arabidopsis sheds new light on T-DNA insertion by illegitimate recombination. T-DNA integration is favoured in plant DNA regions with an A-T-rich content. The formation of a short DNA duplex between the host DNA and the left end of the T-DNA sets the frame for the recombination. The sequence immediately downstream of the plant A-T-rich region is the master element for setting up the DNA duplex, and deletions into the left end of the integrated T-DNA depend on the location of a complementary sequence on the T-DNA. Recombination at the right end of the T-DNA with the host DNA involves another DNA duplex, 2–3 base pairs long, that preferentially includes a G close to the right end of the T-DNA. PMID:12446565

  9. Distribution and characterization of more than 1000 T-DNA tags in the genome of Brachypodium distachyon community standard line Bd21.

    PubMed

    Thole, Vera; Worland, Barbara; Wright, Jonathan; Bevan, Michael W; Vain, Philippe

    2010-08-01

    A collection of 4117 fertile T-DNA lines has been generated by Agrobacterium-mediated transformation of the diploid community standard line Bd21 of Brachypodium distachyon. The regions flanking the T-DNA left and right borders of the first 741 transformed plants were isolated by adapter-ligation PCR and sequenced. A total of 1005 genomic sequences (representing 44.1% of all flanking sequences retrieved) characterized 660 independent T-DNA loci assigned to a unique location in the Brachypodium genome sequence. Seventy-six percent of the fertile plant lines contained at least one anchored T-DNA locus (1.17 loci per tagged line on average). Analysis of the regions flanking both borders of the T-DNA increased the number of T-DNA loci tagged and the number of tagged lines by approximately 50% when compared to a single border analysis. T-DNA integration (2.4 insertions per Mb on average) was proportional to chromosome size, however, varied greatly along each chromosome with often low insertion level around centromeres. The frequency of insertion within transposable elements (5.3%) was fivefold lower than expected if random insertion would have occurred. More than half of the T-DNAs inserted in genic regions. On average, one gene could be tagged for every second fertile plant line produced and more than one plant line out of three contained a T-DNA insertion directly within or 500 bp around the coding sequence. Approximately, 60% of the genes tagged corresponded to expressed genes. The T-DNA lines generated by the BrachyTAG programme are available as a community resource and have been distributed internationally since 2008 via the BrachyTAG.org web site.

  10. Bidirectional promoter trapping T-DNA for insertional mutagenesis in Verticillium dahliae.

    PubMed

    Deng, Sheng; Wang, Cai-yue; Zhang, Xin; Lin, Ling

    2014-07-01

    Transfer DNA (T-DNA)-based random insertional mutagenesis is a universal forward genetic approach for gene identification and cloning in many phytopathogenic fungi. In a large number of randomly selected transformants, screening for mutants with a specific phenotype is laborious, especially for pathogenicity-defective mutants. To accelerate mutant screening and gene identification, a bidirectional promoter-trapping Ti binary vector, 1300-bisGFP-hyg, was constructed and deployed in this study. More than 6000 Verticillium dahliae transformants were obtained by the mediation of Agrobacterium tumefaciens carrying the vector. One thousand randomly selected transformants were cultured on Czapek-Dox and on Czapek-Dox plus cotton root extract media plates. The cultured transformants with green fluorescent protein (GFP) expression or changes in phenotype were selected and used in virulence or promoter-trapping assays. Based on the virulence assay of 60 transformants, the pathogenicity of 17 of these mutants was compromised. Ten pathogenicity-defective mutants were found with GFP expression, and 6 with expression in Czapek-Dox plus cotton root extract media specifically. Using TAIL-PCR (thermal asymmetric interlaced polymerase chain reaction), the T-DNA insertion sites were identified in 8 GFP-expressing transformants, including 5 pathogenicity-defective mutants and 3 unaffected transformants. Promoters of 6 genes were successfully trapped using the T-DNA method in this study. The nonpathogenic transformant 24C9 was the subject of additional investigation. It displayed strong GFP expression on water agar medium supplemented with cotton root extracts and on cotton seedling stems. The results obtained by Southern blot and quantitative real-time PCR confirmed that the transcription level of VdUGPU (encoding UTP-glucose-1-phosphate uridylyltransferase) was significantly reduced owing to T-DNA insertion in the gene promoter region. These results indicate that the bidirectional

  11. The Structural Features of Thousands of T-DNA Insertion Sites Are Consistent with a Double-Strand Break Repair-Based Insertion Mechanism.

    PubMed

    Kleinboelting, Nils; Huep, Gunnar; Appelhagen, Ingo; Viehoever, Prisca; Li, Yong; Weisshaar, Bernd

    2015-11-02

    Transformation by Agrobacterium tumefaciens, an important tool in modern plant research, involves the integration of T-DNA initially present on a plasmid in agrobacteria into the genome of plant cells. The process of attachment of the agrobacteria to plant cells and the transport of T-DNA into the cell and further to the nucleus has been well described. However, the exact mechanism of integration into the host's DNA is still unclear, although several models have been proposed. During confirmation of T-DNA insertion alleles from the GABI-Kat collection of Arabidopsis thaliana mutants, we have generated about 34,000 sequences from the junctions between inserted T-DNA and adjacent genome regions. Here, we describe the evaluation of this dataset with regard to existing models for T-DNA integration. The results suggest that integration into the plant genome is mainly mediated by the endogenous plant DNA repair machinery. The observed integration events showed characteristics highly similar to those of repair sites of double-strand breaks with respect to microhomology and deletion sizes. In addition, we describe unexpected integration events, such as large deletions and inversions at the integration site that are relevant for correct interpretation of results from T-DNA insertion mutants in reverse genetics experiments.

  12. Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition

    PubMed Central

    Kawano, Noriaki; Kiuchi, Fumiyuki; Kawahara, Nobuo; Yoshimatsu, Kayo

    2012-01-01

    The in vitro shoot culture of a T-DNA insertional mutant of Papaver somniferum L. established by the infection of Agrobacterium rhizogenes MAFF03-01724 accumulated thebaine instead of morphine as a major opium alkaloid. To develop a non-narcotic opium poppy and to gain insight into its genetic background, we have transplanted this mutant to soil, and analyzed its alkaloid content along with the manner of inheritance of T-DNA insertion loci among its selfed progenies. In the transplanted T0 primary mutant, the opium (latex) was found to be rich in thebaine (16.3% of dried opium) by HPLC analysis. The analyses on T-DNA insertion loci by inverse PCR, adaptor-ligation PCR, and quantitative real-time PCR revealed that as many as 18 copies of T-DNAs were integrated into a poppy genome in a highly complicated manner. The number of copies of T-DNAs was decreased to seven in the selected T3 progenies, in which the average thebaine content was 2.4-fold that of the wild type plant. This may indicate that the high thebaine phenotype was increasingly stabilized as the number of T-DNA copies was decreased. In addition, by reverse transcription PCR analysis on selected morphine biosynthetic genes, the expression of codeine 6-O-demethylase was clearly shown to be diminished in the T0 in vitro shoot culture, which can be considered as one of the key factors of altered alkaloid composition. PMID:24288085

  13. FLAGdb/FST: a database of mapped flanking insertion sites (FSTs) of Arabidopsis thaliana T-DNA transformants.

    PubMed

    Samson, F; Brunaud, V; Balzergue, S; Dubreucq, B; Lepiniec, L; Pelletier, G; Caboche, M; Lecharny, A

    2002-01-01

    A large collection of T-DNA insertion transformants of Arabidopsis thaliana has been generated at the Institute of Agronomic Research, Versailles, France. The molecular characterisation of the insertion sites is currently performed by sequencing genomic regions flanking the inserted T-DNA (FST). The almost complete sequence of the nuclear genome of A.thaliana provides the framework for organising FSTs in a genome oriented database, FLAGdb/FST (http://genoplante-info.infobiogen.fr). The main scope of FLAGdb/FST is to help biologists to find the FSTs that interrupt the genes in which they are interested. FSTs are anchored to the genome sequences of A.thaliana and positions of both predicted genes and FSTs are shown graphically on sequences. Requests to locate the genomic position of a query sequence are made using BLAST programs. The response delivered by FLAGdb/FST is a graphical representation of the putative FSTs and of predicted genes in a 20 kb region.

  14. The use of T-DNA insertional mutagenesis to improve cellulase production by the thermophilic fungus Humicola insolens Y1.

    PubMed

    Xu, Xinxin; Li, Jinyang; Shi, Pengjun; Ji, Wangli; Liu, Bo; Zhang, Yuhong; Yao, Bin; Fan, Yunliu; Zhang, Wei

    2016-08-10

    Humicola insolens is an excellent producer of pH-neutral active, thermostable cellulases that find many industrial applications. In the present study, we developed an efficient Agrobacterium tumefaciens-mediated transformation system for H. insolens. We transformed plasmids carrying the promoter of the glyceraldehyde-3-phosphate dehydrogenase gene of H. insolens driving the transcription of genes encoding neomycin phosphotransferase, hygromycin B phosphotransferase, and enhanced green fluorescent protein. We optimized transformation efficiency to obtain over 300 transformants/10(6) conidia. T-DNA insertional mutagenesis was employed to generate an H. insolens mutant library, and we isolated a transformant termed T4 with enhanced cellulase and hemicellulase activities. The FPase, endoglucanase, cellobiohydrolase, β-glucosidase, and xylanase activities of T4, measured at the end of fermentation, were 60%, 440%, 320%, 41%, and 81% higher than those of the wild-type strain, respectively. We isolated the sequences flanking the T-DNA insertions and thus identified new genes potentially involved in cellulase and hemicellulase production. Our results show that it is feasible to use T-DNA insertional mutagenesis to identify novel candidate genes involved in cellulase production. This will be valuable when genetic improvement programs seeking to enhance cellulase production are planned, and will also allow us to gain a better understanding of the genetics of the thermophilic fungus H. insolens.

  15. The use of T-DNA insertional mutagenesis to improve cellulase production by the thermophilic fungus Humicola insolens Y1

    PubMed Central

    Xu, Xinxin; Li, Jinyang; Shi, Pengjun; Ji, Wangli; Liu, Bo; Zhang, Yuhong; Yao, Bin; Fan, Yunliu; Zhang, Wei

    2016-01-01

    Humicola insolens is an excellent producer of pH-neutral active, thermostable cellulases that find many industrial applications. In the present study, we developed an efficient Agrobacterium tumefaciens-mediated transformation system for H. insolens. We transformed plasmids carrying the promoter of the glyceraldehyde-3-phosphate dehydrogenase gene of H. insolens driving the transcription of genes encoding neomycin phosphotransferase, hygromycin B phosphotransferase, and enhanced green fluorescent protein. We optimized transformation efficiency to obtain over 300 transformants/106 conidia. T-DNA insertional mutagenesis was employed to generate an H. insolens mutant library, and we isolated a transformant termed T4 with enhanced cellulase and hemicellulase activities. The FPase, endoglucanase, cellobiohydrolase, β-glucosidase, and xylanase activities of T4, measured at the end of fermentation, were 60%, 440%, 320%, 41%, and 81% higher than those of the wild-type strain, respectively. We isolated the sequences flanking the T-DNA insertions and thus identified new genes potentially involved in cellulase and hemicellulase production. Our results show that it is feasible to use T-DNA insertional mutagenesis to identify novel candidate genes involved in cellulase production. This will be valuable when genetic improvement programs seeking to enhance cellulase production are planned, and will also allow us to gain a better understanding of the genetics of the thermophilic fungus H. insolens. PMID:27506519

  16. Characterization of T-DNA insertion mutants with decreased virulence in the entomopathogenic fungus Beauveria bassiana JEF-007.

    PubMed

    Kim, Sihyeon; Lee, Se Jin; Nai, Yu-Shin; Yu, Jeong Seon; Lee, Mi Rong; Yang, Yi-Ting; Kim, Jae Su

    2016-10-01

    The bean bug, Riptortus pedestris, is a major agricultural pest that reduces crop quality and value. Chemical pesticides have contributed to pest management, but resistance to these chemicals has significantly limited their use. Alternative strategies with different modes of action, such as entomopathogenic fungi, are therefore of great interest. Herein, we explored how entomopathogenic fungi can potentially be used to control the bean bug and focused on identifying virulence-related genes. Beauveria bassiana (JEF isolates) were assayed against bean bugs under laboratory conditions. One isolate, JEF-007, showed >80 % virulence by both spray and contact exposure methods. Agrobacterium tumefaciens-mediated transformation (AtMT) of JEF-007 generated 249 random transformants, two of which (B1-06 and C1-49) showed significantly reduced virulence against Tenebrio molitor and R. pedestris immatures. Both species were used for rapid screening of virulence-reduced mutants. The two transformants had different morphologies, conidial production, and thermotolerance than the wild type. To determine the localization of the randomly inserted T-DNA, thermal asymmetric interlaced (TAIL) PCR was conducted and analysis of the two clones found multiple T-DNA insertions (two in B1-06 and three in C1-49). Genes encoding complex I intermediate-associated protein 30 (CIA30) and the autophagy protein (Atg22) were possibly disrupted by the T-DNA insertion and might be involved in the virulence. This work provides a strong platform for future functional genetic studies of bean bug-pathogenic B. bassiana. The genes putatively involved in fungal virulence should be experimentally validated by knockdown in future studies.

  17. Number and accuracy of T-DNA insertions in transgenic banana (Musa spp.) plants characterized by an improved anchored PCR technique.

    PubMed

    Pérez-Hernández, Juan Bernardo; Swennen, Rony; Sági, László

    2006-04-01

    Nineteen transgenic banana plants, produced via Agrobacterium-mediated transformation, were analyzed for the integration of T-DNA border regions using an improved anchored PCR technique. The method described is a relatively fast, three-step procedure (restriction digestion of genomic DNA, ligation of 'vectorette'-type adaptors, and a single round of suppression PCR) for the amplification of specific T-DNA border-containing genomic fragments. Most transgenic plants carried a low number of inserts and the method was suitable for a detailed characterization of the integration events, including T-DNA border integrity as well as the insertion of non-T-DNA vector sequences, which occurred in 26% of the plants. Furthermore, the particular band pattern generated by four enzyme/primer combinations for each individual plant served as a fingerprint, allowing the identification of plants representing identical transformation events. Genomic Southern hybridization and nucleotide sequence analysis of amplification products confirmed the data obtained by anchored PCR. Sequencing of seven right or left border junction regions revealed different T-DNA processing events for each plant, indicating a relatively low frequency of precisely nicked T-DNA integration among the plants studied.

  18. Deep sequencing of the ancestral tobacco species Nicotiana tomentosiformis reveals multiple T-DNA inserts and a complex evolutionary history of natural transformation in the genus Nicotiana.

    PubMed

    Chen, Ke; Dorlhac de Borne, François; Szegedi, Ernö; Otten, Léon

    2014-11-01

    Nicotiana species carry cellular T-DNA sequences (cT-DNAs), acquired by Agrobacterium-mediated transformation. We characterized the cT-DNA sequences of the ancestral Nicotiana tabacum species Nicotiana tomentosiformis by deep sequencing. N. tomentosiformis contains four cT-DNA inserts derived from different Agrobacterium strains. Each has an incomplete inverted-repeat structure. TA is similar to part of the Agrobacterium rhizogenes 1724 mikimopine-type T-DNA, but has unusual orf14 and mis genes. TB carries a 1724 mikimopine-type orf14-mis fragment and a mannopine-agropine synthesis region (mas2-mas1-ags). The mas2' gene codes for an active enzyme. TC is similar to the left part of the A. rhizogenes A4 T-DNA, but also carries octopine synthase-like (ocl) and c-like genes normally found in A. tumefaciens. TD shows a complex rearrangement of T-DNA fragments similar to the right end of the A4 TL-DNA, and including an orf14-like gene and a gene with unknown function, orf511. The TA, TB, TC and TD insertion sites were identified by alignment with N. tabacum and Nicotiana sylvestris sequences. The divergence values for the TA, TB, TC and TD repeats provide an estimate for their relative introduction times. A large deletion has occurred in the central part of the N. tabacum cv. Basma/Xanthi TA region, and another deletion removed the complete TC region in N. tabacum. Nicotiana otophora lacks TA, TB and TD, but contains TC and another cT-DNA, TE. This analysis, together with that of Nicotiana glauca and other Nicotiana species, indicates multiple sequential insertions of cT-DNAs during the evolution of the genus Nicotiana.

  19. Identification of pathogenicity-related genes in the vascular wilt fungus Verticillium dahliae by Agrobacterium tumefaciens-mediated T-DNA insertional mutagenesis.

    PubMed

    Maruthachalam, K; Klosterman, S J; Kang, S; Hayes, R J; Subbarao, K V

    2011-11-01

    Verticillium dahliae is the causal agent of vascular wilt in many economically important crops worldwide. Identification of genes that control pathogenicity or virulence may suggest targets for alternative control methods for this fungus. In this study, Agrobacterium tumefaciens-mediated transformation (ATMT) was applied for insertional mutagenesis of V. dahliae conidia. Southern blot analysis indicated that T-DNAs were inserted randomly into the V. dahliae genome and that 69% of the transformants were the result of single copy T-DNA insertion. DNA sequences flanking T-DNA insertion were isolated through inverse PCR (iPCR), and these sequences were aligned to the genome sequence to identify the genomic position of insertion. V. dahliae mutants of particular interest selected based on culture phenotypes included those that had lost the ability to form microsclerotia and subsequently used for virulence assay. Based on the virulence assay of 181 transformants, we identified several mutant strains of V. dahliae that did not cause symptoms on lettuce plants. Among these mutants, T-DNA was inserted in genes encoding an endoglucanase 1 (VdEg-1), a hydroxyl-methyl glutaryl-CoA synthase (VdHMGS), a major facilitator superfamily 1 (VdMFS1), and a glycosylphosphatidylinositol (GPI) mannosyltransferase 3 (VdGPIM3). These results suggest that ATMT can effectively be used to identify genes associated with pathogenicity and other functions in V. dahliae.

  20. Localization of single-copy T-DNA insertion in transgenic shallots (Allium cepa) by using ultra-sensitive FISH with tyramide signal amplification.

    PubMed

    Khrustaleva, L I; Kik, C

    2001-03-01

    The sensitivity of fluorescence in situ hybridization (FISH) for mapping plant chromosomes of single-copy DNA sequences is limited. We have adapted for plant cytogenetics a new signal-amplification method termed tyramide-FISH (Tyr-FISH). Until present this technique has only been applied to human chromosomes. The method is based on enzymatic deposition of fluorochrome-conjugated tyramide. With Tyr-FISH it was possible to detect target T-DNA sequences on plant metaphase chromosomes as small as 710 bp without using a cooled CCD camera. Short detection time and high sensitivity, in combination with a low background, make the Tyr-FISH method very suitable for routine application in plant cytogenetic research. With Tyr-FISH we analysed the position of T-DNA inserts in transgenic shallots. We found that the inserts were preferentially located in the distal region of metaphase chromosomes. Sequential fluorescence in situ hybridization with a 375 bp satellite sequence suggested that a specific T-DNA insert was located within the satellite sequence hybridization region on a metaphase chromosome. Analysis of less-condensed prophase and interphase chromosomes revealed that the T-DNA was integrated outside the satellite DNA-hybridization region in a more proximal euchromatin region.

  1. Comparison and Validation of Putative Pathogenicity-Related Genes Identified by T-DNA Insertional Mutagenesis and Microarray Expression Profiling in Magnaporthe oryzae

    PubMed Central

    Wáng, Ying; Tan, Qi; Gao, Ying Nv; Li, Yan

    2017-01-01

    High-throughput technologies of functional genomics such as T-DNA insertional mutagenesis and microarray expression profiling have been employed to identify genes related to pathogenicity in Magnaporthe oryzae. However, validation of the functions of individual genes identified by these high-throughput approaches is laborious. In this study, we compared two published lists of genes putatively related to pathogenicity in M. oryzae identified by T-DNA insertional mutagenesis (comprising 1024 genes) and microarray expression profiling (comprising 236 genes), respectively, and then validated the functions of some overlapped genes between the two lists by knocking them out using the method of target gene replacement. Surprisingly, only 13 genes were overlapped between the two lists, and none of the four genes selected from the overlapped genes exhibited visible phenotypic changes on vegetative growth, asexual reproduction, and infection ability in their knockout mutants. Our results suggest that both of the lists might contain large proportions of unrelated genes to pathogenicity and therefore comparing the two gene lists is hardly helpful for the identification of genes that are more likely to be involved in pathogenicity as we initially expected. PMID:28286772

  2. Comparison and Validation of Putative Pathogenicity-Related Genes Identified by T-DNA Insertional Mutagenesis and Microarray Expression Profiling in Magnaporthe oryzae.

    PubMed

    Wang, Ying; Wáng, Ying; Tan, Qi; Gao, Ying Nv; Li, Yan; Bao, Da Peng

    2017-01-01

    High-throughput technologies of functional genomics such as T-DNA insertional mutagenesis and microarray expression profiling have been employed to identify genes related to pathogenicity in Magnaporthe oryzae. However, validation of the functions of individual genes identified by these high-throughput approaches is laborious. In this study, we compared two published lists of genes putatively related to pathogenicity in M. oryzae identified by T-DNA insertional mutagenesis (comprising 1024 genes) and microarray expression profiling (comprising 236 genes), respectively, and then validated the functions of some overlapped genes between the two lists by knocking them out using the method of target gene replacement. Surprisingly, only 13 genes were overlapped between the two lists, and none of the four genes selected from the overlapped genes exhibited visible phenotypic changes on vegetative growth, asexual reproduction, and infection ability in their knockout mutants. Our results suggest that both of the lists might contain large proportions of unrelated genes to pathogenicity and therefore comparing the two gene lists is hardly helpful for the identification of genes that are more likely to be involved in pathogenicity as we initially expected.

  3. Lack of Genotype and Phenotype Correlation in a Rice T-DNA Tagged Line Is Likely Caused by Introgression in the Seed Source

    PubMed Central

    Wei, Fu-Jin; Tsai, Yuan-Ching; Hsu, Yu-Ming; Chen, Yu-An; Huang, Ching-Ting; Wu, Hshin-Ping; Huang, Lin-Tzu; Lai, Ming-Hsin; Kuang, Lin-Yun; Lo, Shuen-Fang; Yu, Su-May; Lin, Yann-Rong; Hsing, Yue-Ie Caroline

    2016-01-01

    Rice (Oryza sativa) is one of the most important crops in the world. Several rice insertional mutant libraries are publicly available for systematic analysis of gene functions. However, the tagging efficiency of these mutant resources–the relationship between genotype and phenotype–is very low. We used whole-genome sequencing to analyze a T-DNA–tagged transformant from the Taiwan Rice Insertional Mutants (TRIM) resource. The phenomics records for M0028590, one of the TRIM lines, revealed three phenotypes–wild type, large grains, and tillering dwarf–in the 12 T1 plants. Using the sequencing data for 7 plants from three generations of this specific line, we demonstrate that introgression from an indica rice variety might occur in one generation before the seed was used for callus generation and transformation of this line. In addition, the large-grain trait came from the GS3 gene of the introgressed region and the tillering dwarf phenotype came from a single nucleotide change in the D17 gene that occurred during the callus induction to regeneration of the transformant. As well, another regenerant showed completely heterozygous single-nucleotide polymorphisms across the whole genome. In addition to the known sequence changes such as T-DNA integration, single nucleotide polymorphism, insertion, deletion, chromosome rearrangement and doubling, spontaneous outcrossing occurred in the rice field may also explain some mutated traits in a tagged mutant population. Thus, the co-segregation of an integration event and the phenotype should be checked when using these mutant populations. PMID:27186981

  4. Cell line fingerprinting using retroelement insertion polymorphism.

    PubMed

    Ustyugova, Svetlana V; Amosova, Anna L; Lebedev, Yuri B; Sverdlov, Eugene D

    2005-04-01

    Human cell lines are an indispensable tool for functional studies of living entities in their numerous manifestations starting with integral complex systems such as signal pathways and networks, regulation of gene ensembles, epigenetic factors, and finishing with pathological changes and impact of artificially introduced elements, such as various transgenes, on the behavior of the cell. Therefore, it is highly desirable to have reliable cell line identification techniques to make sure that the cell lines to be used in experiments are exactly what is expected. To this end, we developed a set of informative markers based on insertion polymorphism of human retroelements (REs). The set includes 47 pairs of PCR primers corresponding to introns of the human genes with dimorphic LINE1 (L1) and Alu insertions. Using locus-specific PCR assays, we have genotyped 10 human cell lines of various origins. For each of these cell lines, characteristic fingerprints were obtained. An estimated probability that two different cell lines possess the same marker genotype is about 10-18. Therefore, the proposed set of markers provides a reliable tool for cell line identification.

  5. The widely used Nicotiana benthamiana 16c line has an unusual T-DNA integration pattern including a transposon sequence

    PubMed Central

    Lorenc, Michał T.; Dudley, Kevin J.; Hellens, Roger P.

    2017-01-01

    Nicotiana benthamiana is employed around the world for many types of research and one transgenic line has been used more extensively than any other. This line, 16c, expresses the Aequorea victoria green fluorescent protein (GFP), highly and constitutively, and has been a major resource for visualising the mobility and actions of small RNAs. Insights into the mechanisms studied at a molecular level in N. benthamiana 16c are likely to be deeper and more accurate with a greater knowledge of the GFP gene integration site. Therefore, using next generation sequencing, genome mapping and local alignment, we identified the location and characteristics of the integrated T-DNA. As suggested from previous molecular hybridisation and inheritance data, the transgenic line contains a single GFP-expressing locus. However, the GFP coding sequence differs from that originally reported. Furthermore, a 3.2 kb portion of a transposon, appears to have co-integrated with the T-DNA. The location of the integration mapped to a region of the genome represented by Nbv0.5scaffold4905 in the www.benthgenome.com assembly, and with less integrity to Niben101Scf03641 in the www.solgenomics.net assembly. The transposon is not endogenous to laboratory strains of N. benthamiana or Agrobacterium tumefaciens strain GV3101 (MP90), which was reportedly used in the generation of line 16c. However, it is present in the popular LBA4404 strain. The integrated transposon sequence includes its 5’ terminal repeat and a transposase gene, and is immediately adjacent to the GFP gene. This unexpected genetic arrangement may contribute to the characteristics that have made the 16c line such a popular research tool and alerts researchers, taking transgenic plants to commercial release, to be aware of this genomic hitchhiker. PMID:28231340

  6. The widely used Nicotiana benthamiana 16c line has an unusual T-DNA integration pattern including a transposon sequence.

    PubMed

    Philips, Joshua G; Naim, Fatima; Lorenc, Michał T; Dudley, Kevin J; Hellens, Roger P; Waterhouse, Peter M

    2017-01-01

    Nicotiana benthamiana is employed around the world for many types of research and one transgenic line has been used more extensively than any other. This line, 16c, expresses the Aequorea victoria green fluorescent protein (GFP), highly and constitutively, and has been a major resource for visualising the mobility and actions of small RNAs. Insights into the mechanisms studied at a molecular level in N. benthamiana 16c are likely to be deeper and more accurate with a greater knowledge of the GFP gene integration site. Therefore, using next generation sequencing, genome mapping and local alignment, we identified the location and characteristics of the integrated T-DNA. As suggested from previous molecular hybridisation and inheritance data, the transgenic line contains a single GFP-expressing locus. However, the GFP coding sequence differs from that originally reported. Furthermore, a 3.2 kb portion of a transposon, appears to have co-integrated with the T-DNA. The location of the integration mapped to a region of the genome represented by Nbv0.5scaffold4905 in the www.benthgenome.com assembly, and with less integrity to Niben101Scf03641 in the www.solgenomics.net assembly. The transposon is not endogenous to laboratory strains of N. benthamiana or Agrobacterium tumefaciens strain GV3101 (MP90), which was reportedly used in the generation of line 16c. However, it is present in the popular LBA4404 strain. The integrated transposon sequence includes its 5' terminal repeat and a transposase gene, and is immediately adjacent to the GFP gene. This unexpected genetic arrangement may contribute to the characteristics that have made the 16c line such a popular research tool and alerts researchers, taking transgenic plants to commercial release, to be aware of this genomic hitchhiker.

  7. High-throughput analysis of T-DNA location and structure using sequence capture

    SciTech Connect

    Inagaki, Soichi; Henry, Isabelle M.; Lieberman, Meric C.; Comai, Luca

    2015-10-07

    Agrobacterium-mediated transformation of plants with T-DNA is used both to introduce transgenes and for mutagenesis. Conventional approaches used to identify the genomic location and the structure of the inserted T-DNA are laborious and high-throughput methods using next-generation sequencing are being developed to address these problems. Here, we present a cost-effective approach that uses sequence capture targeted to the T-DNA borders to select genomic DNA fragments containing T-DNA—genome junctions, followed by Illumina sequencing to determine the location and junction structure of T-DNA insertions. Multiple probes can be mixed so that transgenic lines transformed with different T-DNA types can be processed simultaneously, using a simple, index-based pooling approach. We also developed a simple bioinformatic tool to find sequence read pairs that span the junction between the genome and T-DNA or any foreign DNA. We analyzed 29 transgenic lines of Arabidopsis thaliana, each containing inserts from 4 different T-DNA vectors. We determined the location of T-DNA insertions in 22 lines, 4 of which carried multiple insertion sites. Additionally, our analysis uncovered a high frequency of unconventional and complex T-DNA insertions, highlighting the needs for high-throughput methods for T-DNA localization and structural characterization. Transgene insertion events have to be fully characterized prior to use as commercial products. As a result, our method greatly facilitates the first step of this characterization of transgenic plants by providing an efficient screen for the selection of promising lines.

  8. High-throughput analysis of T-DNA location and structure using sequence capture

    DOE PAGES

    Inagaki, Soichi; Henry, Isabelle M.; Lieberman, Meric C.; ...

    2015-10-07

    Agrobacterium-mediated transformation of plants with T-DNA is used both to introduce transgenes and for mutagenesis. Conventional approaches used to identify the genomic location and the structure of the inserted T-DNA are laborious and high-throughput methods using next-generation sequencing are being developed to address these problems. Here, we present a cost-effective approach that uses sequence capture targeted to the T-DNA borders to select genomic DNA fragments containing T-DNA—genome junctions, followed by Illumina sequencing to determine the location and junction structure of T-DNA insertions. Multiple probes can be mixed so that transgenic lines transformed with different T-DNA types can be processed simultaneously,more » using a simple, index-based pooling approach. We also developed a simple bioinformatic tool to find sequence read pairs that span the junction between the genome and T-DNA or any foreign DNA. We analyzed 29 transgenic lines of Arabidopsis thaliana, each containing inserts from 4 different T-DNA vectors. We determined the location of T-DNA insertions in 22 lines, 4 of which carried multiple insertion sites. Additionally, our analysis uncovered a high frequency of unconventional and complex T-DNA insertions, highlighting the needs for high-throughput methods for T-DNA localization and structural characterization. Transgene insertion events have to be fully characterized prior to use as commercial products. As a result, our method greatly facilitates the first step of this characterization of transgenic plants by providing an efficient screen for the selection of promising lines.« less

  9. A T-DNA Insertion Knockout of the Bifunctional Lysine-Ketoglutarate Reductase/Saccharopine Dehydrogenase Gene Elevates Lysine Levels in Arabidopsis Seeds1

    PubMed Central

    Zhu, Xiaohong; Tang, Guiliang; Granier, Fabienne; Bouchez, David; Galili, Gad

    2001-01-01

    Plants possess both anabolic and catabolic pathways for the essential amino acid lysine (Lys). However, although the biosynthetic pathway was clearly shown to regulate Lys accumulation in plants, the functional significance of Lys catabolism has not been experimentally elucidated. To address this issue, we have isolated an Arabidopsis knockout mutant with a T-DNA inserted into exon 13 of the gene encoding Lys ketoglutarate reductase/saccharopine dehydrogenase. This bifunctional enzyme controls the first two steps of Lys catabolism. The phenotype of the LKR/SDH knockout was indistinguishable from wild-type plants under normal growth conditions, suggesting that Lys catabolism is not an essential pathway under standard growth conditions. However, mature seeds of the knockout mutant over-accumulated Lys compared with wild-type plants. This report provides the first direct evidence for the functional significance of Lys catabolism in regulating Lys accumulation in seeds. Such a knockout mutant may also provide new perspectives to improve the level of the essential amino acid Lys in plant seeds. PMID:11500552

  10. Intronic T-DNA Insertion Renders Arabidopsis opr3 a Conditional Jasmonic Acid-Producing Mutant1[C][W][OA

    PubMed Central

    Chehab, E. Wassim; Kim, Se; Savchenko, Tatyana; Kliebenstein, Daniel; Dehesh, Katayoon; Braam, Janet

    2011-01-01

    Jasmonic acid and its derived metabolites (JAs) orchestrate plant defense against insects and fungi. 12-Oxo-phytodienoic acid (OPDA), a JA precursor, has also been implicated in plant defense. We sought to define JAs and OPDA functions through comparative defense susceptibility characteristics of three Arabidopsis (Arabidopsis thaliana) genotypes: aos, lacking JAs and OPDA; opda reductase3 (opr3), deficient in JA production but can accumulate OPDA; and transgenics that overexpress OPR3. opr3, like aos, is susceptible to cabbage loopers (Trichoplusia ni) but, relative to aos, opr3 has enhanced resistance to a necrotrophic fungus. Gas chromatography-mass spectrometry reveals that opr3 produces OPDA but no detectable JAs following wounding and looper infestation; unexpectedly, substantial levels of JAs accumulate in opr3 upon fungal infection. Full-length OPR3 transcripts accumulate in fungal-infected opr3, potentially through splicing of the T-DNA containing intron. Fungal resistance correlates with levels of JAs not OPDA; therefore, opr3 resistance to some pests is likely due to JA accumulation, and signaling activities ascribed to OPDA should be reassessed because opr3 can produce JAs. Together these data (1) reinforce the primary role JAs play in plant defense against insects and necrotrophic fungi, (2) argue for a reassessment of signaling activities ascribed to OPDA, and (3) provide evidence that mutants with intron insertions can retain gene function. PMID:21487047

  11. Complementation of the embryo-lethal T-DNA insertion mutant of AUXIN-BINDING-PROTEIN 1 (ABP1) with abp1 point mutated versions reveals crosstalk of ABP1 and phytochromes.

    PubMed

    Effendi, Yunus; Ferro, Noel; Labusch, Corinna; Geisler, Markus; Scherer, Günther F E

    2015-01-01

    The function of the extracytoplasmic AUXIN-BINDING-PROTEIN1 (ABP1) is largely enigmatic. We complemented a homozygous T-DNA insertion null mutant of ABP1 in Arabidopsis thaliana Wassilewskia with three mutated and one wild-type (wt) ABP1 cDNA, all tagged C-terminally with a strepII-FLAG tag upstream the KDEL signal. Based on in silico modelling, the abp1 mutants were predicted to have altered geometries of the auxin binding pocket and calculated auxin binding energies lower than the wt. Phenotypes linked to auxin transport were compromised in these three complemented abp1 mutants. Red light effects, such as elongation of hypocotyls in constant red (R) and far-red (FR) light, in white light supplemented by FR light simulating shade, and inhibition of gravitropism by R or FR, were all compromised in the complemented lines. Using auxin- or light-induced expression of marker genes, we showed that auxin-induced expression was delayed already after 10 min, and light-induced expression within 60 min, even though TIR1/AFB or phyB are thought to act as receptors relevant for gene expression regulation. The expression of marker genes in seedlings responding to both auxin and shade showed that for both stimuli regulation of marker gene expression was altered after 10-20 min in the wild type and phyB mutant. The rapidity of expression responses provides a framework for the mechanics of functional interaction of ABP1 and phyB to trigger interwoven signalling pathways.

  12. Selection of T-DNA-tagged male and female gametophytic mutants by segregation distortion in Arabidopsis.

    PubMed Central

    Howden, R; Park, S K; Moore, J M; Orme, J; Grossniklaus, U; Twell, D

    1998-01-01

    As a strategy for the identification of T-DNA-tagged gametophytic mutants, we have used T-DNA insertional mutagenesis based on screening for distorted segregation ratios by antibiotic selection. Screening of approximately 1000 transgenic Arabidopsis families led to the isolation of eight lines showing reproducible segregation ratios of approximately 1:1, suggesting that these lines are putative gametophytic mutants caused by T-DNA insertion at a single locus. Genetic analysis of T-DNA transmission through reciprocal backcrosses with wild type showed severe reductions in genetic transmission of the T-DNA through the male and/or female gametes. Direct evidence for mutant phenotypes in these lines was investigated by DAPI staining of mature pollen grains and by the analysis of seed set and embryo sac morphology in cleared ovules. One line, termed limpet pollen, showed a novel pollen phenotype in that the generative cell failed to migrate inward after pollen mitosis I, such that the generative or sperm cells remained against the pollen wall. Two other lines, andarta and tistrya, were defective in female transmission and showed an early arrest of embryo sac development with the viable megaspore not initiating the nuclear division cycles. These data demonstrate the efficacy of a segregation ratio distortion strategy for the identification of T-DNA-tagged gametophytic mutants in Arabidopsis. PMID:9611178

  13. In-depth molecular and phenotypic characterization in a rice insertion line library facilitates gene identification through reverse and forward genetics approaches.

    PubMed

    Lorieux, Mathias; Blein, Mélisande; Lozano, Jaime; Bouniol, Mathieu; Droc, Gaétan; Diévart, Anne; Périn, Christophe; Mieulet, Delphine; Lanau, Nadège; Bès, Martine; Rouvière, Claire; Gay, Céline; Piffanelli, Pietro; Larmande, Pierre; Michel, Corinne; Barnola, Isabelle; Biderre-Petit, Corinne; Sallaud, Christophe; Perez, Pascual; Bourgis, Fabienne; Ghesquière, Alain; Gantet, Pascal; Tohme, Joe; Morel, Jean Benoit; Guiderdoni, Emmanuel

    2012-06-01

    We report here the molecular and phenotypic features of a library of 31,562 insertion lines generated in the model japonica cultivar Nipponbare of rice (Oryza sativa L.), called Oryza Tag Line (OTL). Sixteen thousand eight hundred and fourteen T-DNA and 12,410 Tos17 discrete insertion sites have been characterized in these lines. We estimate that 8686 predicted gene intervals--i.e. one-fourth to one-fifth of the estimated rice nontransposable element gene complement--are interrupted by sequence-indexed T-DNA (6563 genes) and/or Tos17 (2755 genes) inserts. Six hundred and forty-three genes are interrupted by both T-DNA and Tos17 inserts. High quality of the sequence indexation of the T2 seed samples was ascertained by several approaches. Field evaluation under agronomic conditions of 27,832 OTL has revealed that 18.2% exhibit at least one morphophysiological alteration in the T1 progeny plants. Screening 10,000 lines for altered response to inoculation by the fungal pathogen Magnaporthe oryzae allowed to observe 71 lines (0.7%) developing spontaneous lesions simulating disease mutants and 43 lines (0.4%) exhibiting an enhanced disease resistance or susceptibility. We show here that at least 3.5% (four of 114) of these alterations are tagged by the mutagens. The presence of allelic series of sequence-indexed mutations in a gene among OTL that exhibit a convergent phenotype clearly increases the chance of establishing a linkage between alterations and inserts. This convergence approach is illustrated by the identification of the rice ortholog of AtPHO2, the disruption of which causes a lesion-mimic phenotype owing to an over-accumulation of phosphate, in nine lines bearing allelic insertions.

  14. Inserting Phase Change Lines into Microsoft Excel® Graphs.

    PubMed

    Dubuque, Erick M

    2015-10-01

    Microsoft Excel® is a popular graphing tool used by behavior analysts to visually display data. However, this program is not always friendly to the graphing conventions used by behavior analysts. For example, adding phase change lines has typically been a cumbersome process involving the insertion of line objects that do not move when new data is added to a graph. The purpose of this article is to describe a novel way to add phase change lines that move when new data is added and when graphs are resized.

  15. Proper Angle of Sono-guided Central Venous Line Insertion

    PubMed Central

    Barzegari, Hassan; Forouzan, Arash; Fahimi, Mohammad Ali; Zohrevandi, Behzad; Ghanavati, Mandana

    2016-01-01

    Introduction: Determining the proper angle for inserting central venous catheter (CV line) is of great importance for decreasing the complications and increasing success rate. The present study was designed to determine the proper angle of needle insertion for internal jugular vein catheterization. Methods: In the present case series study, candidate patients for catheterization of the right internal jugular vein under guidance of ultrasonography were studied. At the time of proper placing of the catheter, photograph was taken and Auto Cad 2014 software was used to measure the angles of the needle in the sagittal and axial planes, as well as patient’s head rotation. Result: 114 patients with the mean age of 56.96 ± 14.71 years were evaluated (68.4% male). The most common indications of catheterization were hemodialysis (55.3%) and shock state (24.6%). The mean angles of needle insertion were 102.15 ± 6.80 for axial plane, 36.21 ± 3.12 for sagittal plane and the mean head rotation angle was 40.49 ± 5.09. Conclusion: Based on the results of the present study it seems that CV line insertion under the angles 102.15 ± 6.80 degrees in the axial plane, 36.21 ± 3.12 in the sagittal plane and 40.49 ± 5.09 head rotation yield satisfactory results. PMID:27299146

  16. Inserting epidural patient controlled analgesia into a peripheral venous line.

    PubMed

    2016-01-01

    A case is reported from the Safety Reporting System in Anaesthesia and Resuscitation database. The event occurred in a patient undergoing abdominal surgery in whom an epidural catheter was inserted for analgesia. After the intervention, the patient was transferred to the recovery unit where the patient controlled analgesia (PCA) is programmed. Due to an error, the PCA was connected to a peripheral venous line, which was detected early without harm to the patient. Communication and analysis of this incident served to introduce a new drug delivery protocol through PCA pumps, including the obligation to prescribe the PCA in the electronic system, a dual computerised check immediately before connecting PCA, labelling the medication bag as well as the proximal and distal lines, standardisation of daily visits to patients, and monthly monitoring of results.

  17. Identification of anther-specific gene expression from T-DNA tagging rice.

    PubMed

    Muthukalianan, Gothandam K; Lee, Sanghyun; Yum, Hyunsik; Ku, Sujin; Kwun, Minjung; Kang, Hong Gyu; An, Gynheung; Chung, Yong-Yoon

    2003-02-28

    We have screened a total of 5,500 T-DNA tagging rice lines in which beta-glucuronidase (GUS) gene sequence was randomly inserted as a transgene into the plant genome. Histochemical GUS assays were carried out to select the T-DNA tagging rice lines that show its expression in anther. Of the tagging lines screened, three lines were found to express GUS specifically in the anther that is about 0.05%. Microscopic observation of the anther-expressed lines showed specific expression patterns of GUS in the anther, either gametophytic or sporophytic specificities. Southern blot analysis revealed that the integration copy number of the transgene was 2.3 in average. The detailed expression patterns were analyzed and discussed.

  18. Molecular characterization of transgenic shallots (Allium cepa L.) by adaptor ligation PCR (AL-PCR) and sequencing of genomic DNA flanking T-DNA borders.

    PubMed

    Zheng, S J; Henken, B; Sofiari, E; Jacobsen, E; Krens, F A; Kik, C

    2001-06-01

    Genomic DNA blot hybridization is traditionally used to demonstrate that, via genetic transformation, foreign genes are integrated into host genomes. However, in large genome species, such as Allium cepa L., the use of genomic DNA blot hybridization is pushed towards its limits, because a considerable quantity of DNA is needed to obtain enough genome copies for a clear hybridization pattern. Furthermore, genomic DNA blot hybridization is a time-consuming method. Adaptor ligation PCR (AL-PCR) of genomic DNA flanking T-DNA borders does not have these drawbacks and seems to be an adequate alternative to genomic DNA blot hybridization. Using AL-PCR we proved that T-DNA was integrated into the A. cepa genome of three transgenic lines transformed with Agrobacterium tumefaciens EHA 105 (pCAMBIA 1301). The AL-PCR patterns obtained were specific and reproducible for a given transgenic line. The results showed that T-DNA integration took place and gave insight in the number of T-DNA copies present. Comparison of AL-PCR and previously obtained genomic DNA blot hybridization results pointed towards complex T-DNA integration patterns in some of the transgenic plants. After cloning and sequencing the AL-PCR products, the junctions between plant genomic DNA and the T-DNA insert could be analysed in great detail. For example it was shown that upon T-DNA integration a 66 bp genomic sequence was deleted, and no filler DNA was inserted. Primers located within the left and right flanking genomic DNA in transgenic shallot plants were used to recover the target site of T-DNA integration.

  19. Generation of insect-resistant and glyphosate-tolerant rice by introduction of a T-DNA containing two Bt insecticidal genes and an EPSPS gene*

    PubMed Central

    ZHAO, Qi-chao; LIU, Ming-hong; ZHANG, Xian-wen; LIN, Chao-yang; ZHANG, Qing; SHEN, Zhi-cheng

    2015-01-01

    Insect resistance and glyphosate tolerance have been two of the most important traits in the genetic improvement of various crops. In this study, two Bacillus thuringiensis (Bt) insecticidal genes, Cry1Ac and Cry1Ig, and a modified glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene (G10) were combined into a single transferred DNA (T-DNA) fragment and introduced into rice by Agrobacterium-mediated transformation. A transgenic line with single-copy T-DNA insertion named GAI-14 was found to be highly resistant to striped stem borer and rice leaf roller, and tolerant to glyphosate. Analysis of T-DNA border sequence suggested that the transgenes were inserted at the chromosome 3 and appeared to have not interrupted any known or putative genes. A field trial observed no significant difference in the basic agronomic traits between GAI-14 and the recipient rice. PMID:26465130

  20. Pneumorrhachis Resulting in Transient Paresis after PICC Line Insertion into the Ascending Lumbar Vein.

    PubMed

    Payne, Russell; Sieg, Emily P; Choudhary, Arabinda; Iantosca, Mark

    2016-10-17

    Obtaining intravascular access in the neonatal intensive care unit (NICU) is not only critical but also technically challenging. Malposition of the catheter tip is a known and well-documented complication. Specifically, peripherally inserted central venous catheter (PICC) line insertion into the ascending lumbar vein can lead to neurological dysfunction and, in some cases, even death. We present the first reported case of pneumorrhachis (PR) following PICC line insertion into the ascending lumbar vein. Our patient presented with lower extremity weakness and imaging confirmed the presence of air within the spinal canal. After conservative treatment, the strength deficit resolved and subsequent imaging revealed resolution of the air within the spinal canal. Insertion of central venous catheters into the ascending lumbar vein is a well-documented complication that can lead to neurologic injury and even death. This should be considered in the evaluation of any neonate presenting with an abnormal neurological examination or unexplained change in exam after line insertion.

  1. DEVELOPMENT OF A TOOL TO INSERT ABOMASAL INFUSION LINES INTO DAIRY COWS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A tool was developed to aid in ruminal insertion of abomasal infusion lines into dairy cows. The tool consisted of two pieces cut from polyvinyl chloride pipe. The first piece of pipe, the insertion tool, contained a groove that held the flexible plastic flange that was on the end of the infusion ...

  2. Distribution and characterization of over 1000 T-DNA tags in rice genome.

    PubMed

    Chen, Shuangyan; Jin, Weizheng; Wang, Mingyi; Zhang, Fan; Zhou, Jie; Jia, Qiaojun; Wu, Yunrong; Liu, Feiyan; Wu, Ping

    2003-10-01

    We generated T-DNA insertions throughout the rice genome for saturation mutagenesis. More than 1,000 flanking sequences were mapped on 12 rice chromosomes. Our results showed that T-DNA tags were not randomly spread on rice chromosomes and were preferentially inserted in gene-rich regions. Few insertions (2.4%) were found in repetitive regions. T-DNA insertions in genic (58.1%) and intergenic regions (41.9%) showed a good correlation with the predicted size distribution of these sequences in the rice genome. Whereas, obvious biases were found for the insertions in the 5'- and 3'-regulatory regions outside the coding regions both at 500-bp size and in introns rather than in exons. Such distribution patterns and biases for T-DNA integration in rice are similar to that of the previous report in Arabidopsis, which may result from T-DNA integration mechanism itself. Rice will require approximately the same number of T-DNA insertions for saturation mutagenesis as will Arabidopsis. A database of the T-DNA insertion sites in rice is publicly available at our web site (http://www.genomics.zju.edu.cn/ricetdna).

  3. Cascade of chromosomal rearrangements caused by a heterogeneous T-DNA integration supports the double-strand break repair model for T-DNA integration.

    PubMed

    Hu, Yufei; Chen, Zhiyu; Zhuang, Chuxiong; Huang, Jilei

    2017-02-28

    Transferred DNA (T-DNA) from Agrobacterium tumefaciens can be integrated into the plant genome. The double-strand break repair (DSBR) pathway is a major model for T-DNA integration. From this model, we expect that two ends of a T-DNA molecule would invade into a single DNA double-strand break (DSB) or independent DSBs in the plant genome. We call the later phenomenon a heterogeneous T-DNA integration which has never been observed. In this work, we demonstrated it in an Arabidopsis T-DNA insertion mutant seb19. To resolve the chromosomal structural changes caused by T-DNA integration at both the nucleotide and chromosome levels, we performed inverse PCR, genome resequencing, fluorescence in situ hybridization and linkage analysis. We found, in seb19, a single T-DNA connected two different chromosomal loci and caused complex chromosomal rearrangements. The specific break-junction pattern in seb19 is consistent with the result of heterogeneous T-DNA integration but not of recombination between two T-DNA insertions. We demonstrated that, in seb19, heterogeneous T-DNA integration evoked a cascade of incorrect repair of seven DSBs on chromosome 4 and 5, and then produced translocation, inversion, duplication and deletion. Heterogeneous T-DNA integration supports the DSBR model and suggests that two ends of a T-DNA molecule could be integrated into the plant genome independently. Our results also show a new origin of chromosomal abnormalities. This article is protected by copyright. All rights reserved.

  4. The insertion of chronic indwelling central venous catheters (Hickman lines) in interventional radiology suites.

    PubMed

    Page, A C; Evans, R A; Kaczmarski, R; Mufti, G J; Gishen, P

    1990-08-01

    The insertion of Hickman central venous catheters for chronic venous access is a procedure usually conducted in the operating theatre under local or general anaesthesia. In a prospective study over a one year period we have assessed the feasibility of radiologists inserting central venous catheters for long term access. A subclavicular approach to the subclavian vein with prior digital subtraction angiography or video imaging of the vein was the technique of choice. Thirty-one Hickman catheters were inserted in 21 patients. All but two patients had a haematological malignancy. Ages ranged from 19 to 77 years. The mean time for insertion was 43 min (range 20-80 min). The catheters remained in situ for between 2 days and 242 days with a mean of 86 days. There was one documented line infection; nine patients had episodes of septicaemia with identified organisms, and a further six had pyrexias of unknown origin during the line indwelling period. There were four documented line and or ipsilateral subclavian vein thromboses, and one death occurred within 36 hours of the procedure. We conclude that radiological placement is an excellent alternative to 'blind' surgical placement. Screening during insertion provides immediate facilities for correction of malposition and monitoring of immediate complications. The time taken for catheter insertion did not impede the usual patient throughout in the interventional radiology suite.

  5. Pneumorrhachis Resulting in Transient Paresis after PICC Line Insertion into the Ascending Lumbar Vein

    PubMed Central

    Sieg, Emily P; Choudhary, Arabinda; Iantosca, Mark

    2016-01-01

    Obtaining intravascular access in the neonatal intensive care unit (NICU) is not only critical but also technically challenging. Malposition of the catheter tip is a known and well-documented complication. Specifically, peripherally inserted central venous catheter (PICC) line insertion into the ascending lumbar vein can lead to neurological dysfunction and, in some cases, even death. We present the first reported case of pneumorrhachis (PR) following PICC line insertion into the ascending lumbar vein. Our patient presented with lower extremity weakness and imaging confirmed the presence of air within the spinal canal. After conservative treatment, the strength deficit resolved and subsequent imaging revealed resolution of the air within the spinal canal. Insertion of central venous catheters into the ascending lumbar vein is a well-documented complication that can lead to neurologic injury and even death. This should be considered in the evaluation of any neonate presenting with an abnormal neurological examination or unexplained change in exam after line insertion. PMID:27904815

  6. Comparison of different platelet transfusion thresholds prior to insertion of central lines in patients with thrombocytopenia

    PubMed Central

    Estcourt, Lise J; Desborough, Michael; Hopewell, Sally; Trivella, Marialena; Doree, Carolyn; Stanworth, Simon

    2015-01-01

    This is the protocol for a review and there is no abstract. The objectives are as follows: To assess the effects of different platelet transfusion thresholds prior to the insertion of a central line in patients with thrombocytopenia (low platelet count). PMID:26814707

  7. Odyssey of agrobacterium T-DNA.

    PubMed

    Ziemienowicz, A

    2001-01-01

    Agrobacterium tumefaciens, a plant pathogen, is characterized by the unique feature of interkingdom DNA transfer. This soil bacterium is able to transfer a fragment of its DNA, called T-DNA (transferred DNA), to the plant cell where T-DNA is integrated into the plant genome leading to "genetic colonization" of the host. The fate of T-DNA, its processing, transfer and integration, resembles the journey of Odysseus, although our hero returns from its long trip in a slightly modified form.

  8. Comparison of different platelet transfusion thresholds prior to insertion of central lines in patients with thrombocytopenia

    PubMed Central

    Estcourt, Lise J; Desborough, Michael; Hopewell, Sally; Doree, Carolyn; Stanworth, Simon J

    2016-01-01

    Background Patients with a low platelet count (thrombocytopenia) often require the insertion of central lines (central venous catheters (CVCs)). CVCs have a number of uses; these include: administration of chemotherapy; intensive monitoring and treatment of critically-ill patients; administration of total parenteral nutrition; and long-term intermittent intravenous access for patients requiring repeated treatments. Current practice in many countries is to correct thrombocytopenia with platelet transfusions prior to CVC insertion, in order to mitigate the risk of serious procedure-related bleeding. However, the platelet count threshold recommended prior to CVC insertion varies significantly from country to country. This indicates significant uncertainty among clinicians of the correct management of these patients. The risk of bleeding after a central line insertion appears to be low if an ultrasound-guided technique is used. Patients may therefore be exposed to the risks of a platelet transfusion without any obvious clinical benefit. Objectives To assess the effects of different platelet transfusion thresholds prior to the insertion of a central line in patients with thrombocytopenia (low platelet count). Search methods We searched for randomised controlled trials (RCTs) in CENTRAL (The Cochrane Library 2015, Issue 2), MEDLINE (from 1946), EMBASE (from 1974), the Transfusion Evidence Library (from 1950) and ongoing trial databases to 23 February 2015. Selection criteria We included RCTs involving transfusions of platelet concentrates, prepared either from individual units of whole blood or by apheresis, and given to prevent bleeding in patients of any age with thrombocytopenia requiring insertion of a CVC. Data collection and analysis We used standard methodological procedures expected by The Cochrane Collaboration. Main results One RCT was identified that compared different platelet transfusion thresholds prior to insertion of a CVC in people with chronic liver

  9. Pericardial Effusion and Cardiac Tamponade Following Percutaneously Inserted Central Line Insertion in an Extremely Low-birth-weight Baby: Case Report and Successful Management

    PubMed Central

    Desai, Avinash V.; Kumar, Abnish; Shanbhag, Praful; Shah, Forum

    2017-01-01

    Pericardial effusion and cardiac tamponade are rare complications of percutaneously inserted central lines (PICLs), in extremely low-birth-weight babies. This particular complication carries a high degree of mortality, if not suspected and diagnosed. Bedside echocardiography proves not only diagnostic but also can be lifesaving in these conditions. Here, we wish to report such a baby who suddenly deteriorated and showed cardiovascular instability 2 days after the insertion of PICL. Immediate bedside echocardiography helped pick up the effusion which was drained using a subxiphoid percutaneous approach. PMID:28197055

  10. An evaluation of new and established methods to determine T-DNA copy number and homozygosity in transgenic plants.

    PubMed

    Głowacka, Katarzyna; Kromdijk, Johannes; Leonelli, Lauriebeth; Niyogi, Krishna K; Clemente, Tom E; Long, Stephen P

    2016-04-01

    Stable transformation of plants is a powerful tool for hypothesis testing. A rapid and reliable evaluation method of the transgenic allele for copy number and homozygosity is vital in analysing these transformations. Here the suitability of Southern blot analysis, thermal asymmetric interlaced (TAIL-)PCR, quantitative (q)PCR and digital droplet (dd)PCR to estimate T-DNA copy number, locus complexity and homozygosity were compared in transgenic tobacco. Southern blot analysis and ddPCR on three generations of transgenic offspring with contrasting zygosity and copy number were entirely consistent, whereas TAIL-PCR often underestimated copy number. qPCR deviated considerably from the Southern blot results and had lower precision and higher variability than ddPCR. Comparison of segregation analyses and ddPCR of T1 progeny from 26 T0 plants showed that at least 19% of the lines carried multiple T-DNA insertions per locus, which can lead to unstable transgene expression. Segregation analyses failed to detect these multiple copies, presumably because of their close linkage. This shows the importance of routine T-DNA copy number estimation. Based on our results, ddPCR is the most suitable method, because it is as reliable as Southern blot analysis yet much faster. A protocol for this application of ddPCR to large plant genomes is provided.

  11. Short direct repeats flank the T-DNA on a nopaline Ti plasmid

    PubMed Central

    Yadav, Narendra S.; Vanderleyden, Jos; Bennett, Donald R.; Barnes, Wayne M.; Chilton, Mary-Dell

    1982-01-01

    Crown gall disease results from the insertion of a segment of the Agrobacterium Ti plasmid, called T-DNA, into host plant nuclear DNA. We have subjected to sequence analysis the border regions of pTi T37 (ends of T-DNA) and one left T-DNA/plant DNA border fragment isolated from BT37 tobacco teratoma by molecular cloning. These sequence studies, taken together with published sequence of a right T-DNA/plant DNA border fragment, allowed us to identify the positions of left and right borders at the DNA sequence level. Comparison of left and right border regions of the Ti plasmid revealed a “core” direct repeat of 13 of 14 bases (12 contiguous) precisely at the borders of T-DNA. An extended repeat of 21 of 25 bases overlaps this core repeat. T-DNA on the Ti plasmid exhibits no longer direct or inverted repeats in the border regions, based on Southern hybridization studies. The physical structure of T-DNA differs from that of known prokaryotic and eukaryotic transposable elements but bears a structural resemblance to the prophage of bacteriophage λ. Images PMID:16593241

  12. Corrections to hyperfine splitting and Lamb shift induced by diagrams with second order radiative insertions in the electron line

    SciTech Connect

    Eides, M.I.; Karshenboim, S.G.; Shelyuto, V.A.

    1994-12-31

    Contributions to HFS and to the Lamb shift intervals of order a{sup 2}(Za){sup 5} induced by gauge invariant set of nineteen topologically different graphs with two radiative photons inserted in the electron line are considered. Corrections both to HFS and Lamb shift induced by nine diagrams are calculated in the Fried-Yennie gauge.

  13. Identification of Genomic Insertion and Flanking Sequence of G2-EPSPS and GAT Transgenes in Soybean Using Whole Genome Sequencing Method.

    PubMed

    Guo, Bingfu; Guo, Yong; Hong, Huilong; Qiu, Li-Juan

    2016-01-01

    Molecular characterization of sequence flanking exogenous fragment insertion is essential for safety assessment and labeling of genetically modified organism (GMO). In this study, the T-DNA insertion sites and flanking sequences were identified in two newly developed transgenic glyphosate-tolerant soybeans GE-J16 and ZH10-6 based on whole genome sequencing (WGS) method. More than 22.4 Gb sequence data (∼21 × coverage) for each line was generated on Illumina HiSeq 2500 platform. The junction reads mapped to boundaries of T-DNA and flanking sequences in these two events were identified by comparing all sequencing reads with soybean reference genome and sequence of transgenic vector. The putative insertion loci and flanking sequences were further confirmed by PCR amplification, Sanger sequencing, and co-segregation analysis. All these analyses supported that exogenous T-DNA fragments were integrated in positions of Chr19: 50543767-50543792 and Chr17: 7980527-7980541 in these two transgenic lines. Identification of genomic insertion sites of G2-EPSPS and GAT transgenes will facilitate the utilization of their glyphosate-tolerant traits in soybean breeding program. These results also demonstrated that WGS was a cost-effective and rapid method for identifying sites of T-DNA insertions and flanking sequences in soybean.

  14. Identification of Genomic Insertion and Flanking Sequence of G2-EPSPS and GAT Transgenes in Soybean Using Whole Genome Sequencing Method

    PubMed Central

    Guo, Bingfu; Guo, Yong; Hong, Huilong; Qiu, Li-Juan

    2016-01-01

    Molecular characterization of sequence flanking exogenous fragment insertion is essential for safety assessment and labeling of genetically modified organism (GMO). In this study, the T-DNA insertion sites and flanking sequences were identified in two newly developed transgenic glyphosate-tolerant soybeans GE-J16 and ZH10-6 based on whole genome sequencing (WGS) method. More than 22.4 Gb sequence data (∼21 × coverage) for each line was generated on Illumina HiSeq 2500 platform. The junction reads mapped to boundaries of T-DNA and flanking sequences in these two events were identified by comparing all sequencing reads with soybean reference genome and sequence of transgenic vector. The putative insertion loci and flanking sequences were further confirmed by PCR amplification, Sanger sequencing, and co-segregation analysis. All these analyses supported that exogenous T-DNA fragments were integrated in positions of Chr19: 50543767–50543792 and Chr17: 7980527–7980541 in these two transgenic lines. Identification of genomic insertion sites of G2-EPSPS and GAT transgenes will facilitate the utilization of their glyphosate-tolerant traits in soybean breeding program. These results also demonstrated that WGS was a cost-effective and rapid method for identifying sites of T-DNA insertions and flanking sequences in soybean. PMID:27462336

  15. Proteins encoded by Agrobacterium tumefaciens Ti plasmid DNA (T-DNA) in crown gall tumors

    PubMed Central

    McPherson, Joan C.; Nester, Eugene W.; Gordon, Milton P.

    1980-01-01

    In order to detect proteins that may be produced in crown gall tumors as a result of expression of incorporated Agrobacterium tumefaciens Ti plasmid DNA (T-DNA), we have isolated mRNA complementary to T-DNA and translated this in a protein-synthesizing system derived from wheat germ. mRNA prepared from cultured E1 tumor from Nicotiana tabacum hybridized with HindIII fragment 1 sequences of T-DNA immobilized on cellulose nitrate filters. Two proteins of 30,000 and 16,500 Mr were produced when this selected RNA was released and translated. Other tumor lines from N. tabacum were investigated, and a protein of slightly less than 30,000 Mr was encoded by HindIII fragment 1 sequences of 15955/01 tumor. No products were observed for 15955/1 tumor line, which differs from E1/B6-806 and 15955/01 in that it does not produce octopine. mRNA species of each of the tumor lines hybridized to Bst I fragment 8 sequences of T-DNA and produced a common protein of 15,000 Mr. Because this protein is derived from the region of the T-DNA that is conserved in octopine- and nopaline-type crown gall tumors, it may play a role in oncogenicity. Images PMID:16592819

  16. Agrobacterium T-DNA integration in Arabidopsis is correlated with DNA sequence compositions that occur frequently in gene promoter regions.

    PubMed

    Schneeberger, Richard G; Zhang, Ke; Tatarinova, Tatiana; Troukhan, Max; Kwok, Shing F; Drais, Josh; Klinger, Kevin; Orejudos, Francis; Macy, Kimberly; Bhakta, Amit; Burns, James; Subramanian, Gopal; Donson, Jonathan; Flavell, Richard; Feldmann, Kenneth A

    2005-10-01

    Mobile insertion elements such as transposons and T-DNA generate useful genetic variation and are important tools for functional genomics studies in plants and animals. The spectrum of mutations obtained in different systems can be highly influenced by target site preferences inherent in the mechanism of DNA integration. We investigated the target site preferences of Agrobacterium T-DNA insertions in the chromosomes of the model plant Arabidopsis thaliana. The relative frequencies of insertions in genic and intergenic regions of the genome were calculated and DNA composition features associated with the insertion site flanking sequences were identified. Insertion frequencies across the genome indicate that T-strand integration is suppressed near centromeres and rDNA loci, progressively increases towards telomeres, and is highly correlated with gene density. At the gene level, T-DNA integration events show a statistically significant preference for insertion in the 5' and 3' flanking regions of protein coding sequences as well as the promoter region of RNA polymerase I transcribed rRNA gene repeats. The increased insertion frequencies in 5' upstream regions compared to coding sequences are positively correlated with gene expression activity and DNA sequence composition. Analysis of the relationship between DNA sequence composition and gene activity further demonstrates that DNA sequences with high CG-skew ratios are consistently correlated with T-DNA insertion site preference and high gene expression. The results demonstrate genomic and gene-specific preferences for T-strand integration and suggest that DNA sequences with a pronounced transition in CG- and AT-skew ratios are preferred targets for T-DNA integration.

  17. [Peripherally inserted central catheters (PICC) in onco-hematology. PICC line in onco-hematology].

    PubMed

    Kabsy, Y; Baudin, G; Vinti, H; Novellas, S; Mannone, L; Chevallier, P; Mounier, N

    2010-09-01

    Peripherally inserted central catheters (PICC) have the advantage of limiting the risk of accidents during installation and are easy to remove. Its use in oncology remains debated because of possible infectious complications. We analyzed 52 PICC in patients with hematological tumor from Nice Hospital. An installation failure was noted in 5.8% of cases. After a follow-up of 15 months, the complication rate was 26.9%, mainly mechanical complications: obstruction (13.5%) or accidental removal (9.6%). The organic complications such as infection or thrombophlebitis represented 3.8%. The median duration was 26 days [2-291]. The longest duration was associated with PICC for chemotherapy (median: 58 days). Frequent blood samples (above: 2 week) were associated with lower duration (median: 23 days). In conclusion, PICC represent a simple and effective alternative to intra-venous central devices in onco-hematology. However, physicians have to focus on short-course treatment.

  18. A feasibility study of a PET/MRI insert detector using strip-line and waveform sampling data acquisition

    NASA Astrophysics Data System (ADS)

    Kim, H.; Chen, C.-T.; Eclov, N.; Ronzhin, A.; Murat, P.; Ramberg, E.; Los, S.; Wyrwicz, A. M.; Li, L.; Kao, C.-M.

    2015-06-01

    We are developing a time-of-flight Positron Emission Tomography (PET) detector by using silicon photo-multipliers (SiPM) on a strip-line and high speed waveform sampling data acquisition. In this design, multiple SiPMs are connected on a single strip-line and signal waveforms on the strip-line are sampled at two ends of the strip to reduce readout channels while fully exploiting the fast time response of SiPMs. In addition to the deposited energy and time information, the position of the hit SiPM along the strip-line is determined by the arrival time difference of the waveform. Due to the insensitivity of the SiPMs to magnetic fields and the compact front-end electronics, the detector approach is highly attractive for developing a PET insert system for a magnetic resonance imaging (MRI) scanner to provide simultaneous PET/MR imaging. To investigate the feasibility, experimental tests using prototype detector modules have been conducted inside a 9.4 T small animal MRI scanner (Bruker BioSpec 94/30 imaging spectrometer). On the prototype strip-line board, 16 SiPMs (5.2 mm pitch) are installed on two strip-lines and coupled to 2×8 LYSO scintillators (5.0×5.0×10.0 mm3 with 5.2 mm pitch). The outputs of the strip-line boards are connected to a Domino-Ring-Sampler (DRS4) evaluation board for waveform sampling. Preliminary experimental results show that the effect of interference on the MRI image due to the PET detector is negligible and that PET detector performance is comparable with the results measured outside the MRI scanner.

  19. A feasibility study of a PET/MRI insert detector using strip-line and waveform sampling data acquisition.

    PubMed

    Kim, H; Chen, C-T; Eclov, N; Ronzhin, A; Murat, P; Ramberg, E; Los, S; Wyrwicz, Alice M; Li, Limin; Kao, C-M

    2015-06-01

    We are developing a time-of-flight Positron Emission Tomography (PET) detector by using silicon photo-multipliers (SiPM) on a strip-line and high speed waveform sampling data acquisition. In this design, multiple SiPMs are connected on a single strip-line and signal waveforms on the strip-line are sampled at two ends of the strip to reduce readout channels while fully exploiting the fast time response of SiPMs. In addition to the deposited energy and time information, the position of the hit SiPM along the strip-line is determined by the arrival time difference of the waveform. Due to the insensitivity of the SiPMs to magnetic fields and the compact front-end electronics, the detector approach is highly attractive for developing a PET insert system for a magnetic resonance imaging (MRI) scanner to provide simultaneous PET/MR imaging. To investigate the feasibility, experimental tests using prototype detector modules have been conducted inside a 9.4 Tesla small animal MRI scanner (Bruker BioSpec 94/30 imaging spectrometer). On the prototype strip-line board, 16 SiPMs (5.2 mm pitch) are installed on two strip-lines and coupled to 2 × 8 LYSO scintillators (5.0 × 5.0 × 10.0 mm(3) with 5.2 mm pitch). The outputs of the strip-line boards are connected to a Domino-Ring-Sampler (DRS4) evaluation board for waveform sampling. Preliminary experimental results show that the effect of interference on the MRI image due to the PET detector is negligible and that PET detector performance is comparable with the results measured outside the MRI scanner.

  20. A feasibility study of a PET/MRI insert detector using strip-line and waveform sampling data acquisition

    PubMed Central

    Kim, H.; Chen, C.-T.; Eclov, N.; Ronzhin, A.; Murat, P.; Ramberg, E.; Los, S.; Wyrwicz, Alice M.; Li, Limin; Kao, C.-M.

    2014-01-01

    We are developing a time-of-flight Positron Emission Tomography (PET) detector by using silicon photo-multipliers (SiPM) on a strip-line and high speed waveform sampling data acquisition. In this design, multiple SiPMs are connected on a single strip-line and signal waveforms on the strip-line are sampled at two ends of the strip to reduce readout channels while fully exploiting the fast time response of SiPMs. In addition to the deposited energy and time information, the position of the hit SiPM along the strip-line is determined by the arrival time difference of the waveform. Due to the insensitivity of the SiPMs to magnetic fields and the compact front-end electronics, the detector approach is highly attractive for developing a PET insert system for a magnetic resonance imaging (MRI) scanner to provide simultaneous PET/MR imaging. To investigate the feasibility, experimental tests using prototype detector modules have been conducted inside a 9.4 Tesla small animal MRI scanner (Bruker BioSpec 94/30 imaging spectrometer). On the prototype strip-line board, 16 SiPMs (5.2 mm pitch) are installed on two strip-lines and coupled to 2 × 8 LYSO scintillators (5.0 × 5.0 × 10.0 mm3 with 5.2 mm pitch). The outputs of the strip-line boards are connected to a Domino-Ring-Sampler (DRS4) evaluation board for waveform sampling. Preliminary experimental results show that the effect of interference on the MRI image due to the PET detector is negligible and that PET detector performance is comparable with the results measured outside the MRI scanner. PMID:25937685

  1. Multiple independent defective suppressor-mutator transposon insertions in Arabidopsis: a tool for functional genomics.

    PubMed Central

    Tissier, A F; Marillonnet, S; Klimyuk, V; Patel, K; Torres, M A; Murphy, G; Jones, J D

    1999-01-01

    A new system for insertional mutagenesis based on the maize Enhancer/Suppressor-mutator (En/Spm) element was introduced into Arabidopsis. A single T-DNA construct carried a nonautonomous defective Spm (dSpm) element with a phosphinothricin herbicide resistance (BAR) gene, a transposase expression cassette, and a counterselectable gene. This construct was used to select for stable dSpm transpositions. Treatments for both positive (BAR) and negative selection markers were applicable to soil-grown plants, allowing the recovery of new transpositions on a large scale. To date, a total of 48,000 lines in pools of 50 have been recovered, of which approximately 80% result from independent insertion events. DNA extracted from these pools was used in reverse genetic screens, either by polymerase chain reaction (PCR) using primers from the transposon and the targeted gene or by the display of insertions whereby inverse PCR products of insertions from the DNA pools are spotted on a membrane that is then hybridized with the probe of interest. By sequencing PCR-amplified fragments adjacent to insertion sites, we established a sequenced insertion-site database of 1200 sequences. This database permitted a comparison of the chromosomal distribution of transpositions from various T-DNA locations. PMID:10521516

  2. CACTA-superfamily transposable element is inserted in MYB transcription factor gene of soybean line producing variegated seeds.

    PubMed

    Yan, Fan; Di, Shaokang; Takahashi, Ryoji

    2015-08-01

    The R gene of soybean, presumably encoding a MYB transcription factor, controls seed coat color. The gene consists of multiple alleles, R (black), r-m (black spots and (or) concentric streaks on brown seed), and r (brown seed). This study was conducted to determine the structure of the MYB transcription factor gene in a near-isogenic line (NIL) having r-m allele. PCR amplification of a fragment of the candidate gene Glyma.09G235100 generated a fragment of about 1 kb in the soybean cultivar Clark, whereas a fragment of about 14 kb in addition to fragments of 1 and 1.4 kb were produced in L72-2040, a Clark 63 NIL with the r-m allele. Clark 63 is a NIL of Clark with the rxp and Rps1 alleles. A DNA fragment of 13 060 bp was inserted in the intron of Glyma.09G235100 in L72-2040. The fragment had the CACTA motif at both ends, imperfect terminal inverted repeats (TIR), inverse repetition of short sequence motifs close to the 5' and 3' ends, and a duplication of three nucleotides at the site of integration, indicating that it belongs to a CACTA-superfamily transposable element. We designated the element as Tgm11. Overall nucleotide sequence, motifs of TIR, and subterminal repeats were similar to those of Tgm1 and Tgs1, suggesting that these elements comprise a family.

  3. Application of Discrete Event Control to the Insertion Task of Electric Line Using 6-Link Electro-Hydraulic Manipulators with Dual Arm

    NASA Astrophysics Data System (ADS)

    Ahn, Kyoungkwan; Yokota, Shinichi

    Uninterrupted power supply has become indispensable during the maintenance task of active electric power lines as a result of today's highly information-oriented society and increasing demand of electric utilities. The maintenance task has the risk of electric shock and the danger of falling from high place. Therefore it is necessary to realize an autonomous robot system using electro-hydraulic manipulator because hydraulic manipulators have the advantage of electric insulation. Meanwhile it is relatively difficult to realize autonomous assembly tasks particularly in the case of manipulating flexible objects such as electric lines. In this report, a discrete event control system is introduced for automatic assembly task of electric lines into sleeves as one of a typical task of active electric power lines. In the implementation of a discrete event control system, LVQNN (learning vector quantization neural network) is applied to the insertion task of electric lines to sleeves. In order to apply these proposed control system to the unknown environment, virtual learning data for LVQNN was generated by fuzzy inference. By the experimental results of two types of electric lines and sleeves, these proposed discrete event control and neural network learning algorithm are confirmed very effective to the insertion tasks of electric lines to sleeves as a typical task of active electric power maintenance tasks.

  4. Transcultural Diabetes Nutrition Algorithm (tDNA): Venezuelan Application

    PubMed Central

    Nieto-Martínez, Ramfis; Hamdy, Osama; Marante, Daniel; Inés Marulanda, María; Marchetti, Albert; Hegazi, Refaat A.; Mechanick, Jeffrey I.

    2014-01-01

    Medical nutrition therapy (MNT) is a necessary component of comprehensive type 2 diabetes (T2D) management, but optimal outcomes require culturally-sensitive implementation. Accordingly, international experts created an evidence-based transcultural diabetes nutrition algorithm (tDNA) to improve understanding of MNT and to foster portability of current guidelines to various dysglycemic populations worldwide. This report details the development of tDNA-Venezuelan via analysis of region-specific cardiovascular disease (CVD) risk factors, lifestyles, anthropometrics, and resultant tDNA algorithmic modifications. Specific recommendations include: screening for prediabetes (for biochemical monitoring and lifestyle counseling); detecting obesity using Latin American cutoffs for waist circumference and Venezuelan cutoffs for BMI; prescribing MNT to people with prediabetes, T2D, or high CVD risk; specifying control goals in prediabetes and T2D; and describing regional differences in prevalence of CVD risk and lifestyle. Venezuelan deliberations involved evaluating typical food-based eating patterns, correcting improper dietary habits through adaptation of the Mediterranean diet with local foods, developing local recommendations for physical activity, avoiding stigmatizing obesity as a cosmetic problem, avoiding misuse of insulin and metformin, circumscribing bariatric surgery to appropriate indications, and using integrated health service networks to implement tDNA. Finally, further research, national surveys, and validation protocols focusing on CVD risk reduction in Venezuelan populations are necessary. PMID:24699193

  5. Transcultural diabetes nutrition algorithm (tDNA): Venezuelan application.

    PubMed

    Nieto-Martínez, Ramfis; Hamdy, Osama; Marante, Daniel; Marulanda, María Inés; Marchetti, Albert; Hegazi, Refaat A; Mechanick, Jeffrey I

    2014-04-02

    Medical nutrition therapy (MNT) is a necessary component of comprehensive type 2 diabetes (T2D) management, but optimal outcomes require culturally-sensitive implementation. Accordingly, international experts created an evidence-based transcultural diabetes nutrition algorithm (tDNA) to improve understanding of MNT and to foster portability of current guidelines to various dysglycemic populations worldwide. This report details the development of tDNA-Venezuelan via analysis of region-specific cardiovascular disease (CVD) risk factors, lifestyles, anthropometrics, and resultant tDNA algorithmic modifications. Specific recommendations include: screening for prediabetes (for biochemical monitoring and lifestyle counseling); detecting obesity using Latin American cutoffs for waist circumference and Venezuelan cutoffs for BMI; prescribing MNT to people with prediabetes, T2D, or high CVD risk; specifying control goals in prediabetes and T2D; and describing regional differences in prevalence of CVD risk and lifestyle. Venezuelan deliberations involved evaluating typical food-based eating patterns, correcting improper dietary habits through adaptation of the Mediterranean diet with local foods, developing local recommendations for physical activity, avoiding stigmatizing obesity as a cosmetic problem, avoiding misuse of insulin and metformin, circumscribing bariatric surgery to appropriate indications, and using integrated health service networks to implement tDNA. Finally, further research, national surveys, and validation protocols focusing on CVD risk reduction in Venezuelan populations are necessary.

  6. A T-DNA mutation in the RNA helicase eIF4A confers a dose-dependent dwarfing phenotype in Brachypodium distachyon.

    PubMed

    Vain, Philippe; Thole, Vera; Worland, Barbara; Opanowicz, Magdalena; Bush, Max S; Doonan, John H

    2011-06-01

    In a survey of the BrachyTAG mutant population of Brachypodium distachyon, we identified a line carrying a T-DNA insertion in one of the two eukaryotic initiation factor 4A (eIF4A) genes present in the nuclear genome. The eif4a homozygous mutant plants were slow-growing, and exhibited reduced final plant stature due to a decrease in both cell number and cell size, consistent with roles for eIF4A in both cell division and cell growth. Hemizygous plants displayed a semi-dwarfing phenotype, in which stem length was reduced but leaf length was normal. Linkage between the insertion site and phenotype was confirmed, and we show that the level of eIF4A protein is strongly reduced in the mutant. Transformation of the Brachypodium homozygous mutant with a genomic copy of the Arabidopsis eIF4A-1 gene partially complemented the growth phenotype, indicating that gene function is conserved between mono- and dicotyledonous species. This study identifies eIF4A as a novel dose-dependent regulator of stem elongation, and demonstrates the utility of Brachypodium as a model for grass and cereals research.

  7. Generation and characterization of the Western Regional Research Center brachypodium T-DNA insertional mutant collection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The model grass Brachypodium distachyon (Brachypodium) is an excellent system for studying the basic biology underlying traits relevant to the use of grasses as food, forage and energy crops. To add to the growing collection of Brachypodium resources available to plant scientists, we further optim...

  8. Familial retinoblastoma due to intronic LINE-1 insertion causes aberrant and noncanonical mRNA splicing of the RB1 gene.

    PubMed

    Rodríguez-Martín, Carlos; Cidre, Florencia; Fernández-Teijeiro, Ana; Gómez-Mariano, Gema; de la Vega, Leticia; Ramos, Patricia; Zaballos, Ángel; Monzón, Sara; Alonso, Javier

    2016-05-01

    Retinoblastoma (RB, MIM 180200) is the paradigm of hereditary cancer. Individuals harboring a constitutional mutation in one allele of the RB1 gene have a high predisposition to develop RB. Here, we present the first case of familial RB caused by a de novo insertion of a full-length long interspersed element-1 (LINE-1) into intron 14 of the RB1 gene that caused a highly heterogeneous splicing pattern of RB1 mRNA. LINE-1 insertion was inferred by mRNA studies and full-length sequenced by massive parallel sequencing. Some of the aberrant mRNAs were produced by noncanonical acceptor splice sites, a new finding that up to date has not been described to occur upon LINE-1 retrotransposition. Our results clearly show that RNA-based strategies have the potential to detect disease-causing transposon insertions. It also confirms that the incorporation of new genetic approaches, such as massive parallel sequencing, contributes to characterize at the sequence level these unique and exceptional genetic alterations.

  9. Long Interspersed Element Sequencing (L1-Seq): A Method to Identify Somatic LINE-1 Insertions in the Human Genome

    PubMed Central

    Doucet, Tara T.; Kazazian, Haig H.

    2017-01-01

    L1-seq is a high-throughput sequencing technique which is utilized to identify novel L1 insertions in genomic DNA samples of interest. Using special diagnostic nucleotides unique to the youngest and most active L1 sequence, we can amplify new somatic insertions. This technique has helped to establish the number of L1 insertions present in the general population as well as the variation among individuals with regard to their complement of active L1 elements. More recently, this technique has been employed to assess the level of retrotransposition occurring in various diseases such as cancer. These efforts try to establish a connection between the process of retrotransposition and disease development and/or progression. PMID:26895047

  10. Should nephrologists take a larger role in interventional nephrology, and should central line insertion remain a requirement of nephrology residency training? A debate.

    PubMed

    Mendelssohn, David C

    2015-01-01

    The Canadian Society of Nephrology must soon provide input concerning the future of procedural training in nephrology. While at one time, the ability to insert a central venous catheter (CVC) was an essential skill required by all nephrologists, in 2014, nephrology training and practice has changed in fundamental ways such that it would be both unreasonable, and impractical, to maintain this requirement. Indeed, survey evidence suggests that many current trainees are not achieving this competency. Amongst the reasons that this requirement should be withdrawn include: 1) Not all trainees have the procedural skills to safely learn to insert CVC's. 2) Most nephrologists in training and in practice are intellectually oriented, not procedurally oriented and are not seeking to perform lots of procedures. 3) In most practice settings, interventional radiologists and intensive care doctors perform dialysis line insertions using real time ultrasound guidance frequently, and offer timely, safer, and better service to patients. 4) Most trainees will not enter practice settings where CVC insertion ability is required. 5) Otherwise excellent future trainees may be denied a nephrology certificate of special competence only because they are unable to insert a CVC by the end of their fellowship. 6) Academic nephrology training programs that cannot provide adequate CVC insertion experience to fellows may lose their status as training centres. As a pragmatic way forward, Canadian nephrology training programs must encourage and offer only those nephrology trainees who have the ability and interest in procedural nephrology, a pathway through which they may be provided superb advanced training to become an expert. There is no longer a compelling reason to mandate this for all trainees.

  11. Insertional mutagenesis of genes required for seed development in Arabidopsis thaliana.

    PubMed Central

    McElver, J; Tzafrir, I; Aux, G; Rogers, R; Ashby, C; Smith, K; Thomas, C; Schetter, A; Zhou, Q; Cushman, M A; Tossberg, J; Nickle, T; Levin, J Z; Law, M; Meinke, D; Patton, D

    2001-01-01

    The purpose of this project was to identify large numbers of Arabidopsis genes with essential functions during seed development. More than 120,000 T-DNA insertion lines were generated following Agrobacterium-mediated transformation. Transgenic plants were screened for defective seeds and putative mutants were subjected to detailed analysis in subsequent generations. Plasmid rescue and TAIL-PCR were used to recover plant sequences flanking insertion sites in tagged mutants. More than 4200 mutants with a wide range of seed phenotypes were identified. Over 1700 of these mutants were analyzed in detail. The 350 tagged embryo-defective (emb) mutants identified to date represent a significant advance toward saturation mutagenesis of EMB genes in Arabidopsis. Plant sequences adjacent to T-DNA borders in mutants with confirmed insertion sites were used to map genome locations and establish tentative identities for 167 EMB genes with diverse biological functions. The frequency of duplicate mutant alleles recovered is consistent with a relatively small number of essential (EMB) genes with nonredundant functions during seed development. Other functions critical to seed development in Arabidopsis may be protected from deleterious mutations by extensive genome duplications. PMID:11779812

  12. T-DNA tagging in Brassica napus as an efficient tool for the isolation of new promoters for selectable marker genes.

    PubMed

    Bade, Jacob; van Grinsven, Emiel; Custers, Jerome; Hoekstra, Sietske; Ponstein, Anne

    2003-05-01

    A simple strategy to identify and isolate new promoters suitable for driving the expression of selectable marker genes is described. By employing a Brassica napus hypocotyl transformation protocol and a promoterless gus::nptII tagging construct, a series of 20 kanamycin-resistant tagged lines was produced. Most of the regenerated plants showed hardly any GUS activity in leaf, stem and root tissues. However, expression was readily restored in callus tissue induced on in vitro leaf segments. Genomic sequences upstream of the gus::nptII insertions were isolated via plasmid rescue. Three clones originating from single copy T-DNA lines were selected for further evaluation. The rescued plasmids were cloned as linear fragments in binary vectors and re-transformed to Brassica napus hypocotyl and Solanum tuberosum stem segments. The new sequences maintained their promoter activity, demonstrated by transient and stable GUS activity after transformation. Furthermore, the promoters provided sufficient expression of the nptII gene to yield transgenic plants when using kanamycin as selective agent. Database searching (BLASTN) revealed that the promoters have significant homology with three Arabidopsis BAC clones, one Arabidopsis cDNA and one Brassica napus cDNA. The results presented in this paper illustrate the strength of combined methods for identification, isolation and testing of new plant promoters.

  13. A peripherally inserted central catheter line, inserted the day before surgery, decreases the time from induction to incision for spinal deformity surgery and safely provides central venous access during surgery: a pilot study.

    PubMed

    Stuedemann, Anne E; Schwend, Richard M; Thomas, Valorie K; Leamon, Julia M; Lightner, Tammy S

    2017-02-24

    Pediatric patients undergoing surgery for spinal deformity may benefit from central venous access to provide intraoperative monitoring and fluid resuscitation. For pediatric surgical patients requiring central access, we hypothesized that placing a peripherally inserted central catheter (PICC) line preoperatively should decrease time from induction of anesthesia to incision and result in improved patient safety and decreased operating room charges. This was a retrospective, nonrandomized, and case comparison study. Clinical records of all children with adolescent idiopathic scoliosis or neuromuscular scoliosis treated surgically by the senior author between December 2007 and April 2012 were reviewed. Control group patients had a central venous catheter (CVC) placed by the anesthesiologist after induction of anesthesia. The trial group had a PICC placed under local anesthesia the day before surgery by an experienced vascular access team. The time from induction of anesthesia to the time for the surgical incision was determined for each study group. The CVC line placement charges were determined by the operating room time charges at $214/min. Charges saved were the mean time difference multiplied by the operating room time charge, less the charge for PICC line insertion ($1282). There were 29 neuromuscular patients, the mean age was 13 years (SD: 4 years). The mean time from induction to incision for the PICC group was 91 min [95% confidence interval (CI): 67-115 min] and for the CVC group 113 min (95% CI: 99-127 min, P=0.083). For this mean time difference of 22 min, the estimated cost savings would be $3426 per patient. There were 59 patients with adolescent idiopathic scoliosis, the mean age was 14 years (SD: 2 years). The mean time from induction to incision for the PICC group was 78 min (95% CI: 74-82 min) and for the CVC group 106 min (95% CI: 96-116  min, P≤0.001). For this mean time difference of 28 min, the estimated cost savings would

  14. First corrections to hyperfine splitting and the Lamb shift induced by diagrams with two external photons and second-order radiative insertions in the electron line

    SciTech Connect

    Eides, M.I.; Karshenboim, S.G.; Shelyuto, V.A.

    1994-07-01

    Contributions to hyperfine splitting (HFS) and to the Lamb shift on the order of {alpha}{sup 2}(Z{alpha}){sup 5} induced by graphs with two radiative photons inserted into the electron line are considered. It is shown that this gauge-invariant set of diagrams, which give corrections of the considered order, consists of nineteen topologically different diagrams. Contributions to both HFS and the Lamb shift induced by graphs containing the one-loop electron self-energy as a subgraph and by the graph containing two one-loop vertices are obtained. 15 refs., 3 figs.

  15. Corrections to hyperfine splitting and the Lamb shift due to the insertion of the two-loop electron self-energy with overlapping divergences in the electron line

    SciTech Connect

    Eides, M.I.; Karshenboim, S.G.; Shelyuto, V.A.

    1994-12-01

    Contributions on the order of {alpha}{sup 2}(Z{alpha}){sup 5} to hyperfine splitting (HFS) and the Lamb shift due to the insertion of the two-loop electron self-energy diagram with overlapping divergences in the electron line are considered. An explicit expression for the two-loop self-energy diagram with overlapping divergences is obtained in the Fried-Yennie gauge. Contributions to HFS and the Lamb shift due to the diagram containing this subgraph are calculated. 5 refs., 1 fig.

  16. Transcription Activator-Like Effector Nucleases (TALEN)-Mediated Targeted DNA Insertion in Potato Plants.

    PubMed

    Forsyth, Adrienne; Weeks, Troy; Richael, Craig; Duan, Hui

    2016-01-01

    Targeted DNA integration into known locations in the genome has potential advantages over the random insertional events typically achieved using conventional means of genetic modification. Specifically integrated transgenes are guaranteed to co-segregate, and expression level is more predictable, which makes downstream characterization and line selection more manageable. Because the site of DNA integration is known, the steps to deregulation of transgenic crops may be simplified. Here we describe a method that combines transcription activator-like effector nuclease (TALEN)-mediated induction of double strand breaks (DSBs) and non-autonomous marker selection to insert a transgene into a pre-selected, transcriptionally active region in the potato genome. In our experiment, TALEN was designed to create a DSB in the genome sequence following an endogenous constitutive promoter. A cytokinin vector was utilized for TALENs expression and prevention of stable integration of the nucleases. The donor vector contained a gene of interest cassette and a promoter-less plant-derived herbicide resistant gene positioned near the T-DNA left border which was used to select desired transgenic events. Our results indicated that TALEN induced T-DNA integration occurred with high frequency and resulting events have consistent expression of the gene of interest. Interestingly, it was found that, in most lines integration took place through one sided homology directed repair despite the minimal homologous sequence at the right border. An efficient transient assay for TALEN activity verification is also described.

  17. Transcription Activator-Like Effector Nucleases (TALEN)-Mediated Targeted DNA Insertion in Potato Plants

    PubMed Central

    Forsyth, Adrienne; Weeks, Troy; Richael, Craig; Duan, Hui

    2016-01-01

    Targeted DNA integration into known locations in the genome has potential advantages over the random insertional events typically achieved using conventional means of genetic modification. Specifically integrated transgenes are guaranteed to co-segregate, and expression level is more predictable, which makes downstream characterization and line selection more manageable. Because the site of DNA integration is known, the steps to deregulation of transgenic crops may be simplified. Here we describe a method that combines transcription activator-like effector nuclease (TALEN)-mediated induction of double strand breaks (DSBs) and non-autonomous marker selection to insert a transgene into a pre-selected, transcriptionally active region in the potato genome. In our experiment, TALEN was designed to create a DSB in the genome sequence following an endogenous constitutive promoter. A cytokinin vector was utilized for TALENs expression and prevention of stable integration of the nucleases. The donor vector contained a gene of interest cassette and a promoter-less plant-derived herbicide resistant gene positioned near the T-DNA left border which was used to select desired transgenic events. Our results indicated that TALEN induced T-DNA integration occurred with high frequency and resulting events have consistent expression of the gene of interest. Interestingly, it was found that, in most lines integration took place through one sided homology directed repair despite the minimal homologous sequence at the right border. An efficient transient assay for TALEN activity verification is also described. PMID:27826306

  18. A novel two T-DNA binary vector allows efficient generation of marker-free transgenic plants in three elite cultivars of rice (Oryza sativa L.).

    PubMed

    Breitler, Jean-Christophe; Meynard, Donaldo; Van Boxtel, Jos; Royer, Monique; Bonnot, François; Cambillau, Laurence; Guiderdoni, Emmanuel

    2004-06-01

    A pilot binary vector was constructed to assess the potential of the 2 T-DNA system for generating selectable marker-free progeny plants in three elite rice cultivars (ZhongZuo321, Ariete and Khao Dawk Mali 105) known to exhibit contrasting amenabilities to transformation. The first T-DNA of the vector, delimited by Agrobacterium tumefaciens borders, contains the hygromycin phosphotransferase (hpt) selectable gene and the green fluorescent protein (gfp) reporter gene while the second T-DNA, delimited by Agrobacterium rhizogenes borders, bears the phosphinothricin acetyl transferase (bar) gene, featuring the gene of interest. 82-90% of the hygromycin-resistant primary transformants exhibited tolerance to ammonium glufosinate mediated by the bar gene suggesting very high co-transformation frequency in the three cultivars. All of the regenerated plants were analyzed by Southern blot which confirmed co-integration of the T-DNAs at frequencies consistent with those of co-expression and allowed determination of copy number for each gene as well as detection of two different vector backbone fragments extending between the two T-DNAs. Hygromycin susceptible, ammonium glufosinate tolerant phenotypes represented 14.4, 17.4 and 14.3% of the plants in T1 progenies of ZZ321, Ariete and KDML105 primary transformants, respectively. We developed a statistical model for deducing from the observed copy number of each T-DNA in T0 plants and phenotypic segregations in T1 progenies the most likely constitution and linkage of the T-DNA integration locus. Statistical analysis identified in 40 out of 42 lines a most likely linkage configuration theoretically allowing genetic separation of the two T-DNA types and out segregation of the T-DNA bearing the bar gene. Overall, though improvements of the technology would be beneficial, the 2 T-DNA system appeared to be a useful approach to generate selectable marker-free rice plants with a consistent frequency among cultivars.

  19. Integration of an insertion-type transferred DNA vector from Agrobacterium tumefaciens into the Saccharomyces cerevisiae genome by gap repair.

    PubMed Central

    Risseeuw, E; Franke-van Dijk, M E; Hooykaas, P J

    1996-01-01

    Recently, it was shown that Agrobacterium tumefaciens can transfer transferred DNA (T-DNA) to Saccharomyces cerevisiae and that this T-DNA, when used as a replacement vector, is integrated via homologous recombination into the yeast genome. To test whether T-DNA can be a suitable substrate for integration via the gap repair mechanism as well, a model system developed for detection of homologous recombination events in plants was transferred to S. cerevisiae. Analysis of the yeast transformants revealed that an insertion type T-DNA vector can indeed be integrated via gap repair. Interestingly, the transformation frequency and the type of recombination events turned out to depend strongly on the orientation of the insert between the borders in such an insertion type T-DNA vector. PMID:8816506

  20. Delphi Method Validation of a Procedural Performance Checklist for Insertion of an Ultrasound-Guided Internal Jugular Central Line.

    PubMed

    Hartman, Nicholas; Wittler, Mary; Askew, Kim; Manthey, David

    2016-01-01

    Placement of ultrasound-guided central lines is a critical skill for physicians in several specialties. Improving the quality of care delivered surrounding this procedure demands rigorous measurement of competency, and validated tools to assess performance are essential. Using the iterative, modified Delphi technique and experts in multiple disciplines across the United States, the study team created a 30-item checklist designed to assess competency in the placement of ultrasound-guided internal jugular central lines. Cronbach α was .94, indicating an excellent degree of internal consistency. Further validation of this checklist will require its implementation in simulated and clinical environments.

  1. Plastic pipe insertion

    SciTech Connect

    Diskin, J.

    1987-05-01

    In March 1987 KPL changed all that when the utility inserted 1,000 ft of 16-in. SDR 15.5 Phillips Driscopipe 8000 pipe with a wall thickness of 1.032-in., into an abandoned 24-in. cast-iron line in downtown Kansas City. This is believed to be the largest diameter insert removal job ever done for gas distribution in the U.S. For KPL it was a natural progression from the smaller sizes used earlier. The procedure is the same, and the operation was quick and comparatively simple. Lower construction costs were the bottom line because with insert renewal there is no need to cut up the streets, a major expense in any urban pipeline work. There are other significant costs savings as well because the insert renewal construction process is faster than other techniques.

  2. T-DNA integration in plants results from polymerase-θ-mediated DNA repair.

    PubMed

    van Kregten, Maartje; de Pater, Sylvia; Romeijn, Ron; van Schendel, Robin; Hooykaas, Paul J J; Tijsterman, Marcel

    2016-10-31

    Agrobacterium tumefaciens is a pathogenic bacterium, which transforms plants by transferring a discrete segment of its DNA, the T-DNA, to plant cells. The T-DNA then integrates into the plant genome. T-DNA biotechnology is widely exploited in the genetic engineering of model plants and crops. However, the molecular mechanism underlying T-DNA integration remains unknown(1). Here we demonstrate that in Arabidopsis thaliana T-DNA integration critically depends on polymerase theta (Pol θ). We find that TEBICHI/POLQ mutant plants (which have mutated Pol θ), although susceptible to Agrobacterium infection, are resistant to T-DNA integration. Characterization of >10,000 T-DNA-plant genome junctions reveals a distinct signature of Pol θ action and also indicates that 3' end capture at genomic breaks is the prevalent mechanism of T-DNA integration. The primer-template switching ability of Pol θ can explain the molecular patchwork known as filler DNA that is frequently observed at sites of integration. T-DNA integration signatures in other plant species closely resemble those of Arabidopsis, suggesting that Pol-θ-mediated integration is evolutionarily conserved. Thus, Pol θ provides the mechanism for T-DNA random integration into the plant genome, demonstrating a potential to disrupt random integration so as to improve the quality and biosafety of plant transgenesis.

  3. Effect of loss of T-DNA genes on MIA biosynthetic pathway gene regulation and alkaloid accumulation in Catharanthus roseus hairy roots.

    PubMed

    Taneja, Jyoti; Jaggi, Monika; Wankhede, Dhammaprakash Pandhari; Sinha, Alok Krishna

    2010-10-01

    Hairy roots are generated by integration of T-DNA in host plant genome from root inducing (Ri) plasmid of Agrobacterium rhizogenes and have been utilized for production of secondary metabolites in different plant systems. In Catharanthus roseus, hairy roots are known to show different morphologies, growth patterns, and alkaloid contents. It is also known that during transformation, there is a differential loss of a few T-DNA genes. To decipher the effect of loss of T-DNA genes on the various aspects of hairy roots, ten hairy root clones were analyzed for the presence or absence of T-DNA genes and its implications. It was found that the loss of a few ORFs drastically affects the growth and morphological patterns of hairy roots. The absence of T(R)-DNA from hairy roots revealed increased transcript accumulation and higher alkaloid concentrations, whereas callusing among hairy root lines led to decreased transcript and alkaloid accumulation. Significantly higher expression of MIA biosynthetic pathway genes and low abundance of regulator transcripts in hairy root clones in comparison with non-transformed control roots were also observed. This study indicates that it is not only the integration of T-DNA at certain region of host plant genome but also the presence or absence of important ORFs that affects the expression patterns of MIA biosynthetic pathway genes, regulators, and accumulation of specific alkaloids.

  4. Feeding tube insertion - gastrostomy

    MedlinePlus

    ... tube insertion; G-tube insertion; PEG tube insertion; Stomach tube insertion; Percutaneous endoscopic gastrostomy tube insertion ... and down the esophagus, which leads to the stomach. After the endoscopy tube is inserted, the skin ...

  5. LINE-1 Mediated Insertion into Poc1a (Protein of Centriole 1 A) Causes Growth Insufficiency and Male Infertility in Mice

    PubMed Central

    Geister, Krista A.; Brinkmeier, Michelle L.; Cheung, Leonard Y.; Wendt, Jennifer; Oatley, Melissa J.; Burgess, Daniel L.; Kozloff, Kenneth M.; Cavalcoli, James D.; Oatley, Jon M.; Camper, Sally A.

    2015-01-01

    Skeletal dysplasias are a common, genetically heterogeneous cause of short stature that can result from disruptions in many cellular processes. We report the identification of the lesion responsible for skeletal dysplasia and male infertility in the spontaneous, recessive mouse mutant chagun. We determined that Poc1a, encoding protein of the centriole 1a, is disrupted by the insertion of a processed Cenpw cDNA, which is flanked by target site duplications, suggestive of a LINE-1 retrotransposon-mediated event. Mutant fibroblasts have impaired cilia formation and multipolar spindles. Male infertility is caused by defective spermatogenesis early in meiosis and progressive germ cell loss. Spermatogonial stem cell transplantation studies revealed that Poc1a is essential for normal function of both Sertoli cells and germ cells. The proliferative zone of the growth plate is small and disorganized because chondrocytes fail to re-align after cell division and undergo increased apoptosis. Poc1a and several other genes associated with centrosome function can affect the skeleton and lead to skeletal dysplasias and primordial dwarfisms. This mouse mutant reveals how centrosome dysfunction contributes to defects in skeletal growth and male infertility. PMID:26496357

  6. Regional Fluctuation in the Functional Consequence of LINE-1 Insertion in the Mitf Gene: The Black Spotting Phenotype Arisen from the Mitfmi-bw Mouse Lacking Melanocytes.

    PubMed

    Takeda, Kazuhisa; Hozumi, Hiroki; Ohba, Koji; Yamamoto, Hiroaki; Shibahara, Shigeki

    2016-01-01

    Microphthalmia-associated transcription factor (Mitf) is a key regulator for differentiation of melanoblasts, precursors to melanocytes. The mouse homozygous for the black-eyed white (Mitfmi-bw) allele is characterized by the white-coat color and deafness with black eyes due to the lack of melanocytes. The Mitfmi-bw allele carries LINE-1, a retrotransposable element, which results in the Mitf deficiency. Here, we have established the black spotting mouse that was spontaneously arisen from the homozygous Mitfmi-bw mouse lacking melanocytes. The black spotting mouse shows multiple black patches on the white coat, with age-related graying. Importantly, each black patch also contains hair follicles lacking melanocytes, whereas the white-coat area completely lacks melanocytes. RT-PCR analyses of the pigmented patches confirmed that the LINE-1 insertion is retained in the Mitf gene of the black spotting mouse, thereby excluding the possibility of the somatic reversion of the Mitfmi-bw allele. The immunohistochemical analysis revealed that the staining intensity for beta-catenin was noticeably lower in hair follicles lacking melanocytes of the homozygous Mitfmi-bw mouse and the black spotting mouse, compared to the control mouse. In contrast, the staining intensity for beta-catenin and cyclin D1 was higher in keratinocytes of the black spotting mouse, compared to keratinocytes of the control mouse and the Mitfmi-bw mouse. Moreover, the keratinocyte layer appears thicker in the Mitfmi-bw mouse, with the overexpression of Ki-67, a marker for cell proliferation. We also show that the presumptive black spots are formed by embryonic day 15.5. Thus, the black spotting mouse provides the unique model to explore the molecular basis for the survival and death of developing melanoblasts and melanocyte stem cells in the epidermis. These results indicate that follicular melanocytes are responsible for maintaining the epidermal homeostasis; namely, the present study has provided

  7. Percutaneously inserted central catheter - infants

    MedlinePlus

    PICC - infants; PQC - infants; Pic line - infants; Per-Q cath - infants ... A percutaneously inserted central catheter (PICC) is a long, very thin, soft plastic tube that is put into a small blood vessel. This article addresses PICCs in ...

  8. Facility target insert shielding assessment

    SciTech Connect

    Mocko, Michal

    2015-10-06

    Main objective of this report is to assess the basic shielding requirements for the vertical target insert and retrieval port. We used the baseline design for the vertical target insert in our calculations. The insert sits in the 12”-diameter cylindrical shaft extending from the service alley in the top floor of the facility all the way down to the target location. The target retrieval mechanism is a long rod with the target assembly attached and running the entire length of the vertical shaft. The insert also houses the helium cooling supply and return lines each with 2” diameter. In the present study we focused on calculating the neutron and photon dose rate fields on top of the target insert/retrieval mechanism in the service alley. Additionally, we studied a few prototypical configurations of the shielding layers in the vertical insert as well as on the top.

  9. Use of Random T-DNA Mutagenesis in Identification of Gene UvPRO1, A Regulator of Conidiation, Stress Response, and Virulence in Ustilaginoidea virens

    PubMed Central

    Lv, Bo; Zheng, Lu; Liu, Hao; Tang, Jintian; Hsiang, Tom; Huang, Jinbin

    2016-01-01

    False smut of rice, caused by Ustilaginoidea virens (Cooke) Takahashi (teleomorph: Villosiclava virens), is one of the most important diseases affecting rice worldwide. Agrobacterium tumefaciens-mediated transformation was used to identify functional genes in U. virens. In this study, we selected a single-copy insertion mutant T133 with deficiency in producing conidia by screening the T-DNA insertion mutant library of U. virens. The UvPRO1-deletion mutant was successfully obtained after cloning the targeted gene by analysis of the T-DNA insert site of mutant T133. Further research showed that the UvPRO1 mutant was reduced in growth rate and could not produce conidia in PSB medium, while sensitivities to sodium dodecyl sulfate, Congo red, and hyperosmotic stress increased. Moreover, the UvPRO1 deletion mutant hyphae could extend along the surface of spikelets at 1–3 dpi, but mycelia became shriveled and completely lost the ability to infect spikelets at 4 dpi. The relative expression level of UvPRO1 at 8 dpi was more than twice as high as that at 1–2 dpi. These results suggest that UvPRO1 plays a critical role in hyphal growth and conidiation, as well as in stress response and pathogenesis. These findings provide a novel mode of action for the PRO1 protein in fungi and improve the understanding of the function of UvPRO1 in the life cycle of U. virens. PMID:28082958

  10. Translation Start Sequences Affect the Efficiency of Silencing of Agrobacterium tumefaciens T-DNA Oncogenes1

    PubMed Central

    Lee, Hyewon; Humann, Jodi L.; Pitrak, Jennifer S.; Cuperus, Josh T.; Parks, T. Dawn; Whistler, Cheryl A.; Mok, Machteld C.; Ream, L. Walt

    2003-01-01

    Agrobacterium tumefaciens oncogenes cause transformed plant cells to overproduce auxin and cytokinin. Two oncogenes encode enzymes that convert tryptophan to indole-3-acetic acid (auxin): iaaM (tryptophan mono-oxygenase) and iaaH (indole-3-acetamide hydrolase). A third oncogene (ipt) encodes AMP isopentenyl transferase, which produces cytokinin (isopentenyl-AMP). Inactivation of ipt and iaaM (or iaaH) abolishes tumorigenesis. Because adequate means do not exist to control crown gall, we created resistant plants by introducing transgenes designed to elicit posttranscriptional gene silencing (PTGS) of iaaM and ipt. Transgenes that elicit silencing trigger sequence-specific destruction of the inducing RNA and messenger RNAs with related sequences. Although PTGS has proven effective against a variety of target genes, we found that a much higher percentage of transgenic lines silenced iaaM than ipt, suggesting that transgene sequences influenced the effectiveness of PTGS. Sequences required for oncogene silencing included a translation start site. A transgene encoding a translatable sense-strand RNA from the 5′ end of iaaM silenced the iaaM oncogene, but deletion of the translation start site abolished the ability of the transgene to silence iaaM. Silencing A. tumefaciens T-DNA oncogenes is a new and effective method to produce plants resistant to crown gall disease. PMID:12972655

  11. Final technical report for: Insertional Mutagenesis of Brachypodium distachyon DE-AI02-07ER64452

    SciTech Connect

    John, Vogel P.

    2015-10-29

    Several bioenergy grasses are poised to become a major source of energy in the United States. Despite their increasing importance, we know little about the basic biology underlying the traits that control the utility of grasses as energy crops. Better knowledge of grass biology (e.g. identification of the genes that control cell wall composition, plant architecture, cell size, cell division, reproduction, nutrient uptake, carbon flux, etc.) could be used to design rational strategies for crop improvement and shorten the time required to domesticate these species. The use of an appropriate model system is an efficient way to gain this knowledge. Brachypodium distachyon is a small annual grass with all the attributes needed to be a modern model organism including simple growth requirements, fast generation time, small stature, small genome size and self-fertility. These attributes led to the recommendation in the DOE’s “Breaking the Biological Barriers to Cellulosic Ethanol: A Joint Research Agenda” report to propose developing and using B. distachyon as a model for energy crops to accelerate their domestication. Strategic investments (e.g. genome sequencing) in B. distachyon by the DOE are now bearing fruit and B. distachyon is being used as a model grass by hundreds of laboratories worldwide. Sequence indexed insertional mutants are an extremely powerful tool for both forward and reverse genetics. They allow researchers to order mutants in any gene tagged in the collection by simply emailing a request. The goal of this project was to create a collection of sequence indexed insertional mutants (T-DNA lines) for the model grass Brachypodium distachyon in order to facilitate research by the scientific community. During the course of this grant we created a collection of 23,649 B. distachyon T-DNA lines and identified 26,112 unique insertion sites. The collection can be queried through the project website (http://jgi.doe.gov/our-science/science-programs/plant-genomics/brachypodium/brachypodium-t-dna

  12. Insertion Sequences

    PubMed Central

    Mahillon, Jacques; Chandler, Michael

    1998-01-01

    Insertion sequences (ISs) constitute an important component of most bacterial genomes. Over 500 individual ISs have been described in the literature to date, and many more are being discovered in the ongoing prokaryotic and eukaryotic genome-sequencing projects. The last 10 years have also seen some striking advances in our understanding of the transposition process itself. Not least of these has been the development of various in vitro transposition systems for both prokaryotic and eukaryotic elements and, for several of these, a detailed understanding of the transposition process at the chemical level. This review presents a general overview of the organization and function of insertion sequences of eubacterial, archaebacterial, and eukaryotic origins with particular emphasis on bacterial elements and on different aspects of the transposition mechanism. It also attempts to provide a framework for classification of these elements by assigning them to various families or groups. A total of 443 members of the collection have been grouped in 17 families based on combinations of the following criteria: (i) similarities in genetic organization (arrangement of open reading frames); (ii) marked identities or similarities in the enzymes which mediate the transposition reactions, the recombinases/transposases (Tpases); (iii) similar features of their ends (terminal IRs); and (iv) fate of the nucleotide sequence of their target sites (generation of a direct target duplication of determined length). A brief description of the mechanism(s) involved in the mobility of individual ISs in each family and of the structure-function relationships of the individual Tpases is included where available. PMID:9729608

  13. Plant Enzymes but Not Agrobacterium VirD2 Mediate T-DNA Ligation In Vitro

    PubMed Central

    Ziemienowicz, Alicja; Tinland, Bruno; Bryant, John; Gloeckler, Veronique; Hohn, Barbara

    2000-01-01

    Agrobacterium tumefaciens, a gram-negative soil bacterium, transfers DNA to many plant species. In the plant cell, the transferred DNA (T-DNA) is integrated into the genome. An in vitro ligation-integration assay has been designed to investigate the mechanism of T-DNA ligation and the factors involved in this process. The VirD2 protein, which is produced in Agrobacterium and is covalently attached to T-DNA, did not, under our assay conditions, ligate T-DNA to a model target sequence in vitro. We tested whether plant extracts could ligate T-DNA to target oligonucleotides in our test system. The in vitro ligation-integration reaction did indeed take place in the presence of plant extracts. This reaction was inhibited by dTTP, indicating involvement of a plant DNA ligase. We found that prokaryotic DNA ligases could substitute for plant extracts in this reaction. Ligation of the VirD2-bound oligonucleotide to the target sequence mediated by T4 DNA ligase was less efficient than ligation of a free oligonucleotide to the target. T-DNA ligation mediated by a plant enzyme(s) or T4 DNA ligase requires ATP. PMID:10938108

  14. Distribution of Unlinked Transpositions of a Ds Element from a T-DNA Locus on Tomato Chromosome 4

    PubMed Central

    Briza, J.; Carroll, B. J.; Klimyuk, V. I.; Thomas, C. M.; Jones, D. A.; Jones, JDG.

    1995-01-01

    In maize, receptor sites for unlinked transpositions of Activator (Ac) elements are not distributed randomly. To test whether the same is true in tomato, the receptor sites for a Dissociation (Ds) element derived from Ac, were mapped for 26 transpositions unlinked to a donor T-DNA locus on chromosome 4. Four independent transposed Dss mapped to sites on chromosome 4 genetically unlinked to the donor T-DNA, consistent with a preference for transposition to unlinked sites on the same chromosome as opposed to sites on other chromosomes. There was little preference among the nondonor chromosomes, except perhaps for chromosome 2, which carried seven transposed Dss, but these could not be proven to be independent. However, these data, when combined with those from other studies in tomato examining the distribution of transposed Acs or Dss among nondonor chromosomes, suggest there may be absolute preferences for transposition irrespective of the chromosomal location of the donor site. If true, transposition to nondonor chromosomes in tomato would differ from that in maize, where the preference seems to be determined by the spatial arrangement of chromosomes in the interphase nucleus. The tomato lines carrying Ds elements at known locations are available for targeted transposon tagging experiments. PMID:8536985

  15. Identification of pathogenicity-related genes in the rice pathogen Ustilaginoidea virens through random insertional mutagenesis.

    PubMed

    Yu, Mina; Yu, Junjie; Hu, Jiankun; Huang, Lei; Wang, Yahui; Yin, Xiaole; Nie, Yafeng; Meng, Xiangkun; Wang, Weiduo; Liu, Yongfeng

    2015-03-01

    Agrobacterium tumefaciens-mediated transformation (ATMT) is becoming a popular effective system as an insertional mutagenesis tool in filamentous fungi. To gain more insight into the cellular and molecular mechanisms in the pathogenesis of Ustilaginoidea virens, the causal agent of rice false smut disease, a T-DNA insertion mutant library of U. virens was established using ATMT. We optimized a range of conditions to improve the transformation efficiency. Transformants were most effectively obtained when the optimal co-cultivation time is 72h, with 50μM AS in medium and 100μl A. tumefaciens for co-cultivation, leading to the production of 160-185 hygromycin B resistant transformants per 1×10(5) conidia. Southern blot analysis indicated that 58.14% of transformants had a single T-DNA copy. Among 5600 transformants tested for virulence, 37 mutants with reproducible pathogenic defects were obtained. The flanking sequences of three avirulent tranformants (B20, B1015 and B1465) and two pathogenicity-reduced transformants (B726 and B785) were amplified by high-efficiency thermal asymmetric interlaced PCR. Sequence analyses revealed that single T-DNA insertion in mutant B20 targeted the coding region of a gene encoding a protein highly similar to SUN family protein, and in mutant B726 targeted upstream of a gene with unknown function. The two T-DNA insertion sites in mutant B785 were found in the coding region of a gene encoding C6 transcription factor, but failed in amplified flanking sequence of another T-DNA. Chromosomal rearrangement occurred in the genome of mutant B1016 and B1465 with single T-DNA insertion. Among avirulent mutants, B20 showed altered colony growth and pigmentation. The T-DNA insert in B20 was detected in the coding region of a gene named UvSUN2. Morphophysiological characterization analysis suggested that UvSUN2 might be a virulence factor, and possibly required for proper fungal growth, cell wall construction, and stress responses in U. virens

  16. Function search in a large transcription factor gene family in Arabidopsis: assessing the potential of reverse genetics to identify insertional mutations in R2R3 MYB genes.

    PubMed Central

    Meissner, R C; Jin, H; Cominelli, E; Denekamp, M; Fuertes, A; Greco, R; Kranz, H D; Penfield, S; Petroni, K; Urzainqui, A; Martin, C; Paz-Ares, J; Smeekens, S; Tonelli, C; Weisshaar, B; Baumann, E; Klimyuk, V; Marillonnet, S; Patel, K; Speulman, E; Tissier, A F; Bouchez, D; Jones, J J; Pereira, A; Wisman, E

    1999-01-01

    More than 92 genes encoding MYB transcription factors of the R2R3 class have been described in Arabidopsis. The functions of a few members of this large gene family have been described, indicating important roles for R2R3 MYB transcription factors in the regulation of secondary metabolism, cell shape, and disease resistance, and in responses to growth regulators and stresses. For the majority of the genes in this family, however, little functional information is available. As the first step to characterizing these genes functionally, the sequences of >90 family members, and the map positions and expression profiles of >60 members, have been determined previously. An important second step in the functional analysis of the MYB family, through a process of reverse genetics that entails the isolation of insertion mutants, is described here. For this purpose, a variety of gene disruption resources has been used, including T-DNA-insertion populations and three distinct populations that harbor transposon insertions. We report the isolation of 47 insertions into 36 distinct MYB genes by screening a total of 73 genes. These defined insertion lines will provide the foundation for subsequent detailed functional analyses for the assignment of specific functions to individual members of the R2R3 MYB gene family. PMID:10521515

  17. Dihalocarbene Insertion Experiment

    ERIC Educational Resources Information Center

    Goh, S. H.

    1975-01-01

    Describes the insertion reaction using the insertion of carbenes into carbon-hydrogen bonds as an example. Outlines an experiment that will illustrate dihalocarbene insertions into diisopropyl ether. (GS)

  18. Chest tube insertion

    MedlinePlus

    Chest drainage tube insertion; Insertion of tube into chest; Tube thoracostomy; Pericardial drain ... When your chest tube is inserted, you will lie on your side or sit partly upright, with one arm over your head. Sometimes, ...

  19. Extracellular VirB5 enhances T-DNA transfer from Agrobacterium to the host plant.

    PubMed

    Lacroix, Benoît; Citovsky, Vitaly

    2011-01-01

    VirB5 is a type 4 secretion system protein of Agrobacterium located on the surface of the bacterial cell. This localization pattern suggests a function for VirB5 which is beyond its known role in biogenesis and/or stabilization of the T-pilus and which may involve early interactions between Agrobacterium and the host cell. Here, we identify VirB5 as the first Agrobacterium virulence protein that can enhance infectivity extracellularly. Specifically, we show that elevating the amounts of the extracellular VirB5--by exogenous addition of the purified protein, its overexpression in the bacterium, or transgenic expression in and secretion out of the host cell--enhances the efficiency the Agrobacterium-mediated T-DNA transfer, as measured by transient expression of genes contained on the transferred T-DNA molecule. Importantly, the exogenous VirB5 enhanced transient T-DNA expression in sugar beet, a major crop recalcitrant to genetic manipulation. Increasing the pool of the extracellular VirB5 did not complement an Agrobacterium virB5 mutant, suggesting a dual function for VirB5: in the bacterium and at the bacterium-host cell interface. Consistent with this idea, VirB5 expressed in the host cell, but not secreted, had no effect on the transformation efficiency. That the increase in T-DNA expression promoted by the exogenous VirB5 was not due to its effects on bacterial growth, virulence gene induction, bacterial attachment to plant tissue, or host cell defense response suggests that VirB5 participates in the early steps of the T-DNA transfer to the plant cell.

  20. Enhancing T-DNA Transfer Efficiency in Barley (Hordeum vulgare L.) Cells Using Extracellular Cellulose and Lectin.

    PubMed

    Gürel, Filiz; Uçarlı, Cüneyt; Tufan, Feyza; Kalaskar, Deepak M

    2015-06-01

    A major limitation of transforming barley tissues by Agrobacterium tumefaciens is the low frequency of T-DNA transfer due to recalcitrance of barley as a host. The effect of extracellular cellulose and lectin on Agrobacterium transformation efficiency was investigated in this study. Barley callus cultures were transformed with the AGL1 strain containing the vector pBI121 in the presence of 10 mg mL(-1) cellulose or 0.001, 0.05 and 0.1 mg mL(-1) lectin. Addition of cellulose significantly (P ≤ 0.05) increased the number of GUS spots by 50 % compared to standard conditions in the presence of only 200 μM acetosyringone (AS). Frequency of G418-resistant aggregates on the surfaces of callus cultures was 29 and 71.5 %, following AS and AS + cellulose treatments, respectively, after 4 weeks of selection. Presence of 0.05 or 0.1 mg mL(-1) lectin also increased the number of GUS spots and frequency of G418-resistant cells in the selection period, but the increase in blue spots was not significant. We examined the effect of lectin and cellulose on bacterial attachment to callus tissues. Both cellulose and lectin were found to have a significant positive effect on the numbers of bacteria attached to barley callus. Epifluorescence microscopy revealed that Agrobacterium cells had accumulated in the scaffolds of irregular fibrous cellulose with a mean particle size of 200 μm. Expression of nptII in transformed callus lines confirmed the stable transformation of the gene. Our study showed for the first time the binding of Agrobacterium cells to fibrous cellulose and also demonstrated how polysaccharides and glycoproteins can be used to improve T-DNA transfer in monocotyledon transformation procedures.

  1. Production of marker-free and RSV-resistant transgenic rice using a twin T-DNA system and RNAi.

    PubMed

    Jiang, Yayuan; Sun, Lin; Jiang, Mingsong; Li, Kaidong; Song, Yunzhi; Zhu, Changxiang

    2013-09-01

    A twin T-DNA system is a convenient strategy for creating selectable marker-free transgenic plants. The standard transformation plasmid, pCAMBIA 1300, was modified into a binary vector consisting of two separate T-DNAs, one of which contained the hygromycin phosphotransferase (hpt) marker gene. Using this binary vector, we constructed two vectors that expressed inverted-repeat (IR) structures targeting the rice stripe virus (RSV) coat protein (CP) gene and the special-disease protein (SP) gene. Transgenic rice lines were obtained via Agrobacterium-mediated transformation. Seven independent clones harbouring both the hpt marker gene and the target genes (RSV CP or SP) were obtained in the primary transformants of pDTRSVCP and pDTRSVSP, respectively. The segregation frequencies of the target gene and the marker gene in the T1 plants were 8.72 percent for pDTRSVCP and 12.33 percent for pDTRSVSP. Two of the pDTRSVCP lines and three pDTRSVSP lines harbouring the homozygous target gene, but not the hpt gene, were strongly resistant to RSV. A molecular analysis of the resistant transgenic plants confirmed the stable integration and expression of the target genes. The resistant transgenic plants displayed lower levels of the transgene transcripts and specific small interfering RNAs, suggesting that RNAi induced the viral resistance.

  2. The Agrobacterium tumefaciens virulence D2 protein is responsible for precise integration of T-DNA into the plant genome.

    PubMed Central

    Tinland, B; Schoumacher, F; Gloeckler, V; Bravo-Angel, A M; Hohn, B

    1995-01-01

    The VirD2 protein of Agrobacterium tumefaciens was shown to pilot T-DNA during its transfer to the plant cell nucleus. We analyze here its participation in the integration of T-DNA by using a virD2 mutant. This mutation reduces the efficiency of T-DNA transfer, but the efficiency of integration of T-DNA per se is unaffected. Southern and sequence analyses of integration events obtained with the mutated VirD2 protein revealed an aberrant pattern of integration. These results indicate that the wild-type VirD2 protein participates in ligation of the 5'-end of the T-strand to plant DNA and that this ligation step is not rate limiting for T-DNA integration. Images PMID:7628458

  3. The insertion of a full-length Bos taurus LINE element is responsible for a transcriptional deregulation of the Normande Agouti gene.

    PubMed

    Girardot, Michael; Guibert, Sylvain; Laforet, Marie-Pierre; Gallard, Yves; Larroque, Hélène; Oulmouden, Ahmad

    2006-08-01

    Mammalian pigmentation is controlled by the concerted action of Tyr, Tyrp1 and Dct producing eumelanin and/or pheomelanin in melanocytes. The ratio of these two pigments is determined by the agonist alpha-melanocyte stimulating hormone and the antagonist Agouti protein acting on the Mc1r. Here we show that the Agouti gene is over-expressed in Normande breed compared with Prim'Holstein breed. The Normande cattle have a characteristic coat color phenotype with a variable presence of black (eumelanin) hair over a red/brown background. We have found a previously undescribed full-length L1-BT element inserted in the 5'-genomic sequence of the Agouti gene in Normande cattle which promotes the over-expression of alternative transcripts. The variable expression of the alternative transcript directed by the long interspersed nuclear element promoter may be the origin of the brindle coat color pattern of the Normande breed. This new bovine Agouti allele isolated in Normande breed has been named Abr. Finally, as ectopic over-expression of Agouti in Ay mice is responsible for the obesity syndrome, we discuss the possible consequences of Abr for meat and milk production in cattle.

  4. A design for vertical crossing insertions

    SciTech Connect

    Garren, A.

    1985-10-01

    A crossing insertion designed for an SSC with vertically separated 1-in-1 beam lines is presented in this note. The author supposes that the beam lines consist of separate magnets in separate cryostats separated by about 70 cm. He then describes the design, where vertical separation is done with four vertical dipoles producing a steplike beam line.

  5. β-Thalassemia due to intronic LINE-1 insertion in the β-globin gene (HBB): molecular mechanisms underlying reduced transcript levels of the β-globin(L1) allele.

    PubMed

    Lanikova, Lucie; Kucerova, Jana; Indrak, Karel; Divoka, Martina; Issa, Jean-Pierre; Papayannopoulou, Thalia; Prchal, Josef T; Divoky, Vladimir

    2013-10-01

    We describe the molecular etiology of β(+)-thalassemia that is caused by the insertion of the full-length transposable element LINE-1 (L1) into the intron-2 of the β-globin gene (HBB). The transcript level of the affected β-globin gene was severely reduced. The remaining transcripts consisted of full-length, correctly processed β-globin mRNA and a minute amount of three aberrantly spliced transcripts with a decreased half-life due to activation of the nonsense-mediated decay pathway. The lower steady-state amount of mRNA produced by the β-globin(L1) allele also resulted from a reduced rate of transcription and decreased production of full-length β-globin primary transcripts. The promoter and enhancer sequences of the β-globin(L1) allele were hypermethylated; however, treatment with a demethylating agent did not restore the impaired transcription. A histone deacetylase inhibitor partially reactivated the β-globin(L1) transcription despite permanent β-globin(L1) promoter CpG methylation. This result indicates that the decreased rate of transcription from the β-globin(L1) allele is associated with an altered chromatin structure. Therefore, the molecular defect caused by intronic L1 insertion in the β-globin gene represents a novel etiology of β-thalassemia.

  6. Lines

    ERIC Educational Resources Information Center

    Mires, Peter B.

    2006-01-01

    National Geography Standards for the middle school years generally stress the teaching of latitude and longitude. There are many creative ways to explain the great grid that encircles our planet, but the author has found that students in his college-level geography courses especially enjoy human-interest stories associated with lines of latitude…

  7. [Integration of different T-DNA structures of ACC oxidase gene into carnation genome extended cut flower vase-life differently].

    PubMed

    Yu, Yi-Xun; Bao, Man-Zhu

    2004-09-01

    The cultivar 'Master' of carnation (Dianthus caryophyllus L.) was transformed with four T-DNA structures containing sense, antisense, sense direct repeat and antisense direct repeat gene of ACC oxidase mediated by Agrobacterium tumefaciens. Southern blotting detection showed that foreign gene was integrated into the carnation genome and 14 transgenic lines were obtained. The transgenic plants were transplanted to soil and grew normally in greenhouse. Of the 12 transgenic lines screened, the cut flower vase life of 8 transgenic lines is up to 11 days and the longest one is 12.8 days while the vase life of the control is 5.8 days under 25 degrees C. The vase life of 2 lines out of 3 with single sense ACO gene is same as that of the control, while the vase life of 3 lines out of 4 with single antisense ACO gene is prolonged. The vase life of cut flowers of 5 lines with direct repeat ACO genes is all prolonged by about 6 days, while the vase life of 3 out of 7 lines with single ACO gene is same as that of the control. During the senescence of cut flowers, the ethylene production of the most of the transgenic lines decreased significantly, and the production of ethylene is not detectable in lines T456, T556 and T575. The results of the research demonstrate that antisense foreign gene inhibits expression of endogenesis gene more significantly than sense one. Both sense direct repeat and antisense direct repeat foreign genes can suppress endogenous gene expression more significantly comparing to single foreign genes. The transgenic lines obtained from this research are useful to minimize carnation cut flower transportation and storage expenses.

  8. Grommet Having Metal Insert

    DTIC Science & Technology

    1998-09-28

    axially with respect to the body. The 1 means for releasably securing a tool to the insert comprises 2 female threads formed on an inner surface of the...below 10 the flange 32. These surfaces 34, 36 are threaded ( female 11 threads) so that the end of a tool 38 having male threads can 12 engage the...further includes a rigid insert secured to the body in the 12 centrally located aperture. The insert has female threads formed 13 therein for releasably

  9. Ear tube insertion - slideshow

    MedlinePlus

    ... this page: //medlineplus.gov/ency/presentations/100045.htm Ear tube insertion - series—Normal anatomy To use the ... 4 Overview The eardrum (tympanic membrane) separates the ear canal from the middle ear. Review Date 8/ ...

  10. Core Vessel Insert Handling Robot for the Spallation Neutron Source

    SciTech Connect

    Graves, Van B; Dayton, Michael J

    2011-01-01

    The Spallation Neutron Source provides the world's most intense pulsed neutron beams for scientific research and industrial development. Its eighteen neutron beam lines will eventually support up to twenty-four simultaneous experiments. Each beam line consists of various optical components which guide the neutrons to a particular instrument. The optical components nearest the neutron moderators are the core vessel inserts. Located approximately 9 m below the high bay floor, these inserts are bolted to the core vessel chamber and are part of the vacuum boundary. They are in a highly radioactive environment and must periodically be replaced. During initial SNS construction, four of the beam lines received Core Vessel Insert plugs rather than functional inserts. Remote replacement of the first Core Vessel Insert plug was recently completed using several pieces of custom-designed tooling, including a highly complicated Core Vessel Insert Robot. The design of this tool are discussed.

  11. Albino T-DNA tomato mutant reveals a key function of 1-deoxy-D-xylulose-5-phosphate synthase (DXS1) in plant development and survival.

    PubMed

    García-Alcázar, Manuel; Giménez, Estela; Pineda, Benito; Capel, Carmen; García-Sogo, Begoña; Sánchez, Sibilla; Yuste-Lisbona, Fernando J; Angosto, Trinidad; Capel, Juan; Moreno, Vicente; Lozano, Rafael

    2017-03-28

    Photosynthetic activity is indispensable for plant growth and survival and it depends on the synthesis of plastidial isoprenoids as chlorophylls and carotenoids. In the non-mevalonate pathway (MEP), the 1-deoxy-D-xylulose-5-phosphate synthase 1 (DXS1) enzyme has been postulated to catalyze the rate-limiting step in the formation of plastidial isoprenoids. In tomato, the function of DXS1 has only been studied in fruits, and hence its functional relevance during plant development remains unknown. Here we report the characterization of the wls-2297 tomato mutant, whose severe deficiency in chlorophylls and carotenoids promotes an albino phenotype. Additionally, growth of mutant seedlings was arrested without developing vegetative organs, which resulted in premature lethality. Gene cloning and silencing experiments revealed that the phenotype of wls-2297 mutant was caused by 38.6 kb-deletion promoted by a single T-DNA insertion affecting the DXS1 gene. This was corroborated by in vivo and molecular complementation assays, which allowed the rescue of mutant phenotype. Further characterization of tomato plants overexpressing DXS1 and comparative expression analysis indicate that DXS1 may play other important roles besides to that proposed during fruit carotenoid biosynthesis. Taken together, these results demonstrate that DXS1 is essentially required for the development and survival of tomato plants.

  12. Albino T-DNA tomato mutant reveals a key function of 1-deoxy-D-xylulose-5-phosphate synthase (DXS1) in plant development and survival

    PubMed Central

    García-Alcázar, Manuel; Giménez, Estela; Pineda, Benito; Capel, Carmen; García-Sogo, Begoña; Sánchez, Sibilla; Yuste-Lisbona, Fernando J.; Angosto, Trinidad; Capel, Juan; Moreno, Vicente; Lozano, Rafael

    2017-01-01

    Photosynthetic activity is indispensable for plant growth and survival and it depends on the synthesis of plastidial isoprenoids as chlorophylls and carotenoids. In the non-mevalonate pathway (MEP), the 1-deoxy-D-xylulose-5-phosphate synthase 1 (DXS1) enzyme has been postulated to catalyze the rate-limiting step in the formation of plastidial isoprenoids. In tomato, the function of DXS1 has only been studied in fruits, and hence its functional relevance during plant development remains unknown. Here we report the characterization of the wls-2297 tomato mutant, whose severe deficiency in chlorophylls and carotenoids promotes an albino phenotype. Additionally, growth of mutant seedlings was arrested without developing vegetative organs, which resulted in premature lethality. Gene cloning and silencing experiments revealed that the phenotype of wls-2297 mutant was caused by 38.6 kb-deletion promoted by a single T-DNA insertion affecting the DXS1 gene. This was corroborated by in vivo and molecular complementation assays, which allowed the rescue of mutant phenotype. Further characterization of tomato plants overexpressing DXS1 and comparative expression analysis indicate that DXS1 may play other important roles besides to that proposed during fruit carotenoid biosynthesis. Taken together, these results demonstrate that DXS1 is essentially required for the development and survival of tomato plants. PMID:28350010

  13. Small RNAs Derived from the T-DNA of Agrobacterium rhizogenes in Hairy Roots of Phaseolus vulgaris

    PubMed Central

    Peláez, Pablo; Hernández-López, Alejandrina; Estrada-Navarrete, Georgina; Sanchez, Federico

    2017-01-01

    Agrobacterium rhizogenes is a pathogenic bacteria that causes hairy root disease by transferring bacterial DNA into the plant genome. It is an essential tool for industry and research due to its capacity to produce genetically modified roots and whole organisms. Here, we identified and characterized small RNAs generated from the transfer DNA (T-DNA) of A. rhizogenes in hairy roots of common bean (Phaseolus vulgaris). Distinct abundant A. rhizogenes T-DNA-derived small RNAs (ArT-sRNAs) belonging to several oncogenes were detected in hairy roots using high-throughput sequencing. The most abundant and diverse species of ArT-sRNAs were those of 21- and 22-nucleotides in length. Many T-DNA encoded genes constituted phasiRNA producing loci (PHAS loci). Interestingly, degradome analysis revealed that ArT-sRNAs potentially target genes of P. vulgaris. In addition, we detected low levels of ArT-sRNAs in the A. rhizogenes-induced calli generated at the wound site before hairy root emergence. These results suggest that RNA silencing targets several genes from T-DNA of A. rhizogenes in hairy roots of common bean. Therefore, the role of RNA silencing observed in this study has implications in our understanding and usage of this unique plant-bacteria interaction. PMID:28203245

  14. Small RNAs Derived from the T-DNA of Agrobacterium rhizogenes in Hairy Roots of Phaseolus vulgaris.

    PubMed

    Peláez, Pablo; Hernández-López, Alejandrina; Estrada-Navarrete, Georgina; Sanchez, Federico

    2017-01-01

    Agrobacterium rhizogenes is a pathogenic bacteria that causes hairy root disease by transferring bacterial DNA into the plant genome. It is an essential tool for industry and research due to its capacity to produce genetically modified roots and whole organisms. Here, we identified and characterized small RNAs generated from the transfer DNA (T-DNA) of A. rhizogenes in hairy roots of common bean (Phaseolus vulgaris). Distinct abundant A. rhizogenes T-DNA-derived small RNAs (ArT-sRNAs) belonging to several oncogenes were detected in hairy roots using high-throughput sequencing. The most abundant and diverse species of ArT-sRNAs were those of 21- and 22-nucleotides in length. Many T-DNA encoded genes constituted phasiRNA producing loci (PHAS loci). Interestingly, degradome analysis revealed that ArT-sRNAs potentially target genes of P. vulgaris. In addition, we detected low levels of ArT-sRNAs in the A. rhizogenes-induced calli generated at the wound site before hairy root emergence. These results suggest that RNA silencing targets several genes from T-DNA of A. rhizogenes in hairy roots of common bean. Therefore, the role of RNA silencing observed in this study has implications in our understanding and usage of this unique plant-bacteria interaction.

  15. ALS insertion devices

    NASA Astrophysics Data System (ADS)

    Hoyer, E.; Chin, J.; Halbach, K.; Hassenzahl, W. V.; Humphries, D.; Kincaid, B.; Lancaster, H.; Plate, D.

    1991-08-01

    The Advanced Light Source (ALS), the first US third generation synchrotron radiation source, is currently under construction at the Lawrence Berkeley Laboratory. The low-emittance, 1.5 GeV electron storage ring and the insertion devices are specifically designed to produce high brightness beams in the UV to soft X-Ray range. The planned initial complement of insertion devices includes four 4.6 m long undulators, with period lengths of 3.9 cm, 5.0 cm (2) and 8.0 cm, and a 2.9 m long wiggler of 16 cm period length. Undulator design is well advanced and fabrication has begun on the 5.0 cm and 8.0 cm period length undulators. This paper discusses ALS insertion device requirements; general design philosophy; and design of the magnetic structure, support structure/drive systems, control system and vacuum system.

  16. Identification of pathogenicity-related genes in the vascular wilt fungus verticillium dahliae by agrobacterium tumefaciens-mediated t-DNA insertional mutagenesis.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Verticillium dahliae is the causal agent of vascular wilt in many economically important crops worldwide. Identification of genes that underpin pathogenicity or virulence may suggest targets for alternative control methods for this fungus. In this study, Agrobacterium tumefaciens-mediated transform...

  17. Oxalate-minus mutants of Sclerotinia sclerotiorum via T-DNA insertion accumulate fumarate in culture and retain pathogenicity on plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sclerotinia sclerotiorum is a ubiquitous necrotrophic pathogen capable of infecting over 400 plant species including many economically important crops. Oxalic acid production has been shown in numerous studies to be a pathogenicity factor for S. sclerotiorum through several mechanisms. During our ra...

  18. In planta transformation method for T-DNA transfer in orchids

    NASA Astrophysics Data System (ADS)

    Semiarti, Endang; Purwantoro, Aziz; Mercuriani, Ixora S.; Anggriasari, Anida M.; Jang, Seonghoe; Suhandono, Sony; Machida, Yasunori; Machida, Chiyoko

    2014-03-01

    Transgenic plant technology is an efficient tool to study the function of gene(s) in plant. The most popular and widely used technique is Agrobacterium-mediated transformation in which cocultivation was done by immersing the plant tissues/organ in overnight bacterial cultured for about 30 minutes to one hour under in vitro condition. In this experiment, we developed more easier technique that omitted the in vitro step during cocultivation with Agrobacterium, namely in planta transformation method. Pollinaria (compact pollen mass of orchid) of Phalaenopsis amabilis and Spathoglottis plicata orchids were used as target explants that were immersed into bacterial culture for 30 minutes, then dried up the pollinaria, the transformed pollinaria was used to pollinate orchid flowers. The T-DNA used for this experiments were Ubipro∷PaFT/A. tumefaciens GV3101 for P. amabilis and MeEF1α2 pro∷GUS/ A. tumefaciens LBA 4404 for S.plicata. Seeds that were produced from pollinated flowers were grown onto 10 mg/l hygromicin containing NP (New Phalaenopsis) medium. The existance of transgene in putative transformant protocorm (developing orchid embryo) genome was confirmed using PCR with specific primers of either PaFT or GUS genes. Histochemical GUS assay was also performed to the putative transformants. The result showed that transformation frequencies were 2.1 % in P. amabilis, and 0,53% in S. plicata. These results indicates that in planta transformation method could be used for Agrobacterium-mediated genetic transformation, with advantage easier and more secure work from contaminants than that of the in vitro method.

  19. Thought Insertion Clarified

    PubMed Central

    Ratcliffe, Matthew; Wilkinson, Sam

    2016-01-01

    ‘Thought insertion’ in schizophrenia involves somehow experiencing one’s own thoughts as someone else’s. Some philosophers try to make sense of this by distinguishing between ownership and agency: one still experiences oneself as the owner of an inserted thought but attributes it to another agency. In this paper, we propose that thought insertion involves experiencing thought contents as alien, rather than episodes of thinking. To make our case, we compare thought insertion to certain experiences of ‘verbal hallucination’ and show that they amount to different descriptions of the same phenomenon: a quasi-perceptual experience of thought content. We add that the agency/ownership distinction is unhelpful here. What requires explanation is not why a person experiences a type of intentional state without the usual sense of agency, but why she experiences herself as the agent of one type of intentional state rather than another. We conclude by sketching an account of how this might happen. PMID:28123340

  20. Nucleotide sequences of the Pseudomonas savastanoi indoleacetic acid genes show homology with Agrobacterium tumefaciens T-DNA

    PubMed Central

    Yamada, Tetsuji; Palm, Curtis J.; Brooks, Bob; Kosuge, Tsune

    1985-01-01

    We report the nucleotide sequences of iaaM and iaaH, the genetic determinants for, respectively, tryptophan 2-monooxygenase and indoleacetamide hydrolase, the enzymes that catalyze the conversion of L-tryptophan to indoleacetic acid in the tumor-forming bacterium Pseudomonas syringae pv. savastanoi. The sequence analysis indicates that the iaaM locus contains an open reading frame encoding 557 amino acids that would comprise a protein with a molecular weight of 61,783; the iaaH locus contains an open reading frame of 455 amino acids that would comprise a protein with a molecular weight of 48,515. Significant amino acid sequence homology was found between the predicted sequence of the tryptophan monooxygenase of P. savastanoi and the deduced product of the T-DNA tms-1 gene of the octopine-type plasmid pTiA6NC from Agrobacterium tumefaciens. Strong homology was found in the 25 amino acid sequence in the putative FAD-binding region of tryptophan monooxygenase. Homology was also found in the amino acid sequences representing the central regions of the putative products of iaaH and tms-2 T-DNA. The results suggest a strong similarity in the pathways for indoleacetic acid synthesis encoded by genes in P. savastanoi and in A. tumefaciens T-DNA. Images PMID:16593610

  1. Regulation of oncogene expression in T-DNA-transformed host plant cells.

    PubMed

    Zhang, Yi; Lee, Chil-Woo; Wehner, Nora; Imdahl, Fabian; Svetlana, Veselova; Weiste, Christoph; Dröge-Laser, Wolfgang; Deeken, Rosalia

    2015-01-01

    Virulent Agrobacterium tumefaciens strains integrate their T-DNA into the plant genome where the encoded agrobacterial oncogenes are expressed and cause crown gall disease. Essential for crown gall development are IaaH (indole-3-acetamide hydrolase), IaaM (tryptophan monooxygenase) and Ipt (isopentenyl transferase), which encode enzymes for the biosynthesis of auxin (IaaH, IaaM) and cytokinin (Ipt). Although these oncogenes are well studied as the tumor-inducing principle, nothing is known about the regulation of oncogene expression in plant cells. Our studies show that the intergenic regions (IGRs) between the coding sequences (CDS) of the three oncogenes function as promoters in plant cells. These promoters possess a eukaryotic sequence organization and cis-regulatory elements for the binding of plant transcription factors. WRKY18, WRKY40, WRKY60 and ARF5 were identified as activators of the Ipt promoter whereas IaaH and IaaM is constitutively expressed and no transcription factor further activates their promoters. Consistent with these results, the wrky triple mutant plants in particular, develops smaller crown galls than wild-type and exhibits a reduced Ipt transcription, despite the presence of an intact ARF5 gene. WRKY40 and WRKY60 gene expression is induced by A. tumefaciens within a few hours whereas the ARF5 gene is transcribed later during crown gall development. The WRKY proteins interact with ARF5 in the plant nucleus, but only WRKY40 together with ARF5 synergistically boosts the activation of the Ipt promoter in an auxin-dependent manner. From our data, we propose that A. tumefaciens initially induces WRKY40 gene expression as a pathogen defense response of the host cell. The WRKY protein is recruited to induce Ipt expression, which initiates cytokinin-dependent host cell division. With increasing auxin levels triggered by ubiquitous expression of IaaH and IaaM, ARF5 is activated and interacts with WRKY40 to potentiate Ipt expression and balance

  2. Facilitating the recovery of phenotypically normal transgenic lines in clonal crops: a new strategy illustrated in potato.

    PubMed

    Barrell, Philippa J; Conner, Anthony J

    2011-04-01

    Transgenic plants frequently exhibit altered phenotypes, unrelated to transgene expression, which are attributed to tissue culture-induced variation and/or insertional mutagenesis. Distinguishing between these possibilities has been difficult in clonal crops such as potato, due to their highly heterozygous background and the resulting inherent phenotypic variability associated with segregation. This study reports the use of transgene integration as a molecular marker to trace the clonal origin of single cells in tissue culture. Following transformation, multiple shoots have been regenerated from cell colonies of potato (Solanum tuberosum L.) and Southern analysis used to confirm their derivation from a single transformed cell. Analysis of phenotypic variation in field trials has demonstrated marked differences between these multiple regeneration events, the origin of which must have occurred after T-DNA insertion, and consequently during the tissue culture phase. This result unequivocally demonstrates that somaclonal variation occurs during tissue culture and independent of transgene insertion. Furthermore, the first shoots recovered do not necessarily exhibit less somaclonal variation, since later regeneration events can give rise to plants that are more phenotypically normal. Therefore, when developing transgenic lines for genetic improvement of clonal crops, multiple shoots should be regenerated and evaluated from each transformation event to facilitate the recovery of phenotypically normal transgenic lines.

  3. The LORE1 insertion mutant resource.

    PubMed

    Małolepszy, Anna; Mun, Terry; Sandal, Niels; Gupta, Vikas; Dubin, Manu; Urbański, Dorian; Shah, Niraj; Bachmann, Asger; Fukai, Eigo; Hirakawa, Hideki; Tabata, Satoshi; Nadzieja, Marcin; Markmann, Katharina; Su, Junyi; Umehara, Yosuke; Soyano, Takashi; Miyahara, Akira; Sato, Shusei; Hayashi, Makoto; Stougaard, Jens; Andersen, Stig U

    2016-10-01

    Long terminal repeat (LTR) retrotransposons are closely related to retroviruses, and their activities shape eukaryotic genomes. Here, we present a complete Lotus japonicus insertion mutant collection generated by identification of 640 653 new insertion events following de novo activation of the LTR element Lotus retrotransposon 1 (LORE1) (http://lotus.au.dk). Insertion preferences are critical for effective gene targeting, and we exploit our large dataset to analyse LTR element characteristics in this context. We infer the mechanism that generates the consensus palindromes typical of retroviral and LTR retrotransposon insertion sites, identify a short relaxed insertion site motif, and demonstrate selective integration into CHG-hypomethylated genes. These characteristics result in a steep increase in deleterious mutation rate following activation, and allow LORE1 active gene targeting to approach saturation within a population of 134 682 L. japonicus lines. We suggest that saturation mutagenesis using endogenous LTR retrotransposons with germinal activity can be used as a general and cost-efficient strategy for generation of non-transgenic mutant collections for unrestricted use in plant research.

  4. Inserts Automatically Lubricate Ball Bearings

    NASA Technical Reports Server (NTRS)

    Hager, J. A.

    1983-01-01

    Inserts on ball-separator ring of ball bearings provide continuous film of lubricant on ball surfaces. Inserts are machined or molded. Small inserts in ball pockets provide steady supply of lubricant. Technique is utilized on equipment for which maintenance is often poor and lubrication interval is uncertain, such as household appliances, automobiles, and marine engines.

  5. Cryptococcus neoformans Virulence Gene Discovery through Insertional Mutagenesis

    PubMed Central

    Idnurm, Alexander; Reedy, Jennifer L.; Nussbaum, Jesse C.; Heitman, Joseph

    2004-01-01

    Insertional mutagenesis was applied to Cryptococcus neoformans to identify genes associated with virulence attributes. Using biolistic transformation, we generated 4,300 nourseothricin (NAT)-resistant strains, of which 590 exhibited stable resistance. We focused on mutants with defects in established virulence factors and identified two with reduced growth at 37°C, four with reduced production of the antioxidant pigment melanin, and two with an increased sensitivity to nitric oxide (NO). The NAT insertion and mutant phenotypes were genetically linked in five of eight mutants, and the DNA flanking the insertions was characterized. For the strains with altered growth at 37°C and altered melanin production, mutations were in previously uncharacterized genes, while the two NO-sensitive strains bore insertions in the flavohemoglobin gene FHB1, whose product counters NO stress. Because of the frequent instability of nourseothricin resistance associated with biolistic transformation, Agrobacterium-mediated transformation was tested. This transkingdom DNA delivery approach produced 100% stable nourseothricin-resistant transformants, and three melanin-defective strains were identified from 576 transformants, of which 2 were linked to NAT in segregation analysis. One of these mutants contained a T-DNA insertion in the promoter of the LAC1 (laccase) gene, which encodes a key enzyme required for melanin production, while the second contained an insertion in the promoter of the CLC1 gene, encoding a voltage-gated chloride channel. Clc1 and its homologs are required for ion homeostasis, and in their absence Cu+ transport into the secretory pathway is compromised, depriving laccase and other Cu+-dependent proteins of their essential cofactor. The NAT resistance cassette was optimized for cryptococcal codon usage and GC content and was then used to disrupt a mitogen-activated protein kinase gene, a predicted gene, and two putative chloride channel genes to analyze their

  6. Retrotransposon insertions in the clonal evolution of pancreatic ductal adenocarcinoma.

    PubMed

    Rodić, Nemanja; Steranka, Jared P; Makohon-Moore, Alvin; Moyer, Allison; Shen, Peilin; Sharma, Reema; Kohutek, Zachary A; Huang, Cheng Ran; Ahn, Daniel; Mita, Paolo; Taylor, Martin S; Barker, Norman J; Hruban, Ralph H; Iacobuzio-Donahue, Christine A; Boeke, Jef D; Burns, Kathleen H

    2015-09-01

    Pancreatic ductal adenocarcinoma (PDAC) is typically diagnosed after the disease has metastasized; it is among the most lethal forms of cancer. We recently described aberrant expression of an open reading frame 1 protein, ORF1p, encoded by long interspersed element-1 (LINE-1; L1) retrotransposon, in PDAC. To test whether LINE-1 expression leads to somatic insertions of this mobile DNA, we used a targeted method to sequence LINE-1 insertion sites in matched PDAC and normal samples. We found evidence of 465 somatic LINE-1 insertions in 20 PDAC genomes, which were absent from corresponding normal samples. In cases in which matched normal tissue, primary PDAC and metastatic disease sites were available, insertions were found in primary and metastatic tissues in differing proportions. Two adenocarcinomas secondarily involving the pancreas, but originating in the stomach and duodenum, acquired insertions with a similar discordance between primary and metastatic sites. Together, our findings show that LINE-1 contributes to the genetic evolution of PDAC and suggest that somatic insertions are acquired discontinuously in gastrointestinal neoplasms.

  7. Efforts to deregulate Rainbow papaya in Japan: Molecular Characterization of Transgene and Vector Inserts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transformation plasmid-derived insert number and insert site sequence in 55-1 line papaya derivatives Rainbow and SunUp was determined as part of a larger petition to allow its import into Japan (Suzuki, et al., 2007, 2008). Three insertions were detected by Southern analysis and their correspondin...

  8. Impedance calculation for ferrite inserts

    SciTech Connect

    Breitzmann, S.C.; Lee, S.Y.; Ng, K.Y.; /Fermilab

    2005-01-01

    Passive ferrite inserts were used to compensate the space charge impedance in high intensity space charge dominated accelerators. They study the narrowband longitudinal impedance of these ferrite inserts. they find that the shunt impedance and the quality factor for ferrite inserts are inversely proportional to the imaginary part of the permeability of ferrite materials. They also provide a recipe for attaining a truly passive space charge impedance compensation and avoiding narrowband microwave instabilities.

  9. Homology mapping of T-DNA regions on three Agrobacterium rhizogenes Ri plasmids by electron microscope heteroduplex studies.

    PubMed

    Brevet, J; Tempé, J

    1988-03-01

    Recombinant plasmids carrying segments of the Agrobacterium rhizogenes T-DNA regions of the three Ri plasmids 1855 (TL-DNA only), 8196, and 2659 were used for establishing homology maps by electron microscope examination of heteroduplexes. Plasmid DNA was linearized by digestion with suitable restriction endonucleases in order to generate large T-DNA segments. Heteroduplexes were prepared in 50% formamide and spread under standard conditions. Measurements of double and single strands allowed the drawing of homology maps. The three T-DNAs share mainly two homologous sequences of respectively about 2.5 and 1.5 kb, bracketing a largely nonhomologous central part which is about 5.5 kb long. The T-DNAs from pRi1855 and pRi2659 appear to be more related to each other than to that of pRi8196. With reference to the published nucleotide sequence of the TL-DNA of pRiA4 (probably identical to that of pRi1855), ORFs 8 and 14 seem to be the most conserved sequences of the three T-DNAs. The significance of these conserved sequences is unclear since the genetic loci involved in rhizogenicity of agropine strains identified previously are located in nonhomologous regions.

  10. Mechanisms for Complex Chromosomal Insertions

    PubMed Central

    Szafranski, Przemyslaw; Akdemir, Zeynep Coban; Yuan, Bo; Cooper, Mitchell L.; Magriñá, Maria A.; Bacino, Carlos A.; Lalani, Seema R.; Patel, Ankita; Song, Rodger H.; Bi, Weimin; Cheung, Sau Wai; Carvalho, Claudia M. B.; Lupski, James R.

    2016-01-01

    Chromosomal insertions are genomic rearrangements with a chromosome segment inserted into a non-homologous chromosome or a non-adjacent locus on the same chromosome or the other homologue, constituting ~2% of nonrecurrent copy-number gains. Little is known about the molecular mechanisms of their formation. We identified 16 individuals with complex insertions among 56,000 individuals tested at Baylor Genetics using clinical array comparative genomic hybridization (aCGH) and fluorescence in situ hybridization (FISH). Custom high-density aCGH was performed on 10 individuals with available DNA, and breakpoint junctions were fine-mapped at nucleotide resolution by long-range PCR and DNA sequencing in 6 individuals to glean insights into potential mechanisms of formation. We observed microhomologies and templated insertions at the breakpoint junctions, resembling the breakpoint junction signatures found in complex genomic rearrangements generated by replication-based mechanism(s) with iterative template switches. In addition, we analyzed 5 families with apparently balanced insertion in one parent detected by FISH analysis and found that 3 parents had additional small copy-number variants (CNVs) at one or both sides of the inserting fragments as well as at the inserted sites. We propose that replicative repair can result in interchromosomal complex insertions generated through chromothripsis-like chromoanasynthesis involving two or three chromosomes, and cause a significant fraction of apparently balanced insertions harboring small flanking CNVs. PMID:27880765

  11. Imaging of the complications of peripherally inserted central venous catheters.

    PubMed

    Amerasekera, S S H; Jones, C M; Patel, R; Cleasby, M J

    2009-08-01

    Peripherally inserted central catheters (PICC) are widely used to provide central venous access, often in chronically ill patients with long-term intravenous access requirements. There are a number of significant complications related to both insertion and maintenance of PICC lines, including catheter malposition, migration, venous thrombosis, and line fracture. The incidence of these complications is likely to rise as the number of patients undergoing intravenous outpatient therapy increases, with a corresponding rise in radiologist input. This paper provides an overview of the relevant peripheral and central venous anatomy, including anatomical variations, and outlines the complications of PICC lines. Imaging examples demonstrate the range of radiological findings seen in these complications.

  12. Agrobacterium tumefaciens-mediated transformation as an efficient tool for insertional mutagenesis of Cercospora zeae-maydis.

    PubMed

    Lu, Yuanyuan; Xiao, Shuqin; Wang, Fen; Sun, Jiaying; Zhao, Likun; Yan, Libin; Xue, Chunsheng

    2017-02-01

    An efficient Agrobacterium tumefaciens-mediated transformation (ATMT) approach was developed for the plant pathogenic fungus, Cercospora zeae-maydis, which is the causative agent of gray leaf spot in maize. The transformation was evaluated with five parameters to test the efficiencies of transformation. Results showed that spore germination time, co-cultivation temperature and time were the significant influencing factors in all parameters. Randomly selected transformants were confirmed and the transformants were found to be mitotically stable, with single-copy T-DNA integration in the genome. T-DNA flanking sequences were cloned by thermal asymmetric interlaced PCR. Thus, the ATMT approach is an efficient tool for insertional mutagenesis of C. zeae-maydis.

  13. Sink Inserts for Flood Prevention

    NASA Astrophysics Data System (ADS)

    Fleming, Fraser F.; Bodnar, Daniel J.; Hardesty, David L.

    2004-09-01

    A simple, inexpensive insert is described for preventing flooding in lab sinks. The insert is essentially a tube with slots cut into the side that fits snugly into the drain outlet, preventing water buildup and providing additional drainage sites to avoid constriction by small lab items and paper towels.

  14. Cloning of insertion site flanking sequence and construction of transfer DNA insert mutant library in Stylosanthes colletotrichum.

    PubMed

    Chen, Helong; Hu, Caiping; Yi, Kexian; Huang, Guixiu; Gao, Jianming; Zhang, Shiqing; Zheng, Jinlong; Liu, Qiaolian; Xi, Jingen

    2014-01-01

    Stylosanthes sp. is the most important forage legume in tropical areas worldwide. Stylosanthes anthracnose, which is mainly caused by Colletotrichum gloeosporioides, is a globally severe disease in stylo production. Little progress has been made in anthracnose molecular pathogenesis research. In this study, Agrobacterium tumefaciens-mediated transformation was used to transform Stylosanthes colletotrichum strain CH008. The major factors of the genetic transformation system of S. colletotrichum were optimized as follows: A. tumefaciens' AGL-1 concentration (OD(600)), 0.8; concentration of Colletotrichum conidium, 1 × 10(6) conidia/mL; acetosyringone concentration, 100 mmol/L; induction time, 6 h; co-culture temperature, 25 °C; and co-culture time, 3 d. Thus, the transformation efficiency was increased to 300-400 transformants per 106 conidia. Based on the optimized system, a mutant library containing 4616 mutants was constructed, from which some mutants were randomly selected for analysis. Results show that the mutants were single copies that could be stably inherited. The growth rate, spore amount, spore germination rate, and appressorium formation rate in some mutants were significantly different from those in the wild-type strain. We then selected the most appropriate method for the preliminary screening and re-screening of each mutant's pathogenic defects. We selected 1230 transformants, and obtained 23 strains with pathogenic defects, namely, 18 strains with reduced pathogenicity and five strains with lost pathogenicity. Thermal asymmetric interlaced PCR was used to identify the transfer DNA (T-DNA) integration site in the mutant that was coded 2430, and a sequence of 476 bp was obtained. The flanking sequence of T-DNA was compared with the Colletotrichum genome by BLAST, and a sequence of 401 bp was found in Contig464 of the Colletotrichum genome. By predicting the function of the flanking sequence, we discovered that T-DNA insertion in the promoter region

  15. Cloning of Insertion Site Flanking Sequence and Construction of Transfer DNA Insert Mutant Library in Stylosanthes Colletotrichum

    PubMed Central

    Huang, Guixiu; Gao, Jianming; Zhang, Shiqing; Zheng, Jinlong; Liu, Qiaolian; Xi, Jingen

    2014-01-01

    Stylosanthes sp. is the most important forage legume in tropical areas worldwide. Stylosanthes anthracnose, which is mainly caused by Colletotrichum gloeosporioides, is a globally severe disease in stylo production. Little progress has been made in anthracnose molecular pathogenesis research. In this study, Agrobacterium tumefaciens-mediated transformation was used to transform Stylosanthes colletotrichum strain CH008. The major factors of the genetic transformation system of S. colletotrichum were optimized as follows: A. tumefaciens’ AGL-1 concentration (OD600), 0.8; concentration of Colletotrichum conidium, 1×106 conidia/mL; acetosyringone concentration, 100 mmol/L; induction time, 6 h; co-culture temperature, 25°C; and co-culture time, 3 d. Thus, the transformation efficiency was increased to 300–400 transformants per 106 conidia. Based on the optimized system, a mutant library containing 4616 mutants was constructed, from which some mutants were randomly selected for analysis. Results show that the mutants were single copies that could be stably inherited. The growth rate, spore amount, spore germination rate, and appressorium formation rate in some mutants were significantly different from those in the wild-type strain. We then selected the most appropriate method for the preliminary screening and re-screening of each mutant’s pathogenic defects. We selected 1230 transformants, and obtained 23 strains with pathogenic defects, namely, 18 strains with reduced pathogenicity and five strains with lost pathogenicity. Thermal asymmetric interlaced PCR was used to identify the transfer DNA (T-DNA) integration site in the mutant that was coded 2430, and a sequence of 476 bp was obtained. The flanking sequence of T-DNA was compared with the Colletotrichum genome by BLAST, and a sequence of 401 bp was found in Contig464 of the Colletotrichum genome. By predicting the function of the flanking sequence, we discovered that T-DNA insertion in the promoter region of

  16. Beamline Insertions Manager at Jefferson Lab

    SciTech Connect

    Johnson, Michael C.

    2015-09-01

    The beam viewer system at Jefferson Lab provides operators and beam physicists with qualitative and quantitative information on the transverse electron beam properties. There are over 140 beam viewers installed on the 12 GeV CEBAF accelerator. This paper describes an upgrade consisting of replacing the EPICS-based system tasked with managing all viewers with a mixed system utilizing EPICS and high-level software. Most devices, particularly the beam viewers, cannot be safely inserted into the beam line during high-current beam operations. Software is partly responsible for protecting the machine from untimely insertions. The multiplicity of beam-blocking and beam-vulnerable devices motivates us to try a data-driven approach. The beamline insertions application components are centrally managed and configured through an object-oriented software framework created for this purpose. A rules-based engine tracks the configuration and status of every device, along with the beam status of the machine segment containing the device. The application uses this information to decide on which device actions are allowed at any given time.

  17. Tool Removes Coil-Spring Thread Inserts

    NASA Technical Reports Server (NTRS)

    Collins, Gerald J., Jr.; Swenson, Gary J.; Mcclellan, J. Scott

    1991-01-01

    Tool removes coil-spring thread inserts from threaded holes. Threads into hole, pries insert loose, grips insert, then pulls insert to thread it out of hole. Effects essentially reverse of insertion process to ease removal and avoid further damage to threaded inner surface of hole.

  18. Sensitive detection for coralyne and mercury ions based on homo-A/T DNA by exonuclease signal amplification.

    PubMed

    Huang, Hailiang; Shi, Shuo; Zheng, Xuyue; Yao, Tianming

    2015-09-15

    Based on specific homo-A/T DNA binding properties, a strategy for coralyne and mercury ions detection was realised by exonuclease-aided signal amplification. Coralyne could specifically bind homo-A DNA and protect it from the hydrolysis of exonuclease I. The coralyne-protected DNA was subsequently used as a trigger strand to hydrolyze DNA2 in exonuclease-aided signal amplification process. Thiazole orange was used to quantify the remainder DNA2. Under the optimal condition, the fluorescence intensity was linearly proportional to the concentration of coralyne in the range of 0.2-100 nM with a limit of detection (LOD) of 0.31 nM, which presented the lowest LOD for coralyne among all reported. With homo-T and Hg(2+) taking the place of homo-A DNA and coralyne, respectively, the system could also be used for Hg(2+) detection. The experiments in real samples also showed good results. This method was label-free, low-cost, easy-operating and highly repeatable for the detection of coralyne and mercury ions. It could also be extended to detect various analytes, such as other metal ions, proteins and small molecules by using appropriate aptamers.

  19. Gene Insertion Patterns and Sites

    NASA Astrophysics Data System (ADS)

    Vain, Philippe; Thole, Vera

    During the past 25 years, the molecular analysis of transgene insertion patterns and sites in plants has greatly contributed to our understanding of the mechanisms underlying transgene integration, expression, and stability in the nuclear genome. Molecular characterization is also an essential step in the safety assessment of genetically modified crops. This chapter describes the standard experimental procedures used to analyze transgene insertion patterns and loci in cereals and grasses transformed using Agrobacterium tumefaciens or direct transfer of DNA. Methods and protocols enabling the determination of the number and configuration of transgenic loci via a combination of inheritance studies, polymerase chain reaction, and Southern analyses are presented. The complete characterization of transgenic inserts in plants is, however, a holistic process relying on a wide variety of experimental approaches. In this chapter, these additional approaches are not detailed but references to relevant bibliographic records are provided.

  20. Templated Sequence Insertion Polymorphisms in the Human Genome

    PubMed Central

    Onozawa, Masahiro; Aplan, Peter D.

    2016-01-01

    Templated Sequence Insertion Polymorphism (TSIP) is a recently described form of polymorphism recognized in the human genome, in which a sequence that is templated from a distant genomic region is inserted into the genome, seemingly at random. TSIPs can be grouped into two classes based on nucleotide sequence features at the insertion junctions; Class 1 TSIPs show features of insertions that are mediated via the LINE-1 ORF2 protein, including (1) target-site duplication (TSD), (2) polyadenylation 10–30 nucleotides downstream of a “cryptic” polyadenylation signal, and (3) preference for insertion at a 5′-TTTT/A-3′ sequence. In contrast, class 2 TSIPs show features consistent with repair of a DNA double-strand break (DSB) via insertion of a DNA “patch” that is derived from a distant genomic region. Survey of a large number of normal human volunteers demonstrates that most individuals have 25–30 TSIPs, and that these TSIPs track with specific geographic regions. Similar to other forms of human polymorphism, we suspect that these TSIPs may be important for the generation of human diversity and genetic diseases. PMID:27900318

  1. Templated sequence insertion polymorphisms in the human genome

    NASA Astrophysics Data System (ADS)

    Onozawa, Masahiro; Aplan, Peter

    2016-11-01

    Templated Sequence Insertion Polymorphism (TSIP) is a recently described form of polymorphism recognized in the human genome, in which a sequence that is templated from a distant genomic region is inserted into the genome, seemingly at random. TSIPs can be grouped into two classes based on nucleotide sequence features at the insertion junctions; Class 1 TSIPs show features of insertions that are mediated via the LINE-1 ORF2 protein, including 1) target-site duplication (TSD), 2) polyadenylation 10-30 nucleotides downstream of a “cryptic” polyadenylation signal, and 3) preference for insertion at a 5’-TTTT/A-3’ sequence. In contrast, class 2 TSIPs show features consistent with repair of a DNA double-strand break via insertion of a DNA “patch” that is derived from a distant genomic region. Survey of a large number of normal human volunteers demonstrates that most individuals have 25-30 TSIPs, and that these TSIPs track with specific geographic regions. Similar to other forms of human polymorphism, we suspect that these TSIPs may be important for the generation of human diversity and genetic diseases.

  2. Construction and uses of a new transposable element whose insertion is able to produce gene fusions with the neomycin-phosphotransferase-coding region of Tn903.

    PubMed

    Ratet, P; Richaud, F

    1986-01-01

    We describe the construction of a transposable element derived from the Mu phage that upon insertion is able to create a gene fusion between the region of Tn903 coding for neomycin phosphotransferase (NPT I), which confers resistance to aminoglycosides including kanamycin (KmR), neomycin and G418, and the control elements of the gene where the insertion occurs. A chloramphenicol (Cm) transacetylase gene (cat) that confers resistance to Cm is present in the transposon so that transposition events can be monitored even when no active fusions with the nptI coding region occur. The transposase gene is deleted and, therefore, this transposon is perfectly stable upon insertion. The properties of this new transposable element were studied by obtaining gene fusions between the Escherichia coli L-arabinose operon and 'nptI gene. In some of them the KmR phenotype is induced by arabinose. Insertions of this element in cloned fragments of the T-DNA region of Agrobacterium rhizogenes were also isolated. Some of them confer a KmR phenotype upon its E. coli carriers, which indicates that portions of the T-DNA are expressed in these cells.

  3. Characterization of insertion sites in Rainbow papaya, the first commercialized transgenic tree-fruit crop

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inserts and insert sites in transgenic, commercial papaya line 55-1 derivatives Rainbow and SunUp were characterized as part of a petition to Japan to allow import of fresh fruit of these cultivars from the U.S. and to provide data for a larger study aimed at understanding the global impact of DNA t...

  4. Ultrasound guided spine needle insertion

    NASA Astrophysics Data System (ADS)

    Chen, Elvis C. S.; Mousavi, Parvin; Gill, Sean; Fichtinger, Gabor; Abolmaesumi, Purang

    2010-02-01

    An ultrasound (US) guided, CT augmented, spine needle insertion navigational system is introduced. The system consists of an electromagnetic (EM) sensor, an US machine, and a preoperative CT volume of the patient anatomy. Three-dimensional (3D) US volume is reconstructed intraoperatively from a set of two-dimensional (2D) freehand US slices, and is coregistered with the preoperative CT. This allows the preoperative CT volume to be used in the intraoperative clinical coordinate. The spatial relationship between the patient anatomy, surgical tools, and the US transducer are tracked using the EM sensor, and are displayed with respect to the CT volume. The pose of the US transducer is used to interpolate the CT volume, providing the physician with a 2D "x-ray vision" to guide the needle insertion. Many of the system software components are GPU-accelerated, allowing real-time performance of the guidance system in a clinical setting.

  5. Insertion device calculations with mathematica

    SciTech Connect

    Carr, R.; Lidia, S.

    1995-02-01

    The design of accelerator insertion devices such as wigglers and undulators has usually been aided by numerical modeling on digital computers, using code in high level languages like Fortran. In the present era, there are higher level programming environments like IDL{reg_sign}, MatLab{reg_sign}, and Mathematica{reg_sign} in which these calculations may be performed by writing much less code, and in which standard mathematical techniques are very easily used. The authors present a suite of standard insertion device modeling routines in Mathematica to illustrate the new techniques. These routines include a simple way to generate magnetic fields using blocks of CSEM materials, trajectory solutions from the Lorentz force equations for given magnetic fields, Bessel function calculations of radiation for wigglers and undulators and general radiation calculations for undulators.

  6. Inserting Agility in System Development

    DTIC Science & Technology

    2012-07-01

    Agile IT Acquisition, IT Box, Scrum Inserting Agility in System Development Matthew R. Kennedy and Lt Col Dan Ward, USAF With the fast-paced nature...1,700 individuals and 71 countries, found Scrum and eXtreme Programming to be the most widely followed method- ologies (VersionOne, 2007). Other...University http://www.dau.mil 259 Defense ARJ, July 2012, Vol. 19 No. 3 : 249–264 Scrum Scrum is a framework used for project management, which is

  7. Clostridium perfringens gas gangrene at a wrist intravenous line insertion.

    PubMed

    Determann, Catherine; Walker, Craig Andrew

    2013-10-09

    A patient admitted to the intensive care unit for management of hypotension following a multiple medications overdose subsequently deteriorated rapidly with sepsis. A cannula site was noted to be bruised, swollen and erythematous and the X-ray demonstrated gas sitting within the tissues surrounding the metacarpal bones. The patient was referred to the orthopaedic surgeons and quickly taken for debridement of the affected area and fasciotomies of the forearm. Microbiological investigation confirmed Clostridium perfringens to be present in multiple fluid samples taken from the affected site.

  8. Pneumothorax as a complication of central venous catheter insertion.

    PubMed

    Tsotsolis, Nikolaos; Tsirgogianni, Katerina; Kioumis, Ioannis; Pitsiou, Georgia; Baka, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Rapti, Aggeliki; Trakada, Georgia; Katsikogiannis, Nikolaos; Tsakiridis, Kosmas; Karapantzos, Ilias; Karapantzou, Chrysanthi; Barbetakis, Nikos; Zissimopoulos, Athanasios; Kuhajda, Ivan; Andjelkovic, Dejan; Zarogoulidis, Konstantinos; Zarogoulidis, Paul

    2015-03-01

    The central venous catheter (CVC) is a catheter placed into a large vein in the neck [internal jugular vein (IJV)], chest (subclavian vein or axillary vein) or groin (femoral vein). There are several situations that require the insertion of a CVC mainly to administer medications or fluids, obtain blood tests (specifically the "central venous oxygen saturation"), and measure central venous pressure. CVC usually remain in place for a longer period of time than other venous access devices. There are situations according to the drug administration or length of stay of the catheter that specific systems are indicated such as; a Hickman line, a peripherally inserted central catheter (PICC) line or a Port-a-Cath may be considered because of their smaller infection risk. Sterile technique is highly important here, as a line may serve as a port of entry for pathogenic organisms, and the line itself may become infected with organisms such as Staphylococcus aureus and coagulase-negative Staphylococci. In the current review we will present the complication of pneumothorax after CVC insertion.

  9. Pneumothorax as a complication of central venous catheter insertion

    PubMed Central

    Tsotsolis, Nikolaos; Tsirgogianni, Katerina; Kioumis, Ioannis; Pitsiou, Georgia; Baka, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Rapti, Aggeliki; Trakada, Georgia; Katsikogiannis, Nikolaos; Tsakiridis, Kosmas; Karapantzos, Ilias; Karapantzou, Chrysanthi; Barbetakis, Nikos; Zissimopoulos, Athanasios; Kuhajda, Ivan; Andjelkovic, Dejan; Zarogoulidis, Konstantinos

    2015-01-01

    The central venous catheter (CVC) is a catheter placed into a large vein in the neck [internal jugular vein (IJV)], chest (subclavian vein or axillary vein) or groin (femoral vein). There are several situations that require the insertion of a CVC mainly to administer medications or fluids, obtain blood tests (specifically the “central venous oxygen saturation”), and measure central venous pressure. CVC usually remain in place for a longer period of time than other venous access devices. There are situations according to the drug administration or length of stay of the catheter that specific systems are indicated such as; a Hickman line, a peripherally inserted central catheter (PICC) line or a Port-a-Cath may be considered because of their smaller infection risk. Sterile technique is highly important here, as a line may serve as a port of entry for pathogenic organisms, and the line itself may become infected with organisms such as Staphylococcus aureus and coagulase-negative Staphylococci. In the current review we will present the complication of pneumothorax after CVC insertion. PMID:25815301

  10. Large-scale mapping of transposable element insertion sites using digital encoding of sample identity.

    PubMed

    Gohl, Daryl M; Freifeld, Limor; Silies, Marion; Hwa, Jennifer J; Horowitz, Mark; Clandinin, Thomas R

    2014-03-01

    Determining the genomic locations of transposable elements is a common experimental goal. When mapping large collections of transposon insertions, individualized amplification and sequencing is both time consuming and costly. We describe an approach in which large numbers of insertion lines can be simultaneously mapped in a single DNA sequencing reaction by using digital error-correcting codes to encode line identity in a unique set of barcoded pools.

  11. A coumaroyl-ester-3-hydroxylase Insertion Mutant Reveals the Existence of Nonredundant meta-Hydroxylation Pathways and Essential Roles for Phenolic Precursors in Cell Expansion and Plant Growth1[W][OA

    PubMed Central

    Abdulrazzak, Nawroz; Pollet, Brigitte; Ehlting, Jürgen; Larsen, Kim; Asnaghi, Carole; Ronseau, Sebastien; Proux, Caroline; Erhardt, Mathieu; Seltzer, Virginie; Renou, Jean-Pierre; Ullmann, Pascaline; Pauly, Markus; Lapierre, Catherine; Werck-Reichhart, Danièle

    2006-01-01

    Cytochromes P450 monooxygenases from the CYP98 family catalyze the meta-hydroxylation step in the phenylpropanoid biosynthetic pathway. The ref8 Arabidopsis (Arabidopsis thaliana) mutant, with a point mutation in the CYP98A3 gene, was previously described to show developmental defects, changes in lignin composition, and lack of soluble sinapoyl esters. We isolated a T-DNA insertion mutant in CYP98A3 and show that this mutation leads to a more drastic inhibition of plant development and inhibition of cell growth. Similar to the ref8 mutant, the insertion mutant has reduced lignin content, with stem lignin essentially made of p-hydroxyphenyl units and trace amounts of guaiacyl and syringyl units. However, its roots display an ectopic lignification and a substantial proportion of guaiacyl and syringyl units, suggesting the occurrence of an alternative CYP98A3-independent meta-hydroxylation mechanism active mainly in the roots. Relative to the control, mutant plantlets produce very low amounts of sinapoyl esters, but accumulate flavonol glycosides. Reduced cell growth seems correlated with alterations in the abundance of cell wall polysaccharides, in particular decrease in crystalline cellulose, and profound modifications in gene expression and homeostasis reminiscent of a stress response. CYP98A3 thus constitutes a critical bottleneck in the phenylpropanoid pathway and in the synthesis of compounds controlling plant development. CYP98A3 cosuppressed lines show a gradation of developmental defects and changes in lignin content (40% reduction) and structure (prominent frequency of p-hydroxyphenyl units), but content in foliar sinapoyl esters is similar to the control. The purple coloration of their leaves is correlated to the accumulation of sinapoylated anthocyanins. PMID:16377748

  12. Field errors in hybrid insertion devices

    SciTech Connect

    Schlueter, R.D.

    1995-02-01

    Hybrid magnet theory as applied to the error analyses used in the design of Advanced Light Source (ALS) insertion devices is reviewed. Sources of field errors in hybrid insertion devices are discussed.

  13. Insert metering plates for gas turbine nozzles

    DOEpatents

    Burdgick, Steven S.; Itzel, Gary; Chopra, Sanjay; Abuaf, Nesim; Correia, Victor H.

    2004-05-11

    The invention comprises a metering plate which is assembled to an impingement insert for use in the nozzle of a gas turbine. The metering plate can have one or more metering holes and is used to balance the cooling flow within the nozzle. A metering plate with multiple holes reduces static pressure variations which result from the cooling airflow through the metering plate. The metering plate can be assembled to the insert before or after the insert is inserted into the nozzle.

  14. Electromagnetically tracked placement of a peripherally inserted central catheter

    NASA Astrophysics Data System (ADS)

    Sacolick, Laura; Patel, Neilesh; Tang, Jonathan; Levy, Elliot; Cleary, Kevin R.

    2004-05-01

    This paper describes a computer program to utilize electromagnetic tracking guidance during insertion of peripherally inserted central catheters. Placement of a Peripherally Inserted Central Catheter (PICC) line is a relatively simple, routine procedure in which a catheter is inserted into the veins of the lower arm and threaded up the arm to the vena cava to sit just above the heart. However, the procedure requires x-ray verification of the catheter position and is usually done under continuous fluoroscopic guidance. The computer program is designed to replace fluoroscopic guidance in this procedure and make PICC line placement a bedside procedure. This would greatly reduce the time and resources dedicated to this procedure. The physician first goes through a quick registration procedure to register the patient space with the computer screen coordinates. Once registration is completed, the program provides a continuous, real-time display of the position of the catheter tip overlaid on an x-ray image of the patient on an adjacent computer screen. Both the position and orientation of the catheter tip is shown. The display is very similar to that shown when using fluoroscopy.

  15. JT/LJT connector insert material evaluation

    SciTech Connect

    Baca, J.R.F.

    1991-10-01

    Different insert (insulator) materials are undergoing evaluation to replace the Fiberite E-3938 BE96 material currently used. Also being evaluated is the reconfiguration of the insert and metal shell-edge geometries for the purpose of reducing the alleged interference principally responsible for insert damage.

  16. Placement of a peripherally inserted central catheter into the azygous vein

    SciTech Connect

    Franklin, Iain Gilmore, Christopher

    2015-06-15

    Peripherally inserted central catheters (PICC) are used for a variety of infusion therapies. They are indicated in patients requiring long-term venous access. Incorrect positioning of the insertion of a PICC line is one of the known complications when inserting the device in clinical practice. Radiographers once performing imaging will commonly check if the tip of a PICC has entered the superior vena cava. This case study will report on a lesser known incorrect placement of a PICC line into the azygous vein and how this can be detected on radiographic imaging. This outcome for the patient can be detrimental as it has an increased risk of perforation, thrombus, and fistula formation.

  17. Exchange of peripherally inserted central catheters is associated with an increased risk for bloodstream infection.

    PubMed

    McCoy, Michael; Bedwell, Susan; Noori, Shahab

    2011-06-01

    It is not uncommon that the peripherally inserted central catheter (PICC) needs to be replaced either due to blockage or migration to a peripheral position. In such circumstances, there are two methods of PICC placement: new-site insertion and exchange by using the old PICC as a guide wire. Our objective was to investigate risk of infection associated with the exchange method. In this retrospective study, data on all PICC insertions in the neonatal intensive care unit in 2004 to 2008 were obtained. In the population who needed removal of existing PICC and insertion of a new one, we compared central line-associated bloodstream infection (CLABSI) within 1 week of insertion between the two insertion methods. Of 1148 PICC insertions reviewed, 164 (103 new-site and 61 exchange insertions) were performed after removal of a blocked/malpositioned PICC and therefore comprised the study population. The rate of CLABSI was higher in the exchange method (9.8% versus 1%, P < 0.007). After adjusting for the confounders, the odds for CLABSI within 7 days of PICC insertion was higher with the exchange method (odds ratio 25.2, 95% confidence interval: 2.17 to 292.98; P = 0.01). In infants, insertion of PICCs using the exchange method carries an increased risk of bloodstream infection.

  18. Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) Furnace Development

    NASA Technical Reports Server (NTRS)

    Crouch, Myscha; Carswell, William; Farmer, Jeff; Rose, Fred; Tidwell, Paul

    2000-01-01

    This paper presents, in viewgraph form, QMI (Quench Module Insert) and DMI (Diffusion Module Insert) furnace development. The topics include: 1) Furnace Module in Rack; 2) Quench Module Insert; 3) QMI in MSL Core; 4) Diffusion Module Insert; 5) QMI; and 6) QMI Development and Testing.

  19. Focus on peripherally inserted central catheters in critically ill patients

    PubMed Central

    Cotogni, Paolo; Pittiruti, Mauro

    2014-01-01

    associated with each type of CVAD (CICCs or PICCs). Second, an inappropriate CVAD choice and, particularly, an inadequate insertion technique are relevant-and often not recognized-potential risk factors for complications in critically ill patients. We strongly believe that all healthcare professionals involved in the choice, insertion or management of CVADs in critically ill patients should know all potential risk factors of complications. This knowledge may minimize complications and guarantee longevity to the CVAD optimizing the risk/benefit ratio of CVAD insertion and use. Proper management of CVADs in critical care saves lines and lives. Much evidence from the medical literature and from the clinical practice supports our belief that, compared to CICCs, the so-called power-injectable peripherally inserted central catheters are a good alternative choice in critical care. PMID:25374804

  20. Mars Observer Orbit Insertion Briefing

    NASA Technical Reports Server (NTRS)

    1993-01-01

    Steve Wall is the host of this video entitled, "Return to the Red Planet". Live animation of the Mars Observer orbiting Mars is presented. Steve Wall explains the spacecraft insertion maneuver and also explains the purpose for the Mars Observer launch. Live coverage of the Cape Canaveral launch of the Mars Observer is also presented. Suzanne Dodd, Chief of the Mission Planning team describes the burn start and how the spacecraft will be captured by Mars' gravity. Glenn Cunningham, Mars Observer Project Manager, gives background information on the Mars Observer and describes the organizations behind the Mars Observer Spacecraft, such as the Deep Space Network, the Mission Operation Support Office, Science Investigators, the Flight Engineering Office, Operations Office, and the Ground Data System Office. Dr. William Piotrowski, Acting Director, Solar System Exploration Division, NASA, talks about the purpose of the Mars Pathfinder which is to develop the technology and systems for landing small science packages on Mars. Mr. Roger Gibbs, Former Mars Observer Spacecraft Systems Engineer, tells us how the Mars Observer was built and describes the structural elements on the Mars Observer. The 11-month cruise period for the spacecraft is given by Joseph Beerer, Manager of the Engineering office. The thrust for the Mars Orbit Insertion is described by Ronald Klemetson, Technical Manager, Propulsion Subsystem Jet Propulsion Laboratory (JPL). George Chen, Lead Engineer Attitude and Articulation Subsystem Spacecraft Team, explains the importance of the attitude control engines on the Spacecraft. Marvin Traxler, Manager of Tracking and Data Acquisition, describes how searching for a signal from the Mars Observer works. See NONP-NASA-VT-2000081555 for a continuation of this discussion with Marvin Traxler.

  1. Formation of nanofilms on cell surfaces to improve the insertion efficiency of a nanoneedle into cells

    SciTech Connect

    Amemiya, Yosuke; Kawano, Keiko; Matsusaki, Michiya; Akashi, Mitsuru; Nakamura, Noriyuki; Nakamura, Chikashi

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer We examined the insertion efficiency of nanoneedles into fibroblast and neural cells. Black-Right-Pointing-Pointer Nanofilms formed on cell surfaces improved the insertion efficiency of nanoneedles. Black-Right-Pointing-Pointer Nanofilms improved the insertion efficiency even in Y27632-treated cells. -- Abstract: A nanoneedle, an atomic force microscope (AFM) tip etched to 200 nm in diameter and 10 {mu}m in length, can be inserted into cells with the aid of an AFM and has been used to introduce functional molecules into cells and to analyze intracellular information with minimal cell damage. However, some cell lines have shown low insertion efficiency of the nanoneedle. Improvement in the insertion efficiency of a nanoneedle into such cells is a significant issue for nanoneedle-based cell manipulation and analysis. Here, we have formed nanofilms composed of extracellular matrix molecules on cell surfaces and found that the formation of the nanofilms improved insertion efficiency of a nanoneedle into fibroblast and neural cells. The nanofilms were shown to improve insertion efficiency even in cells in which the formation of actin stress fibers was inhibited by the ROCK inhibitor Y27632, suggesting that the nanofilms with the mesh structure directly contributed to the improved insertion efficiency of a nanoneedle.

  2. Characterization of Arabidopsis 6-phosphogluconolactonase T-DNA insertion mutants reveals an essential role for the oxidative section of the plastidic pentose phosphate pathway in plant growth and development.

    PubMed

    Xiong, Yuqing; DeFraia, Christopher; Williams, Donna; Zhang, Xudong; Mou, Zhonglin

    2009-07-01

    Arabidopsis PGL1, PGL2, PGL4 and PGL5 are predicted to encode cytosolic isoforms of 6-phosphogluconolactonase (6PGL), whereas PGL3 is predicted to encode a 6PGL that has been shown to localize in both plastids and peroxisomes. Therefore, 6PGL may exist in the cytosol, plastids and peroxisomes. However, the function of 6PGL in these three subcellular locations has not been well defined. Here we show that PGL3 is essential, whereas PGL1, PGL2 and PGL5 are individually dispensable for plant growth and development. Knockdown of PGL3 in the pgl3 mutant leads to a dramatic decrease in plant size, a significant increase in total glucose-6-phosphate dehydrogenase activity and a marked decrease in cellular redox potential. Interestingly, the pgl3 plants exhibit constitutive pathogenesis-related gene expression and enhanced resistance to Pseudomonas syringae pv. maculicola ES4326 and Hyaloperonospora arabidopsidis Noco2. We found that although pgl3 does not spontaneously accumulate elevated levels of free salicylic acid (SA), the constitutive defense responses in pgl3 plants are almost completely suppressed by the npr1 and sid2/eds16/ics1 mutations, suggesting that the pgl3 mutation activates NPR1- and SID2/EDS16/ICS1-dependent defense responses. We demonstrate that plastidic (not peroxisomal) localization and 6PGL activity of the PGL3 protein are essential for complementing all pgl3 phenotypes, indicating that the oxidative section of the plastidic pentose phosphate pathway (PPP) is required for plant normal growth and development. Thus, pgl3 provides a useful tool not only for defining the role of the PPP in different subcellular compartments, but also for dissecting the SA/NPR1-mediated signaling pathway.

  3. Insertion devices at the advanced photon source

    SciTech Connect

    Moog, E.R.

    1996-07-01

    The insertion devices being installed at the Advanced Photon Source cause the stored particle beam to wiggle, emitting x-rays with each wiggle. These x-rays combine to make an intense beam of radiation. Both wiggler and undulator types of insertion devices are being installed; the characteristics of the radiation produced by these two types of insertion devices are discussed, along with the reasons for those characteristics.

  4. Nozzle insert for mixed mode fuel injector

    DOEpatents

    Lawrence, Keith E.

    2006-11-21

    A fuel injector includes a homogenous charge nozzle outlet set and a conventional nozzle outlet set controlled respectively, by first and second needle valve members. The homogeneous charged nozzle outlet set is defined by a nozzle insert that is attached to an injector body, which defines the conventional nozzle outlet set. The nozzle insert is a one piece metallic component with a large diameter segment separated from a small diameter segment by an annular engagement surface. One of the needle valve members is guided on an outer surface of the nozzle insert, and the nozzle insert has an interference fit attachment to the injector body.

  5. Properties of a symmetric RHIC insertion

    SciTech Connect

    Lee, S.Y.

    1991-07-01

    This report evaluates the lattice functions of the symmetric insertion proposed by A.G. Ruggiero for the RHIC insertion. The crossing geometry, Inner and Outer matching sections, and chromatic properties are studied in detail. Some properties of the missing dipole dispersion correction scheme are also discussed. We found that the chromatic properties of the symmetric insertion is not better than the antisymmetric insertion. The problem is that the four family sextupole correction scheme seems not able to improve the chromatic distortion. Analytic understanding of the failure of the four family sextupole correction scheme will be very useful. 9 figs., 1 tab.

  6. Multipurpose Transposon-Insertion Libraries in Yeast.

    PubMed

    Kumar, Anuj

    2016-06-01

    Libraries of transposon-insertion alleles constitute powerful and versatile tools for large-scale analysis of yeast gene function. Transposon-insertion libraries are constructed most simply through mutagenesis of a plasmid-based genomic DNA library; modification of the mutagenizing transposon by incorporation of yeast selectable markers, recombination sites, and an epitope tag enables the application of insertion alleles for phenotypic screening and protein localization. In particular, yeast genomic DNA libraries have been mutagenized with modified bacterial transposons carrying the URA3 marker, lox recombination sites, and sequence encoding multiple copies of the hemagglutinin (HA) epitope. Mutagenesis with these transposons has yielded a large resource of insertion alleles affecting nearly 4000 yeast genes in total. Through well-established protocols, these insertion libraries can be introduced into the desired strain backgrounds and the resulting insertional mutants can be screened or systematically analyzed. Relative to alternative methods of UV irradiation or chemical mutagenesis, transposon-insertion alleles can be easily identified by PCR-based approaches or high-throughput sequencing. Transposon-insertion libraries also provide a cost-effective alternative to targeted deletion approaches, although, in contrast to start-codon to stop-codon deletions, insertion alleles might not represent true null-mutants. For protein-localization studies, transposon-insertion alleles can provide encoded epitope tags in-frame with internal codons; in many cases, these transposon-encoded epitope tags can provide a more accurate localization for proteins in which terminal sequences are crucial for intracellular targeting. Thus, overall, transposon-insertion libraries can be used quickly and economically and have a particular utility in screening for desired phenotypes and localization patterns in nonstandard genetic backgrounds.

  7. Turbine vane segment and impingement insert configuration for fail-safe impingement insert retention

    DOEpatents

    Burdgick, Steven Sebastian; Kellock, Iain Robertson

    2003-05-13

    An impingement insert sleeve is provided that is adapted to be disposed in a coolant cavity defined through a stator vane. The insert has a generally open inlet end and first and second pairs of diametrically opposed side walls, and at least one fail-safe tab defined at a longitudinal end of the insert for limiting radial displacement of the insert with respect to the stator vane.

  8. Central Solenoid Insert Technical Specification

    SciTech Connect

    Martovetsky, Nicolai N; Smirnov, Alexandre

    2011-09-01

    The US ITER Project Office (USIPO) is responsible for the ITER central solenoid (CS) contribution to the ITER project. The Central Solenoid Insert (CSI) project will allow ITER validation the appropriate lengths of the conductors to be used in the full-scale CS coils under relevant conditions. The ITER Program plans to build and test a CSI to verify the performance of the CS conductor. The CSI is a one-layer solenoid with an inner diameter of 1.48 m and a height of 4.45 m between electric terminal ends. The coil weight with the terminals is approximately 820 kg without insulation. The major goal of the CSI is to measure the temperature margin of the CS under the ITER direct current (DC) operating conditions, including determining sensitivity to load cycles. Performance of the joints, ramp rate sensitivity, and stability against thermal or electromagnetic disturbances, electrical insulation, losses, and instrumentation are addressed separately and therefore are not major goals in this project. However, losses and joint performance will be tested during the CSI testing campaign. The USIPO will build the CSI that will be tested at the Central Solenoid Model Coil (CSMC) Test Facility at the Japan Atomic Energy Agency (JAEA), Naka, Japan. The industrial vendors (the Suppliers) will report to the USIPO (the Company). All approvals to proceed will be issued by the Company, which in some cases, as specified in this document, will also require the approval of the ITER Organization. Responsibilities and obligations will be covered by respective contracts between the USIPO, called Company interchangeably, and the industrial Prime Contractors, called Suppliers. Different stages of work may be performed by more than one Prime Contractor, as described in this specification. Technical requirements of the contract between the Company and the Prime Contractor will be covered by the Fabrication Specifications developed by the Prime Contractor based on this document and approved by

  9. Insertion sequence elements in Lactococcus garvieae.

    PubMed

    Eraclio, Giovanni; Ricci, Giovanni; Fortina, Maria Grazia

    2015-01-25

    Insertion sequences are the simplest intracellular Mobile Genetic Elements which can occur in very high numbers in prokaryotic genomes, where they play an important evolutionary role by promoting genome plasticity. As such, the studies on the diversity and distribution of insertion sequences in genomes not yet investigated can contribute to improve the knowledge on a bacterial species and to identify new transposable elements. The present work describes the occurrence of insertion sequences in Lactococcus garvieae, an opportunistic emerging zoonotic and human pathogen, also associated with different food matrices. To date, no insertion elements have been described for L. garvieae in the IS element database. The analysis of the twelve published L. garvieae genomes identified 15 distinct insertion sequences that are members of the IS3, IS982, IS6, IS21 and IS256 families, including five new elements. Most of the insertion sequences in L. garvieae show substantial homology to the Lactococcus lactis elements, suggesting the movement of IS between these two species phylogenetically closely related. ISLL6 elements belonging to IS3 family were most abundant, with several copies distributed in 9 of the 12 genomes analyzed. An alignment analysis of two complete genomes carrying multi-copies of this insertion sequence indicates a possible involvement of ISLL6 in chromosomal rearrangement.

  10. Sequential cooling insert for turbine stator vane

    DOEpatents

    Jones, Russell B; Krueger, Judson J; Plank, William L

    2014-04-01

    A sequential impingement cooling insert for a turbine stator vane that forms a double impingement for the pressure and suction sides of the vane or a triple impingement. The insert is formed from a sheet metal formed in a zigzag shape that forms a series of alternating impingement cooling channels with return air channels, where pressure side and suction side impingement cooling plates are secured over the zigzag shaped main piece. Another embodiment includes the insert formed from one or two blocks of material in which the impingement channels and return air channels are machined into each block.

  11. Elliptically polarizing adjustable phase insertion device

    DOEpatents

    Carr, Roger

    1995-01-01

    An insertion device for extracting polarized electromagnetic energy from a beam of particles is disclosed. The insertion device includes four linear arrays of magnets which are aligned with the particle beam. The magnetic field strength to which the particles are subjected is adjusted by altering the relative alignment of the arrays in a direction parallel to that of the particle beam. Both the energy and polarization of the extracted energy may be varied by moving the relevant arrays parallel to the beam direction. The present invention requires a substantially simpler and more economical superstructure than insertion devices in which the magnetic field strength is altered by changing the gap between arrays of magnets.

  12. Insertion Profiles of 4 Headless Compression Screws

    PubMed Central

    Hart, Adam; Harvey, Edward J.; Lefebvre, Louis-Philippe; Barthelat, Francois; Rabiei, Reza; Martineau, Paul A.

    2013-01-01

    Purpose In practice, the surgeon must rely on screw position (insertion depth) and tactile feedback from the screwdriver (insertion torque) to gauge compression. In this study, we identified the relationship between interfragmentary compression and these 2 factors. Methods The Acutrak Standard, Acutrak Mini, Synthes 3.0, and Herbert-Whipple implants were tested using a polyurethane foam scaphoid model. A specialized testing jig simultaneously measured compression force, insertion torque, and insertion depth at half-screw-turn intervals until failure occurred. Results The peak compression occurs at an insertion depth of −3.1 mm, −2.8 mm, 0.9 mm, and 1.5 mm for the Acutrak Mini, Acutrak Standard, Herbert-Whipple, and Synthes screws respectively (insertion depth is positive when the screw is proud above the bone and negative when buried). The compression and insertion torque at a depth of −2 mm were found to be 113 ± 18 N and 0.348 ± 0.052 Nm for the Acutrak Standard, 104 ± 15 N and 0.175 ± 0.008 Nm for the Acutrak Mini, 78 ± 9 N and 0.245 ± 0.006 Nm for the Herbert-Whipple, and 67 ± 2N, 0.233 ± 0.010 Nm for the Synthes headless compression screws. Conclusions All 4 screws generated a sizable amount of compression (> 60 N) over a wide range of insertion depths. The compression at the commonly recommended insertion depth of −2 mm was not significantly different between screws; thus, implant selection should not be based on compression profile alone. Conically shaped screws (Acutrak) generated their peak compression when they were fully buried in the foam whereas the shanked screws (Synthes and Herbert-Whipple) reached peak compression before they were fully inserted. Because insertion torque correlated poorly with compression, surgeons should avoid using tactile judgment of torque as a proxy for compression. Clinical relevance Knowledge of the insertion profile may improve our understanding of the implants, provide a better basis for comparing screws

  13. Sequential cooling insert for turbine stator vane

    SciTech Connect

    Jones, Russel B; Krueger, Judson J; Plank, William L

    2014-11-04

    A sequential impingement cooling insert for a turbine stator vane that forms a double impingement for the pressure and suction sides of the vane or a triple impingement. The insert is formed from a sheet metal formed in a zigzag shape that forms a series of alternating impingement cooling channels with return air channels, where pressure side and suction side impingement cooling plates are secured over the zigzag shaped main piece. Another embodiment includes the insert formed from one or two blocks of material in which the impingement channels and return air channels are machined into each block.

  14. Screening insertion libraries for mutations in many genes simultaneously using DNA microarrays

    PubMed Central

    Mahalingam, Ramamurthy; Fedoroff, Nina

    2001-01-01

    We describe a method to screen pools of DNA from multiple transposon lines for insertions in many genes simultaneously. We use thermal asymmetric interlaced–PCR, a hemispecific PCR amplification protocol that combines nested, insertion-specific primers with degenerate primers, to amplify DNA flanking the transposons. In reconstruction experiments with previously characterized Arabidopsis lines carrying insertions of the maize Dissociation (Ds) transposon, we show that fluorescently labeled, transposon-flanking fragments overlapping ORFs hybridize to cognate expressed sequence tags (ESTs) on a DNA microarray. We further show that insertions can be detected in DNA pools from as many as 100 plants representing different transposon lines and that all of the tested, transposon-disrupted genes whose flanking fragments can be amplified individually also can be detected when amplified from the pool. The ability of a transposon-flanking fragment to hybridize declines rapidly with decreasing homology to the spotted DNA fragment, so that only ESTs with >90% homology to the transposon-disrupted gene exhibit significant cross-hybridization. Because thermal asymmetric interlaced–PCR fragments tend to be short, use of the present method favors recovery of insertions in and near genes. We apply the technique to screening pools of new Ds lines using cDNA microarrays containing ESTs for ≈1,000 stress-induced and -repressed Arabidopsis genes. PMID:11416215

  15. Modeling and characterization of partially inserted electrical connector faults

    NASA Astrophysics Data System (ADS)

    Tokgöz, ćaǧatay; Dardona, Sameh; Soldner, Nicholas C.; Wheeler, Kevin R.

    2016-03-01

    Faults within electrical connectors are prominent in avionics systems due to improper installation, corrosion, aging, and strained harnesses. These faults usually start off as undetectable with existing inspection techniques and increase in magnitude during the component lifetime. Detection and modeling of these faults are significantly more challenging than hard failures such as open and short circuits. Hence, enabling the capability to locate and characterize the precursors of these faults is critical for timely preventive maintenance and mitigation well before hard failures occur. In this paper, an electrical connector model based on a two-level nonlinear least squares approach is proposed. The connector is first characterized as a transmission line, broken into key components such as the pin, socket, and connector halves. Then, the fact that the resonance frequencies of the connector shift as insertion depth changes from a fully inserted to a barely touching contact is exploited. The model precisely captures these shifts by varying only two length parameters. It is demonstrated that the model accurately characterizes a partially inserted connector.

  16. Insertable fluid flow passage bridgepiece and method

    DOEpatents

    Jones, Daniel O.

    2000-01-01

    A fluid flow passage bridgepiece for insertion into an open-face fluid flow channel of a fluid flow plate is provided. The bridgepiece provides a sealed passage from a columnar fluid flow manifold to the flow channel, thereby preventing undesirable leakage into and out of the columnar fluid flow manifold. When deployed in the various fluid flow plates that are used in a Proton Exchange Membrane (PEM) fuel cell, bridgepieces of this invention prevent mixing of reactant gases, leakage of coolant or humidification water, and occlusion of the fluid flow channel by gasket material. The invention also provides a fluid flow plate assembly including an insertable bridgepiece, a fluid flow plate adapted for use with an insertable bridgepiece, and a method of manufacturing a fluid flow plate with an insertable fluid flow passage bridgepiece.

  17. Utility Bill Insert for Wastewater Services

    EPA Pesticide Factsheets

    Intended for use by wastewater and water supply utilities, one side of the utility bill insert has information for customers that discharge to sanitary sewer systems; the other side is for customers with septic systems.

  18. Peripherally inserted central catheter - dressing change

    MedlinePlus

    PICC - dressing change ... You have a peripherally inserted central catheter (PICC). This is a tube that goes into a vein in your arm. It carries nutrients and medicines into your body. It may also ...

  19. Small Unit Space Transport and Insertion (SUSTAIN)

    DTIC Science & Technology

    2009-01-01

    Small Unit Space Transport and Insertion ( SUSTAIN ) Study prepared for: LTC Paul E. Damphousse, USMC National Security Space Office Chief of...failing to comply with a collection of information if it does not display a currently valid OMB control number. 1. REPORT DATE 2009 2. REPORT TYPE N/A...3. DATES COVERED - 4. TITLE AND SUBTITLE Small Unit Space Transport and Insertion ( SUSTAIN ) 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM

  20. Efficient instruction sequencing with Inline Target Insertion

    NASA Technical Reports Server (NTRS)

    Hwu, Wen-Mei W.; Chang, Pohua P.

    1992-01-01

    Inline target insertion, a specific compiler and pipeline implementation method for delayed branches with squashing, is defined. The method is shown to offer two important features not discovered in previous studies. First, branches inserted into branch slots are correctly executed. Second, the execution returns correctly from interrupts or exceptions with only one program counter. These two features result in better performance and less software/hardware complexity than conventional delayed branching mechanisms.

  1. Shrink-Fit Solderable Inserts Seal Hermetically

    NASA Technical Reports Server (NTRS)

    Croucher, William C.

    1992-01-01

    Shrink-fit stainless-steel insert in aluminum equipment housing allows electrical connectors to be replaced by soldering, without degrading hermeticity of housing or connector. Welding could destroy electrostatic-sensitive components and harm housing and internal cables. Steel insert avoids problems because connector soldered directly to it rather than welded to housing. Seals between flange and housing, and between connector and flange resistant to leaks, even after mechanical overloading and thermal shocking.

  2. Z-2 Threaded Insert Design and Testing

    NASA Technical Reports Server (NTRS)

    Ross, Amy; Rhodes, Richard; Jones, Robert J.; Graziosi, David; Ferl, Jinny; Sweeny, Mitch; Scarborough, Stephen

    2016-01-01

    NASA's Z-2 prototype space suit contains several components fabricated from an advanced hybrid composite laminate consisting of IM10 carbon fiber and fiber glass. One requirement was to have removable, replaceable helicoil inserts to which other suit components would be fastened. An approach utilizing bonded in inserts with helicoils inside of them was implemented. During initial assembly, cracking sounds were heard followed by the lifting of one of the blind inserts out of its hole when the screws were torqued. A failure investigation was initiated to understand the mechanism of the failure. Ultimately, it was determined that the pre-tension caused by torqueing the fasteners is a much larger force than induced from the pressure loads of the suit which was not considered in the insert design. Bolt tension is determined by dividing the torque on the screw by a k value multiplied by the thread diameter of the bolt. The k value is a factor that accounts for friction in the system. A common value used for k for a non-lubricated screw is 0.2. The k value can go down by as much as 0.1 if the screw is lubricated which means for the same torque, a much larger tension could be placed on the bolt and insert. This paper summarizes the failure investigation that was performed to identify the root cause of the suit failure and details how the insert design was modified to resist a higher pull out tension.

  3. Strength of inserts in titanium alloy machining

    NASA Astrophysics Data System (ADS)

    Kozlov, V.; Huang, Z.; Zhang, J.

    2016-04-01

    In this paper, a stressed state of a non-worn cutting wedge in a machined titanium alloy (Ti6Al2Mo2Cr) is analyzed. The distribution of contact loads on the face of a cutting tool was obtained experimentally with the use of a ‘split cutting tool’. Calculation of internal stresses in the indexable insert made from cemented carbide (WC8Co) was carried out with the help of ANSYS 14.0 software. Investigations showed that a small thickness of the cutting insert leads to extremely high compressive stresses near the cutting edge, stresses that exceed the ultimate compressive strength of cemented carbide. The face and the base of the insert experience high tensile stresses, which approach the ultimate tensile strength of cemented carbide and increase a probability of cutting insert destruction. If the thickness of the cutting insert is bigger than 5 mm, compressive stresses near the cutting edge decrease, and tensile stresses on the face and base decrease to zero. The dependences of the greatest normal and tangential stresses on thickness of the cutting insert were found. Abbreviation and symbols: m/s - meter per second (cutting speed v); mm/r - millimeter per revolution (feed rate f); MPa - mega Pascal (dimension of specific contact loads and stresses); γ - rake angle of the cutting tool [°] α - clearance angle of the sharp cutting tool [°].

  4. Rectal suppository: commonsense and mode of insertion.

    PubMed

    Abd-el-Maeboud, K H; el-Naggar, T; el-Hawi, E M; Mahmoud, S A; Abd-el-Hay, S

    1991-09-28

    Rectal suppository is a well-known form of medication and its use is increasing. The commonest shape is one with an apex (pointed end) tapering to a base (blunt end). Because of a general lack of information about mode of insertion, we asked 360 lay subjects (Egyptians and non-Egyptians) and 260 medical personnel (physicians, pharmacists, and nurses) by questionnaire which end they inserted foremost. Apart from 2 individuals, all subjects suggested insertion with the apex foremost. Commonsense was the most frequent basis for this practice (86.9% of lay subjects and 84.6% of medical personnel) followed by information from a relative, a friend, or medical personnel, or from study at medical school. Suppository insertion with the base or apex foremost was compared in 100 subjects (60 adults, 40 infants and children). Retention with the former method was more easily achieved in 98% of the cases, with no need to introduce a finger in the anal canal (1% vs 83%), and lower expulsion rate (0% vs 3%). The designer of the "torpedo-shaped" suppository suggested its insertion with apex foremost. Our data suggest that a suppository is better inserted with the base foremost. Reversed vermicular contractions or pressure gradient of the anal canal might press it inwards.

  5. Transposable elements: insertion pattern and impact on gene expression evolution in hominids.

    PubMed

    Warnefors, Maria; Pereira, Vini; Eyre-Walker, Adam

    2010-08-01

    Transposable elements (TEs) can affect the regulation of nearby genes through several mechanisms. Here, we examine to what extent recent TE insertions have contributed to the evolution of gene expression in hominids. We compare expression levels of human and chimpanzee orthologs and detect a weak increase in expression divergence (ED) for genes with species-specific TE insertions compared with unaffected genes. However, we show that genes with TE insertions predating the human-chimpanzee split also exhibit a similar increase in ED and therefore conclude that the increase is not due to the transcriptional influence of the TEs. These results are further confirmed by lineage-specific analysis of ED, using rhesus macaque as an outgroup: Human-chimpanzee ortholog pairs, where one ortholog has suffered TE insertion but not the other, do not show increased ED along the lineage where the insertion occurred, relative to the other lineage. We also show that genes with recent TE insertions tend to produce more alternative transcripts but find no evidence that the TEs themselves promote transcript diversity. Finally, we observe that TEs are enriched upstream relative to downstream of genes and show that this is due to insertional bias, rather than selection, because this bias is only observed in genes expressed in the germ line. This provides an alternative neutral explanation for the accumulation of TEs in upstream sequences.

  6. Peptide partitioning properties from direct insertion studies

    SciTech Connect

    Ulmschneider, Martin; Smith, Jeremy C; Ulmschneider, Jakob

    2010-06-01

    Partitioning properties of polypeptides are at the heart of biological membrane phenomena and their precise quantification is vital for ab-initio structure prediction and the accurate simulation of membrane protein folding and function. Recently the cellular translocon machinery has been employed to determine membrane insertion propensities and transfer energetics for a series of polyleucine segments embedded in a carrier sequence. We show here that the insertion propensity, pathway, and transfer energetics into synthetic POPC bilayers can be fully described by direct atomistic peptide partitioning simulations. The insertion probability as a function of peptide length follows two-state Boltzmann statistics, in agreement with the experiments. The simulations expose a systematic offset between translocon-mediated and direct insertion free energies. Compared to the experiment the insertion threshold is shifted toward shorter peptides by 2 leucine residues. The simulations reveal many hitherto unknown atomic-resolution details about the partitioning process and promise to provide a powerful tool for urgently needed calibration of lipid parameters to match experimentally observed peptide transfer energies.

  7. Nozzle cavity impingement/area reduction insert

    DOEpatents

    Yu, Yufeng Phillip; Itzel, Gary Michael; Osgood, Sarah Jane

    2002-01-01

    A turbine vane segment is provided that has inner and outer walls spaced from one another, a vane extending between the inner and outer walls and having leading and trailing edges and pressure and suction sides, the vane including discrete leading edge, intermediate, aft and trailing edge cavities between the leading and trailing edges and extending lengthwise of the vane for flowing a cooling medium; and an insert sleeve within at least one of the cavities and spaced from interior wall surfaces thereof. The insert sleeve has an inlet for flowing the cooling medium into the insert sleeve and has impingement holes defined in first and second walls thereof that respectively face the pressure and suction sides of the vane. The impingement holes of at least one of those first and second walls are defined along substantially only a first, upstream portion thereof, whereby the cooling flow is predominantly impingement cooling along a first region of the insert wall corresponding to the first, upstream portion and the cooling flow is predominantly convective cooling along a second region corresponding to a second, downstream portion of the at least one wall of the insert sleeve.

  8. Reactivity of methacrylates in insertion polymerization.

    PubMed

    Rünzi, Thomas; Guironnet, Damien; Göttker-Schnetmann, Inigo; Mecking, Stefan

    2010-11-24

    Polymerization of ethylene by complexes [{(P^O)PdMe(L)}] (P^O = κ(2)-(P,O)-2-(2-MeOC(6)H(4))(2)PC(6)H(4)SO(3))) affords homopolyethylene free of any methyl methacrylate (MMA)-derived units, even in the presence of substantial concentrations of MMA. In stoichiometric studies, reactive {(P^O)Pd(Me)L} fragments generated by halide abstraction from [({(P^O)Pd(Me)Cl}μ-Na)(2)] insert MMA in a 1,2- as well as 2,1-mode. The 1,2-insertion product forms a stable five-membered chelate by coordination of the carbonyl group. Thermodynamic parameters for MMA insertion are ΔH(++) = 69.0(3.1) kJ mol(-1) and ΔS(++) = -103(10) J mol(-1) K(-1) (total average for 1,2- and 2,1-insertion), in comparison to ΔH(++) = 48.5(3.0) kJ mol(-1) and ΔS(++) = -138(7) J mol(-1) K(-1) for methyl acrylate (MA) insertion. These data agree with an observed at least 10(2)-fold preference for MA incorporation vs MMA incorporation (not detected) under polymerization conditions. Copolymerization of ethylene with a bifunctional acrylate-methacrylate monomer yields linear polyethylenes with intact methacrylate substituents. Post-polymerization modification of the latter was exemplified by free-radical thiol addition and by cross-metathesis.

  9. Interactive simulation of needle insertion models.

    PubMed

    DiMaio, Simon P; Salcudean, Septimiu E

    2005-07-01

    A novel interactive virtual needle insertion simulation is presented. The simulation models are based on measured planar tissue deformations and needle insertion forces. Since the force-displacement relationship is only of interest along the needle shaft, a condensation technique is shown to reduce the computational complexity of linear simulation models significantly. As the needle penetrates or is withdrawn from the tissue model, the boundary conditions that determine the tissue and needle motion change. Boundary condition and local material coordinate changes are facilitated by fast low-rank matrix updates. A large-strain elastic needle model is coupled to the tissue models to account for needle deflection and bending during simulated insertion. A haptic environment, based on these novel interactive simulation techniques, allows users to manipulate a three-degree-of-freedom virtual needle as it penetrates virtual tissue models, while experiencing steering torques and lateral needle forces through a planar haptic interface.

  10. Rack Insertion End Effector (RIEE) automation

    NASA Technical Reports Server (NTRS)

    Malladi, Narasimha

    1993-01-01

    NASA is developing a mechanism to manipulate and insert Racks into the Space Station Logistic modules. The mechanism consists of the following: a base with three motorized degrees of freedom, a 3 section motorized boom that goes from 15 to 44 feet in length, and a Rack Insertion End Effector (RIEE) with 5 hand wheels for precise alignment. The robotics section was tasked with the automation of the RIEE unit. In this report, for the automation of the RIEE unit, application of the Perceptics Vision System was conceptually developed to determine the position and orientation of the RIEE relative to the logistic module, and a MathCad program is written to display the needed displacements for precise alignment and final insertion of the Rack. The uniqueness of this report is that the whole report is in fact a MathCad program including text, derivations, and executable equations with example inputs and outputs.

  11. Elliptically polarizing adjustable phase insertion device

    DOEpatents

    Carr, R.

    1995-01-17

    An insertion device for extracting polarized electromagnetic energy from a beam of particles is disclosed. The insertion device includes four linear arrays of magnets which are aligned with the particle beam. The magnetic field strength to which the particles are subjected is adjusted by altering the relative alignment of the arrays in a direction parallel to that of the particle beam. Both the energy and polarization of the extracted energy may be varied by moving the relevant arrays parallel to the beam direction. The present invention requires a substantially simpler and more economical superstructure than insertion devices in which the magnetic field strength is altered by changing the gap between arrays of magnets. 3 figures.

  12. Study of Uranium Oxide Insertion Compounds

    DTIC Science & Technology

    1993-01-01

    band gap (AE) for U0 3 polymorphs is in the range 2.52 - 2.68 eV (20,325 - • 21,616 cm-1) and the activation energy (Es) for U30 8 is 1.10 eV (8,872...with uranium ions and move by a phonon- activated 0 hopping mechanism. Electrons introduced into the uranium oxide during insertion A - Aj" + e’u (1.7...insertion into a-U 30 8 at - 25"C in 1M LiBF4 in propylene carbonate /1,2- dimethoxyethane Lithium insertion into a-U 30 8 causes very little change in

  13. Genetic transformation of Colletotrichum truncatum associated with anthracnose disease of chili by random insertional mutagenesis.

    PubMed

    Auyong, Adelene Shu Mei; Ford, Rebecca; Taylor, Paul William James

    2012-08-01

    An Agrobacterium tumefaciens -mediated transformation (ATMT) system was successfully developed for Colletotrichum truncatum, the causal agent of chili anthracnose. A. tumefaciens carrying a hygromycin phosphotransferase gene (hph) and a green fluorescent protein (gfp) gene was used to transform the conidiospores of two C. truncatum pathotypes F8-3B and BRIP26974. Optimum transformation efficiency was obtained when equal volumes of A. tumefaciens strain AGL1 carrying either pJF1 or pPK2 binary vector was used to transform C. truncatum conidiospores at 10(6) /ml and co-cultivated at 24 °C for three days. Southern blot analysis indicated that 87.5% of the transformants contained randomly inserted, single copies of the T-DNA. Infection and colonisation of chili fruit at the mature red stage with F8-3B-GFP and BRIP26974-GFP confirmed the maintenance of virulence within these transformed pathotypes. In situ studies of infection and colonisation of the susceptible genotype fruit using fluorescent microscopy and transformed isolates of C. truncatum expressing GFP revealed that the pathogen was able to colonise healthy fruit tissue intercellularly in an endophytic manner without producing secondary biotrophic infection structures. The developed transformation system will be used to study the function of pathogenicity genes in C. truncatum using both forward and reverse genetics approaches.

  14. Mosaic retroposon insertion patterns in placental mammals.

    PubMed

    Churakov, Gennady; Kriegs, Jan Ole; Baertsch, Robert; Zemann, Anja; Brosius, Jürgen; Schmitz, Jürgen

    2009-05-01

    One and a half centuries after Charles Darwin and Alfred Russel Wallace outlined our current understanding of evolution, a new scientific era is dawning that enables direct observations of genetic variation. However, pure sequence-based molecular attempts to resolve the basal origin of placental mammals have so far resulted only in apparently conflicting hypotheses. By contrast, in the mammalian genomes where they were highly active, the insertion of retroelements and their comparative insertion patterns constitute a neutral, virtually homoplasy-free archive of evolutionary histories. The "presence" of a retroelement at an orthologous genomic position in two species indicates their common ancestry in contrast to its "absence" in more distant species. To resolve the placental origin controversy we extracted approximately 2 million potentially phylogenetically informative, retroposon-containing loci from representatives of the major placental mammalian lineages and found highly significant evidence challenging all current single hypotheses of their basal origin. The Exafroplacentalia hypothesis (Afrotheria as the sister group to all remaining placentals) is significantly supported by five retroposon insertions, the Epitheria hypothesis (Xenarthra as the sister group to all remaining placentals) by nine insertion patterns, and the Atlantogenata hypothesis (a monophyletic clade comprising Xenarthra and Afrotheria as the sister group to Boreotheria comprising all remaining placentals) by eight insertion patterns. These findings provide significant support for a "soft" polytomy of the major mammalian clades. Ancestral successive hybridization events and/or incomplete lineage sorting associated with short speciation intervals are viable explanations for the mosaic retroposon insertion patterns of recent placental mammals and for the futile search for a clear root dichotomy.

  15. Mosaic retroposon insertion patterns in placental mammals

    PubMed Central

    Churakov, Gennady; Kriegs, Jan Ole; Baertsch, Robert; Zemann, Anja; Brosius, Jürgen; Schmitz, Jürgen

    2009-01-01

    One and a half centuries after Charles Darwin and Alfred Russel Wallace outlined our current understanding of evolution, a new scientific era is dawning that enables direct observations of genetic variation. However, pure sequence-based molecular attempts to resolve the basal origin of placental mammals have so far resulted only in apparently conflicting hypotheses. By contrast, in the mammalian genomes where they were highly active, the insertion of retroelements and their comparative insertion patterns constitute a neutral, virtually homoplasy-free archive of evolutionary histories. The “presence” of a retroelement at an orthologous genomic position in two species indicates their common ancestry in contrast to its “absence” in more distant species. To resolve the placental origin controversy we extracted ∼2 million potentially phylogenetically informative, retroposon-containing loci from representatives of the major placental mammalian lineages and found highly significant evidence challenging all current single hypotheses of their basal origin. The Exafroplacentalia hypothesis (Afrotheria as the sister group to all remaining placentals) is significantly supported by five retroposon insertions, the Epitheria hypothesis (Xenarthra as the sister group to all remaining placentals) by nine insertion patterns, and the Atlantogenata hypothesis (a monophyletic clade comprising Xenarthra and Afrotheria as the sister group to Boreotheria comprising all remaining placentals) by eight insertion patterns. These findings provide significant support for a “soft” polytomy of the major mammalian clades. Ancestral successive hybridization events and/or incomplete lineage sorting associated with short speciation intervals are viable explanations for the mosaic retroposon insertion patterns of recent placental mammals and for the futile search for a clear root dichotomy. PMID:19261842

  16. True anteroposterior view pedicle screw insertion technique

    PubMed Central

    Bai, Jia-yue; Zhang, Wei; An, Ji-long; Sun, Ya-peng; Ding, Wen-yuan; Shen, Yong

    2016-01-01

    Background The wide use of minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) surgery in the treatment of degenerative disc disease of lumbar spine in spinal surgery highlights the gradual decrease in the use of traditional pedicle screw insertion technology. This study aims to analyze the accuracy of the true anteroposterior view pedicle screw insertion technique in MIS-TLIF surgery, compare it with conventional pedicle screw insertion technology, and discuss its clinical application value. Methods Fifty-two patients undergoing true anteroposterior view (group A) and 87 patients undergoing conventional pedicle screw insertion (group B) were diagnosed with lumbar disc herniation or lumbar spinal stenosis. Time for screw placement, intraoperative irradiation exposure, accuracy rate of pedicle screw insertion, and incidence of neurovascular injury were compared between the two groups. Results The time for screw placement and intraoperative irradiation exposure was significantly less in group A. Penetration rates of the paries lateralis of vertebral pedicle, medial wall of vertebral pedicle, and anterior vertebral wall were 1.44%, 0%, and 2.40%, respectively, all of which were significantly lower than that in group B. No additional serious complications caused by the placement of screw were observed during the follow-up period in patients in group A, but two patients with medial penetration underwent revision for unbearable radicular pain. Conclusion The application of true anteroposterior view pedicle screw insertion technique in MIS-TLIF surgery shortens time for screw placement and reduces the intraoperative irradiation exposure along with a higher accuracy rate of screw placement, which makes it a safe, accurate, and efficient technique. PMID:27418828

  17. Circular permutant GFP insertion folding reporters

    SciTech Connect

    Waldo, Geoffrey S.; Cabantous, Stephanie

    2013-04-16

    Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.

  18. Circular permutant GFP insertion folding reporters

    DOEpatents

    Waldo, Geoffrey S.; Cabantous, Stephanie

    2008-06-24

    Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.

  19. Circular permutant GFP insertion folding reporters

    DOEpatents

    Waldo, Geoffrey S; Cabantous, Stephanie

    2013-02-12

    Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.

  20. Circular permutant GFP insertion folding reporters

    DOEpatents

    Waldo, Geoffrey S.; Cabantous, Stephanie

    2011-06-14

    Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.

  1. The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes.

    PubMed Central

    Bellen, Hugo J; Levis, Robert W; Liao, Guochun; He, Yuchun; Carlson, Joseph W; Tsang, Garson; Evans-Holm, Martha; Hiesinger, P Robin; Schulze, Karen L; Rubin, Gerald M; Hoskins, Roger A; Spradling, Allan C

    2004-01-01

    The Berkeley Drosophila Genome Project (BDGP) strives to disrupt each Drosophila gene by the insertion of a single transposable element. As part of this effort, transposons in >30,000 fly strains were localized and analyzed relative to predicted Drosophila gene structures. Approximately 6300 lines that maximize genomic coverage were selected to be sent to the Bloomington Stock Center for public distribution, bringing the size of the BDGP gene disruption collection to 7140 lines. It now includes individual lines predicted to disrupt 5362 of the 13,666 currently annotated Drosophila genes (39%). Other lines contain an insertion at least 2 kb from others in the collection and likely mutate additional incompletely annotated or uncharacterized genes and chromosomal regulatory elements. The remaining strains contain insertions likely to disrupt alternative gene promoters or to allow gene misexpression. The expanded BDGP gene disruption collection provides a public resource that will facilitate the application of Drosophila genetics to diverse biological problems. Finally, the project reveals new insight into how transposons interact with a eukaryotic genome and helps define optimal strategies for using insertional mutagenesis as a genomic tool. PMID:15238527

  2. The BDGP gene disruption project: Single transposon insertions associated with 40 percent of Drosophila genes

    SciTech Connect

    Bellen, Hugo J.; Levis, Robert W.; Liao, Guochun; He, Yuchun; Carlson, Joseph W.; Tsang, Garson; Evans-Holm, Martha; Hiesinger, P. Robin; Schulze, Karen L.; Rubin, Gerald M.; Hoskins, Roger A.; Spradling, Allan C.

    2004-01-13

    The Berkeley Drosophila Genome Project (BDGP) strives to disrupt each Drosophila gene by the insertion of a single transposable element. As part of this effort, transposons in more than 30,000 fly strains were localized and analyzed relative to predicted Drosophila gene structures. Approximately 6,300 lines that maximize genomic coverage were selected to be sent to the Bloomington Stock Center for public distribution, bringing the size of the BDGP gene disruption collection to 7,140 lines. It now includes individual lines predicted to disrupt 5,362 of the 13,666 currently annotated Drosophila genes (39 percent). Other lines contain an insertion at least 2 kb from others in the collection and likely mutate additional incompletely annotated or uncharacterized genes and chromosomal regulatory elements. The remaining strains contain insertions likely to disrupt alternative gene promoters or to allow gene mis-expression. The expanded BDGP gene disruption collection provides a public resource that will facilitate the application of Drosophila genetics to diverse biological problems. Finally, the project reveals new insight into how transposons interact with a eukaryotic genome and helps define optimal strategies for using insertional mutagenesis as a genomic tool.

  3. PBmice: an integrated database system of piggyBac (PB) insertional mutations and their characterizations in mice

    PubMed Central

    Sun, Ling V.; Jin, Ke; Liu, Yiming; Yang, Wenwei; Xie, Xing; Ye, Lin; Wang, Li; Zhu, Lin; Ding, Sheng; Su, Yi; Zhou, Jie; Han, Min; Zhuang, Yuan; Xu, Tian; Wu, Xiaohui; Gu, Ning; Zhong, Yang

    2008-01-01

    DNA transposon piggyBac (PB) is a newly established mutagen for large-scale mutagenesis in mice. We have designed and implemented an integrated database system called PBmice (PB Mutagenesis Information CEnter) for storing, retrieving and displaying the information derived from PB insertions (INSERTs) in the mouse genome. This system is centered on INSERTs with information including their genomic locations and flanking genomic sequences, the expression levels of the hit genes, and the expression patterns of the trapped genes if a trapping vector was used. It also archives mouse phenotyping data linked to INSERTs, and allows users to conduct quick and advanced searches for genotypic and phenotypic information relevant to a particular or a set of INSERT(s). Sequence-based information can be cross-referenced with other genomic databases such as Ensembl, BLAST and GBrowse tools used in PBmice offer enhanced search and display for additional information relevant to INSERTs. The total number and genomic distribution of PB INSERTs, as well as the availability of each PB insertional LINE can also be viewed with user-friendly interfaces. PBmice is freely available at http://www.idmshanghai.cn/PBmice or http://www.scbit.org/PBmice/. PMID:17932058

  4. LINE-ABOVE-GROUND ATTENUATOR

    DOEpatents

    Wilds, R.B.; Ames, J.R.

    1957-09-24

    The line-above-ground attenuator provides a continuously variable microwave attenuator for a coaxial line that is capable of high attenuation and low insertion loss. The device consists of a short section of the line-above- ground plane type transmission lime, a pair of identical rectangular slabs of lossy material like polytron, whose longitudinal axes are parallel to and indentically spaced away from either side of the line, and a geared mechanism to adjust amd maintain this spaced relationship. This device permits optimum fineness and accuracy of attenuator control which heretofore has been difficult to achieve.

  5. 21 CFR 886.5420 - Contact lens inserter/remover.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Contact lens inserter/remover. 886.5420 Section... (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5420 Contact lens inserter/remover. (a) Identification. A contact lens inserter/remover is a handheld device intended to insert or...

  6. 21 CFR 886.5420 - Contact lens inserter/remover.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Contact lens inserter/remover. 886.5420 Section... (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5420 Contact lens inserter/remover. (a) Identification. A contact lens inserter/remover is a handheld device intended to insert or...

  7. 21 CFR 886.5420 - Contact lens inserter/remover.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Contact lens inserter/remover. 886.5420 Section... (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5420 Contact lens inserter/remover. (a) Identification. A contact lens inserter/remover is a handheld device intended to insert or...

  8. 21 CFR 886.5420 - Contact lens inserter/remover.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Contact lens inserter/remover. 886.5420 Section... (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5420 Contact lens inserter/remover. (a) Identification. A contact lens inserter/remover is a handheld device intended to insert or...

  9. 21 CFR 886.5420 - Contact lens inserter/remover.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Contact lens inserter/remover. 886.5420 Section... (CONTINUED) MEDICAL DEVICES OPHTHALMIC DEVICES Therapeutic Devices § 886.5420 Contact lens inserter/remover. (a) Identification. A contact lens inserter/remover is a handheld device intended to insert or...

  10. Patients' experiences of the PICC insertion procedure.

    PubMed

    Nicholson, Jackie; Davies, Louise

    Peripherally inserted central catheters (PICCs) are a type of central venous access device used to deliver a variety of intravenous therapies, including chemotherapy. PICCs may be placed by interventional radiologists, anaesthetists or, as is increasingly common, by specialist nurses in the hospital setting. However, little is known about how patients feel regarding the PICC insertion procedure. The aim of this study was to interview patients who had undergone a recent PICC insertion in the chemotherapy day unit to identify their experiences. On analysis of the qualitative data obtained from the semi-structured interview, five themes emerged: the context of cancer; expectations; levels of pain and anxiety; coping strategies; and explanation. The findings of this study support some previously described elements of procedural experiences; however, new understanding has provided implications for practice in the areas of expectations, allaying anxiety levels, supporting individual coping strategies and providing explanation. The major limitation of the study was the homogenous sample of oncology patients with a clear link between the patient experience of the PICC insertion and the context of cancer. The main recommendation for further research would be to repeat this study with a broader patient population.

  11. Thermal Performance of the XRS Helium Insert

    NASA Technical Reports Server (NTRS)

    Breon, Susan R.; DiPirro, Michael J.; Tuttle, James G.; Shirron, Peter J.; Warner, Brent A.; Boyle, Robert F.; Canavan, Edgar R.

    1999-01-01

    The X-Ray Spectrometer (XRS) is an instrument on the Japanese Astro-E satellite, scheduled for launch early in the year 2000. The XRS Helium Insert comprises a superfluid helium cryostat, an Adiabatic Demagnetization Refrigerator (ADR), and the XRS calorimeters with their cold electronics. The calorimeters are capable of detecting X-rays over the energy range 0.1 to 10 keV with a resolution of 12 eV. The Helium Insert completed its performance and verification testing at Goddard in January 1999. It was shipped to Japan, where it has been integrated with the neon dewar built by Sumitomo Heavy Industries. The Helium Insert was given a challenging lifetime requirement of 2.0 years with a goal of 2.5 years. Based on the results of the thermal performance tests, the predicted on-orbit lifetime is 2.6 years with a margin of 30%. This is the result of both higher efficiency in the ADR cycle and the low temperature top-off, more than compensating for an increase in the parasitic heat load. This paper presents a summary of the key design features and the results of the thermal testing of the XRS Helium Insert.

  12. Insertion Loss of Personal Protective Clothing

    SciTech Connect

    Shull D.J.; Biesel, V.B.; Cunefare, K.A.

    1999-05-13

    'The use of personal protective clothing that covers the head is a common practice in many industries. Such personal protective clothing will impact the sound pressure level and the frequency content of sounds to which the wearer will be exposed. The use of such clothing, then, may impact speech and alarm audibility. A measure of the impact of such clothing is its insertion loss. Insertion loss measurements were performed on four types of personal protective clothing in use by Westinghouse Savannah River Company personnel which utilize cloth and plastic hood configurations to protect the head. All clothing configurations tested at least partially cover the ears. The measurements revealed that insertion loss of the items tested was notable at frequencies above 1000 Hz only and was a function of material stiffness and acoustic flanking paths to the ear. Further, an estimate of the clothing''s noise reduction rating reveals poor performance in that regard, even though the insertion loss of the test articles was significant at frequencies at and above 1000 Hz.'

  13. Frequency, Gradience, and Variation in Consonant Insertion

    ERIC Educational Resources Information Center

    An, Young-ran

    2010-01-01

    This dissertation addresses the extent to which linguistic behavior can be described in terms of the projection of patterns from existing lexical items, through an investigation of Korean reduplication. Korean has a productive pattern of reduplication in which a consonant is inserted in a vowel-initial base, illustrated by forms such as "alok"--"t…

  14. Peripherally inserted central catheters. Intravenous Nurses Society.

    PubMed

    1997-01-01

    The Intravenous Nurses Society (INS) recognizes the need for uniform terminology for peripherally inserted central catheters (PICCs) to encourage standardization for indications, care, and maintenance strategies for these devices. It also recognizes the need for recommendations regarding the choice, use, management, and discontinuation of PICCs to promote positive patient outcomes and enhance patient comfort, safety, and satisfaction.

  15. Lymphatic Leak Complicating Central Venous Catheter Insertion

    SciTech Connect

    Barnacle, Alex M. Kleidon, Tricia M.

    2005-12-15

    Many of the risks associated with central venous access are well recognized. We report a case of inadvertent lymphatic disruption during the insertion of a tunneled central venous catheter in a patient with raised left and right atrial pressures and severe pulmonary hypertension, which led to significant hemodynamic instability. To our knowledge, this rare complication is previously unreported.

  16. Embedded Multiprocessor Technology for VHSIC Insertion

    NASA Technical Reports Server (NTRS)

    Hayes, Paul J.

    1990-01-01

    Viewgraphs on embedded multiprocessor technology for VHSIC insertion are presented. The objective was to develop multiprocessor system technology providing user-selectable fault tolerance, increased throughput, and ease of application representation for concurrent operation. The approach was to develop graph management mapping theory for proper performance, model multiprocessor performance, and demonstrate performance in selected hardware systems.

  17. Inserting new technology into small missions

    NASA Technical Reports Server (NTRS)

    Deutsch, L. J.

    2001-01-01

    Part of what makes small missions small is that they have less money. Executing missions at low cost implies extensive use of cost sharing with other missions or use of existing solutions. Luckily, there are methods for creating new technology and inserting it into faster-better-cheaper missions.

  18. Consolidation and disposal of PWR fuel inserts

    SciTech Connect

    Wakeman, B.H. )

    1992-08-01

    Design and licensing of the Surry Power Station Independent Spent Fuel Storage Installation was initiated in 1982 by Virginia Power as part of a comprehensive strategy to increase spent fuel storage capacity at the Station. Designed to use large, metal dry storage casks, the Surry Installation will accommodate 84 such casks with a total storage capacity of 811 MTU of spent pressurized water reactor fuel assemblies. Virginia Power provided three storage casks for testing at the Idaho National Engineerinq Laboratory's Test Area North and the testing results have been published by the Electric Power Research Institute. Sixty-nine spent fuel assemblies were transported in truck casks from the Surry Power Station to Test Area North for testing in the three casks. Because of restrictions imposed by the cask testing equipment at Test Area North, the irradiated insert components stored in these fuel assemblies at Surry were removed prior to transport of the fuel assemblies. Retaining these insert components proved to be a problem because of a shortage of spent fuel assemblies in the spent fuel storage pool that did not already contain insert components. In 1987 Virginia Power contracted with Chem-Nuclear Systems, Inc. to process and dispose of 136 irradiated insert components consisting of 125 burnable poison rod assemblies, 10 thimble plugging devices and 1 part-length rod cluster control assembly. This work was completed in August and September 1987, culminating in the disposal at the Barnwell, SC low-level radioactive waste facility of two CNS 3-55 liners containing the consolidated insert components.

  19. Toolbox for mobile-element insertion detection on cancer genomes.

    PubMed

    Lee, Wan-Ping; Wu, Jiantao; Marth, Gabor T

    2015-01-01

    Mobile elements constitute greater than 45% of the human genome as a result of repeated insertion events during human genome evolution. Although most of mobile elements are fixed within the human population, some elements (including ALU, long interspersed elements (LINE) 1 (L1), and SVA) are still actively duplicating and may result in life-threatening human diseases such as cancer, motivating the need for accurate mobile-element insertion (MEI) detection tools. We developed a software package, TANGRAM, for MEI detection in next-generation sequencing data, currently serving as the primary MEI detection tool in the 1000 Genomes Project. TANGRAM takes advantage of valuable mapping information provided by our own MOSAIK mapper, and until recently required MOSAIK mappings as its input. In this study, we report a new feature that enables TANGRAM to be used on alignments generated by any mainstream short-read mapper, making it accessible for many genomic users. To demonstrate its utility for cancer genome analysis, we have applied TANGRAM to the TCGA (The Cancer Genome Atlas) mutation calling benchmark 4 dataset. TANGRAM is fast, accurate, easy to use, and open source on https://github.com/jiantao/Tangram.

  20. Toolbox for mobile-element insertion detection on cancer genomes.

    PubMed

    Lee, Wan-Ping; Wu, Jiantao; Marth, Gabor T

    2014-01-01

    Mobile elements constitute greater than 45% of the human genome as a result of repeated insertion events during human genome evolution. Although most of mobile elements are fixed within the human population, some elements (including ALU, long interspersed elements (LINE) 1 (L1), and SVA) are still actively duplicating and may result in life-threatening human diseases such as cancer, motivating the need for accurate mobile-element insertion (MEI) detection tools. We developed a software package, TANGRAM, for MEI detection in next-generation sequencing data, currently serving as the primary MEI detection tool in the 1000 Genomes Project. TANGRAM takes advantage of valuable mapping information provided by our own MOSAIK mapper, and until recently required MOSAIK mappings as its input. In this study, we report a new feature that enables TANGRAM to be used on alignments generated by any mainstream short-read mapper, making it accessible for many genomic users. To demonstrate its utility for cancer genome analysis, we have applied TANGRAM to the TCGA (The Cancer Genome Atlas) mutation calling benchmark 4 dataset. TANGRAM is fast, accurate, easy to use, and open source on https://github.com/jiantao/Tangram.

  1. Toolbox for Mobile-Element Insertion Detection on Cancer Genomes

    PubMed Central

    Lee, Wan-Ping; Wu, Jiantao; Marth, Gabor T

    2015-01-01

    Mobile elements constitute greater than 45% of the human genome as a result of repeated insertion events during human genome evolution. Although most of mobile elements are fixed within the human population, some elements (including ALU, long interspersed elements (LINE) 1 (L1), and SVA) are still actively duplicating and may result in life-threatening human diseases such as cancer, motivating the need for accurate mobile-element insertion (MEI) detection tools. We developed a software package, TANGRAM, for MEI detection in next-generation sequencing data, currently serving as the primary MEI detection tool in the 1000 Genomes Project. TANGRAM takes advantage of valuable mapping information provided by our own MOSAIK mapper, and until recently required MOSAIK mappings as its input. In this study, we report a new feature that enables TANGRAM to be used on alignments generated by any mainstream short-read mapper, making it accessible for many genomic users. To demonstrate its utility for cancer genome analysis, we have applied TANGRAM to the TCGA (The Cancer Genome Atlas) mutation calling benchmark 4 dataset. TANGRAM is fast, accurate, easy to use, and open source on https://github.com/jiantao/Tangram. PMID:25931804

  2. Distal Insertions of the Biceps Femoris

    PubMed Central

    Branch, Eric A.; Anz, Adam W.

    2015-01-01

    Background: Avulsion of the biceps femoris from the fibula and proximal tibia is encountered in clinical practice. While the anatomy of the primary posterolateral corner structures has been qualitatively and quantitatively described, a quantitative analysis regarding the insertions of the biceps femoris on the fibula and proximal tibia is lacking. Purpose: To quantitatively assess the insertions of the biceps femoris, fibular collateral ligament (FCL), and anterolateral ligament (ALL) on the fibula and proximal tibia as well as establish relationships among these structures and to pertinent surgical anatomy. Study Design: Descriptive laboratory study. Methods: Dissections were performed on 12 nonpaired, fresh-frozen cadaveric specimens identifying the biceps femoris, FCL, and ALL, and their insertions on the proximal tibia and fibula. The footprint areas, orientations, and distances from relevant osseous landmarks were measured using a 3-dimensional coordinate measurement device. Results: Dissection produced 6 easily identifiable and reproducible anatomic footprints. Tibial footprints included the insertion of the ALL and an insertion of the biceps femoris (TBF). Fibular footprints included the insertion of the FCL, a distal insertion of the biceps femoris (DBF), a medial footprint of the biceps femoris (MBF), and a proximal footprint of the biceps femoris (PBF). The mean area of these footprints (95% CI) was as follows: ALL, 53.0 mm2 (38.4-67.6); TBF, 93.9 mm2 (72.0-115.8); FCL, 86.8 mm2 (72.3-101.2); DBF, 119 mm2 (91.1-146.9); MBF, 46.8 mm2 (29.0-64.5); and PBF, 215 mm2 (192.4-237.5). The mean distance (95% CI) from the Gerdy tubercle to the center of the ALL footprint was 24.3 mm (21.6-27.0) and to the center of the TBF was 22.5 mm (21.0-24.0). The center of the DBF was 8.68 mm (7.0-10.3) from the anterior border of the fibula, the center of the FCL was 14.6 mm (12.5-16.7) from the anterior border of the fibula and 20.7 mm (19.0-22.4) from the tip of the fibular

  3. Effect of Off-Axis Screw Insertion, Insertion Torque, and Plate Contouring on Locked Screw Strength

    PubMed Central

    Gallagher, Bethany; Silva, Matthew J.; Ricci, William M.

    2015-01-01

    Objectives This study quantifies the effects of insertion torque, off-axis screw angulation, and plate contouring on the strength of locking plate constructs. Methods Groups of locking screws (n = 6–11 screws) were inserted at 50%, 100%, 150%, and 200% of the manufacturer-recommended torque (3.2 Nm) into locking compression plates at various angles: orthogonal (control), 5-degree angle off-axis, and 10-degree angle off-axis. Screws were loaded to failure by a transverse force (parallel to the plate) either in the same (“+”) or opposite direction (“−”) of the initial screw angulation. Separately, locking plates were bent to 5 and 10-degree angles, with the bend apex at a screw hole. Locking screws inserted orthogonally into the apex hole at 100% torque were loaded to failure. Results Orthogonal insertion resulted in the highest average load to failure, 2577 ± 141 N (range, 2413–2778 N), whereas any off-axis insertion significantly weakened constructs (165–1285 N, at 100% torque) (P < 0.05). For “+” loading, torque beyond 100% did not increase strength, but 50% torque reduced screw strength (P < 0.05). Loading in the “−” direction consistently resulted in higher strengths than “+” loading (P < 0.05). Plate contouring of 5-degree angle did not significantly change screw strength compared with straight plates but contouring of 10-degree angle significantly reduced load to failure (P < 0.05). Conclusions To maximize the screw plate interface strength, locking screws should be inserted without cross-threading. The mechanical stability of locked screws is significantly compromised by loose insertion, off-axis insertion, or severe distortion of the locking mechanism. PMID:24343255

  4. Descemet's Stripping Automated Endothelial Keratoplasty Tissue Insertion Devices

    PubMed Central

    Khan, Salman Nasir; Shiakolas, Panos S.; Mootha, Venkateswara Vinod

    2015-01-01

    This review study provides information regarding the construction, design, and use of six commercially available endothelial allograft insertion devices applied for Descemet's stripping automated endothelial keratoplasty (DSAEK). We also highlight issues being faced in DSAEK and discuss the methods through which medical devices such as corneal inserters may alleviate these issues. Inserter selection is of high importance in the DSAEK procedure since overcoming the learning curve associated with the use of an insertion device is a time and energy consuming process. In the present review, allograft insertion devices were compared in terms of design, construction material, insertion technique, dimensions, incision requirements and endothelial cell loss to show their relative merits and capabilities based on available data in the literature. Moreover, the advantages/disadvantages of various insertion devices used for allograft insertion in DSAEK are reviewed and compared. The information presented in this review can be utilized for better selection of an insertion device for DSAEK. PMID:27051492

  5. Perception and Action in Teleoperated Needle Insertion

    PubMed Central

    Nisky, Ilana; Pressman, Assaf; Pugh, Carla M.; Mussa-Ivaldi, Ferdinando A.; Karniel, Amir

    2015-01-01

    We studied the effect of delay on perception and action in contact with a force field that emulates elastic soft tissue with a rigid nonlinear boundary. Such field is similar to forces exerted on a needle during teleoperated needle insertion. We found that delay causes motor underestimation of the stiffness of this nonlinear soft tissue, without perceptual change. These experimental results are supported by simulation of a simplified mechanical model of the arm and neural controller, and a model for perception of stiffness, which is based on regression in the force-position space. In addition, we show that changing the gain of the teleoperation channel cancels the motor effect of delay without adding perceptual distortion. We conclude that it is possible to achieve perceptual and motor transparency in virtual one-dimensional remote needle insertion task. PMID:26379813

  6. A small, insertable oven for boronization

    SciTech Connect

    Brouchous, D.A.; Diebold, D.A.; Doczy, M.L.

    1996-04-01

    A small insertable oven for benchmarking the boronizing characteristics of solid compounds, such as decaborane and carborane, has been developed for the Phaedrus-T tokamak. Assembly and installation of the oven are relatively easy as the oven design utilizes a Langmuir probe drive assembly, which is standard equipment on most tokamaks and allows the oven to be inserted into the tokamak without requiring a vent. Films deposited by heating carborane into the vapor state with the oven are found to be spatially nonuniform in both thickness and in the ratio of boron to carbon as compared to films deposited with trimethylboron, a gaseous compound. Overall plasma performance is not found to be greatly affected by whether decaborane, carborane or trimethylboron is used for boronization in Phaedrus-T. {copyright} {ital 1996 American Institute of Physics.} {lt}ii;010512{gt}

  7. INSERTION DEVICE ACTIVITIES FOR NSLS-II.

    SciTech Connect

    TANABE,T.; HARDER, D.A.; HULBERT, S.; RAKOWSKI, G.; SKARITKA, J.

    2007-06-25

    National Synchrotron Light Source-II (NSLS-II) will be a medium energy storage ring of 3GeV electron beam energy with sub-nm.rad horizontal emittance and top-off capability at 500mA. Damping wigglers will be used not only to reduce the beam emittance but also used as broadband sources for users. Cryo-Permanent Magnet Undulators (CPMUs) are considered for hard X-ray linear device, and permanent magnet based elliptically polarized undulators (EPUs) for variable polarization devices for soft X-ray. 6T superconducting wiggler with minimal fan angle will be installed in the second phase as well as quasi-periodic EPU for VUV and possibly high-temperature superconducting undulator. R&D plans have been established to pursue the performance enhancement of the baseline devices and to design new types of insertion devices. A new insertion device development laboratory will also be established.

  8. Novel Landmark for Cervical Pedicle Screw Insertion Point from Computed Tomography-Based Study

    PubMed Central

    2017-01-01

    Study Design Cross-sectional study. Purpose The purpose of this study was to evaluate a novel landmark for the cervical pedicle screw insertion point. Overview of Literature To improve the accuracy of pedicle screw placement, several studies have employed the lateral mass, lateral vertical notch, and/or inferior articular process as landmarks; however, we often encounter patients in whom we cannot identify accurate insertion points for pedicle screws using these landmarks because of degenerative changes in the facet joints. The superomedial edge of the lamina is less affected by degenerative changes, and we hypothesized that it could be a new landmark for identifying an accurate cervical pedicle screw insertion point. Methods A total of 327 consecutive patients, who had undergone neck computed tomographic scanning for determination of neck disease in our institute, were included in the study. At first, the line was drawn parallel to the superior border of the pedicle in the sagittal plane and parallel to the vertical body in the coronal plane. The line was moved downward in 1-mm increments to the inferior border of the pedicle. We determined whether the line passing through the superomedial edge of the lamina (termed the “N-line”) was located between the superior and inferior borders of the pedicle in the sagittal plane. Results The percentages of N-lines located between the superior and inferior borders of the pedicle were 100% at C3, 100% at C4, 99% at C5, 96% at C6, and 97% at C7. The lower cervical spine has the higher N-line location. Conclusions The N-line was frequently located at the level of the pedicle of each cervical spine in the sagittal plane. The superomedial edge of the lamina could be a new landmark for the insertion point of the cervical pedicle screw. PMID:28243374

  9. Umbilical cord insertion: normal location in neonates.

    PubMed

    Salgado, L; Paz, J E

    1992-12-01

    The distances between the xyphoid appendix and the insertion point of the umbilical cord (XU) and between the xyphoid appendix and upper edge of the pubic symphysis (XP) were measured in 201 newborn infants. The mean ratio XU/XP was 0.62 (S.D. 0.044) with no differences between sexes nor correlations with weight or length. Ratios lying between 0.53 and 0.71 can be considered as within the normal range.

  10. J85 Rejuvenation Through Technology Insertion

    DTIC Science & Technology

    2000-10-01

    and Sabre 75 business addition to military production, the J85 was jets . Number Model Produced Aircraft Type(s) Engine Type Thrust (lbs) J85-GE-4 740...REJUVENATION THROUGH TECHNOLOGY INSERTION T.A. Brisken, P.N. Howell, A.C. Ewing Military Engines Operation GE Aircraft Engines 1 Neumann Way Cincinnati...OH 45215, USA Summary thrust to weight ratio turbojet engines with potential application to early cruise missiles and drones. The history of the

  11. Thermally Activated Retainers For Insertion In Gaps

    NASA Technical Reports Server (NTRS)

    Grimaldi, Margaret E.; Hartz, Leslie S.

    1992-01-01

    Mechanical retainers of new type for use with gap filler easy to install and to attach themselves securely. Concept based on shape-memory properties of metal alloy Nitinol, alloy of nickel and titanium. Retainers conceived for use with Space Shuttle insulating tiles but used on other assemblies of blocks or tiles configured similarly. Tabs bent outward made flush by cooling below memory transition temperature. After insertion in gap and reheating, tabs spring outward.

  12. Anionic phospholipids modulate peptide insertion into membranes.

    PubMed

    Liu, L P; Deber, C M

    1997-05-06

    While the insertion of a hydrophobic peptide or membrane protein segment into the bilayer can be spontaneous and driven mainly by the hydrophobic effect, anionic lipids, which comprise ca. 20% of biological membranes, provide a source of electrostatic attractions for binding of proteins/peptides into membranes. To unravel the interplay of hydrophobicity and electrostatics in the binding of peptides into membranes, we designed peptides de novo which possess the typical sequence Lys-Lys-Ala-Ala-Ala-X-Ala-Ala-Ala-Ala-Ala-X-Ala-Ala-Trp-Ala-Ala-X-Ala-Al a-Ala-Lys-Lys-Lys-Lys-amide, where X residues correspond to "guest" residues which encompass a range of hydrophobicity (Leu, Ile, Gly, and Ser). Circular dichroism spectra demonstrated that peptides were partially (40-90%) random in aqueous buffer but were promoted to form 100% alpha-helical structures by anionic lipid micelles. In neutral lipid micelles, only the relatively hydrophobic peptides (X = L and I) spontaneously adopted the alpha-helical conformation, but when 25% of negatively charged lipids were mixed in to mimic the content of anionic lipids in biomembranes, the less hydrophobic (X = S and G) peptides then formed alpha-helical conformations. Consistent with these findings, fluorescence quenching by the aqueous-phase quencher iodide indicated that in anionic (dimyristoylphosphatidylglycerol) vesicles, the peptide Trp residue was buried in the lipid vesicle hydrophobic core, while in neutral (dimyristoylphosphatidylcholine) vesicles, only hydrophobic (X = L and I) peptides were shielded from the aqueous solution. Trp emission spectra of peptides in the presence of phospholipids doxyl-labeled at the 5-, 7-, 10-, 12-, and 16-fatty acid positions implied not only a transbilayer orientation for inserted peptides but also that mixed peptide populations (transbilayer + surface-associated) may arise. Overall results suggest that for hydrophobic peptides with segmental threshold hydrophobicity below that which

  13. Probe Insertion Apparatus with Inflatable Seal

    NASA Technical Reports Server (NTRS)

    Trimarchi, Paul A.

    1991-01-01

    A sealing apparatus for inserting a probe into a pressure vessel having an elongated opening includes Ii pair of resiliently defQrmable seals opposingly disposed in sealing engagement with each other. A retainer is connected to the pressure vessel around the elongated opening and holds the pair of seals rigidly to the pressure vessel. A wedge is engageable with the pair of seals and carries the probe, for longitudinally translating the probe in the pressure vessel.

  14. Inserting new technology into small missions

    NASA Technical Reports Server (NTRS)

    Deutsch, L. J.

    2001-01-01

    Part of what makes small missions small is that they have less money. Executing missions at low cost implies extensive use of cost sharing with other missions or use of existing solutions. However, in order to create many small missions, new technology must be developed, applied, and assimilated. Luckily, there are methods for creating new technology and inserting it into faster-better-cheaper (FBC) missions.

  15. Rack Insertion End Effector (RIEE) guidance

    NASA Technical Reports Server (NTRS)

    Malladi, Narasimha S.

    1994-01-01

    NASA-KSC has developed a mechanism to handle and insert Racks into the Space Station Logistic Modules. This mechanism consists of a Base with 3 motorized degrees of freedom, a 3 section motorized Boom that goes from 15 to 44 feet in length, and a Rack Insertion End Effector (RIEE) with 5 hand wheels for precise alignment. During the 1993 NASA-ASEE Summer Faculty Fellowship Program at KSC, I designed an Active Vision (Camera) Arrangement and developed an algorithm to determine (1) the displacements required by the Room for its initial positioning and (2) the rotations required at the five hand-wheels of the RIEE, for the insertion of the Rack, using the centroids fo the Camera Images of the Location Targets in the Logistic Module. Presently, during the summer of '94, I completed the preliminary design of an easily portable measuring instrument using encoders to obtain the 3-Dimensional Coordinates of Location Targets in the Logistics Module relative to the RIEE mechanism frame. The algorithm developed in '93 can use the output of this instrument also. Simplification of the '93 work and suggestions for the future work are discussed.

  16. App-assisted external ventricular drain insertion.

    PubMed

    Eftekhar, Behzad

    2016-09-01

    The freehand technique for insertion of an external ventricular drain (EVD) is based on fixed anatomical landmarks and does not take individual variations into consideration. A patient-tailored approach based on augmented-reality techniques using devices such as smartphones can address this shortcoming. The Sina neurosurgical assist (Sina) is an Android mobile device application (app) that was designed and developed to be used as a simple intraoperative neurosurgical planning aid. It overlaps the patient's images from previously performed CT or MRI studies on the image seen through the device camera. The device is held by an assistant who aligns the images and provides information about the relative position of the target and EVD to the surgeon who is performing EVD insertion. This app can be used to provide guidance and continuous monitoring during EVD placement. The author describes the technique of Sina-assisted EVD insertion into the frontal horn of the lateral ventricle and reports on its clinical application in 5 cases as well as the results of ex vivo studies of ease of use and precision. The technique has potential for further development and use with other augmented-reality devices.

  17. DEVICE FOR CONTROLLING INSERTION OF ROD

    DOEpatents

    Beaty, B.J.

    1958-10-14

    A device for rapidly inserting a safety rod into a nuclear reactor upon a given signal or in the event of a power failure in order to prevent the possibility of extensive damage caused by a power excursion is described. A piston is slidably mounted within a vertical cylinder with provision for an electromagnetic latch at the top of the cylinder. This assembly, with a safety rod attached to the piston, is mounted over an access port to the core region of the reactor. The piston is normally latched at the top of the cylinder with the safety rod clear of the core area, however, when the latch is released, the piston and rod drop by their own weight to insert the rod. Vents along the side of the cylinder permit the escape of the air entrapped under the piston over the greater part of the distance, however, at the end of the fall the entrapped air is compressed thereby bringing the safety rod gently to rest, thus providing for a rapid automatic insertion of the rod with a minimum of structural shock.

  18. Force measurement of insertion of cochlear implant electrode arrays in-vitro: Comparison of surgeon to automated insertion tool

    PubMed Central

    Majdani, O; Hussong, A; Rau, TS; Wittkopf, J; Lenarz, T; Labadie, RF

    2016-01-01

    Conclusions We have demonstrated that an automated insertion tool (a.k.a. robot) can be used to duplicate a complex surgical motion in inserting cochlear implant electrode arrays via the “advance-off-stylet” technique (AOS). As compared to human operators, the forces generated by the robot were slightly larger but the robot was more reliable (i.e. less force maxima). Objectives We present force data collected during cochlear implant electrode insertion by human operators and by an automated insertion tool (a.k.a. robot). Methods Using a three-dimensional, anatomically-correct, translucent model of the scala tympani chamber of the cochlea, cochlear implant electrodes were inserted either by one of three surgeons (26 insertions) or by the robotic insertion tool (8 insertions). Force was recorded using a load beam cell calibrated for expected forces of less than 0.1 Newtons. The insertions were also videotaped to allow correlation of force with depth of penetration into the cochlea and speed of insertion. Results Average insertion force by the surgeons was 0.004±0.001N and for the insertion tool 0.005±0.014N (p < 0.00001, Student’s t-test). While the average insertion force of the automated tool was larger than that of the surgeons, the surgeons did have intermittent peaks during the AOS component of the insertion (between 120° and 200°). PMID:19484593

  19. Epidural insertion simulator of higher insertion resistance & drop rate after puncture.

    PubMed

    Naemura, K; Sakai, A; Hayashi, T; Saito, H

    2008-01-01

    Accidents such as dural puncture remain one of the problems of epidural anesthesia, and unskilled doctors can repeat such accidents. The purpose of the current research was to provide a new simulator for epidural insertion training. No reference data regarding the resistance force used when inserting a needle into patients have been reported. A comparative study was conducted to aid in the development of a new simulator. Pork loin (n=5) were employed as a substitute for patients. Thickness was set at 2 cm so as to improve the reproducibility. The authors took the conventional simulator apart, and picked a block as an analogue of muscle and ligamentum flavum. A new simulator was made of a melamine foam resin block and a latex rubber sheet. An epidural needle fixed on a motorized stage was inserted at the speed of 2 mm per second. The reaction force was measured while the needle was inserted into each specimen. Waveform of the pork loin exhibited two slopes of different inclines up to peaks and then falls after puncture. The conventional simulator showed a simple increase up to peak and a slow fall after puncture. In contrast, the new simulator showed two slopes up to peak and then a sudden fall after puncture. The insertion resistances were 2.5 N/s for the porcine, 0.8 N/s for the conventional and 2.1 N/s for the new simulator. The drop rates were 5 N/s for the porcine, 0.6 N/s for the conventional and 24 N/s for the new simulator. The higher insertion resistance and drop rate for the new simulator than the conventional simulator will be suitable for epidural insertion training.

  20. Insertional Mutation on Mouse Chromosome 18 with Vestibular and Craniofacial Abnormalities

    PubMed Central

    Ting, C. N.; Kohrman, D.; Burgess, D. L.; Boyle, A.; Altschuler, R. A.; Gholizadeh, G.; Samuelson, L. C.; Jang, W.; Meisler, M. H.

    1994-01-01

    A dominant mutation was generated in transgenic mice as a consequence of insertional mutation. Heterozygous mice from transgenic line 9257 (Tg(9257)) are hyperactive with bidirectional circling behavior and have a distinctive facial appearance due to hypoplasia of the nasal bone. Morphological analysis of the inner ear revealed asymmetric abnormalities of the horizontal canal and flattening or invagination of the crista ampullaris, which can account for the circling behavior. The sensory epithelium appeared to be normal. The transgene insertion site was localized by in situ hybridization to the B1 band of mouse chromosome 18. Genetic mapping in an interspecific backcross demonstrated the gene order centromere--Tg(9257)--8.8 +/- 3.4--Grl-1, Egr-1, Fgf-1, Apc--14.7 +/- 4.3--Pdgfr. The phenotype and the mapping data suggest that the transgene may be inserted at the Twirler locus. Homozygosity for the transgene results in prenatal lethality, but compound heterozygotes carrying the Tw allele and the transgene are viable. The function of the closely linked ataxia locus is not disrupted by the transgene insertion. This insertional mutant will provide molecular access to genes located in the Twirler region of mouse chromosome 18. PMID:7511123

  1. Estimation of insertion depth angle based on cochlea diameter and linear insertion depth: a prediction tool for the CI422.

    PubMed

    Franke-Trieger, Annett; Mürbe, Dirk

    2015-11-01

    Beside the cochlear size, the linear insertion depth (LID) influences the insertion depth angle of cochlear implant electrode arrays. For the specific implant CI422 the recommended LID is not fixed but can vary continuously between 20 and 25 mm. In the current study, the influence of cochlea size and LID on the final insertion depth angle was investigated to develop a prediction tool for the insertion depth angle by means of cochlea diameter and LID. Preoperative estimation of insertion depth angles might help surgeons avoid exceeding an intended insertion depth, especially with respect to low-frequency residual hearing preservation. Postoperative, high-resolution 3D-radiographs provided by Flat Panel Computed Volume Tomography (FPCT) were used to investigate the insertion depth angle in 37 CI422 recipients. Furthermore, the FPCT images were used to measure linear insertion depth and diameter of the basal turn of the cochlea. A considerable variation of measured insertion depth angles ranging from 306° to 579° was identified. The measured linear insertion depth ranged from -18.6 to 26.2 mm and correlated positively with the insertion depth angle. The cochlea diameter ranged from 8.11 to 10.42 mm and correlated negatively with the insertion depth angle. The results suggest that preoperatively measured cochlea diameter combined with the option of different array positions by means of LID may act as predictors for the final insertion depth angle.

  2. Commercial Generic Bioprocessing Apparatus Science Insert - 03

    NASA Technical Reports Server (NTRS)

    Moreno, Nancy; Stodieck, Louis; Cushing, Paula; Stowe, Mark; Hamilton, Mary Ann; Werner, Ken

    2008-01-01

    Commercial Generic Bioprocessing Apparatus Science Insert - 03 (CSI-03) is the third set of investigations in the CSI program series. The CSI program provides the K-12 community opportunities to utilize the unique microgravity environment of the International Space Station as part of the regular classroom to encourage learning and interest in science, technology, engineering and math. CSI-03 will examine the complete life cycle of the painted lady butterfly and the ability of an orb weaving spider to spin a web, eat and remain healthy in space.

  3. Phoenix's Probe Inserted in Martian Soil

    NASA Technical Reports Server (NTRS)

    2008-01-01

    The Phoenix Mars lander's robotic-arm camera took this image of the spacecraft's thermal and electrical-conductivity probe (TECP) inserted into Martian soil on day 149 of the mission. Phoenix landed on Mars' northern plains on May 25, 2008, landing.

    The robotic-arm camera acquired this image at 16:02:41 local solar time. The camera pointing was elevation -72.6986 degrees and azimuth 2.1093 degrees.

    The Phoenix mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  4. Compiler-assisted static checkpoint insertion

    NASA Technical Reports Server (NTRS)

    Long, Junsheng; Fuchs, W. K.; Abraham, Jacob A.

    1992-01-01

    This paper describes a compiler-assisted approach for static checkpoint insertion. Instead of fixing the checkpoint location before program execution, a compiler enhanced polling mechanism is utilized to maintain both the desired checkpoint intervals and reproducible checkpoint 1ocations. The technique has been implemented in a GNU CC compiler for Sun 3 and Sun 4 (Sparc) processors. Experiments demonstrate that the approach provides for stable checkpoint intervals and reproducible checkpoint placements with performance overhead comparable to a previously presented compiler assisted dynamic scheme (CATCH) utilizing the system clock.

  5. Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) Furnace Development

    NASA Technical Reports Server (NTRS)

    Crouch, Myscha R.; Carswell, William E.; Farmer, Jeff; Rose, Fred; Tidwell, Paul H., II

    2000-01-01

    The Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) are microgravity furnaces under development at Marshall Space Flight Center. The furnaces are being developed for the first Materials Science Research Rack (MSRR-1) of the Materials Science Research Facility (MSRF), one of the first International Space Station (ISS) scientific payloads. QMI is a Bridgman furnace with quench capability for studying interface behavior during directional solidification of metallic and alloy materials. DMI will be a Bridgman-Stockbarger furnace to study diffusion processes in semiconductors. The design for each insert, both QMI and DMI, is driven by specific science, operations and safety requirements, as well as by constraints arising from resource limitations, such as volume, mass and power. Preliminary QMI analysis and testing indicates that the design meets these requirements.

  6. Insertion torque versus mechanical resistance of mini-implants inserted in different cortical thickness

    PubMed Central

    Santos, Renata de Faria; Ruellas, Antonio Carlos de Oliveira; Fernandes, Daniel Jogaib; Elias, Carlos Nelson

    2014-01-01

    Objective This study aimed to measure insertion torque, tip mechanical resistance to fracture and transmucosal neck of mini-implants (MI) (Conexão Sistemas de PróteseT), as well as to analyze surface morphology. Methods Mechanical tests were carried out to measure the insertion torque of MIs in different cortical thicknesses, and tip mechanical resistance to fracture as well as transmucosal neck of MIs. Surface morphology was assessed by scanning electron microscopy (SEM) before and after the mechanical tests. Results Values of mechanical resistance to fracture (22.14 N.cm and 54.95 N.cm) were higher and statistically different (P < 0.05) from values of insertion torque for 1-mm (7.60 N.cm) and 2-mm (13.27 N.cm) cortical thicknesses. Insertion torque was statistically similar (P > 0.05) to torsional fracture in the tip of MI (22.14 N.cm) when 3 mm cortical thickness (16.11 N.cm) and dense bone (23.95 N.cm) were used. Torsional fracture of the transmucosal neck (54.95 N.cm) was higher and statistically different (P < 0.05) from insertion torsional strength in all tested situations. SEM analysis showed that the MIs had the same smooth surface when received from the manufacturer and after the mechanical tests were performed. Additionally, no significant marks resulting from the manufacturing process were observed. Conclusion All mini-implants tested presented adequate surface morphology. The resistance of mini-implants to fracture safely allows placement in 1 and 2-mm cortical thickness. However, in 3-mm cortical thickness and dense bones, pre-drilling with a bur is recommended before insertion. PMID:25162571

  7. Horizontally separated 1-in-1 crossing insertions

    SciTech Connect

    Syphers, M.J.

    1985-10-01

    Previous to this workshop, realistic lattices have been developed for vertically separated l-in-l (e.g., D.E. Johnson, A.A. Garren) and 2-in-1 (e.g., S. Heifets) magnets as well as for horizontally separated 2-in-l magnets (e.g., SSC RDS). Bringing together the widely separated ({approximately}60-70 cm) beams in a reasonable length of tunnel and keeping the dispersion zero at the interaction point has been difficult in the vertical l-in-l case. Most designs have required spacial 2-in-1 quadrupoles near the interaction point where the beams are separated by 15 cm or less. It is not clear that such magnets, as dictated by some of these lattice designs, can easily be built. The purpose of this exercise is to provide a crossing insertion for a realistic lattice which involves horizontally separated l-in-l magnets. The following horizontal crossing insertions, which incorporate the dispersion suppressors and phase trombones into the major arcs, need no special 2-in-1 magnets near the interaction point. The dispersion at the IP created by the horizontal crossing can be cancelled by the dispersion suppressor and one set of triplets.

  8. Radiological Insertion and Management of Peritoneovenous Shunt

    SciTech Connect

    Bratby, M. J.; Hussain, F. F. Lopez, A. J.

    2007-06-15

    The purpose of the study was to report our experience of the management of complications following the insertion of a peritoneovenous shunt for intractable malignant ascites. From June 1999 to January 2006, 26 patients underwent insertion of a peritoneovenous shunt for ascites by interventional radiologists. We have used ultrasound and shuntography to assist in the diagnosis of the cause of shunt blockage. Successful techniques for the restoration of the shunt function include port- pumping, stripping of any fibrin sheath, and revision of either the venous or peritoneal catheter. The procedure was initially successful in all patients with continued patency until death in 17. A further four patients are still alive with a functioning shunt. There was one rapid postprocedure death resulting from pulmonary edema. Two patients developed pneumothorax, managed successfully with either a chest drain or aspiration. Shunt dysfunction occurred eight times in seven patients. There were five successful revisions in four patients. Overall, shunt patency has been maintained in 80.1% of patients. Shunt dysfunction is seen in a significant number of patients, but successful revision of the shunt can be achieved in the majority.

  9. Model of a Hollow Cathode Insert Plasma

    NASA Technical Reports Server (NTRS)

    Mikellides, Ioannis G.; Katz, Ira; Goebel, Dan M.; Polk, James E.

    2004-01-01

    A 2-D axisymmetric fluid model of the plasma in the insert region of a hollow cathode is presented. The level of sophistication included in the model is motivated in part by the need to determine quantitatively plasma fluxes to the emitter surface. The ultimate goal is to assess whether plasma effects can degrade the life of impregnated inserts beyond those documented throughout the 30-50 year history of vacuum cathode technologies. Results from simulations of a 1.2-cm diameter cathode operating at a discharge current of 25 A, and a gas flow rate of 5 sccm, suggest that approximately 10 A of electron current, and 3.5 A of ion current return to the emitter surface. The total emitted electron current computed by the model is about 35 A. Comparisons with plasma measurements suggest that anomalous heating of the plasma due to two-stream instabilities is possible near the orifice region. Solution to the heavy species energy equation, with classical transport and no viscous effects, predicts heavy species temperatures as high as 2640 K.

  10. Insertional mutagenesis and illegitimate recombination in mycobacteria.

    PubMed Central

    Kalpana, G V; Bloom, B R; Jacobs, W R

    1991-01-01

    Mycobacteria, particularly Mycobacterium tuberculosis, Mycobacterium leprae, and Mycobacterium avium, are major pathogens of man. Although insertional mutagenesis has been an invaluable genetic tool for analyzing the mechanisms of microbial pathogenesis, it has not yet been possible to apply it to the mycobacteria. To overcome intrinsic difficulties in directly manipulating the genetics of slow-growing mycobacteria, including M. tuberculosis and bacille Calmette-Guérin (BCG) vaccine strains, we developed a system for random shuttle mutagenesis. A genomic library of Mycobacterium smegmatis was subjected to transposon mutagenesis with Tn5 seq1, a derivative of Tn5, in Escherichia coli and these transposon-containing recombinant plasmids were reintroduced into mycobacterial chromosomes by homologous recombination. This system has allowed us to isolate several random auxotrophic mutants of M. smegmatis. To extend this strategy to M. tuberculosis and BCG, targeted mutagenesis was performed using a cloned BCG methionine gene that was subjected to Tn5 seq1 mutagenesis in E. coli and reintroduced into the mycobacteria. Surprisingly for prokaryotes, both BCG and M. tuberculosis were found to incorporate linear DNA fragments into illegitimate sites throughout the mycobacterial genomes at a frequency of 10(-5) to 10(-4) relative to the number of transformants obtained with autonomously replicating vectors. Thus the efficient illegitimate recombination of linear DNA fragments provides the basis for an insertional mutagenesis system for M. tuberculosis and BCG. Images PMID:2052623

  11. Lithium Insertion Chemistry of Some Iron Vanadates

    SciTech Connect

    Patoux, Sebastien; Richardson, Thomas J.

    2007-02-02

    Lithium insertion into various iron vanadates has been investigated. Fe{sub 2}V{sub 4}O{sub 13} and Fe{sub 4}(V{sub 2}O{sub 7}){sub 3} {center_dot} 3H{sub 2}O have discharge capacities approaching 200 mAh/g above 2.0 V vs. Li{sup +}/Li. Although the potential profiles change significantly between the first and subsequent discharges, capacity retention is unexpectedly good. Other phases, structurally related to FeVO{sub 4}, containing copper and/or sodium ions were also studied. One of these, {beta}-Cu{sub 3}Fe{sub 4}(VO{sub 4}){sub 6}, reversibly consumes almost 10 moles of electrons per formula unit (ca. 240 mAh g{sup -1}) between 3.6 and 2.0 V vs. Li{sup +}/Li, in a non-classical insertion process. It is proposed that both copper and vanadium are electrochemically active, whereas iron(III) reacts to form LiFe{sup III}O{sub 2}. The capacity of the Cu{sub 3}Fe{sub 4}(VO{sub 4}){sub 6}/Li system is nearly independent of cycling rate, stabilizing after a few cycles at 120-140 mAh g{sup -1}. Iron vanadates exhibit better capacities than their phosphate analogues, whereas the latter display more constant discharge potentials.

  12. CRUCIBLE LINING METHOD

    DOEpatents

    Bone, W.H.; Schmidt, W.W.

    1958-11-01

    A method is presented for forming refractory liners in cylindrical reaction vessels used for the reductlon of uranium tetrafluoride to metallic uranium. A preliminary form, having positioning lugs attached thereto, is inserted into the reaction vessel and the refractory powder, usually CaO, is put in the annular space between the form and the inner wall of the reaction vessel. A jolting table is used to compact this charge of liner material ln place, and after thls has been done, the preliminary form is removed and the flnal form or plug is lnserted without disturbing the partially completed lining. The remainder of the lining charge is then introduced and compacted by jolting, after which the form is removed.

  13. QuickMap: a public tool for large-scale gene therapy vector insertion site mapping and analysis.

    PubMed

    Appelt, J-U; Giordano, F A; Ecker, M; Roeder, I; Grund, N; Hotz-Wagenblatt, A; Opelz, G; Zeller, W J; Allgayer, H; Fruehauf, S; Laufs, S

    2009-07-01

    Several events of insertional mutagenesis in pre-clinical and clinical gene therapy studies have created intense interest in assessing the genomic insertion profiles of gene therapy vectors. For the construction of such profiles, vector-flanking sequences detected by inverse PCR, linear amplification-mediated-PCR or ligation-mediated-PCR need to be mapped to the host cell's genome and compared to a reference set. Although remarkable progress has been achieved in mapping gene therapy vector insertion sites, public reference sets are lacking, as are the possibilities to quickly detect non-random patterns in experimental data. We developed a tool termed QuickMap, which uniformly maps and analyzes human and murine vector-flanking sequences within seconds (available at www.gtsg.org). Besides information about hits in chromosomes and fragile sites, QuickMap automatically determines insertion frequencies in +/- 250 kb adjacency to genes, cancer genes, pseudogenes, transcription factor and (post-transcriptional) miRNA binding sites, CpG islands and repetitive elements (short interspersed nuclear elements (SINE), long interspersed nuclear elements (LINE), Type II elements and LTR elements). Additionally, all experimental frequencies are compared with the data obtained from a reference set, containing 1 000 000 random integrations ('random set'). Thus, for the first time a tool allowing high-throughput profiling of gene therapy vector insertion sites is available. It provides a basis for large-scale insertion site analyses, which is now urgently needed to discover novel gene therapy vectors with 'safe' insertion profiles.

  14. Recessed impingement insert metering plate for gas turbine nozzles

    DOEpatents

    Itzel, Gary Michael; Burdgick, Steven Sebastian

    2002-01-01

    An impingement insert sleeve is provided that is adapted to be disposed in a coolant cavity defined through a stator vane. The insert has a generally open inlet end and first and second diametrically opposed, perforated side walls. A metering plate having at least one opening defined therethrough for coolant flow is mounted to the side walls to generally transverse a longitudinal axis of the insert, and is disposed downstream from said inlet end. The metering plate improves flow distribution while reducing ballooning stresses within the insert and allowing for a more flexible insert attachment.

  15. Insertion of coherence requests for debugging a multiprocessor

    DOEpatents

    Blumrich, Matthias A.; Salapura, Valentina

    2010-02-23

    A method and system are disclosed to insert coherence events in a multiprocessor computer system, and to present those coherence events to the processors of the multiprocessor computer system for analysis and debugging purposes. The coherence events are inserted in the computer system by adding one or more special insert registers. By writing into the insert registers, coherence events are inserted in the multiprocessor system as if they were generated by the normal coherence protocol. Once these coherence events are processed, the processing of coherence events can continue in the normal operation mode.

  16. Method Of Wire Insertion For Electric Machine Stators

    DOEpatents

    Brown, David L; Stabel, Gerald R; Lawrence, Robert Anthony

    2005-02-08

    A method of inserting coils in slots of a stator is provided. The method includes interleaving a first set of first phase windings and a first set of second phase windings on an insertion tool. The method also includes activating the insertion tool to radially insert the first set of first phase windings and the first set of second phase windings in the slots of the stator. In one embodiment, interleaving the first set of first phase windings and the first set of second phase windings on the insertion tool includes forming the first set of first phase windings in first phase openings defined in the insertion tool, and forming the first set of second phase windings in second phase openings defined in the insertion tool.

  17. The rice miniature inverted repeat transposable element mPing is an effective insertional mutagen in soybean.

    PubMed

    Hancock, C Nathan; Zhang, Feng; Floyd, Kristen; Richardson, Aaron O; Lafayette, Peter; Tucker, Donna; Wessler, Susan R; Parrott, Wayne A

    2011-10-01

    Insertional mutagenesis of legume genomes such as soybean (Glycine max) should aid in identifying genes responsible for key traits such as nitrogen fixation and seed quality. The relatively low throughput of soybean transformation necessitates the use of a transposon-tagging strategy where a single transformation event will produce many mutations over a number of generations. However, existing transposon-tagging tools being used in legumes are of limited utility because of restricted transposition (Ac/Ds: soybean) or the requirement for tissue culture activation (Tnt1: Medicago truncatula). A recently discovered transposable element from rice (Oryza sativa), mPing, and the genes required for its mobilization, were transferred to soybean to determine if it will be an improvement over the other available transposon-tagging tools. Stable transformation events in soybean were tested for mPing transposition. Analysis of mPing excision at early and late embryo developmental stages revealed increased excision during late development in most transgenic lines, suggesting that transposition is developmentally regulated. Transgenic lines that produced heritable mPing insertions were identified, with the plants from the highest activity line producing at least one new insertion per generation. Analysis of the mPing insertion sites in the soybean genome revealed that features displayed in rice were retained including transposition to unlinked sites and a preference for insertion within 2.5 kb of a gene. Taken together these findings indicate that mPing has the characteristics necessary for an effective transposon-tagging resource.

  18. Intermetallic insertion anodes for lithium batteries.

    SciTech Connect

    Thackeray, M. M.; Vaughey, J.; Johnson, C. S.; Kepler, K. D.

    1999-11-12

    Binary intermetallic compounds containing lithium, or lithium alloys, such as Li{sub x}Al, Li{sub x}Si and Li{sub x}Sn have been investigated in detail in the past as negative electrode materials for rechargeable lithium batteries. It is generally acknowledged that the major limitation of these systems is the large volumetric expansion that occurs when lithium reacts with the host metal. Such large increases in volume limit the practical use of lithium-tin electrodes in electrochemical cells. It is generally recognized that metal oxide electrodes, MO{sub y}, in lithium-ion cells operate during charge and discharge by means of a reversible lithium insertion/extraction process, and that the cells offer excellent cycling behavior when the crystallographic changes to the unit cell parameters and unit cell volume of the Li{sub x}MO{sub y} electrode are kept to a minimum. An excellent example of such an electrode is the spinel Li{sub 4}Ti{sub 5}O{sub 12}, which maintains its cubic symmetry without any significant change to the lattice parameter (and hence unit cell volume) during lithium insertion to the rock-salt composition Li{sub 7}Ti{sub 5}O{sub 12}. This spinel electrode is an example of a ternary Li{sub x}MO{sub y} system in which a binary MO{sub y} framework provides a stable host structure for lithium. With this approach, the authors have turned their attention to exploring ternary intermetallic systems Li{sub x}MM{prime} in the hope of finding a system that is not subject to the high volumetric expansion that typifies many binary systems. In this paper, the authors present recent data of their investigations of lithium-copper-tin and lithium-indium-antimonide electrodes in lithium cells. The data show that lithium can be inserted reversibly into selected intermetallic compounds with relatively small expansion of the lithiated intermetallic structures.

  19. Single connector provides safety fuses for multiple lines

    NASA Technical Reports Server (NTRS)

    Weber, G. J.

    1966-01-01

    Fuse-bearing sleeve which is inserted between the male and female members of a multiple-line connector contains a safety fuse for each pin of the connector assembly. The sleeve is easily and quickly opened for fuse replacement.

  20. Impact of Orthodontic Decompensation on Bone Insertion

    PubMed Central

    Guedes, Fabio Pinto; Capelozza Filho, Leopoldino; Garib, Daniela Gamba; Nary Filho, Hugo; Borgo, Evandro José; Cardoso, Mauricio de Almeida

    2014-01-01

    There has always been concern in determining the relationship between orthodontic tooth movement and the consequent biological costs to the periodontium and tooth root. The possibility of evaluating the tooth and bone morphology by CBCT allows more accurate analysis of qualitative and quantitative aspects of these processes. This paper presents a case report of a 20-year-old male patient with Class III malocclusion and hyperdivergent facial pattern, who was surgically treated. A significant amount of labial movement of mandibular incisors was performed during orthodontic treatment before surgery. CBCT was used for evaluation of buccal and lingual bone plates before and after tooth decompensation. The changes in the bone insertion level of maxillary and mandibular incisors in the present case encourage a reflection on the treatment protocol in individuals with dentoskeletal discrepancies. PMID:25436157

  1. Polymorphic Alu insertions among Mayan populations.

    PubMed

    Herrera, R J; Rojas, D P; Terreros, M C

    2007-01-01

    The Mayan homeland within Mesoamerica spans five countries: Belize, El Salvador, Guatemala, Honduras and Mexico. There are indications that the people we call the Maya migrated from the north to the highlands of Guatemala as early as 4000 B.C. Their existence was village-based and agricultural. The culture of these Preclassic Mayans owes much to the earlier Olmec civilization, which flourished in the southern portion of North America. In this study, four different Mayan groups were examined to assess their genetic variability. Ten polymorphic Alu insertion (PAI) loci were employed to ascertain the genetic affinities among these Mayan groups. North American, African, European and Asian populations were also examined as reference populations. Our results suggest that the Mayan groups examined in this study are not genetically homogeneous.

  2. INSERTION DEVICE UPGRADE PLANS AT THE NSLS.

    SciTech Connect

    TANABE, T.; BLEDNYKH, A.; HARDER, D.; LEHECKA, M.; RAKOWSKY, G.; SKARITKA, J.

    2005-05-16

    This paper describes plans to upgrade insertion devices (IDs) at the National Synchrotron Light Source (NSLS), Brookhaven National Laboratory, U.S.A. The aging wiggler (W120) at X25 is being replaced by a 1 m long in-vacuum mini-gap undulator (MGU-X25) optimized for a dedicated macromolecular crystallography program. A new, 1/3 m long, undulator (MGU or SCU-X9), will be installed between a pair of RF cavities at X9, and will serve a new beamline dedicated for small angle x-ray scattering (SAXS). Both IDs will have provision for cryocooling the NdFeB hybrid arrays to 150K to raise the field and K-value and to obtain better spectral coverage. Design issues of the devices and other considerations, especially magnetic measurement at low temperature, will be discussed.

  3. Regulatory perspective on incomplete control rod insertions

    SciTech Connect

    Chatterton, M.

    1997-01-01

    The incomplete control rod insertions experienced at South Texas Unit 1 and Wolf Creek are of safety concern to the NRC staff because they represent potential precursors to loss of shutdown margin. Even before it was determined if these events were caused by the control rods or by the fuel there was an apparent correlation of the problem with high burnup fuel. It was determined that there was also a correlation between high burnup and high drag forces as well as with rod drop time histories and lack of rod recoil. The NRC staff initial actions were aimed at getting a perspective on the magnitude of the problem as far as the number of plants and the amount of fuel that could be involved, as well as the safety significance in terms of shutdown margin. As tests have been performed and data has been analyzed the focus has shifted more toward understanding the problem and the ways to eliminate it. At this time the staff`s understanding of the phenomena is that it was a combination of factors including burnup, power history and temperature. The problem appears to be very sensitive to these factors, the interaction of which is not clearly understood. The model developed by Westinghouse provides a possible explanation but there is not sufficient data to establish confidence levels and sensitivity studies involving the key parameters have not been done. While several fixes to the problem have been discussed, no definitive fixes have been proposed. Without complete understanding of the phenomena, or fixes that clearly eliminate the problem the safety concern remains. The safety significance depends on the amount of shutdown margin lost due to incomplete insertion of the control rods. Were the control rods to stick high in the core, the reactor could not be shutdown by the control rods and other means such as emergency boration would be required.

  4. Chromosomal rearrangements associated with LINE elements in the mouse genome.

    PubMed Central

    Shyman, S; Weaver, S

    1985-01-01

    Two segments of DNA that have apparently inserted in the interval between the two adult beta-globin genes in BALB/c (Hbbd haplotype) but not in C57B1/10 (Hbbs haplotype) mouse strains have been described (1). These putative insertions, each about 1000 bp in length, mapped near a repetitive element. To determine the precise position of these alleged insertions, their target sites, and the nature of their boundaries, we cloned and sequenced the appropriate regions of both chromosomes. One of the two segments is not an insertion but rather a region between two independently integrated L1 repetitive elements (LINEs) (2), one in Hbbd and the other in the Hbbs chromosome. The other segment is an insertion of 940 bp which is located within the L1 element in the Hbbd chromosome. This insert is unusual in that it exists in only one copy in the BALB/c genome. PMID:2991852

  5. Neonatal peripherally inserted central catheters: recommendations for prevention of insertion and postinsertion complications.

    PubMed

    Paulson, Pamela R; Miller, Kellee M

    2008-01-01

    Peripherally inserted central catheters (PICCs) continue to be necessary in neonatal care. They benefit many premature infants and those needing long-term intravenous access. An experienced inserter, early recognition of PICC candidates, early PICC placement, knowledge of anatomy, and correct choice of vein all increase placement success. As with any invasive procedure, there are risks. These include pain, difficulty advancing the catheter, damage to vessels, catheter malposition, and bleeding. Utilizing assessment skills, following the product manufacturer's instructions, and carefully placing the catheter should minimize most of these risks. Additional risks include postinsertion complications such as occlusions, thrombosis, catheter failure, infection, and catheter malposition. Proper nursing care--which includes controlling infection, properly securing the catheter, and changing the dressing as needed--is key to preventing complications and maintaining the PICC until treatment has been completed.

  6. Comparison of Postoperative Drain Insertion versus No Drain Insertion in Thyroidectomies

    PubMed Central

    Al-Habsi, Asma S.; Al-Sulaimani, Al-Anood K.; Taqi, Kadhim M.; Al-Qadhi, Hani A.

    2016-01-01

    Objectives A thyroidectomy is a frequently performed surgical procedure which can result in life-threatening complications. The insertion of a drain after a thyroidectomy has been suggested to prevent such complications. This study aimed to evaluate the use of surgical drains following thyroidectomies in relation to postoperative complications and mass sizes. Methods This retrospective case-control study included all thyroidectomies conducted at the Sultan Qaboos University Hospital, Muscat, Oman, from January 2011 to December 2013. Length of hospital stay, readmission, postoperative complications and mass size were evaluated. Results During the study period, 250 surgeries were carried out on 241 patients. The majority of patients were female (87.2%). Drains were inserted postoperatively after 202 surgeries (80.8%) compared to 48 surgeries (19.2%) without drains. A total of 32 surgeries (12.8%) were conducted on patients with thyroid masses <1 cm, 138 (55.2%) on those with masses between 1–4 cm and 80 (32.0%) on those with masses >4 cm. The association between drain use and mass size was not significant (P = 0.439). Although postoperative complications were more prevalent in patients with drains, the relationship between these factors was not significant (P >0.050). Length of hospital stay was significantly longer among patients with postoperative drains (P <0.010). Conclusion The routine insertion of drains after thyroid surgeries was found to result in longer hospital stays and did not reduce rates of post-thyroidectomy complications. Thyroid mass size should not be used as an indicator for the insertion of a drain after thyroidectomy. PMID:28003893

  7. Large insert environmental genomic library production.

    PubMed

    Taupp, Marcus; Lee, Sangwon; Hawley, Alyse; Yang, Jinshu; Hallam, Steven J

    2009-09-23

    The vast majority of microbes in nature currently remain inaccessible to traditional cultivation methods. Over the past decade, culture-independent environmental genomic (i.e. metagenomic) approaches have emerged, enabling researchers to bridge this cultivation gap by capturing the genetic content of indigenous microbial communities directly from the environment. To this end, genomic DNA libraries are constructed using standard albeit artful laboratory cloning techniques. Here we describe the construction of a large insert environmental genomic fosmid library with DNA derived from the vertical depth continuum of a seasonally hypoxic fjord. This protocol is directly linked to a series of connected protocols including coastal marine water sampling [1], large volume filtration of microbial biomass [2] and a DNA extraction and purification protocol [3]. At the outset, high quality genomic DNA is end-repaired with the creation of 5 -phosphorylated blunt ends. End-repaired DNA is subjected to pulsed-field gel electrophoresis (PFGE) for size selection and gel extraction is performed to recover DNA fragments between 30 and 60 thousand base pairs (Kb) in length. Size selected DNA is purified away from the PFGE gel matrix and ligated to the phosphatase-treated blunt-end fosmid CopyControl vector pCC1 (EPICENTRE http://www.epibio.com/item.asp?ID=385). Linear concatemers of pCC1 and insert DNA are subsequently headfull packaged into phage particles by lambda terminase, with subsequent infection of phage-resistant E. coli cells. Successfully transduced clones are recovered on LB agar plates under antibiotic selection and archived in 384-well plate format using an automated colony picking robot (Qpix2, GENETIX). The current protocol draws from various sources including the CopyControl Fosmid Library Production Kit from EPICENTRE and the published works of multiple research groups [4-7]. Each step is presented with best practice in mind. Whenever possible we highlight subtleties

  8. Threaded insert for compact cryogenic-capable pressure vessels

    DOEpatents

    Espinosa-Loza, Francisco; Ross, Timothy O.; Switzer, Vernon A.; Aceves, Salvador M.; Killingsworth, Nicholas J.; Ledesma-Orozco, Elias

    2015-06-16

    An insert for a cryogenic capable pressure vessel for storage of hydrogen or other cryogenic gases at high pressure. The insert provides the interface between a tank and internal and external components of the tank system. The insert can be used with tanks with any or all combinations of cryogenic, high pressure, and highly diffusive fluids. The insert can be threaded into the neck of a tank with an inner liner. The threads withstand the majority of the stress when the fluid inside the tank that is under pressure.

  9. Phoenix Conductivity Probe Inserted into Martian Soil

    NASA Technical Reports Server (NTRS)

    2008-01-01

    NASA's Phoenix Mars Lander inserted the four needles of its thermal and conductivity probe into Martian soil during the 98th Martian day, or sol, of the mission and left it in place until Sol 99 (Sept. 4, 2008).

    The Robotic Arm Camera on Phoenix took this image on the morning of Sol 99 while the probe's needles were in the ground. The science team informally named this soil target 'Gandalf.'

    The thermal and conductivity probe measures how fast heat and electricity move from one needle to an adjacent one through the soil or air between the needles. Conductivity readings can be indicators about water vapor, water ice and liquid water.

    The probe is part of Phoenix's Microscopy, Electrochemistry and Conductivity suite of instruments.

    The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  10. IYA2009 newspaper insert in your community

    NASA Astrophysics Data System (ADS)

    Harvey, J.

    2008-06-01

    The Gemini PIO (Public Information Officer) offers suggestions on how to approach your local newspaper about a newspaper insert for your community being published during IYA2009. Local government support, articles by astronomers, advertisers, and appointing someone within your organisation to manage the content will be discussed. We will explain the timeline required, the number of personnel hours required, developmental stages and income a local newspaper would have to generate to produce a quality, table-top supplement. In 2003, over 30,000 copies of Stars over Mauna Kea, a special supplement in tabloid format were produced and distributed in the local newspapers in Hilo, Hawaii. The publication, 48 pages in total, featured profiles of the observatories on Mauna Kea, stories about the geology and legends of Mauna Kea, and historical information about the evolution of astronomy in Hawaii. In additionthe publication included a series of essays titled In their own words. These articles were written by key members of the astronomy community. Sixty thousand copies of Stars over Mauna Kea II were printed as a follow-up to the first edition in 2005. This second edition included an article on the `Imiloa Astronomy Education Center, explanations about the types of telescopes on Mauna Kea and columns written by scientists about the fascinating and significant discoveries made on Hawaii. Personal stories about careers in astronomy were also highlighted. In Chile, a similar eight-page supplement, featuring Gemini, CTIO and SOAR telescopes, was published in 2005 and 5000 copies were distributed throughout the country.

  11. Insertion devices for DORIS III (invited)

    NASA Astrophysics Data System (ADS)

    Pflüger, J.

    1992-01-01

    Recently, a major reconstruction of the electron storage ring DORIS II, the DORIS III project, has been completed [W. Brefeld, H. Nesemann, and J. Rossbach, Proceedings of the European Particle Accelerator Conference, Rome (World Scientific, Singapore, 1988), p. 2389]. Figure 1 shows an overview of the new ring. Originally DORIS II had a twofold symmetry. In part C each of the two dipole magnets adjacing to the 65-m-long straight section was replaced by three corresponding weaker ones. In this way a total of seven straight sections for insertion devices are provided. Six of them are 4-m long and the one in the center is only 2.7-m long. After extensive discussions with the user groups involved, four x-ray wigglers, one asymmetric hybrid structure, one x-ray undulator, and one XUV multiple undulator of the revolver type have been proposed for six of the sections [J. Pflüger and P. Gurtler, Nucl. Instrum. Methods A 287, 628 (1990)]. One section is presently still free. All devices are either in construction or have already been completed and installed. In this contribution the mechanical and magnetic design of these devices will be described. Results of magnetic measurements of those devices which are already completed will be given in more detail.

  12. Replicative resolution of integron cassette insertion

    PubMed Central

    Loot, Céline; Ducos-Galand, Magaly; Escudero, José Antonio; Bouvier, Marie; Mazel, Didier

    2012-01-01

    Site-specific recombination catalyzed by tyrosine recombinases follows a common pathway consisting of two consecutive strand exchanges. The first strand exchange generates a Holliday junction (HJ), which is resolved by a second strand exchange. In integrons, attC sites recombine as folded single-stranded substrates. Only one of the two attC site strands, the bottom one, is efficiently bound and cleaved by the integrase during the insertion of gene cassettes at the double-stranded attI site. Due to the asymmetry of this complex, a second strand exchange on the attC bottom strand (bs) would form linearized abortive recombination products. We had proposed that HJ resolution would rely on an uncharacterized mechanism, probably replication. Using an attC site carried on a plasmid with each strand specifically tagged, we followed the destiny of each strand after recombination. We demonstrated that only one strand, the one carrying the attC bs, is exchanged. Furthermore, we show that the recombination products contain the attC site bs and its entire de novo synthesized complementary strand. Therefore, we demonstrate the replicative resolution of single-strand recombination in integrons and rule out the involvement of a second strand exchange of any kind in the attC × attI reaction. PMID:22740653

  13. Inferences on the role of insertion in a mutation accumulation experiment with Drosophila melanogaster using RAPDs.

    PubMed

    Salgado, C; Nieto, B; Toro, M A; López-Fanjul, C; García-Dorado, A

    2005-01-01

    The genetic variability for RAPDs band pattern was studied in a set of 157 mutation accumulation (MA) lines of Drosophila melanogaster. These MA lines were derived from the same isogenic base population and subsequently maintained by full-sib mating during 132 generations. The ancestral pattern of the original isogenic base can be unambiguously established as the consensus pattern of the MA lines and, because these lines are expected to be homozygous, dominance for band pattern is not a concern. Only repeatable changes in band pattern were considered. The number of ancestral bands detected implies that nine-nucleotide targets are enough for repeatable PCR amplification. Compared with the ancestral pattern, one MA line lost one band and two MA lines gained a new one. These results can be accounted for by the insertion of transposable elements occurring at a rate 0.07 < i < 0.21 per whole haploid genome and generation. This range is typical for Drosophila and consistent with the previously observed mobility for the roo family, supporting the generality of previous estimates of spontaneous mutation rates for morphological and fitness traits based on these MA lines. The sequence of one of the new bands suggests that the Idefix family is also active in the lines.

  14. Peripherally inserted central venous catheter safety in burn care: a single-center retrospective cohort review.

    PubMed

    Austin, Ryan E; Shahrokhi, Shahriar; Bolourani, Siavash; Jeschke, Marc G

    2015-01-01

    The use of peripherally inserted central catheter (PICC) line for central venous access in thermally injured patients has increased in recent years despite a lack of evidence regarding safety in this patient population. A recent survey of invasive catheter practices among 44 burn centers in the United States found that 37% of burn units use PICC lines as part of their treatment protocol. The goal of this study was to compare PICC-associated complication rates with the existing literature in both the critical care and burn settings. The methodology involved is a single institution retrospective cohort review of patients who received a PICC line during admission to a regional burn unit between 2008 and 2013. Fifty-three patients were identified with a total of seventy-three PICC lines. The primary outcome measurement for this study was indication for PICC line discontinuation. The most common reason for PICC line discontinuation was that the line was no longer indicated (45.2%). Four cases of symptomatic upper extremity deep vein thrombosis (5.5%) and three cases of central line-associated bloodstream infection (4.3%, 2.72 infections per 1000 line days) were identified. PICC lines were in situ an average of 15 days (range 1 to 49 days). We suggest that PICC line-associated complication rates are similar to those published in the critical care literature. Though these rates are higher than those published in the burn literature, they are similar to central venous catheter-associated complication rates. While PICC lines can be a useful resource in the treatment of the thermally injured patient, they are associated with significant and potentially fatal risks.

  15. Roles of carboxyl groups in the transmembrane insertion of peptides

    PubMed Central

    Barrera, Francisco N.; Weerakkody, Dhammika; Anderson, Michael; Andreev, Oleg A.; Reshetnyak, Yana K.; Engelman, Donald M.

    2011-01-01

    We have used the pHLIP® peptide to study the roles of carboxyl groups in transmembrane peptide insertion. The pH (low) insertion peptide (pHLIP) binds to the surface of a lipid bilayer as a disordered peptide at neutral pH, and when the pH is lowered it inserts across the membrane to form a transmembrane helix. Peptide insertion is reversed when the pH is raised above the characteristic pKa (6.0). A key event facilitating the membrane insertion is the protonation of aspartic (Asp) and/or glutamic (Glu) acid residues, since at neutral pH their negatively charged side chains hinder membrane insertion. In order to gain mechanistic understanding, we studied the membrane insertion and exit of a series of pHLIP variants where the four Asp residues were sequentially mutated to nonacidic residues, including histidine (His). Our results show that the presence of His residues does not prevent the pH-dependent peptide membrane insertion at ~pH 4 driven by the protonation of carboxyl groups at the inserting end of the peptide. A further pH drop leads to the protonation of His residues in the TM part of peptide, which induces peptide exit from the bilayer. We also find that the number of ionizable residues that undergo a change in protonation during membrane insertion correlates with the pH-dependent insertion into and exit from the lipid bilayer, and that cooperativity increases with their number. We expect that our understanding will be used to improve the targeting of acidic diseased tissue by pHLIP peptides. PMID:21888917

  16. Genome sequence of the basal haplorrhine primate Tarsius syrichta reveals unusual insertions

    PubMed Central

    Schmitz, Jürgen; Noll, Angela; Raabe, Carsten A.; Churakov, Gennady; Voss, Reinhard; Kiefmann, Martin; Rozhdestvensky, Timofey; Brosius, Jürgen; Baertsch, Robert; Clawson, Hiram; Roos, Christian; Zimin, Aleksey; Minx, Patrick; Montague, Michael J.; Wilson, Richard K.; Warren, Wesley C.

    2016-01-01

    Tarsiers are phylogenetically located between the most basal strepsirrhines and the most derived anthropoid primates. While they share morphological features with both groups, they also possess uncommon primate characteristics, rendering their evolutionary history somewhat obscure. To investigate the molecular basis of such attributes, we present here a new genome assembly of the Philippine tarsier (Tarsius syrichta), and provide extended analyses of the genome and detailed history of transposable element insertion events. We describe the silencing of Alu monomers on the lineage leading to anthropoids, and recognize an unexpected abundance of long terminal repeat-derived and LINE1-mobilized transposed elements (Tarsius interspersed elements; TINEs). For the first time in mammals, we identify a complete mitochondrial genome insertion within the nuclear genome, then reveal tarsier-specific, positive gene selection and posit population size changes over time. The genomic resources and analyses presented here will aid efforts to more fully understand the ancient characteristics of primate genomes. PMID:27708261

  17. [Umbilical and peripherally inserted venous central catheterization of the newborn].

    PubMed

    Bouissou, A; Rakza, T; Storme, L; Lafarghe, A; Fily, A; Diependaele, J-F; Dalmas, S

    2008-09-01

    Umbilical venous and peripherally inserted venous central catheters are widely used to perfuse low-weight preterm and term newborns in intensive care units. This catheter must be inserted carefully and monitored rigorously to prevent complications. This paper develops today's knowledge on the use and complications in the newborn population.

  18. 21 CFR 310.515 - Patient package inserts for estrogens.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 5 2010-04-01 2010-04-01 false Patient package inserts for estrogens. 310.515 Section 310.515 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) DRUGS FOR HUMAN USE NEW DRUGS Requirements for Specific New Drugs or Devices § 310.515 Patient package inserts for estrogens....

  19. Interface study of insertion layers in organic semiconductor devices

    NASA Astrophysics Data System (ADS)

    Ding, Huanjun; Irfan, "; So, Franky; Gao, Yongli

    2009-08-01

    Inserting an ultra-thin interlayer has been an important means in modifying the performance of organic semiconductor devices. Using photoemission and inverse photoemission spectroscopy (UPS, XPS and IPES), we have investigated the electronic structure of a number of insertion layers widely used in organic semiconductor devices. We found that inserting alkali metal compound thin layers such as LiF between the electron transport layer (ETL) and the cathode can induce energy level shift in the ETL that reduces the electron injection barrier. The reduction is attributed to the release of the alkali metal that n-doped the ETL, and as such it depends on the cathode material deposited on top of the insertion layer. For thin metal oxide insertion layers, such as MoO3, between the anode and the hole transport layer (HTL), reduction of the hole injection barrier is also observed. This reduction, however, is due to the large workfunction of the oxide that subsequently moves the highest occupied molecular orbital (HOMO) toward the anode Fermi level. Effects of other insertion layers, such as metal insertion layer in organic bistable device (OBD) and organic insertion layer in bipolar organic thin film transistors (OTFT) will also be discussed.

  20. Combined plasma and thermal hollow cathode insert model

    NASA Technical Reports Server (NTRS)

    Katz, Ira; Polk, James E.; Mikellides, Ionnis G.; Goebel, Dan m.; Hornbeck, Sarah E.

    2005-01-01

    In this paper, we present the first results from a Hollow Cathode Thermal (HCThermal) model that uses the spatially distributed plasma fluxes calculated by the InsertRegion of an Orificed Cathode (IROrCa2D) code as the heat source to predict the hollow cathode and insert temperatures.

  1. Piezo-Driven Vibrating Insertion Device for Microelectrode Array

    NASA Astrophysics Data System (ADS)

    Noda, Takahiro; Kanzaki, Ryohei; Takahashi, Hirokazu

    Microelectrode arrays are commonly used to measure neural activities in the brain, and arrays with some 100 electrodes are commercially available to date. However, insertion of a dense grid array deforms the brain, resulting in deterioration of the measurements. In order to overcome this problem, we propose a piezo-driven vibrating insertion device to reduce the insertion-induced deformation of the brain. We attempted under various conditions to insert the array into an agarose substrate, whose hardness was adjusted to that of the cerebral cortex of rats. Our experiments demonstrated that inverse-sawtooth vibration reduced the insertion-induced deformation of the substrate in proportion to the logarithm of an upstroke velocity when the velocity was higher than 10 mm/s, and vibrating insertion of the maximum velocity at 36.7 mm/s reduced the deformation by up to 40% as compared to insertion without vibration. In addition, we tested the vibrating insertion device in an electrophysiological experiment in the rat auditory cortex in vivo, and successfully measured tone-evoked neuronal activities.

  2. Load beam unit replaceable inserts for dry coal extrusion pumps

    DOEpatents

    Saunders, Timothy; Brady, John D.

    2012-11-13

    A track assembly for a particulate material extrusion pump according to an exemplary aspect of the present disclosure includes a link assembly with a roller bearing. An insert mounted to a load beam located such that the roller bearing contacts the insert.

  3. Cruciate retaining and cruciate substituting ultra-congruent insert

    PubMed Central

    Deledda, Davide; Rosso, Federica; Ratto, Nicola; Bruzzone, Matteo; Bonasia, Davide Edoardo; Rossi, Roberto

    2016-01-01

    The posterior cruciate ligament (PCL) conservation and the polyethylene insert constraint in total knee arthroplasty (TKA) are still debated. The PCL is one of the primary stabilizers of the joint, but cruciate retaining (CR) implants have the disadvantage of a difficult balancing of the PCL. Postero-stabilized (PS) implants were introduced to reduce this problem. However, also the PS implants have some disadvantages, due to the cam-mechanism, such as high risk of cam-mechanism polyethylene wear. To minimize the polyethylene wear of the cam-mechanism and the bone sacrifice due to the intercondylar box, different types of inserts were developed, trying to increase the implant conformity and to reduce stresses on the bone-implant interface. In this scenario ultra-congruent (UC) inserts were developed. Those inserts are characterized by a high anterior wall and a deep-dished plate. This conformation should guarantee a good stability without the posterior cam. Few studies on both kinematic and clinical outcomes of UC inserts are available. Clinical and radiological outcomes, as well as kinematic data are similar between UC mobile bearing (MB) and standard PS MB inserts at short to mid-term follow-up. In this manuscript biomechanics and clinical outcomes of UC inserts will be described, and they will be compared to standard PS or CR inserts. PMID:26855938

  4. Z-2 Threaded Insert Design and Testing Abstract

    NASA Technical Reports Server (NTRS)

    Rhodes, RIchard; Graziosi, Dave; Jones, Bobby; Ferl, Jinny; Scarborough, Steve; Sweeney, Mitch

    2016-01-01

    The Z-2 Prototype Planetary Extravehicular Space Suit Assembly is a continuation of NASA's Z series of spacesuits. The Z-2 is another step in the NASA's technology development roadmap leading to human exploration of the Martian surface. To meet a more challenging set of requirements than previous suit systems standard design features, such as threaded inserts, have been re-analyzed and improved. NASA's Z-2 prototype space suit contains several components fabricated from an advanced hybrid composite laminate consisting of IM10 carbon fiber and fiber glass. One requirement NASA levied on the suit composites was the ability to have removable, replaceable helicoil inserts to which other suit components would be fastened. An approach utilizing bonded in inserts with helicoils inside of them was implemented. The design of the interface flanges of the composites allowed some of the inserts to be a "T" style insert that was installed through the entire thickness of the laminate. The flange portion of the insert provides a mechanical lock as a redundancy to the adhesive aiding in the pullout load that the insert can withstand. In some locations it was not possible to utilize at "T" style insert and a blind insert was used instead. These inserts rely completely on the bond strength of the adhesive to resist pullout. It was determined during the design of the suit that the inserts did not need to withstand loads induced from pressure cycling but instead tension induced from torqueing the screws to bolt on hardware which creates a much higher stress on them. Bolt tension is determined by dividing the torque on the screw by a k value multiplied by the thread diameter of the bolt. The k value is a factor that accounts for friction in the system. A common value used for k for a non-lubricated screw is 0.2. The k value can go down by as much as 0.1 if the screw is lubricated which means for the same torque, a much larger tension could be placed on the bolt and insert. This paper

  5. Measurement of Vein Diameter for Peripherally Inserted Central Catheter (PICC) Insertion: An Observational Study.

    PubMed

    Sharp, Rebecca; Cummings, Melita; Childs, Jessie; Fielder, Andrea; Mikocka-Walus, Antonina; Grech, Carol; Esterman, Adrian

    2015-01-01

    Choosing an appropriately sized vein reduces the risk of venous thromboembolism associated with peripherally inserted central catheters. This observational study described the diameters of the brachial, basilic, and cephalic veins and determined the effect of patient factors on vein size. Ultrasound was used to measure the veins of 176 participants. Vein diameter was similar in both arms regardless of hand dominance and side. Patient factors-including greater age, height, and weight, as well as male gender-were associated with increased vein diameter. The basilic vein tended to have the largest diameter statistically. However, this was the case in only 55% of patients.

  6. Intraaortic balloon pump insertion through the subclavian artery. Subclavian artery insertion of IABP.

    PubMed

    Marcu, Constantin B; Donohue, Thomas J; Ferneini, Antoine; Ghantous, Andre E

    2006-04-01

    The intraaortic balloon pump (IABP) is frequently used in the management of cardiac failure in the setting of myocardial infarction or as a bridge for coronary revascularisation surgery. The IABP is usually inserted through the femoral artery. Occasionally severe aorto-iliac occlusive disease prevents the retrograde passage of the balloon, in which case an anterograde route, usually through the ascending aorta is used. We describe four patients in whom an IABP was placed through the subclavian artery by the joint efforts of cardiologists and vascular surgeons.

  7. Small genomic insertions form enhancers that misregulate oncogenes

    PubMed Central

    Abraham, Brian J.; Hnisz, Denes; Weintraub, Abraham S.; Kwiatkowski, Nicholas; Li, Charles H.; Li, Zhaodong; Weichert-Leahey, Nina; Rahman, Sunniyat; Liu, Yu; Etchin, Julia; Li, Benshang; Shen, Shuhong; Lee, Tong Ihn; Zhang, Jinghui; Look, A. Thomas; Mansour, Marc R.; Young, Richard A.

    2017-01-01

    The non-coding regions of tumour cell genomes harbour a considerable fraction of total DNA sequence variation, but the functional contribution of these variants to tumorigenesis is ill-defined. Among these non-coding variants, somatic insertions are among the least well characterized due to challenges with interpreting short-read DNA sequences. Here, using a combination of Chip-seq to enrich enhancer DNA and a computational approach with multiple DNA alignment procedures, we identify enhancer-associated small insertion variants. Among the 102 tumour cell genomes we analyse, small insertions are frequently observed in enhancer DNA sequences near known oncogenes. Further study of one insertion, somatically acquired in primary leukaemia tumour genomes, reveals that it nucleates formation of an active enhancer that drives expression of the LMO2 oncogene. The approach described here to identify enhancer-associated small insertion variants provides a foundation for further study of these abnormalities across human cancers. PMID:28181482

  8. Gas turbine nozzle vane insert and methods of installation

    DOEpatents

    Miller, William John; Predmore, Daniel Ross; Placko, James Michael

    2002-01-01

    A pair of hollow elongated insert bodies are disposed in one or more of the nozzle vane cavities of a nozzle stage of a gas turbine. Each insert body has an outer wall portion with apertures for impingement-cooling of nozzle wall portions in registration with the outer wall portion. The insert bodies are installed into the cavity separately and spreaders flex the bodies toward and to engage standoffs against wall portions of the nozzle whereby the designed impingement gap between the outer wall portions of the insert bodies and the nozzle wall portions is achieved. The spreaders are secured to the inner wall portions of the insert bodies and the bodies are secured to one another and to the nozzle vane by welding or brazing.

  9. Smart structures for shock wave attenuation using ER inserts

    NASA Astrophysics Data System (ADS)

    Kim, Jaehwan; Kim, Jung-Yup; Choi, Seung-Bok; Kim, Kyung-Su

    2001-08-01

    This Paper demonstrates the possibility of shock wave attenuation propagating through a smart structure that incorporates ER insert. The wave transmission of ER inserted beam is theoretically derived using Mead & Markus model and the theoretical results are compared with the finite element analysis results. To experimentally verify the shock wave attenuation, ER insert in an aluminum plate is made and two piezoceramic disks are used as transmitter and receiver of the wave. The transmitter sends a sine pulse signal such that a component of shock wave travels through the plate structure and the receiver gets the transmitted wave signal. Wave propagation of the ER insert can be adjusted by changing the applied electric field on the ER insert. Details of the experiment are addressed and the possibility of shock wave attenuation is experimentally verified. This kind of smart structure can be used for warship and submarine hull structures to protect fragile and important equipment.

  10. Method for improving the durability of ion insertion materials

    DOEpatents

    Lee, Se-Hee; Tracy, C. Edwin; Cheong, Hyeonsik M.

    2002-01-01

    The invention provides a method of protecting an ion insertion material from the degradative effects of a liquid or gel-type electrolyte material by disposing a protective, solid ion conducting, electrically insulating, layer between the ion insertion layer and the liquid or gel-type electrolyte material. The invention further provides liquid or gel-type electrochemical cells having improved durability having a pair of electrodes, a pair of ion insertion layers sandwiched between the pair of electrodes, a pair of solid ion conducting layers sandwiched between the ion insertion layers, and a liquid or gel-type electrolyte material disposed between the solid ion conducting layers, where the solid ion conducting layer minimizes or prevents degradation of the faces of the ion insertion materials facing the liquid or gel-type electrolyte material. Electrochemical cells of this invention having increased durability include secondary lithium batteries and electrochromic devices.

  11. Detection for processing history of seam insertion and contrast enhancement

    NASA Astrophysics Data System (ADS)

    Li, Jianwei; Zhao, Yao; Ni, Rongrong

    2014-11-01

    With the development of manipulations techniques of digital images, digital image forensic technology is becoming more and more necessary. However, the determination of processing history of multi-operation is still a challenge problem. In this paper, we improve the traditional seam insertion algorithm, and propose corresponding detection method. Then an algorithm that focuses on detecting the processing history of seam insertion and contrast enhancement is proposed, which can be widely used in practical image forgery. Based on comprehensive analysis, we have discovered the inherent relationship between seam insertion and contrast enhancement. Different orders of processing make different impacts on images. By using the newly proposed algorithm, both contrast enhancement followed by seam insertion and seam insertion followed by contrast enhancement can be detected correctly. Plenty of experiments have been implemented to prove the accuracy.

  12. A retroelement modifies pre-mRNA splicing: the murine Glrb(spa) allele is a splicing signal polymorphism amplified by long interspersed nuclear element insertion.

    PubMed

    Becker, Kristina; Braune, Marlen; Benderska, Natalya; Buratti, Emanuele; Baralle, Francisco; Villmann, Carmen; Stamm, Stefan; Eulenburg, Volker; Becker, Cord-Michael

    2012-09-07

    The glycine receptor-deficient mutant mouse spastic carries a full-length long interspersed nuclear element (LINE1) retrotransposon in intron 6 of the glycine receptor β subunit gene, Glrb(spa). The mutation arose in the C57BL/6J strain and is associated with skipping of exon 6 or a combination of the exons 5 and 6, thus resulting in a translational frameshift within the coding regions of the GlyR β subunit. The effect of the Glrb(spa) LINE1 insertion on pre-mRNA splicing was studied using a minigene approach. Sequence comparison as well as motif prediction and mutational analysis revealed that in addition to the LINE1 insertion the inactivation of an exonic splicing enhancer (ESE) within exon 6 is required for skipping of exon 6. Reconstitution of the ESE by substitution of a single residue was sufficient to prevent exon skipping. In addition to the ESE, two regions within the 5' and 3' UTR of the LINE1 were shown to be critical determinants for exon skipping, indicating that LINE1 acts as efficient modifier of subtle endogenous splicing phenotypes. Thus, the spastic allele of the murine glycine receptor β subunit gene is a two-hit mutation, where the hypomorphic alteration in an ESE is amplified by the insertion of a LINE1 element in the adjacent intron. Conversely, the LINE1 effect on splicing may be modulated by individual polymorphisms, depending on the insertional environment within the host genome.

  13. Morphometric analysis of the seventh cervical vertebra for pedicle screw insertion

    PubMed Central

    Liao, Wensheng; Guo, Liangbing; Bao, Heng; Wang, Limin

    2015-01-01

    Background: Anatomy of the pedicles of the seventh cervical vertebra (C7) at the cervicothoracic junction is different from other cervical vertebrae. Fixation of C7 is required during cervical vertebra and upper thoracic injuries in clinical practice. However, the typical pedicle screw insertion methods may have problems in clinical practice based on the anatomical features of C7. This study is to explore a new pedicle screw insertion technique for C7 and to provide anatomical and radiographic basis for clinical application. Materials and Methods: C7 vertebral specimens from six human cadavers were observed for the relative position between the posterior bony landmark and the pedicle projection. Computed tomography (CT) was performed for 30 patients with cervical spondylosis (26–61 years old, mean age was 42.3 years old). The CT scan data were processed by Mimics 8.1 software for associated parameter measurement. Appropriate screw entry points (Eps) and insertion angles were selected. A total of 12 pedicle screws were inserted and then observed. The six specimens were observed after inserting the screw using this method. The junction site of the middle 1/3 and outer 1/3 segment of line G [The junction between point A (the intersection point of the superior margin of the lamina of C7 and the medial margin of the superior articular process) and point B (the intersection point of the lateral margin of the inferior articular process and the transverse process)] was taken as the Ep. The screw insertion direction parallel horizontally to the upper terminal lamina of C7 and the sagittal angle was between 35° and 45°. Results: Gross and imaging observations revealed that pedicle projection was on the line (line G) between point A (the intersection point of the superior margin of the lamina of C7 and the medial margin of the superior articular process) and point B (the intersection point of the lateral margin of the inferior articular process and the transverse process

  14. Service development of a nurse-led community-based PICC insertion service.

    PubMed

    Bedford, Eva; Waterhouse, Dee

    2017-01-26

    Patients receiving intravenous therapy require reliable venous access. Typically patients with poor peripheral access or requiring long-term treatment from an outpatient antibiotic therapy (OPAT) service need to receive secondary care input for safe central line placement, and radiological confirmation of the correct line tip placement where necessary, if treatment is to proceed as planned. Technological developments that enable accurate ultrasound-guided vein selection and electrocardiograph (ECG)-guided central line tip placement have eliminated the need for radiological or fluoroscopic confirmation of correct tip placement for peripherally placed central catheters (PICCs). This article outlines the development of an out-of-hospital nurse-led PICC insertion service using the Sherlock 3CG(®) Tip Confirmation System (C.R Bard) to meet the needs of patients requiring long-term intravenous treatment from an OPAT service, and its impact on reducing treatment delays and the need for secondary care intervention.

  15. Method of joining a vane cavity insert to a nozzle segment of a gas turbine

    SciTech Connect

    Burdgick, Steven Sebastian

    2002-01-01

    An insert containing apertures for impingement cooling a nozzle vane of a nozzle segment in a gas turbine is inserted into one end of the vane. The leading end of the insert is positioned slightly past a rib adjacent the opposite end of the vane through which the insert is inserted. The end of the insert is formed or swaged into conformance with the inner margin of the rib. The insert is then brazed or welded to the rib.

  16. The genomic landscape of polymorphic human nuclear mitochondrial insertions

    PubMed Central

    Dayama, Gargi; Emery, Sarah B.; Kidd, Jeffrey M.; Mills, Ryan E.

    2014-01-01

    The transfer of mitochondrial genetic material into the nuclear genomes of eukaryotes is a well-established phenomenon that has been previously limited to the study of static reference genomes. The recent advancement of high throughput sequencing has enabled an expanded exploration into the diversity of polymorphic nuclear mitochondrial insertions (NumtS) within human populations. We have developed an approach to discover and genotype novel Numt insertions using whole genome, paired-end sequencing data. We have applied this method to a thousand individuals in 20 populations from the 1000 Genomes Project and other datasets and identified 141 new sites of Numt insertions, extending our current knowledge of existing NumtS by almost 20%. We find that recent Numt insertions are derived from throughout the mitochondrial genome, including the D-loop, and have integration biases that differ in some respects from previous studies on older, fixed NumtS in the reference genome. We determined the complete inserted sequence for a subset of these events and have identified a number of nearly full-length mitochondrial genome insertions into nuclear chromosomes. We further define their age and origin of insertion and present an analysis of their potential impact to ongoing studies of mitochondrial heteroplasmy and disease. PMID:25348406

  17. Toward automated cochlear implant insertion using tubular manipulators

    NASA Astrophysics Data System (ADS)

    Granna, Josephine; Rau, Thomas S.; Nguyen, Thien-Dang; Lenarz, Thomas; Majdani, Omid; Burgner-Kahrs, Jessica

    2016-03-01

    During manual cochlear implant electrode insertion the surgeon is at risk to damage the intracochlear fine-structure, as the electrode array is inserted through a small opening in the cochlea blindly with little force-feedback. This paper addresses a novel concept for cochlear electrode insertion using tubular manipulators to reduce risks of causing trauma during insertion and to automate the insertion process. We propose a tubular manipulator incorporated into the electrode array composed of an inner wire within a tube, both elastic and helically shaped. It is our vision to use this manipulator to actuate the initially straight electrode array during insertion into the cochlea by actuation of the wire and tube, i.e. translation and slight axial rotation. In this paper, we evaluate the geometry of the human cochlea in 22 patient datasets in order to derive design requirements for the manipulator. We propose an optimization algorithm to automatically determine the tube set parameters (curvature, torsion, diameter, length) for an ideal final position within the cochlea. To prove our concept, we demonstrate that insertion can be realized in a follow-the-leader fashion for 19 out of 22 cochleas. This is possible with only 4 different tube/wire sets.

  18. Isthmus-guided cortical bone trajectory for pedicle screw insertion.

    PubMed

    Iwatsuki, Koichi; Yoshimine, Toshiki; Ohnishi, Yu-ichiro; Ninomiya, Kosi; Ohkawa, Toshika

    2014-08-01

    Herein is described cortical bone trajectory (CBT), a new path for pedicle screw insertion for lumbar vertebral fusion. Because the points of insertion are under the end of the inferior articular process, and because the screws are inserted toward the lateral side, there is less soft tissue development than with the conventional technique; the CBT technique therefore enables less invasive surgery than the conventional technique. However, it has some drawbacks. For example, in the original CBT approach, the points of insertion are in the vicinity of the end of the inferior articular process. Because this joint has been destroyed in many patients who have indications for intervertebral fusion surgery, it is sometimes difficult to use it as a reference point for screw insertion location. With severe lateral slippage, the screw insertion site can become significantly dislocated sideways, with possible resultant damaging to the spinal canal and/or nerve root. The CBT technique here involved inserting the screws while keeping clear of the intervertebral foramen with the assistance of side view X-ray fluoroscopy and using the end of the inferior articular process and the isthmus as points of reference for screw location.

  19. World lines.

    PubMed

    Waser, Jürgen; Fuchs, Raphael; Ribicić, Hrvoje; Schindler, Benjamin; Blöschl, Günther; Gröller, Eduard

    2010-01-01

    In this paper we present World Lines as a novel interactive visualization that provides complete control over multiple heterogeneous simulation runs. In many application areas, decisions can only be made by exploring alternative scenarios. The goal of the suggested approach is to support users in this decision making process. In this setting, the data domain is extended to a set of alternative worlds where only one outcome will actually happen. World Lines integrate simulation, visualization and computational steering into a single unified system that is capable of dealing with the extended solution space. World Lines represent simulation runs as causally connected tracks that share a common time axis. This setup enables users to interfere and add new information quickly. A World Line is introduced as a visual combination of user events and their effects in order to present a possible future. To quickly find the most attractive outcome, we suggest World Lines as the governing component in a system of multiple linked views and a simulation component. World Lines employ linking and brushing to enable comparative visual analysis of multiple simulations in linked views. Analysis results can be mapped to various visual variables that World Lines provide in order to highlight the most compelling solutions. To demonstrate this technique we present a flooding scenario and show the usefulness of the integrated approach to support informed decision making.

  20. Industrial stator vane with sequential impingement cooling inserts

    DOEpatents

    Jones, Russell B; Fedock, John A; Goebel, Gloria E; Krueger, Judson J; Rawlings, Christopher K; Memmen, Robert L

    2013-08-06

    A turbine stator vane for an industrial engine, the vane having two impingement cooling inserts that produce a series of impingement cooling from the pressure side to the suction side of the vane walls. Each insert includes a spar with a row of alternating impingement cooling channels and return air channels extending in a radial direction. Impingement cooling plates cover the two sides of the insert and having rows of impingement cooling holes aligned with the impingement cooling channels and return air openings aligned with the return air channel.

  1. CT fluoroscopic guided insertion of inferior vena cava filters.

    PubMed

    Ignotus, P; Wetton, C; Berry, J

    2006-03-01

    The value and use of inferior vena cava (IVC) filters is well documented and has been growing since the first reported filter placement in 1973 and the first percutaneous insertion in 1982. Access routes now include both jugular veins, both ante-cubital veins and both femoral veins. However, all insertions require some form of imaging, usually fluoroscopy, to identify the location of the filter with respect to the IVC and the renal veins. We describe two cases where the patients' weight was significantly greater than the weight limit of the angiography table, necessitating insertion under CT fluoroscopic guidance.

  2. A Multiple RCC Device for Polygonal Peg Insertion

    NASA Astrophysics Data System (ADS)

    Cheng, Chi-Cheng; Chen, Gin-Shan

    This paper proposes a new passive compliant center device, the Multiple Remote Center Compliance (MRCC), for assembly tasks. The MRCC introduces a new azimuthal compliance over traditional passive compliance mechanisms that can effectively compensate the peg's orientation deviation for polygonal insertions. Besides, a special feature of the adjustable compliance provides capability to overcome the gravity effect. Non-vertical insertions therefore become possible. A spring-supported object in space is also adopted for stability analysis of this compliant device. Actual experimental assembly processes demonstrate promising results on polygonal insertions in both traditional top-down and horizontal directions.

  3. International Workshop on Magnetic Measurements of Insertion Devices

    SciTech Connect

    Not Available

    1993-10-01

    The International Workshop on Magnetic Measurements of Insertion Devices was held at the Advanced Photon Source, Argonne National Laboratory, on September 28--29, 1993. The workshop brought together scientists and engineers from Europe, Japan, and the United States to discuss the following topics: Special techniques for magnetic measurements of insertion devices, magnetic tolerances of the insertion devices for third generation synchrotron radiation sources, methods for and accuracy of the multipole moments measurements, magnetic sensors, among other topics. The workshop included thirteen presentations that are collected in this volume.

  4. Positioning long lines: contrast versus plain radiography

    PubMed Central

    Reece, A; Ubhi, T; Craig, A; Newell, S

    2001-01-01

    AIM—To assess the value of contrast versus plain radiography in determining radio-opaque long line tip position in neonates.
METHODS—In a prospective study, plain radiography was performed after insertion of radio-opaque long lines. If the line tip was not visible on the plain film, a second film with contrast was obtained in an attempt to visualise the tip.
RESULTS—Sixty eight lines were inserted during the study period, 62 of which were included in the study. In 31, a second radiographic examination with contrast was necessary to determine position of the tip. In 29 of these, the line tip was clearly visualised with contrast. On two occasions, the line tip could not be seen because the contrast had filled the vein and obscured the tip from view. Eight of the lines that required a second radiograph with contrast were repositioned.
CONCLUSION—Intravenous contrast should be routinely used in the assessment of long line position in the neonate.

 PMID:11207231

  5. Effect of insertion speed on tissue response and insertion mechanics of a chronically implanted silicon-based neural probe.

    PubMed

    Welkenhuysen, M; Andrei, A; Ameye, L; Eberle, W; Nuttin, B

    2011-11-01

    In this study, the effect of insertion speed on long-term tissue response and insertion mechanics was investigated. A dummy silicon parylene-coated probe was used in this context and implanted in the rat brain at 10 μm/s (n = 6) or 100 μm/s (n = 6) to a depth of 9 mm. The insertion mechanics were assessed by the dimpling distance, and the force at the point of penetration, at the end of the insertion phase, and after a 3-min rest period in the brain. After 6 weeks, the tissue response was evaluated by estimating the amount of gliosis, inflammation, and neuronal cell loss with immunohistochemistry. No difference in dimpling, penetration force, or the force after a 3-min rest period in the brain was observed. However, the force at the end of the insertion phase was significantly higher when inserting the probes at 100 μm/s compared to 10 μm/s. Furthermore, an expected tissue response was seen with an increase of glial and microglial reactivity around the probe. This reaction was similar along the entire length of the probe. However, evidence for a neuronal kill zone was observed only in the most superficial part of the implant. In this region, the lesion size was also greatest. Comparison of the tissue response between insertion speeds showed no differences.

  6. Physics-based RF/microwave characterization of wave interactions within electrical connectors with partial insertion faults

    NASA Astrophysics Data System (ADS)

    Tokgöz, Çaǧatay; Dardona, Sameh

    2016-09-01

    Electrical failures in avionics systems may result from connector faults. If fault precursors are not detected in advance, they may lead to hard failures such as open and short circuits that could ultimately result in fire or loss of flight critical systems. Therefore, It is crucial to detect, locate, and characterize fault precursors for timely preventive maintenance and mitigation before hard failures occur. In this paper, a physics-based connector model consisting of multiple coaxial line sections with different characteristic impedances and lengths is proposed. Method of Moments (MoM) analyses were performed using commercial electromagnetic simulation software, FEKO, for transverse electric and magnetic (TEM) wave propagation through a connector. The physical parameters of the connector were optimized to match the measured S parameters for multiple insertion depths. The proposed models represent the connector for multiple insertion depths by varying only two length parameters at a time while other parameters are fixed. Insertion depth-dependent resonant frequency shifts observed during measurement are also captured by the model over the full range of fully inserted to barely touching contacts. Hence, the models provide accurate representations of the connector and properly detect precursors to partial insertion faults.

  7. A possible aid in targeted insertion of large DNA elements by CRISPR/Cas in mouse zygotes.

    PubMed

    Nakao, Harumi; Harada, Takeshi; Nakao, Kazuki; Kiyonari, Hiroshi; Inoue, Kenichi; Furuta, Yasuhide; Aiba, Atsu

    2016-02-01

    The CRISPR/Cas system has rapidly emerged recently as a new tool for genome engineering, and is expected to allow for controlled manipulation of specific genomic elements in a variety of species. A number of recent studies have reported the use of CRISPR/Cas for gene disruption (knockout) or targeted insertion of foreign DNA elements (knock-in). Despite the ease of simple gene knockout and small insertions or nucleotide substitutions in mouse zygotes by the CRISPR/Cas system, targeted insertion of large DNA elements remains an apparent challenge. Here the generation of knock-in mice with successful targeted insertion of large donor DNA elements ranged from 3.0 to 7.1 kb at the ROSA26 locus using the CRISPR/Cas system was achieved. Multiple independent knock-in founder mice were obtained by injection of hCas9 mRNA/sgRNA/donor vector mixtures into the cytoplasm of C57BL/6N zygotes when the injected zygotes were treated with an inhibitor of actin polymerization, cytochalasin. Successful germ line transmission of three of these knock-in alleles was also confirmed. The results suggested that treatment of zygotes with actin polymerization inhibitors following microinjection could be a viable method to facilitate targeted insertion of large DNA elements by the CRISPR/Cas system, enabling targeted knock-in readily attainable in zygotes.

  8. Mining for single nucleotide polymorphisms and insertions/deletions in maize expressed sequence tag data.

    PubMed

    Batley, Jacqueline; Barker, Gary; O'Sullivan, Helen; Edwards, Keith J; Edwards, David

    2003-05-01

    We have developed a computer based method to identify candidate single nucleotide polymorphisms (SNPs) and small insertions/deletions from expressed sequence tag data. Using a redundancy-based approach, valid SNPs are distinguished from erroneous sequence by their representation multiple times in an alignment of sequence reads. A second measure of validity was also calculated based on the cosegregation of the SNP pattern between multiple SNP loci in an alignment. The utility of this method was demonstrated by applying it to 102,551 maize (Zea mays) expressed sequence tag sequences. A total of 14,832 candidate polymorphisms were identified with an SNP redundancy score of two or greater. Segregation of these SNPs with haplotype indicates that candidate SNPs with high redundancy and cosegregation confidence scores are likely to represent true SNPs. This was confirmed by validation of 264 candidate SNPs from 27 loci, with a range of redundancy and cosegregation scores, in four inbred maize lines. The SNP transition/transversion ratio and insertion/deletion size frequencies correspond to those observed by direct sequencing methods of SNP discovery and suggest that the majority of predicted SNPs and insertion/deletions identified using this approach represent true genetic variation in maize.

  9. Monoaxial external fixation of the calcaneus: An anatomical study assessing the safety of monoaxial pin insertion.

    PubMed

    Thomson, Callum M; Esparon, Tom; Rea, Paul M; Jamal, Bilal

    2016-10-01

    The use of external fixation for intra-articular calcaneal fractures is increasing in popularity. Studies have shown fine wire and monoaxial external fixation to be a viable surgical alternative to more invasive methods of open reduction and internal fixation of the calcaneus. However, there is an absence of literature that quantifies the risk of pin insertion for monoaxial fixation. This study aimed to determine the safety of inserting monoaxial pins within the calcaneus to house the Orthofix Calcaneal Mini-Fixator. Five formalin embalmed cadaveric ankle and lower leg specimens were inserted with six monoaxial pins. Careful dissection then revealed the presence of the tendons of peroneus longus and brevis, the sural nerve and the small saphenous vein in relation to these pins. Measurements from each pin to each of these structures were made as the structures transected lines drawn from each pin to two palpable bony landmarks: the inferior tip of the lateral malleolus and the posterosuperior calcaneus. In doing this, the risk posed by each pin could be evaluated. We found that two particular pins, those used to hold the articular surface of the subtalar joint in a reduced position, posed a larger risk of injury to surrounding structures than the remaining pins. These findings therefore suggest that monoaxial fixation of the calcaneus using a six pin approach is a relatively safe method of rectifying calcaneal fractures and thus may serve as a welcome alternative to other methods of calcaneal fixation.

  10. The use of a newspaper insertion to promote DIY testing of vision in India

    PubMed Central

    Murthy, G; Gupta, S. K.; Dada, V. K.; Pant, T. D.; Savita, C.; Sanga, L.; Neena, J.

    2001-01-01

    BACKGROUND—The mass media have the potential to motivate people to participate in self appraisal of their own health status. An innovative communication package was designed to help people to examine vision at home. The impact of publishing the "do it yourself" (DIY) kit in Indian newspapers was evaluated.
METHODS—A pretested bilingual vision testing kit was published in three newspapers. The kit comprised four tumbling Es corresponding to 6/12 line of Snellen's optotypes. Directions on using the kit were enclosed. 3 -7 days after publication of the kit, a telephone survey of newspaper readers was undertaken to evaluate the impact and cost effectiveness.
RESULTS—603 people were contacted over the telephone. 125 (20.73%) subscribed to the newspaper carrying the DIY insertion. 43.2% (54) noticed the insertion of which 88.89% (48) read the enclosed instructions carefully. 58.33% respondents felt sufficiently motivated to contact an ophthalmologist. Graduates had a 3.83 times higher probability of reading the communication insertion compared with others. Differences in relation to other demographic variables were not statistically significant.
CONCLUSIONS—Newspapers are an excellent medium for communicating self appraisal kits for vision testing. The medium is cost effective and has significant reach in the urban agglomerates of India.

 PMID:11466254

  11. LiPS-A3S, a human genomic site for robust expression of inserted transgenes

    PubMed Central

    Kotini, Andriana G; Sadelain, Michel; Papapetrou, Eirini P

    2016-01-01

    Transgenesis of human pluripotent stem cells (hPSCs) can enable and empower a variety of studies in stem cell research, including lineage tracing and functional genetics studies. While in recent years much progress has been made in the development of tools for gene targeting, little attention has been given to the identification of sites in the human genome where transgenes can be inserted and reliably expressed. In order to find human genomic sites capable of supporting long-term and high-level transgene expression in hPSCs, we performed a lentiviral screen in human induced pluripotent stem cells (iPSCs). We isolated 40 iPSC clones each harboring a single vector copy and characterized the level of transgene expression afforded by each unique integration site. We selected one clone, LiPS-A3 with an integration site in chromosome 15 maintaining robust expression without silencing and demonstrate that different transgenes can be inserted therein rapidly and efficiently through recombinase-mediated cassette exchange (RMCE). The LiPS-A3 line can greatly facilitate the insertion of reporter and other genes in hPSCs. Targeting transgenes in the LiPS-A3S genomic locus can find broad applications in stem cell research and possibly cell and gene therapy. PMID:27898090

  12. A novel insertion mutation on exon 20 of epidermal growth factor receptor, conferring resistance to erlotinib.

    PubMed

    Khan, Nawazish A; Mirshahidi, Saied; Mirshahidi, Hamid R

    2014-05-01

    The epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein tyrosine kinase receptor. The small-molecule tyrosine kinase receptor inhibitors (TKIs) are in clinical use to treat non-small cell lung cancer with EGFR mutations. Variable tumor responses to erlotinib and gefitinib have been observed. The response to these TKIs varies by the type of EGFR mutations found in the tumor. The deletion on exon 19 and the L858R substitution on exon 21 constitute the most frequent mutations and are known to show good response to TKIs. However, mutations on exon 20 are less common and seem to respond poorly to TKIs. In clinical settings, the reported response of exon 20 mutations to reversible TKIs (both gefitinib and erlotinib) remains inconstant. The type of coexisting mutation seems to affect the response of these insertions to TKIs. We herein present a case of disease progression despite the use of erlotinib in a female patient who had a novel insertion mutation on exon 20. Our patient was a never-smoker and was identified to have a Pro772_His773insGlnCysPro mutation on exon 20. She had previously been treated with cisplatin and gemcitabine and then with carboplatin and pemetrexed. She was treated with erlotinib upon intolerance to second-line chemotherapy and did not respond. Our patient had a novel insertion mutation on exon 20, which was found to be resistant to erlotinib.

  13. 33. Launch Control Center, close view of launch key inserted ...

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

    33. Launch Control Center, close view of launch key inserted in the launch panel. Lyon - Whiteman Air Force Base, Oscar O-1 Minuteman Missile Alert Facility, Southeast corner of Twelfth & Vendenberg Avenues, Knob Noster, Johnson County, MO

  14. Interior, detail closeup shot of window with stained glass inserts ...

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

    Interior, detail closeup shot of window with stained glass inserts in top southeast room taken from ther west - J. Weingartner & Son Cigar Factory, 414 East Walnut Street, North Wales, Montgomery County, PA

  15. Membrane protein insertion: mixing eukaryotic and prokaryotic concepts.

    PubMed

    Schleiff, Enrico; Soll, Jürgen

    2005-11-01

    Proteins are translocated across or inserted into membranes by machines that are composed of soluble and membrane-anchored subunits. The molecular action of these machines and their evolutionary origin are at present the focus of intense research. For instance, our understanding of the mode of insertion of beta-barrel membrane proteins into the outer membrane of endosymbiotically derived organelles has increased rapidly during the past few years. In particular, the identification of the Omp85/YaeT-involving pathways in Neisseria meningitidis, Escherichia coli and cyanobacteria, and homologues of Omp85/YaeT in chloroplasts and mitochondria, has provided new clues about the ancestral beta-barrel protein insertion pathway. This review focuses on recent advances in the elucidation of the evolutionarily conserved concepts that underlie the translocation and insertion of beta-barrel membrane proteins.

  16. Spontaneous Radiation Emission from Short, High Field Strength Insertion Devices

    SciTech Connect

    Geoffrey Krafft

    2005-09-15

    Since the earliest papers on undulaters were published, it has been known how to calculate the spontaneous emission spectrum from ''short'' undulaters when the magnetic field strength parameter is small compared to unity, or in ''single'' frequency sinusoidal undulaters where the magnetic field strength parameter is comparable to or larger than unity, but where the magnetic field amplitude is constant throughout the undulater. Fewer general results have been obtained in the case where the insertion device is both short, i.e., the magnetic field strength parameter changes appreciably throughout the insertion device, and the magnetic field strength is high enough that ponderomotive effects, radiation retardation, and harmonic generation are important physical phenomena. In this paper a general method is presented for calculating the radiation spectrum for short, high-field insertion devices. It is used to calculate the emission from some insertion device designs of recent interest.

  17. Pressure molding of powdered materials improved by rubber mold insert

    NASA Technical Reports Server (NTRS)

    1964-01-01

    Pressure molding tungsten microspheres is accomplished by applying hydraulic pressure to a silicone rubber mold insert with several barrel shaped chambers which is placed in a steel die cavity. This technique eliminates castings containing shear fractures.

  18. Development of Magnet Technologies for HTS Insert Coils

    NASA Astrophysics Data System (ADS)

    Wesche, Rainer; Uglietti, Davide; Bruzzone, Pierluigi; March, Stephen; Marinucci, Claudio; Stepanov, Boris; Glowa, Natalia

    An existing Nb3Sn laboratory magnet generating a magnetic field of 12 T is intended to be upgraded to 16 T by means of the use of a high temperature superconductor (HTS) insert coil. An outline design of the HTS insert coil is presented. In the design, the aspects of the maximum achievable operation current, the required copper cross-section to ensure a hot spot temperature below 200 K and the resulting forces and stresses have been considered. The length of the insert coil has been selected in such a way that the field uniformity will be better than 1% within a sphere of 3 cm diameter. The protection of the whole magnet system (LTS & HTS insert) is briefly described.

  19. The use of dimorphic Alu insertions in human DNA fingerprinting

    SciTech Connect

    Novick, G.E.; Gonzalez, T.; Garrison, J.; Novick, C.C.; Herrera, R.J.; Batzer, M.A.; Deininger, P.L.

    1992-12-04

    We have characterized certain Human Specific Alu Insertions as either dimorphic (TPA25, PV92, APO), sightly dimorphic (C2N4 and C4N4) or monomorphic (C3N1, C4N6, C4N2, C4N5, C4N8), based on studies of Caucasian, Asian, American Black and African Black populations. Our approach is based upon: (1) PCR amplification using primers directed to the sequences that flank the site of insertion of the different Alu elements studied; (2) gel electrophoresis and scoring according to the presence or absence of an Alu insertion in one or both homologous chromosomes; (3) allelic frequencies calculated and compared according to Hardy-Weinberg equilibrium. Our DNA fingerprinting procedure using PCR amplification of dimorphic Human Specific Alu insertions, is stable enough to be used not only as a tool for genetic mapping but also to characterize populations, study migrational patterns and track the inheritance of human genetic disorders.

  20. 21 CFR 529.1940 - Progesterone intravaginal inserts.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... postpartum estrus in suckled beef cows; and for advancement of first pubertal estrus in replacement beef... postpartum. Do not use an insert more than once. To prevent the potential transmission of venereal...

  1. 21 CFR 529.1940 - Progesterone intravaginal inserts.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... postpartum estrus in suckled beef cows; and for advancement of first pubertal estrus in replacement beef... postpartum. Do not use an insert more than once. To prevent the potential transmission of venereal...

  2. 21 CFR 529.1940 - Progesterone intravaginal inserts.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... postpartum estrus in suckled beef cows; and for advancement of first pubertal estrus in replacement beef... postpartum. Do not use an insert more than once. To prevent the potential transmission of venereal...

  3. 21 CFR 529.1940 - Progesterone intravaginal inserts.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... postpartum estrus in suckled beef cows; and for advancement of first pubertal estrus in replacement beef... genital tracts. Do not use in beef cows that are fewer than 20 days postpartum. Do not use an insert...

  4. Bougie insertion: A common practice with underestimated dangers

    PubMed Central

    Theodorou, D.; Doulami, G.; Larentzakis, A.; Almpanopoulos, K.; Stamou, K.; Zografos, G.; Menenakos, E.

    2011-01-01

    Introduction Esophageal perforation after bariatric operations is rare. We report two cases of esophageal perforation after bariatric operations indicating the dangers of a common practice – like insertion of esophageal tubes – and we describe our management of that complication. Presentation of case A 56 year old woman who underwent laparoscopic sleeve gastrectomy and a 41 year old woman who underwent laparoscopic adjustable gastric banding respectively. In both operations a bougie has been used and led to esophageal perforation. Discussion The insertion of bougie and especially of inflated bougie is a common practice. It is an invasive procedure that in most cases is performed by the anesthesiologist team. Conclusion Bougie insertion is an invasive procedure with risks and should always be attempted under direct supervision of surgical team or should be inserted by a surgeon. PMID:22288051

  5. Metal Insertion in a Microporous Metal-Organic Framework Lined with 2,2'-Bipyridine

    SciTech Connect

    Bloch, Eric D.; Britt, David; Lee, Chain; Doonan, Christian J.; Uribe-Romo, Fernando J.; Furukawa, Hiroyasu; Long, Jeffrey R.; Yaghi, Omar M.

    2010-10-20

    Reaction of AlCl3·6H2O with 2,2'-bipyridine-5,5'-dicarboxylic acid (H2bpydc) affords Al(OH)(bpydc) (1, MOF-253), the first metal-organic framework with open 2,2'-bipyridine (bpy) coordination sites. The material displays a BET surface area of 2160 m2/g and readily complexes metals to afford, for example, 1·xPdCl2 (x = 0.08, 0.83) and 1·0.97Cu(BF4)2. EXAFS spectroscopy performed on 1·0.83PdCl2 reveals the expected square planar coordination geometry, matching the structure of the model complex (bpy)PdCl2. Finally, significantly, the selectivity factor for binding CO2 over N2 under typical flue gas conditions is observed to increase from 2.8 in 1 to 12 in 1·0.97Cu(BF4)2.

  6. MTR MAIN FLOOR. MEN DEMONSTRATE INSERTION OF DUMMY PLUG INTO ...

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

    MTR MAIN FLOOR. MEN DEMONSTRATE INSERTION OF DUMMY PLUG INTO AN MTR BEAM HOLE. ONE MAN CHECKS RADIATION LEVEL AT THE END OF THE UNIVERSAL COFFIN, WHILE ANOTHER USES TOOL TO INSERT PLUG INTO HOLE THROUGH COFFIN. MEN WEAR "ANTI-C" (ANTI-CONTAMINATION) CLOTHING. INL NEGATIVE NO. 6198. R.G. Larsen, Photographer, 6/27/1952 - Idaho National Engineering Laboratory, Test Reactor Area, Materials & Engineering Test Reactors, Scoville, Butte County, ID

  7. Impediments to rapid insertion of innovative displays and peripherals

    NASA Astrophysics Data System (ADS)

    Nicholson, Gail

    2012-06-01

    In order to optimize system performance and minimize cost for a system to fill capability gaps, an improvement to rapid insertion of innovative display and peripheral technology is required to take advantage of human-machine intersections. Current approaches to testing and integration impedes successful rapid insertion of innovative technology for new systems and incremental upgrades. Considerations to innovative displays and peripherals must occur further to the left of the lifecycle to be successful and key integration areas must be address for success.

  8. Evaluation of preferable insertion routes for esophagogastroduodenoscopy using ultrathin endoscopes

    PubMed Central

    Ono, Satoshi; Niimi, Keiko; Fujishiro, Mitsuhiro; Takahashi, Yu; Sakaguchi, Yoshiki; Nakayama, Chiemi; Minatsuki, Chihiro; Matsuda, Rie; Hirayama-Asada, Itsuko; Tsuji, Yosuke; Mochizuki, Satoshi; Kodashima, Shinya; Yamamichi, Nobutake; Ozeki, Atsuko; Matsumoto, Lumine; Ohike, Yumiko; Yamazaki, Tsutomu; Koike, Kazuhiko

    2014-01-01

    AIM: To evaluate the discomfort associated with esophagogastroduodenoscopy (EGD) using an ultrathin endoscope through different insertion routes. METHODS: This study (January 2012-March 2013) included 1971 consecutive patients [male/female (M/F), 1158/813, 57.5 ± 11.9 years] who visited a single institute for annual health checkups. Transnasal EGD was performed in 1394 patients and transoral EGD in 577. EGD-associated discomfort was assessed using a visual analog scale score (VAS score: 0-10). RESULTS: Multivariate analysis revealed gender (M vs F: 4.02 ± 2.15 vs 5.06 ± 2.43) as the only independent predictor of the VAS score in 180 patients who underwent EGD for the first time; whereas it revealed gender (M vs F 3.60 ± 2.20 vs 4.84 ± 2.37), operator, age group (A: < 39 years; B: 40-49 years; C: 50-59 years; D: 60-69 years; E: > 70 years; A/B/C/D/E: 4.99 ± 2.32/4.34 ± 2.49/4.19 ± 2.31/3.99 ± 2.27/3.63 ± 2.31), and type of insertion as independent predictors in the remaining patients. Subanalysis for gender, age group, and insertion route revealed that the VAS score decreased with age regardless of gender and insertion route, was high in female patients regardless of age and insertion route, and was low in males aged over 60 years who underwent transoral insertion. CONCLUSION: Although comprehensive analysis revealed that the insertion route may not be an independent predictor of the VAS score, transoral insertion may reduce EGD-associated discomfort in elderly patients. PMID:24803817

  9. Mechanism for electrochemical hydrogen insertion in carbonaceous materials

    NASA Astrophysics Data System (ADS)

    Qu, Deyang

    The mechanism for safe and reversible storage of hydrogen in porous carbonaceous materials by electrochemical decomposition of water in alkaline electrolyte is proposed. Atomic H was found to be inserted into the microdomains of defective graphene layers. Hydrogen storage capacity increases with increasing interlayer distance between carbon sheets. Hydrogen insertion in carbonaceous materials occurs at ambient conditions. Static potential acts as an electrochemical valve which can retain the hydrogen in the carbon structure, thus preventing leakage during storage.

  10. Evolution models with base substitutions, insertions, deletions, and selection

    NASA Astrophysics Data System (ADS)

    Saakian, D. B.

    2008-12-01

    The evolution model with parallel mutation-selection scheme is solved for the case when selection is accompanied by base substitutions, insertions, and deletions. The fitness is assumed to be either a single-peak function (i.e., having one finite discontinuity) or a smooth function of the Hamming distance from the reference sequence. The mean fitness is calculated exactly in large-genome limit. In the case of insertions and deletions the evolution characteristics depend on the choice of reference sequence.

  11. Sensorless Motion Planning for Medical Needle Insertion in Deformable Tissues

    PubMed Central

    Alterovitz, Ron; Goldberg, Kenneth Y.; Pouliot, Jean; Hsu, I-Chow

    2009-01-01

    Minimally invasive medical procedures such as biopsies, anesthesia drug injections, and brachytherapy cancer treatments require inserting a needle to a specific target inside soft tissues. This is difficult because needle insertion displaces and deforms the surrounding soft tissues causing the target to move during the procedure. To facilitate physician training and preoperative planning for these procedures, we develop a needle insertion motion planning system based on an interactive simulation of needle insertion in deformable tissues and numerical optimization to reduce placement error. We describe a 2-D physically based, dynamic simulation of needle insertion that uses a finite-element model of deformable soft tissues and models needle cutting and frictional forces along the needle shaft. The simulation offers guarantees on simulation stability for mesh modications and achieves interactive, real-time performance on a standard PC. Using texture mapping, the simulation provides visualization comparable to ultrasound images that the physician would see during the procedure. We use the simulation as a component of a sensorless planning algorithm that uses numerical optimization to compute needle insertion offsets that compensate for tissue deformations. We apply the method to radioactive seed implantation during permanent seed prostate brachytherapy to minimize seed placement error. PMID:19126473

  12. Polyubiquitin insertions and the phylogeny of Cercozoa and Rhizaria.

    PubMed

    Bass, David; Moreira, David; López-García, Purificación; Polet, Stephane; Chao, Ema E; von der Heyden, Sophie; Pawlowski, Jan; Cavalier-Smith, Thomas

    2005-08-01

    A single or double amino acid insertion at the monomer-monomer junction of the universal eukaryotic protein polyubiquitin is unique to Cercozoa and Foraminifera, closely related 'core' phyla in the protozoan infrakingdom Rhizaria. We screened 11 other candidate rhizarians for this insertion: Radiozoa (polycystine and acantharean radiolaria), a 'microheliozoan', and Apusozoa; all lack it, supporting suggestions that Foraminifera are more closely related to Cercozoa than either is to other eukaryotes. The insertion's size was ascertained for 12 additional Cercozoa to help resolve their basal branching order. The earliest branching Cercozoa generally have a single amino acid insertion, like all Foraminifera, but a large derived clade consisting of all Monadofilosa except Metopion, Helk-esimastix, and Cercobodo agilis has two amino acids, suggesting one doubling event and no reversions to a single amino acid. Metromonas and Sainouron, cercozoans of uncertain position, have a double insertion, suggesting that they belong in Monadofilosa. An alternative interpretation, suggested by the higher positions for Metopion and Cercobodo on Bayesian trees compared with most distance trees, cannot be ruled out, i.e. that the second insertion took place earlier, in the ancestral filosan, and was followed by three independent reversions to a single amino acid in Chlorarachnea, Metopion and Cercobodo.

  13. Sensorless motion planning for medical needle insertion in deformable tissues.

    PubMed

    Alterovitz, Ron; Goldberg, Kenneth Y; Pouliot, Jean; Hsu, I-Chow Joe

    2009-03-01

    Minimally invasive medical procedures such as biopsies, anesthesia drug injections, and brachytherapy cancer treatments require inserting a needle to a specific target inside soft tissues. This is difficult because needle insertion displaces and deforms the surrounding soft tissues causing the target to move during the procedure. To facilitate physician training and preoperative planning for these procedures, we develop a needle insertion motion planning system based on an interactive simulation of needle insertion in deformable tissues and numerical optimization to reduce placement error. We describe a 2-D physically based, dynamic simulation of needle insertion that uses a finite-element model of deformable soft tissues and models needle cutting and frictional forces along the needle shaft. The simulation offers guarantees on simulation stability for mesh modifications and achieves interactive, real-time performance on a standard PC. Using texture mapping, the simulation provides visualization comparable to ultrasound images that the physician would see during the procedure. We use the simulation as a component of a sensorless planning algorithm that uses numerical optimization to compute needle insertion offsets that compensate for tissue deformations. We apply the method to radioactive seed implantation during permanent seed prostate brachytherapy to minimize seed placement error.

  14. Evaluation of a high resolution silicon PET insert module

    NASA Astrophysics Data System (ADS)

    Grkovski, Milan; Brzezinski, Karol; Cindro, Vladimir; Clinthorne, Neal H.; Kagan, Harris; Lacasta, Carlos; Mikuž, Marko; Solaz, Carles; Studen, Andrej; Weilhammer, Peter; Žontar, Dejan

    2015-07-01

    Conventional PET systems can be augmented with additional detectors placed in close proximity of the region of interest. We developed a high resolution PET insert module to evaluate the added benefit of such a combination. The insert module consists of two back-to-back 1 mm thick silicon sensors, each segmented into 1040 1 mm2 pads arranged in a 40 by 26 array. A set of 16 VATAGP7.1 ASICs and a custom assembled data acquisition board were used to read out the signal from the insert module. Data were acquired in slice (2D) geometry with a Jaszczak phantom (rod diameters of 1.2-4.8 mm) filled with 18F-FDG and the images were reconstructed with ML-EM method. Both data with full and limited angular coverage from the insert module were considered and three types of coincidence events were combined. The ratio of high-resolution data that substantially improves quality of the reconstructed image for the region near the surface of the insert module was estimated to be about 4%. Results from our previous studies suggest that such ratio could be achieved at a moderate technological expense by using an equivalent of two insert modules (an effective sensor thickness of 4 mm).

  15. [Duration and complications following grommet insertion in childhood].

    PubMed

    Fiebach, A; Matschke, R G

    1987-02-01

    We report 1000 insertions of ventilation tubes in 534 children for secretory otitis media (SOM) within a period of 6 years. In 77.5% of the cases, the air-bone gap was greater than 20 dB. The grommets are allowed to undergo spontaneous expulsion which happened in 319 ears, about 7 months after insertion. SOM recurred in 32.6% of the cases once, in 5.0% of the cases twice, and in 1.9% three times, requiring re-insertion of grommets. Recurrence appeared between 4 and 65 months after the first insertion. In three cases perforations of the tympanic membrane persisted and required tympanoplasty. Tympanometric examination and pure tone audiometry are necessary, in addition to pure tone audiometry to follow up successfully ventilated middle ears and to show recurrence as early as possible. Pre-existing but unsuspected sensorineural hearing loss was discovered in 10 cases. The deprivation of neural auditory pathways in early childhood and the possible retardation of speech development caused by SOM are mentioned. Normal hearing is the most important goal of therapy in children suffering from SOM, and insertion of ventilation tubes is the preferred method of achieving that goal safely. Adenoidectomy and treatment of upper airway infections are indispensable parts of the therapy of SOM. Regular follow up after insertion of grommets and even after their expulsion is necessary because of the high rate of late recurrence of SOM. Induction of cholesteatoma by grommets was not observed.

  16. Retrotransposon-Induced Heterochromatin Spreading in the Mouse Revealed by Insertional Polymorphisms

    PubMed Central

    Rebollo, Rita; Karimi, Mohammad M.; Bilenky, Misha; Gagnier, Liane; Miceli-Royer, Katharine; Zhang, Ying; Goyal, Preeti; Keane, Thomas M.; Jones, Steven; Hirst, Martin; Lorincz, Matthew C.; Mager, Dixie L.

    2011-01-01

    The “arms race” relationship between transposable elements (TEs) and their host has promoted a series of epigenetic silencing mechanisms directed against TEs. Retrotransposons, a class of TEs, are often located in repressed regions and are thought to induce heterochromatin formation and spreading. However, direct evidence for TE–induced local heterochromatin in mammals is surprisingly scarce. To examine this phenomenon, we chose two mouse embryonic stem (ES) cell lines that possess insertionally polymorphic retrotransposons (IAP, ETn/MusD, and LINE elements) at specific loci in one cell line but not the other. Employing ChIP-seq data for these cell lines, we show that IAP elements robustly induce H3K9me3 and H4K20me3 marks in flanking genomic DNA. In contrast, such heterochromatin is not induced by LINE copies and only by a minority of polymorphic ETn/MusD copies. DNA methylation is independent of the presence of IAP copies, since it is present in flanking regions of both full and empty sites. Finally, such spreading into genes appears to be rare, since the transcriptional start sites of very few genes are less than one Kb from an IAP. However, the B3galtl gene is subject to transcriptional silencing via IAP-induced heterochromatin. Hence, although rare, IAP-induced local heterochromatin spreading into nearby genes may influence expression and, in turn, host fitness. PMID:21980304

  17. A Missing Guide Wire After Placement of Peripherally Inserted Central Venous Catheter

    PubMed Central

    Kashif, Muhammad; Hashmi, Hafiz; Jadhav, Preeti; Khaja, Misbahuddin

    2016-01-01

    Patient: Male, 50 Final Diagnosis: Retained guidewire removal by interventional radiology Symptoms: Swelling Medication: — Clinical Procedure: Fluoroscopic retrieval of the guidewire Specialty: Critical Care Medicine Objective: Unusual setting of medical care Background: Central venous catheterization is a common tool used in critically ill patients to monitor central venous pressure and administer fluids and medications such as vasopressors. Here we present a case of a missing guide wire after placement of peripherally inserted central catheter (PICC), which was incidentally picked up by bedside ultrasound in the intensive care unit. Case Report: A 50-year-old Hispanic male was admitted to the intensive care unit for alcohol intoxication. He was managed for septic shock and required placement of a peripherally inserted central line in his left upper extremity for antibiotics and vasopressor administration. A bedside ultrasound performed by the intensivist to evaluate upper extremity swelling revealed a foreign body in the left arm. Percutaneous procedure by Interventional radiologist was required for retrieval of the guidewire. Conclusions: Guide wire related complications are rarely reported, but are significantly associated with mortality and morbidity. The use of ultrasound guidance placement of PICC lines decreases the risk of complications, provides better optimal vein selection, and enhances success. PMID:27920421

  18. Identification of a new insertion in exon 20 of EGFR in a woman with NSCLC.

    PubMed

    Zupa, Angela; Vita, Giulia; Landriscina, Matteo; Possidente, Luciana; Aieta, Michele; Tartarone, Alfredo; Improta, Giuseppina

    2012-12-01

    Mutations of epidermal growth factor receptor 1 (EGFR) gene occur in about 15 % of all NSCLCs in Western Europe and are frequently located in exons 19 and 21, being associated with high sensitivity to EGFR tyrosine kinase inhibitors (TKIs). By contrast, exon 20 insertions account for up to 10 % of all EGFR mutations and are correlated to EGFR TKI resistance. Herein, we describe a novel mutation in EGFR exon 20 in a female non-smoker bearing a lung adenocarcinoma, characterized by the insertion of a nucleotide triplet GTT, which translates into a protein with an additional Valine between Proline 772 and Histidine 773 (p.P772_H773insV-c.2316_2317insGTT). The patient was treated with cisplatin/pemetrexed 1st-line and docetaxel 2nd-line chemotherapies, reporting a prolonged disease stabilization of 25 months. The identification and the biological and clinical characterization of novel EGFR mutations represent a prerequisite for their wide use as predictive biomarkers for personalized therapy in NSCLC.

  19. Characterization of a novel promoter insertion in the c-rel locus.

    PubMed Central

    Kabrun, N; Bumstead, N; Hayman, M J; Enrietto, P J

    1990-01-01

    Avian leukosis virus (ALV)-induced neoplasias are commonly found associated with integrations of proviral DNA in proximity to the myc gene. However, studies suggest that other genetic events are necessary for the complete neoplastic phenotype. A cell line (HP46) derived from an ALV-induced tumor has been analyzed and found to contain, in addition to an alteration in the myc gene, a promoter insertion in the c-rel locus. Both loci expressed large amounts of mRNA coding for their respective proteins. Several rel-related transcripts were expressed in the HP46 line, and four rel-related proteins of lower molecular weight than the wild-type p68c-rel product were detected. At least two of these transcripts contained U5 long terminal repeat sequences on the 5' end of the RNA. Structural data suggest that the messages may have evolved by an alternative splicing mechanism. This is the first example of a promoter insertion in the c-rel locus, a gene whose viral counterpart v-rel is responsible for the induction of lymphoid tumors. Images PMID:2167440

  20. Lithium insertion in carbonaceous materials containing silicon

    NASA Astrophysics Data System (ADS)

    Wilson, Alfred Macdonald

    Three different series of silicon-containing carbonaceous materials were synthesized for use as anodes in lithium ion cells. Disordered (or pregraphitic) carbons containing nanodispersed silicon were prepared by the chemical vapour deposition (CVD) of various chlorosilanes (SiClsb4, (CHsb3)sb2Clsb2Si, and (CHsb3)sb3ClSi) with benzene in two different apparatuses. Silicon oxycarbide glasses were synthesized by the pyrolysis of over 50 silicon-containing polymers at various temperatures, although the principal materials in the study were prepared at 1000sp°C. Finally, materials which we believe to be similar to disordered carbons containing nanodispersed silicon were prepared by the pyrolysis of various blends of pitches with polysilanes. Powder X-ray diffraction was used to learn about the structure of all the materials made. Thermal gravimetric analysis was used to determine the silicon content in the CVD materials and, when coupled to a residual gas analyzer, to study the decomposition process of the polymers. Near edge X-ray absorption spectroscopy measurements of the silicon L- and K-edges of CVD materials and the silicon K-edges of silicon oxycarbides were used to learn about local chemical environments of the silicon atoms. Lithium metal electrochemical test cells of the silicon-containing CVD materials showed larger capacities (up to 500 mAh/g) than pure carbons prepared in the same way (˜300 mAh/g). The additional capacity was observed to be centered near 0.4 V on charge, the average voltage observed for the removal of lithium from a silicon-lithium alloy. Chemical analysis showed that the stoichiometries of materials made by polymer pyrolysis were distributed over a well-defined region in the Si-O-C Gibbs phase diagram. An interesting series of materials is found near the line in the Si-O-C Gibbs triangle connecting carbon to SiOsb{1.3}. Lithium metal electrochemical test cells made using all the silicon oxycarbides synthesized showed that a stoichiometry

  1. Comparison of large-insert, small-insert and pyrosequencing libraries for metagenomic analysis.

    PubMed

    Danhorn, Thomas; Young, Curtis R; DeLong, Edward F

    2012-11-01

    The development of DNA sequencing methods for characterizing microbial communities has evolved rapidly over the past decades. To evaluate more traditional, as well as newer methodologies for DNA library preparation and sequencing, we compared fosmid, short-insert shotgun and 454 pyrosequencing libraries prepared from the same metagenomic DNA samples. GC content was elevated in all fosmid libraries, compared with shotgun and 454 libraries. Taxonomic composition of the different libraries suggested that this was caused by a relative underrepresentation of dominant taxonomic groups with low GC content, notably Prochlorales and the SAR11 cluster, in fosmid libraries. While these abundant taxa had a large impact on library representation, we also observed a positive correlation between taxon GC content and fosmid library representation in other low-GC taxa, suggesting a general trend. Analysis of gene category representation in different libraries indicated that the functional composition of a library was largely a reflection of its taxonomic composition, and no additional systematic biases against particular functional categories were detected at the level of sequencing depth in our samples. Another important but less predictable factor influencing the apparent taxonomic and functional library composition was the read length afforded by the different sequencing technologies. Our comparisons and analyses provide a detailed perspective on the influence of library type on the recovery of microbial taxa in metagenomic libraries and underscore the different uses and utilities of more traditional, as well as contemporary 'next-generation' DNA library construction and sequencing technologies for exploring the genomics of the natural microbial world.

  2. A short insertion mutation disrupts genesis of miR-16 and causes increased body weight in domesticated chicken

    PubMed Central

    Jia, Xinzheng; Lin, Huiran; Nie, Qinghua; Zhang, Xiquan; Lamont, Susan J.

    2016-01-01

    Body weight is one of the most important quantitative traits with high heritability in chicken. We previously mapped a quantitative trait locus (QTL) for body weight by genome-wide association study (GWAS) in an F2 chicken resource population. To identify the causal mutations linked to this QTL, expression profiles were determined on livers of high-weight and low-weight chicken lines by microarray. Combining the expression pattern with SNP effects by GWAS, miR-16 was identified as the most likely potential candidate with a 3.8-fold decrease in high-weight lines. Re-sequencing revealed that a 54-bp insertion mutation in the upstream region of miR-15a-16 displayed high allele frequencies in high-weight commercial broiler line. This mutation resulted in lower miR-16 expression by introducing three novel splicing sites instead of the missing 5′ terminal splicing of mature miR-16. Elevating miR-16 significantly inhibited DF-1 chicken embryo cell proliferation, consistent with a role in suppression of cellular growth. The 54-bp insertion was significantly associated with increased body weight, bone size and muscle mass. Also, the insertion mutation tended towards fixation in commercial broilers (Fst > 0.4). Our findings revealed a novel causative mutation for body weight regulation that aids our basic understanding of growth regulation in birds. PMID:27808177

  3. [The PICC line, a new approach for venous access].

    PubMed

    Levigoureux, E; Charbonnel, J-F; Latour, J-F; Rosay, H

    2013-03-01

    Peripheral Inserted Central Catheter (PICC) line is a peripherally inserted central catheter. This implantable medical device is placed into a peripheral vein of the arm in order to obtain an intravenous central access. This device can find its use in various applications like intravenous delivery of parenteral nutrition, anticancer agents and antibiotics, as well as for blood sampling. PICC line is not widely used in medical practice because it remains largely unknown. The aim of this review is thus to introduce PICC line to the medical and scientific community. First, we will approach its insertion and maintenance of the dressing. We will then detail the benefits and drawbacks associated with its use, and finally discuss its position with regards to the other central venous access available.

  4. Patterns of Transposable Element Expression and Insertion in Cancer

    PubMed Central

    Clayton, Evan A.; Wang, Lu; Rishishwar, Lavanya; Wang, Jianrong; McDonald, John F.; Jordan, I. King

    2016-01-01

    Human transposable element (TE) activity in somatic tissues causes mutations that can contribute to tumorigenesis. Indeed, TE insertion mutations have been implicated in the etiology of a number of different cancer types. Nevertheless, the full extent of somatic TE activity, along with its relationship to tumorigenesis, have yet to be fully explored. Recent developments in bioinformatics software make it possible to analyze TE expression levels and TE insertional activity directly from transcriptome (RNA-seq) and whole genome (DNA-seq) next-generation sequence data. We applied these new sequence analysis techniques to matched normal and primary tumor patient samples from the Cancer Genome Atlas (TCGA) in order to analyze the patterns of TE expression and insertion for three cancer types: breast invasive carcinoma, head and neck squamous cell carcinoma, and lung adenocarcinoma. Our analysis focused on the three most abundant families of active human TEs: Alu, SVA, and L1. We found evidence for high levels of somatic TE activity for these three families in normal and cancer samples across diverse tissue types. Abundant transcripts for all three TE families were detected in both normal and cancer tissues along with an average of ~80 unique TE insertions per individual patient/tissue. We observed an increase in L1 transcript expression and L1 insertional activity in primary tumor samples for all three cancer types. Tumor-specific TE insertions are enriched for private mutations, consistent with a potentially causal role in tumorigenesis. We used genome feature analysis to investigate two specific cases of putative cancer-causing TE mutations in further detail. An Alu insertion in an upstream enhancer of the CBL tumor suppressor gene is associated with down-regulation of the gene in a single breast cancer patient, and an L1 insertion in the first exon of the BAALC gene also disrupts its expression in head and neck squamous cell carcinoma. Our results are consistent with

  5. LMA Supreme insertion by novices in manikins and patients.

    PubMed

    Howes, B W; Wharton, N M; Gibbison, B; Cook, T M

    2010-04-01

    The LMA Supreme has been suggested for use in emergency situations by medical personnel with no experience in endotracheal intubation. We evaluated the LMA Supreme when inserted by non-anaesthetists, firstly in a manikin and then in patients. Fifty airway novices inserted a LMA Supreme in a manikin without any complications so we proceeded to the patient phase. Fifty airway novices inserted the LMA Supreme in anaesthetised patients undergoing elective surgery. First time insertion success rate was 86% and overall insertion success rate was 100%. Mechanical ventilation was successful in all cases. Median (IQR [range]) time to establish an airway was 34 s (26-40 [18-145] s). Median (IQR [range]) pharyngeal seal pressure was 23 cmH(2)O (19-28 [13-40] cmH(2)O). There were no important complications. Results are consistent with previous studies of use of the LMA Supreme by airway experts. We conclude that the LMA supreme is suitable for use by airway novices. Further research is needed before it may be recommended for cardiopulmonary resuscitation and emergency airway use.

  6. Quantitative assessment of insertion sequence impact on bacterial genome architecture

    PubMed Central

    Bishop, Brian; Wright, Meredith S.

    2016-01-01

    Insertion sequence (IS) elements are important mediators of genome plasticity and can lead to phenotypic changes with evolutionary significance. In multidrug-resistant Acinetobacter baumannii and Klebsiella pneumoniae, IS elements have contributed significantly to the mobilization of genes that encode resistance to antimicrobial drugs. A systematic analysis of IS elements is needed for a more comprehensive understanding of their evolutionary impact. We developed a computational approach (ISseeker) to annotate IS elements in draft genome assemblies and applied the method to analysis of IS elements in all publicly available A. baumannii(>1000) and K. pneumoniae(>800) genome sequences, in a phylogenetic context. Most IS elements in A. baumanniigenomes are species-specific ISAba elements, whereas K. pneumoniaegenomes contain significant numbers of both ISKpn elements and elements that are found throughout the Enterobacteriaceae. A. baumanniigenomes have a higher density of IS elements than K. pneumoniae, averaging ~33 vs ~27 copies per genome. In K. pneumoniae, several insertion sites are shared by most genomes in the ST258 clade, whereas in A. baumannii, different IS elements are abundant in different phylogenetic groups, even among closely related Global Clone 2 strains. IS elements differ in the distribution of insertion locations relative to genes, with some more likely to disrupt genes and others predominantly in intergenic regions. Several genes and intergenic regions had multiple independent insertion events, suggesting that those events may confer a selective advantage. Genome- and taxon-wide characterization of insertion locations revealed that IS elements have been active contributors to genome diversity in both species.

  7. Seamless lesion insertion in digital mammography: methodology and reader study

    NASA Astrophysics Data System (ADS)

    Pezeshk, Aria; Petrick, Nicholas; Sahiner, Berkman

    2016-03-01

    Collection of large repositories of clinical images containing verified cancer locations is costly and time consuming due to difficulties associated with both the accumulation of data and establishment of the ground truth. This problem poses a significant challenge to the development of machine learning algorithms that require large amounts of data to properly train and avoid overfitting. In this paper we expand the methods in our previous publications by making several modifications that significantly increase the speed of our insertion algorithms, thereby allowing them to be used for inserting lesions that are much larger in size. These algorithms have been incorporated into an image composition tool that we have made publicly available. This tool allows users to modify or supplement existing datasets by seamlessly inserting a real breast mass or micro-calcification cluster extracted from a source digital mammogram into a different location on another mammogram. We demonstrate examples of the performance of this tool on clinical cases taken from the University of South Florida Digital Database for Screening Mammography (DDSM). Finally, we report the results of a reader study evaluating the realism of inserted lesions compared to clinical lesions. Analysis of the radiologist scores in the study using receiver operating characteristic (ROC) methodology indicates that inserted lesions cannot be reliably distinguished from clinical lesions.

  8. Some physical and chemical indices of clique-inserted lattices

    NASA Astrophysics Data System (ADS)

    Zhang, Zuhe

    2013-10-01

    The operation of replacing every vertex of an r-regular lattice H by a complete graph of order r is called clique-insertion, and the resulting lattice is called the clique-inserted lattice of H. For any given r-regular lattice, applying this operation iteratively, an infinite family of r-regular lattices is generated. Some interesting lattices including the 3-12-12 lattice can be constructed this way. In this paper, we recall the relationship between the spectra of an r-regular lattice and that of its clique-inserted lattice, and investigate the graph energy and resistance distance statistics. As an application, the asymptotic energy per vertex and average resistance distance of the 3-12-12 and 3-6-24 lattices are computed. We also give formulae expressing the numbers of spanning trees and dimer coverings of the kth iterated clique-inserted lattices in terms of those of the original one. Moreover, we show that new families of expander graphs can be constructed from the known ones by clique-insertion.

  9. Customizable engineered blood vessels using 3D printed inserts.

    PubMed

    Pinnock, Cameron B; Meier, Elizabeth M; Joshi, Neeraj N; Wu, Bin; Lam, Mai T

    2016-04-15

    Current techniques for tissue engineering blood vessels are not customizable for vascular size variation and vessel wall thickness. These critical parameters vary widely between the different arteries in the human body, and the ability to engineer vessels of varying sizes could increase capabilities for disease modeling and treatment options. We present an innovative method for producing customizable, tissue engineered, self-organizing vascular constructs by replicating a major structural component of blood vessels - the smooth muscle layer, or tunica media. We utilize a unique system combining 3D printed plate inserts to control construct size and shape, and cell sheets supported by a temporary fibrin hydrogel to encourage cellular self-organization into a tubular form resembling a natural artery. To form the vascular construct, 3D printed inserts are adhered to tissue culture plates, fibrin hydrogel is deposited around the inserts, and human aortic smooth muscle cells are then seeded atop the fibrin hydrogel. The gel, aided by the innate contractile properties of the smooth muscle cells, aggregates towards the center post insert, creating a tissue ring of smooth muscle cells. These rings are then stacked into the final tubular construct. Our methodology is robust, easily repeatable and allows for customization of cellular composition, vessel wall thickness, and length of the vessel construct merely by varying the size of the 3D printed inserts. This platform has potential for facilitating more accurate modeling of vascular pathology, serving as a drug discovery tool, or for vessel repair in disease treatment.

  10. EGFR exon 20 insertion mutation in Japanese lung cancer.

    PubMed

    Sasaki, Hidefumi; Endo, Katsuhiko; Takada, Minoru; Kawahara, Masaaki; Kitahara, Naoto; Tanaka, Hisaichi; Okumura, Meinoshin; Matsumura, Akihide; Iuchi, Keiji; Kawaguchi, Tomoya; Kawano, Osamu; Yukiue, Haruhiro; Yokoyama, Tomoki; Yano, Motoki; Fujii, Yoshitaka

    2007-12-01

    Mutations of the epidermal growth factor receptor (EGFR) gene have been reported in non-small cell lung cancer (NSCLC), especially in female, never smoker patients with adenocarcinoma. Some common somatic mutations in EGFR, including deletion mutations in exon 19 and leucine to arginine substitution at amino acid position 858 (L858R) in exon 21, have been examined for their ability to predict sensitivity to gefitinib or erlotinib. On the other hand, previous report has shown that the insertion mutation at exon 20 is related to gefitinib resistance. We investigated the exon 20 EGFR mutation statuses in 322 surgically treated non-small cell lung cancer cases. Two hundred and five adenocarcinoma cases were included. The presence or absence of EGFR mutations of kinase domains was analyzed by direct sequences. EGFR insertion mutations at exon 20 were found from 7 of 322 (2.17%) lung cancer patients. We also detected the 18 deletion type mutations in exon 19, and 25 L858R type mutations in exon 21. There was a tendency towards higher exon 20 insertion ratio in never smoker (never smoker 4.4% versus smoker 1.3%, p=0.0996) and female (female 4.5% versus male 1.3%, p=0.0917). Two exon 20 insertion cases were treated with gefitinib and failed to response. EGFR insertion mutation in exon 20 could not be ignored from Japanese lung cancers.

  11. Recombination of an intrachromosomal paracentric insertion of chromosome 3

    SciTech Connect

    Best, R.G.; Burnett, W.J.; Brock, J.K.

    1994-09-01

    Cytogenetic studies were initiated on a newborn female due to multiple congenital anomalies including microcephaly, clinodactyly, abnormal positioning of hands, left facial palsy, heart defect, sacral dimple, and facial dysmorphic features. Facial features were described as low set rotated ears, nystagmus, and a small, flattened nose. A structural rearrangement of the long arm of chromosome 3 was observed with a complex banding pattern. Study of parental chromosomes revealed a normal male pattern for the father, and an intrachromosomal insertion on the long arm of chromosome 3 for the mother described as 46,XX,dir ins(3)(q21q23q26.2). Further characterization of the proband`s structurally abnormal chromosome 3 revealed a karyotype best described as: 46,XX,rec(3),dupq23{r_arrow}q26.2::q21{r_arrow}q23,dir ins(3)(q21q23q26.2), which is a partial duplication of both the inserted segment as well as the intervening segment between the inserted segment and the insertion site. This would appear to be the result of a three-strand double cross-over within the insertion loop. Molecular cytogenetic studies are presently underway to further elucidate chromosome structure of the proband and her mother.

  12. Direct insertion of metal ions into tetrasulfonated phthalocyanine

    SciTech Connect

    Linkous, C.A.; Shea, C.E.; Jaber, M.R.A. )

    1989-07-01

    A series of metal salts were reacted with aqueous solutions of the tetrasulfonated free base phthalocyanine (H{sub 2}TsPc) in an attempt to prepare the corresponding metal TsPc derivative by direct insertion instead of the usual template reaction in a urea melt. Spectrophotometric analysis showed that Ni{sup 2+}, Cu{sup 2+}, Zn{sup 2+}, Co{sup 2+}, Fe{sup 2+}, Mn{sup 2+}, Pd{sup 2+}, Cd{sup 2+}, and Sn{sup 2+} could be successfully inserted; higher oxidation states as well as Pb{sup 2+} caused precipitation. Reducing species such as Cr{sup 2+} and V{sup 2+} reduced H{sub 2}TsPc but did not insert; Sn{sup 2+} inserted with formation of an air-sensitive intermediate species. Mg{sup 2+}, VO{sup 2+}, Pt{sup 2+}, and Hg{sup 2+} were unreactive. It is recommended that insertion may be the preferable method for synthesis of the MTsPc's listed above when air sensitivity, reaction time, and microscale are important issues in an overall synthetic scheme. 24 refs., 4 figs.

  13. Insertion of liquid crystal molecules into hydrocarbon monolayers

    SciTech Connect

    Popov, Piotr Mann, Elizabeth K.; Lacks, Daniel J.; Jákli, Antal

    2014-08-07

    Atomistic molecular dynamics simulations were carried out to investigate the molecular mechanisms of vertical surface alignment of liquid crystals. We study the insertion of nCB (4-Cyano-4{sup ′}-n-biphenyl) molecules with n = 0,…,6 into a bent-core liquid crystal monolayer that was recently found to provide good vertical alignment for liquid crystals. The results suggest a complex-free energy landscape for the liquid crystal within the layer. The preferred insertion direction of the nCB molecules (core or tail first) varies with n, which can be explained by entropic considerations. The role of the dipole moments was found to be negligible. As vertical alignment is the leading form of present day liquid crystal displays (LCD), these results will help guide improvement of the LCD technology, as well as lend insight into the more general problem of insertion of biological and other molecules into lipid and surfactant layers.

  14. Genetic aspects of targeted insertion mutagenesis in yeasts.

    PubMed

    Klinner, U; Schäfer, B

    2004-05-01

    Targeted insertion mutagenesis is a main molecular tool of yeast science initially applied in Saccharomyces cerevisiae. The method was extended to fission yeast Schizosaccharomyces pombe and to "non-conventional" yeast species, which show specific properties of special interest to both basic and applied research. Consequently, the behaviour of such non-Saccharomyces yeasts is reviewed against the background of the knowledge of targeted insertion mutagenesis in S. cerevisiae. Data of homologous integration efficiencies obtained with circular, ends-in or ends-out vectors in several yeasts are compared. We follow details of targeted insertion mutagenesis in order to recognize possible rate-limiting steps. The route of the vector to the target and possible mechanisms of its integration into chromosomal genes are considered. Specific features of some yeast species are discussed. In addition, similar approaches based on homologous recombination that have been established for the mitochondrial genome of S. cerevisiae are described.

  15. Synchronized monochromator and insertion device energy scans at SLS

    SciTech Connect

    Krempasky, J.; Flechsig, U.; Korhonen, T.; Zimoch, D.; Quitmann, Ch.; Nolting, F.

    2010-06-23

    Synchronous monochromator and insertion device energy scans were implemented at the Surfaces/Interfaces:Microscopy (SIM) beamline in order to provide the users fast X-ray magnetic dichroism studies (XMCD). A simple software control scheme is proposed based on a fast monochromator run-time energy readback which quickly updates the insertion device requested energy during an on-the-fly X-ray absorption scan (XAS). In this scheme the Plain Grating Monochromator (PGM) motion control, being much slower compared with the insertion device (APPLE-II type undulator), acts as a 'master' controlling the undulator 'slave' energy position. This master-slave software implementation exploits EPICS distributed device control over computer network and allows for a quasi-synchronous motion control combined with data acquisition needed for the XAS or XMCD experiment.

  16. Synchronized monochromator and insertion device energy scans at SLS

    NASA Astrophysics Data System (ADS)

    Krempaský, J.; Flechsig, U.; Korhonen, T.; Zimoch, D.; Quitmann, Ch.; Nolting, F.

    2010-06-01

    Synchronous monochromator and insertion device energy scans were implemented at the Surfaces/Interfaces:Microscopy (SIM) beamline in order to provide the users fast X-ray magnetic dichroism studies (XMCD). A simple software control scheme is proposed based on a fast monochromator run-time energy readback which quickly updates the insertion device requested energy during an on-the-fly X-ray absorption scan (XAS). In this scheme the Plain Grating Monochromator (PGM) motion control, being much slower compared with the insertion device (APPLE-II type undulator), acts as a "master" controlling the undulator "slave" energy position. This master-slave software implementation exploits EPICS distributed device control over computer network and allows for a quasi-synchronous motion control combined with data acquisition needed for the XAS or XMCD experiment.

  17. Polyglutamine expansion in huntingtin increases its insertion into lipid bilayers.

    PubMed

    Kegel, Kimberly B; Schewkunow, Vitali; Sapp, Ellen; Masso, Nicholas; Wanker, Erich E; DiFiglia, Marian; Goldmann, Wolfgang H

    2009-09-25

    An expanded polyglutamine (Q) tract (>37Q) in huntingtin (htt) causes Huntington disease. Htt associates with membranes and polyglutamine expansion in htt may alter membrane function in Huntington disease through a mechanism that is not known. Here we used differential scanning calorimetry to examine the effects of polyQ expansion in htt on its insertion into lipid bilayers. We prepared synthetic lipid vesicles composed of phosphatidylcholine and phosphatidylethanolamine and tested interactions of htt amino acids 1-89 with 20Q, 32Q or 53Q with the vesicles. GST-htt1-89 with 53Q inserted into synthetic lipid vesicles significantly more than GST-htt1-89 with 20Q or 32Q. We speculate that by inserting more into cell membranes, mutant huntingtin could increase disorder within the lipid bilayer and thereby disturb cellular membrane function.

  18. Tool Life Detecting System Using Damage Sensor-Integrated Insert

    NASA Astrophysics Data System (ADS)

    Sekiya, Katsuhiko; Yamane, Yasuo; Torimoto, Ayumu

    In-process tool life detecting system has been developed. In order to detect tool life, we monitored breakage of an electrical conductive thin film band of titanium nitride (TiN) on flank faces of a ceramic insert that was an insulation material. The conductive band was parallel to the cutting edge and was broken by fracture of the cutting edge or excessive tool wear. A coil and a capacitor were connected in series to the band on an insert to make a series resonance circuit in a tool holder for turning, or in an arbor for face milling. A detecting coil with high frequency oscillator was coupled to the coil in the holder/arbor electromagnetically. An electromagnetic induction between two coils was used to take the signal from the insert. The system developed in this study showed good stability against electric noises radiated from electrical devices of machine tools.

  19. Molecular and genetic characterization of an Alcaligenes eutrophus insertion element.

    PubMed Central

    Kung, S S; Chen, J; Chow, W Y

    1992-01-01

    An insertion element, ISAE1, was discovered during the molecular analysis of mutants defective in the autotrophic growth (Aut-) of Alcaligenes eutrophus H1-4, a mitomycin C-generated derivative of strain H1. ISAE1 is 1,313 bp long, has 12-bp nearly perfect inverted terminal repeats, and contains an open reading frame that has a coding capacity of 408 amino acids. Direct repeats of 8 bp were generated by insertion of ISAE1 into chromosomes or plasmids. Most insertion were found in the AT-rich target sites. The distribution of ISAE1 is limited to A. eutrophus H1 (ATCC 17698) and H16 (ATCC 17699). Variants with newly transposed copies of ISAE1 could be isolated at an elevated frequency by changing the growth conditions. Images PMID:1334068

  20. Gene Insertion Into Genomic Safe Harbors for Human Gene Therapy

    PubMed Central

    Papapetrou, Eirini P; Schambach, Axel

    2016-01-01

    Genomic safe harbors (GSHs) are sites in the genome able to accommodate the integration of new genetic material in a manner that ensures that the newly inserted genetic elements: (i) function predictably and (ii) do not cause alterations of the host genome posing a risk to the host cell or organism. GSHs are thus ideal sites for transgene insertion whose use can empower functional genetics studies in basic research and therapeutic applications in human gene therapy. Currently, no fully validated GSHs exist in the human genome. Here, we review our formerly proposed GSH criteria and discuss additional considerations on extending these criteria, on strategies for the identification and validation of GSHs, as well as future prospects on GSH targeting for therapeutic applications. In view of recent advances in genome biology, gene targeting technologies, and regenerative medicine, gene insertion into GSHs can potentially catalyze nearly all applications in human gene therapy. PMID:26867951

  1. Optimized production and concentration of lentiviral vectors containing large inserts.

    PubMed

    al Yacoub, Nadya; Romanowska, Malgorzata; Haritonova, Natalie; Foerster, John

    2007-07-01

    Generation of high titer lentiviral stocks and efficient virus concentration are central to maximize the utility of lentiviral technology. Here we evaluate published protocols for lentivirus production on a range of transfer vectors differing in size (7.5-13.2 kb). We present a modified virus production protocol robustly yielding useful titers (up to 10(7)/ml) for a range of different transfer vectors containing packaging inserts up to 7.5 kb. Moreover, we find that virus recovery after concentration by ultracentrifugation depends on the size of the packaged inserts, heavily decreasing for large packaged inserts. We describe a fast (4 h) centrifugation protocol at reduced speed allowing high virus recovery even for large and fragile lentivirus vectors. The protocols outlined in the current report should be useful for many labs interested in producing and concentrating high titer lentiviral stocks.

  2. Epigenetic Regulation of Intronic Transgenes in Arabidopsis

    PubMed Central

    Osabe, Kenji; Harukawa, Yoshiko; Miura, Saori; Saze, Hidetoshi

    2017-01-01

    Defense mechanisms of plant genomes can epigenetically inactivate repetitive sequences and exogenous transgenes. Loss of mutant phenotypes in intronic T-DNA insertion lines by interaction with another T-DNA locus, termed T-DNA suppression, has been observed in Arabidopsis thaliana, although the molecular basis of establishment and maintenance of T-DNA suppression is poorly understood. Here we show that maintenance of T-DNA suppression requires heterochromatinisation of T-DNA sequences and the nuclear proteins, INCREASED IN BONSAI METHYLATION 2 (IBM2) and ENHANCED DOWNY MILDEW 2 (EDM2), which prevent ectopic 3′ end processing of mRNA in atypically long introns containing T-DNA sequences. Initiation of T-DNA suppression is mediated by the canonical RdDM pathway after hybridisation of two T-DNA strains, accompanied by DNA hypermethylation of T-DNA sequences in the F1 generation. Our results reveal the presence of a genome surveillance mechanism through genome hybridisation that masks repetitive DNAs intruding into transcription units. PMID:28338020

  3. A collection of enhancer trap insertional mutants for functional genomics in tomato.

    PubMed

    Pérez-Martín, Fernando; Yuste-Lisbona, Fernando J; Pineda, Benito; Angarita-Díaz, María Pilar; García-Sogo, Begoña; Antón, Teresa; Sánchez, Sibilla; Giménez, Estela; Atarés, Alejandro; Fernández-Lozano, Antonia; Ortíz-Atienza, Ana; García-Alcázar, Manuel; Castañeda, Laura; Fonseca, Rocío; Capel, Carmen; Goergen, Geraldine; Sánchez, Jorge; Quispe, Jorge L; Capel, Juan; Angosto, Trinidad; Moreno, Vicente; Lozano, Rafael

    2017-03-19

    With the completion of genome sequencing projects, the next challenge is to close the gap between gene annotation and gene functional assignment. Genomic tools to identify gene functions are based on the analysis of phenotypic variations between a wild-type and its mutant; hence, mutant collections are a valuable resource. In this sense, T-DNA collections allow for an easy and straightforward identification of the tagged gene, serving as the basis of both forward and reverse genetic strategies. This study reports on the phenotypic and molecular characterization of an enhancer trap T-DNA collection in tomato (Solanum lycopersicum L.), which has been produced by Agrobacterium-mediated transformation using a binary vector bearing a minimal promoter fused to the uidA reporter gene. Two genes have been isolated from different T-DNA mutants, one of these genes codes for a UTP-glucose-1-phosphate uridylyltransferase involved in programmed cell death and leaf development, which means a novel gene function reported in tomato. Together, our results support that enhancer trapping is a powerful tool to identify novel genes and regulatory elements in tomato and that this T-DNA mutant collection represents a highly valuable resource for functional analyses in this fleshy-fruited model species. This article is protected by copyright. All rights reserved.

  4. Rhizobium phaseoli symbiotic mutants with transposon Tn5 insertions.

    PubMed Central

    Noel, K D; Sanchez, A; Fernandez, L; Leemans, J; Cevallos, M A

    1984-01-01

    Rhizobium phaseoli CFN42 DNA was mutated by random insertion of Tn5 from suicide plasmid pJB4JI to obtain independently arising strains that were defective in symbiosis with Phaseolus vulgaris but grew normally outside the plant. When these mutants were incubated with the plant, one did not initiate visible nodule tissue (Nod-), seven led to slow nodule development (Ndv), and two led to superficially normal early nodule development but lacked symbiotic nitrogenase activity (Sna-). The Nod- mutant lacked the large transmissible indigenous plasmid pCFN42d that has homology to Klebsiella pneumoniae nitrogenase (nif) genes. The other mutants had normal plasmid content. In the two Sna- mutants and one Ndv mutant, Tn5 had inserted into plasmid pCFN42d outside the region of nif homology. The insertions of the other Ndv mutants were apparently in the chromosome. They were not in plasmids detected on agarose gels, and, in contrast to insertions on indigenous plasmids, they were transmitted in crosses to wild-type strain CFN42 at the same frequency as auxotrophic markers and with the same enhancement of transmission by conjugation plasmid R68.45. In these Ndv mutants the Tn5 insertions were the same as or very closely linked to mutations causing the Ndv phenotype. However, in two mutants with Tn5 insertions on plasmid pCFN42d, an additional mutation on the same plasmid, rather than Tn5, was responsible for the Sna- or Ndv phenotype. When plasmid pJB4JI was transferred to two other R. phaseoli strains, analysis of symbiotic mutants was complicated by Tn5-containing deleted forms of pJB4JI that were stably maintained. Images PMID:6325385

  5. The heterozygous abp1/ABP1 insertional mutant has defects in functions requiring polar auxin transport and in regulation of early auxin-regulated genes.

    PubMed

    Effendi, Yunus; Rietz, Steffen; Fischer, Urs; Scherer, Günther F E

    2011-01-01

    AUXIN-BINDING PROTEIN 1 (ABP1) is not easily accessible for molecular studies because the homozygous T-DNA insertion mutant is embryo-lethal. We found that the heterozygous abp1/ABP1 insertion mutant has defects in auxin physiology-related responses: higher root slanting angles, longer hypocotyls, agravitropic roots and hypocotyls, aphototropic hypocotyls, and decreased apical dominance. Heterozygous plants flowered earlier than wild-type plants under short-day conditions. The length of the main root, the lateral root density and the hypocotyl length were little altered in the mutant in response to auxin. Compared to wild-type plants, transcription of early auxin-regulated genes (IAA2, IAA11, IAA13, IAA14, IAA19, IAA20, SAUR9, SAUR15, SAUR23, GH3.5 and ABP1) was less strongly up-regulated in the mutant by 0.1, 1 and 10 μm IAA. Surprisingly, ABP1 was itself an early auxin-up-regulated gene. IAA uptake into the mutant seedlings during auxin treatments was indistinguishable from wild-type. Basipetal auxin transport in young roots was slower in the mutant, indicating a PIN2/EIR1 defect, while acropetal transport was indistinguishable from wild-type. In the eir1 background, three of the early auxin-regulated genes tested (IAA2, IAA13 and ABP1) were more strongly induced by 1 μm IAA in comparison to wild-type, but eight of them were less up-regulated in comparison to wild-type. Similar but not identical disturbances in regulation of early auxin-regulated genes indicate tight functional linkage of ABP1 and auxin transport regulation. We hypothesize that ABP1 is involved in the regulation of polar auxin transport, and thus affects local auxin concentration and early auxin gene regulation. In turn, ABP1 itself is under the transcriptional control of auxin.

  6. Rectal fist insertion. An unusual form of sexual behavior.

    PubMed

    Shook, L L; Whittle, R; Rose, E F

    1985-12-01

    Rectal fist insertion (fist fucking) is an uncommon and potentially dangerous sexual practice. This is usually a homosexual activity, but can also be a heterosexual or an autoerotic practice. One known death has been reported associated with rectal fist insertion, in which the complications of anal and colonic tears and bleeding had occurred (see Editor's note). The possibility of drug overdose is also probable, as drugs and alcohol are commonly introduced into the rectum to promote sphincter relaxation and to ease the discomfort of anal dilatation.

  7. Rays inserting method (RIM) to design dielectric optical devices

    NASA Astrophysics Data System (ADS)

    Taskhiri, Mohammad Mahdi; Khalaj Amirhosseini, Mohammad

    2017-01-01

    In this article, a novel approach, called Rays Inserted Method (RIM), is introduced to design dielectric optical devices. In this approach, some rays are inserted between two ends of desired device and then the refractive index of the points along the route of rays are obtained. The validity of the introduced approach is verified by designing three types of optical devices, i.e. power splitter, bend, and flat lens. The results are confirmed with numerical simulations by the means of FDTD scheme at the frequency of 100 GHz.

  8. Synthesis and characterization of conducting polymer inserted carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Choi, A. Jeong; Nam, Young Woo; Park, Yung Woo

    2008-03-01

    The carbon nanotubes filled with the photo-conducting polymer poly(N-vinyl carbazole) and the conducting polymer polypyrrole were prepared by polymerizing the monomers inside the nanotubes using the supercritical carbon dioxide. The endohedral nanotubes were characterized by HRTEM and ^1H NMR, which confirmed that the inserted material was indeed the conducting polymer [1]. I-V characteristics of the polymer inserted carbon nanotubes are presented. [1] Johannes Steinmetz, Soyoung Kwon, Hyun-Jung Lee, Edy Abou-Hamad, Robert Almairac, Christophe Goze-Bac, Hwayong Kim, Yung-Woo Park,, Chem. Phys. Lett., 431, 139 (2006)

  9. Method of modifying a volume mesh using sheet insertion

    SciTech Connect

    Borden, Michael J.; Shepherd, Jason F.

    2006-08-29

    A method and machine-readable medium provide a technique to modify a hexahedral finite element volume mesh using dual generation and sheet insertion. After generating a dual of a volume stack (mesh), a predetermined algorithm may be followed to modify (refine) the volume mesh of hexahedral elements. The predetermined algorithm may include the steps of locating a sheet of hexahedral mesh elements, determining a plurality of hexahedral elements within the sheet to refine, shrinking the plurality of elements, and inserting a new sheet of hexahedral elements adjacently to modify the volume mesh. Additionally, another predetermined algorithm using mesh cutting may be followed to modify a volume mesh.

  10. Advanced photon source experience with vacuum chambers for insertion devices

    SciTech Connect

    Hartog, P.D.; Grimmer, J.; Xu, S.; Trakhtenberg, E.; Wiemerslage, G.

    1997-08-01

    During the last five years, a new approach to the design and fabrication of extruded aluminum vacuum chambers for insertion devices was developed at the Advanced Photon Source (APS). With this approach, three different versions of the vacuum chamber, with vertical apertures of 12 mm, 8 mm, and 5 mm, were manufactured and tested. Twenty chambers were installed into the APS vacuum system. All have operated with beam, and 16 have been coupled with insertion devices. Two different vacuum chambers with vertical apertures of 16 mm and 11 mm were developed for the BESSY-II storage ring and 3 of 16 mm chambers were manufactured.

  11. Dosimetry of portable image intensifiers in pacemaker insertions.

    PubMed

    Hayt, D B; Malsky, S J; Blatt, C J; Greenberg, E I; Pochaczevsky, R; Simon, D F; Perez, L A

    1975-01-01

    With the increased utilization of portable image intensifiers for pacemaker insertions, an investigation of the radiation hazards to operative personnel was undertaken. The personnel were tested first utilizing a Rando phantom and later during actual pacemaker insertions. Studies of medical personnel show the critical organs to be the thyroid (approximately 15mR per procedure) and the lens of the eye (approximately 13mR per procedure). While the MPD/week was not exceeded, the radiation dosage was substantial and could increase under operating conditions other than those measured. Recommendations to insure safer operation of this equipment are presented.

  12. 20 TeV collider lattices with low-. beta. insertions

    SciTech Connect

    Garren, A.A.

    1983-08-01

    A lattice containing insertions designed for collisions of 20 TeV proton beams at crossing points having beta values of two meters or less is presented. The machine would use high-field double bore superconducting magnets, with opposite focusing action on the two beams passing through each quadrupole. Hence the focusing pattern in the insertions is antisymmetric about the crossings. The beams, separated by 16 cms in the arcs are made colinear by dipoles common to both beams and then focused to the low-..beta.. collision points by quadrupole triplets. A similar machine design for pp collisions is also included.

  13. Insertion of lithium into electrochromic devices after completion

    DOEpatents

    Berland, Brian Spencer; Lanning, Bruce Roy; Frey, Jonathan Mack; Barrett, Kathryn Suzanne; DuPont, Paul Damon; Schaller, Ronald William

    2015-12-22

    The present disclosure describes methods of inserting lithium into an electrochromic device after completion. In the disclosed methods, an ideal amount of lithium can be added post-fabrication to maximize or tailor the free lithium ion density of a layer or the coloration range of a device. Embodiments are directed towards a method to insert lithium into the main device layers of an electrochromic device as a post-processing step after the device has been manufactured. In an embodiment, the methods described are designed to maximize the coloration range while compensating for blind charge loss.

  14. Neutron reflectometry of supported hybrid bilayers with inserted peptide

    PubMed Central

    Smith, Matthew B.; McGillivray, Duncan J.; Genzer, Jan; Lösche, Mathias; Kilpatrick, Peter K.

    2011-01-01

    The insertion of a synthetic amphiphilic, α-helical peptide into a supported hybrid bilayer membrane (HBM) was studied by neutron reflectometry to elucidate the resulting nanostructure. The HBM consisted of a self-assembled monolayer of perdeuterated octadecanethiol on gold and an overlying leaflet of acyl-deuterated phosphatidylcholine (d-DMPC). Using contrast variation, several reflectivity spectra were recorded for each step of film fabrication, and simultaneously modeled. This analysis indicated that peptide insertion into the DMPC lipid leaflet is the likeliest mode of incorporation. PMID:21274414

  15. Technology Insertion and Management: Options for the Canadian Forces

    DTIC Science & Technology

    2010-01-01

    enhancement” of system performance and functionality in existing or deployed defence systems by the utilization of a new or improved technology. Importantly...du fardeau logistique si l’insertion technologique est mise en œuvre de façon efficace. La réduction des coûts de propriété d’un système de défense...powerful Honeywell engines that improve fuel efficiency and enhance lift performance by 3900lbs. [7] Horizontal technology insertion (HTI) refers to

  16. Definition of Metrics to Evaluate Cochlear Array Insertion Forces Performed with Forceps, Insertion Tool, or Motorized Tool in Temporal Bone Specimens

    PubMed Central

    Ferrary, Evelyne; Sterkers, Olivier

    2014-01-01

    Introduction. In order to achieve a minimal trauma to the inner ear structures during array insertion, it would be suitable to control insertion forces. The aim of this work was to compare the insertion forces of an array insertion into anatomical specimens with three different insertion techniques: with forceps, with a commercial tool, and with a motorized tool. Materials and Methods. Temporal bones have been mounted on a 6-axis force sensor to record insertion forces. Each temporal bone has been inserted, with a lateral wall electrode array, in random order, with each of the 3 techniques. Results. Forceps manual and commercial tool insertions generated multiple jerks during whole length insertion related to fits and starts. On the contrary, insertion force with the motorized tool only rose at the end of the insertion. Overall force momentum was 1.16 ± 0.505 N (mean ± SD, n = 10), 1.337 ± 0.408 N (n = 8), and 1.573 ± 0.764 N (n = 8) for manual insertion with forceps and commercial and motorized tools, respectively. Conclusion. Considering force momentum, no difference between the three techniques was observed. Nevertheless, a more predictable force profile could be observed with the motorized tool with a smoother rise of insertion forces. PMID:25126565

  17. Molecular cytogenetic analysis of feline leukemia virus insertions in cat lymphoid tumor cells.

    PubMed

    Fujino, Yasuhito; Satoh, Hitoshi; Ohno, Koichi; Tsujimoto, Hajime

    2010-02-01

    This study was conducted to map the acquired proviral insertions in the chromosomal genome of feline lymphoid tumors induced by feline leukemia virus (FeLV). Chromosome specimens of the lymphoid tumor-derived cell lines and normal cat lymphocytes were subjected to fluorescence in situ hybridization and tyramide signal amplification, using an exogenous FeLV-A genome as a probe. Specific hybridization signals were detected only on the metaphase chromosomes of the tumor cells. Poisson's distribution-based statistics indicated that 6 chromosomal loci in each cell line showed FeLV integration. In the examination of metaphase chromosomes of FL-74, FT-1 and KO-1 cells, significant signals were detected on B2p15-p14, B2q11, D1p14, E1p14-p13, E1q12 and F2q16; A2p23-p22, B2p15-p14, B4p15-p14, D4q23-q24, E1p14-p13 and E2p13-p12; and A2p22, A3q22, B1p13, B1q13, D1p13 and D3p15-p14, respectively. Consistently, Southern blot hybridization using an FeLV LTR-U3 probe specific for exogenous FeLV revealed the presence of at least 6 copies of exogenous FeLV proviruses at different integration sites in each cell line. These results indicate that there may be common FeLV integration sites at least in A2p22 and B2p15-p14. The cytogenetic analysis used in this study can promptly screen FeLV insertions and provide tags for identifying the novel common integration site.

  18. The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes.

    PubMed Central

    Spradling, A C; Stern, D; Beaton, A; Rhem, E J; Laverty, T; Mozden, N; Misra, S; Rubin, G M

    1999-01-01

    A fundamental goal of genetics and functional genomics is to identify and mutate every gene in model organisms such as Drosophila melanogaster. The Berkeley Drosophila Genome Project (BDGP) gene disruption project generates single P-element insertion strains that each mutate unique genomic open reading frames. Such strains strongly facilitate further genetic and molecular studies of the disrupted loci, but it has remained unclear if P elements can be used to mutate all Drosophila genes. We now report that the primary collection has grown to contain 1045 strains that disrupt more than 25% of the estimated 3600 Drosophila genes that are essential for adult viability. Of these P insertions, 67% have been verified by genetic tests to cause the associated recessive mutant phenotypes, and the validity of most of the remaining lines is predicted on statistical grounds. Sequences flanking >920 insertions have been determined to exactly position them in the genome and to identify 376 potentially affected transcripts from collections of EST sequences. Strains in the BDGP collection are available from the Bloomington Stock Center and have already assisted the research community in characterizing >250 Drosophila genes. The likely identity of 131 additional genes in the collection is reported here. Our results show that Drosophila genes have a wide range of sensitivity to inactivation by P elements, and provide a rationale for greatly expanding the BDGP primary collection based entirely on insertion site sequencing. We predict that this approach can bring >85% of all Drosophila open reading frames under experimental control. PMID:10471706

  19. External Tank (ET) Bipod Fitting Bolted Attachment Locking Insert Performance

    NASA Technical Reports Server (NTRS)

    Larsen, Curtis E.; Wilson, Tim R.; Elliott, Kenny B.; Raju, Ivatury S.; McManamen, John

    2008-01-01

    Following STS-107, the External Tank (ET) Project implemented corrective actions and configuration changes at the ET bipod fitting. Among the corrective actions, the existing bolt lock wire which provided resistance to potential bolt rotation was removed. The lock wire removal was because of concerns with creating voids during foam application and potential for lock wire to become debris. The bolts had been previously lubricated to facilitate assembly but, because of elimination of the lock wire, the ET Project wanted to enable the locking feature of the insert. Thus, the lubrication was removed from bolt threads and instead applied to the washer under the bolt head. Lubrication is necessary to maximize joint pre-load while remaining within the bolt torque specification. The locking feature is implemented by thread crimping in at four places in the insert. As the bolt is torqued into the insert the bolt threads its way past the crimped parts of the insert. This provides the locking of the bolt, as torque is required to loosen the joint after clamping.

  20. Selective insertion of sulfur dioxide reduction intermediates on graphene oxide.

    PubMed

    Humeres, Eduardo; Debacher, Nito A; Smaniotto, Alessandra; de Castro, Karen M; Benetoli, Luís O B; de Souza, Eduardo P; Moreira, Regina de F P M; Lopes, Cristiane N; Schreiner, Wido H; Canle, Moisés; Santaballa, J Arturo

    2014-04-22

    Graphite microparticles (d50 6.20 μm) were oxidized by strong acids, and the resultant graphite oxide was thermally exfoliated to graphene oxide sheets (MPGO, C/O 1.53). Graphene oxide was treated with nonthermal plasma under a SO2 atmosphere at room temperature. The XPS spectrum showed that SO2 was inserted only as the oxidized intermediate at 168.7 eV in the S 2p region. Short thermal shocks at 600 and 400 °C, under an Ar atmosphere, produced reduced sulfur and carbon dioxide as shown by the XPS spectrum and TGA analysis coupled to FTIR. MPGO was also submitted to thermal reaction with SO2 at 630 °C, and the XPS spectrum in the S 2p region at 164.0 eV showed that this time only the nonoxidized episulfide intermediate was inserted. Plasma and thermal treatment produced a partial reduction of MPGO. The sequence of thermal reaction followed by plasma treatment inserted both sulfur intermediates. Because oxidized and nonoxidized intermediates have different reactivities, this selective insertion would allow the addition of selective types of organic fragments to the surface of graphene oxide.

  1. Insertion Success of the Laryngeal Tube in Emergency Airway Management.

    PubMed

    Bernhard, Michael; Gries, André; Ramshorn-Zimmer, Alexandra; Wenzel, Volker; Hossfeld, Bjoern

    2016-01-01

    Background. Emergency airway management (AM) is a priority when resuscitating critically ill or severely injured patients. The goal of this study was to determine the success rates of LT insertion during AM. Methods. Studies that included LT first-pass insertion (FPI) and overall-pass insertion (OPI) success by emergency medical services and in-hospital providers performing AM for emergency situations as well as for scheduled surgery published until July 2014 were searched systematically in Medline. Results. Data of 36 studies (n = 1,897) reported a LT FPI success by physicians of 82.5% with an OPI success of 93.6% (p < 0.001). A cumulative analysis of all 53 studies (n = 3,600) led to FPI and OPI success of 80.1% and 92.6% (p < 0.001), respectively. The results of 26 studies (n = 2,159) comparing the LT with the laryngeal mask airway (LMA) demonstrated a FPI success of 77.0 versus 78.7% (p = 0.36) and an OPI success of 92.2 versus 97.7% (p < 0.001). Conclusion. LT insertion failed in the first attempt in one out of five patients, with an overall failure rate in one out of 14 patients. When compared with the LT, the LMA had a cumulative 5.5% better OPI success rate.

  2. Insertion of short transmembrane helices by the Sec61 translocon.

    PubMed

    Jaud, Simon; Fernández-Vidal, Mónica; Nilsson, Ingmarie; Meindl-Beinker, Nadja M; Hübner, Nadja C; Tobias, Douglas J; von Heijne, Gunnar; White, Stephen H

    2009-07-14

    The insertion efficiency of transmembrane (TM) helices by the Sec61 translocon depends on helix amino acid composition, the positions of the amino acids within the helix, and helix length. We have used an in vitro expression system to examine systematically the insertion efficiency of short polyleucine segments (L(n), n = 4 ... 12) flanked at either end by 4-residue sequences of the form XXPX-L(n)-XPXX with X = G, N, D, or K. Except for X = K, insertion efficiency (p) is <10% for n < 8, but rises steeply to 100% for n = 12. For X = K, p is already close to 100% for n = 10. A similar pattern is observed for synthetic peptides incorporated into oriented phospholipid bilayer arrays, consistent with the idea that recognition of TM segments by the translocon critically involves physical partitioning of nascent peptide chains into the lipid bilayer. Molecular dynamics simulations suggest that insertion efficiency is determined primarily by the energetic cost of distorting the bilayer in the vicinity of the TM helix. Very short lysine-flanked leucine segments can reduce the energetic cost by extensive hydrogen bonding with water and lipid phosphate groups (snorkeling) and by partial unfolding.

  3. Large scale screen for transposon insertions into cloned genes.

    PubMed Central

    Hamilton, B A; Palazzolo, M J; Chang, J H; VijayRaghavan, K; Mayeda, C A; Whitney, M A; Meyerowitz, E M

    1991-01-01

    We describe a method of screening for transposon insertions in or near Drosophila loci that correspond to cloned DNA sequences. We mobilize a modified P element transposon that carries a bacterial plasmid origin of replication and a drug-resistance marker. The genomic sequences flanking each transposon insertion site can then be rescued as a plasmid in Escherichia coli. Libraries of such plasmids, representing pools of transposon-mutagenized individuals, are used as hybridization probes against cloned sequences to determine whether a transposon has inserted next to a particular site in the genome. The number of loci that can be screened simultaneously by this procedure is quite large. We have screened an array of cDNA clones representing almost 700 distinct loci against libraries representing 760 mutagenized flies, and we obtained hybridization signals to 7 different cDNAs. Three of these events have been analyzed in detail and represent genuine insertions near genomic sequences that correspond to the cDNAs. Images PMID:1849274

  4. Postplacental or delayed levonorgestrel intrauterine device insertion and breastfeeding duration

    PubMed Central

    Chen, Beatrice A.; Reeves, Matthew F.; Creinin, Mitchell D.; Schwarz, E. Bimla

    2011-01-01

    BACKGROUND The objective was to assess the effect of timing of postpartum levonorgestrel-releasing IUD insertion on breastfeeding continuation. STUDY DESIGN Women interested in using a levonorgestrel IUD postpartum were randomized to immediate postplacental insertion (postplacental group) or insertion 6–8 weeks after vaginal delivery (delayed group). Duration and exclusivity of breastfeeding were assessed at 6–8 weeks, 3 months, and 6 months postpartum. Only women who received an IUD were included in this analysis. RESULTS Breastfeeding was initiated by 32/50 (64%) of women receiving a postplacental IUD and 27/46 (58.7%) of women receiving a delayed IUD (p=0.59). More women in the delayed group compared to the postplacental group continued to breastfeed at 6–8 weeks (16/46 vs 15/50, p=0.62), 3 months (13/46 vs 7/50, p=0.13), and 6 months postpartum (11/46 vs 3/50, p=0.02). The results did not differ when only women who initiated breastfeeding or only primiparous women with no prior breastfeeding experience were analyzed. CONCLUSIONS Immediate postplacental insertion of the levonorgestrel IUD is associated with shorter duration of breastfeeding and less exclusive breastfeeding. Further studies are needed of the effects of early initiation of progestin-only methods on women’s lactation experience. PMID:22018124

  5. Folding and insertion thermodynamics of the transmembrane WALP peptide

    SciTech Connect

    Bereau, Tristan; Bennett, W. F. Drew; Pfaendtner, Jim; Deserno, Markus; Karttunen, Mikko

    2015-12-28

    The anchor of most integral membrane proteins consists of one or several helices spanning the lipid bilayer. The WALP peptide, GWW(LA){sub n} (L)WWA, is a common model helix to study the fundamentals of protein insertion and folding, as well as helix-helix association in the membrane. Its structural properties have been illuminated in a large number of experimental and simulation studies. In this combined coarse-grained and atomistic simulation study, we probe the thermodynamics of a single WALP peptide, focusing on both the insertion across the water-membrane interface, as well as folding in both water and a membrane. The potential of mean force characterizing the peptide’s insertion into the membrane shows qualitatively similar behavior across peptides and three force fields. However, the Martini force field exhibits a pronounced secondary minimum for an adsorbed interfacial state, which may even become the global minimum—in contrast to both atomistic simulations and the alternative PLUM force field. Even though the two coarse-grained models reproduce the free energy of insertion of individual amino acids side chains, they both underestimate its corresponding value for the full peptide (as compared with atomistic simulations), hinting at cooperative physics beyond the residue level. Folding of WALP in the two environments indicates the helix as the most stable structure, though with different relative stabilities and chain-length dependence.

  6. Folding and insertion thermodynamics of the transmembrane WALP peptide

    NASA Astrophysics Data System (ADS)

    Bereau, Tristan; Bennett, W. F. Drew; Pfaendtner, Jim; Deserno, Markus; Karttunen, Mikko

    2015-12-01

    The anchor of most integral membrane proteins consists of one or several helices spanning the lipid bilayer. The WALP peptide, GWW(LA)n (L)WWA, is a common model helix to study the fundamentals of protein insertion and folding, as well as helix-helix association in the membrane. Its structural properties have been illuminated in a large number of experimental and simulation studies. In this combined coarse-grained and atomistic simulation study, we probe the thermodynamics of a single WALP peptide, focusing on both the insertion across the water-membrane interface, as well as folding in both water and a membrane. The potential of mean force characterizing the peptide's insertion into the membrane shows qualitatively similar behavior across peptides and three force fields. However, the Martini force field exhibits a pronounced secondary minimum for an adsorbed interfacial state, which may even become the global minimum—in contrast to both atomistic simulations and the alternative PLUM force field. Even though the two coarse-grained models reproduce the free energy of insertion of individual amino acids side chains, they both underestimate its corresponding value for the full peptide (as compared with atomistic simulations), hinting at cooperative physics beyond the residue level. Folding of WALP in the two environments indicates the helix as the most stable structure, though with different relative stabilities and chain-length dependence.

  7. New transposon delivery plasmids for insertional mutagenesis in Bacillus anthracis

    PubMed Central

    Wilson, Adam C.; Perego, Marta; Hoch, James A.

    2007-01-01

    Two new transposon delivery vector systems utilizing Mariner and mini-Tn10 transposons have been developed for in vivo insertional mutagenesis in Bacillus anthracis and other compatible Gram-positive species. The utility of both systems was directly demonstrated through the mutagenesis of a widely used B. anthracis strain. PMID:17931726

  8. Observations on rotating needle insertions using a brachytherapy robot

    NASA Astrophysics Data System (ADS)

    Meltsner, M. A.; Ferrier, N. J.; Thomadsen, B. R.

    2007-09-01

    A robot designed for prostate brachytherapy implantations has the potential to greatly improve treatment success. Much of the research in robotic surgery focuses on measuring accuracy. However, there exist many factors that must be optimized before an analysis of needle placement accuracy can be determined. Some of these parameters include choice of the needle type, insertion velocity, usefulness of the rotating needle and rotation speed. These parameters may affect the force at which the needle interacts with the tissue. A reduction in force has been shown to decrease the compression of the prostate and potentially increase the accuracy of seed position. Rotating the needle as it is inserted may reduce frictional forces while increasing accuracy. However, needle rotations are considered to increase tissue damage due to the drilling nature of the insertion. We explore many of the factors involved in optimizing a brachytherapy robot, and the potential effects each parameter may have on the procedure. We also investigate the interaction of rotating needles in gel and suggest the rotate-cannula-only method of conical needle insertion to minimize any tissue damage while still maintaining the benefits of reduced force and increased accuracy.

  9. Spline modelling electron insert factors using routine measurements.

    PubMed

    Biggs, S; Sobolewski, M; Murry, R; Kenny, J

    2016-01-01

    There are many methods available to predict electron output factors; however, many centres still measure the factors for each irregular electron field. Creating an electron output factor prediction model that approaches measurement accuracy--but uses already available data and is simple to implement--would be advantageous in the clinical setting. This work presents an empirical spline model for output factor prediction that requires only the measured factors for arbitrary insert shapes. Equivalent ellipses of the insert shapes are determined and then parameterised by width and ratio of perimeter to area. This takes into account changes in lateral scatter, bremsstrahlung produced in the insert material, and scatter from the edge of the insert. Agreement between prediction and measurement for the 12 MeV validation data had an uncertainty of 0.4% (1SD). The maximum recorded deviation between measurement and prediction over the range of energies was 1.0%. The validation methodology showed that one may expect an approximate uncertainty of 0.5% (1SD) when as little as eight data points are used. The level of accuracy combined with the ease with which this model can be generated demonstrates its suitability for clinical use. Implementation of this method is freely available for download at https://github.com/SimonBiggs/electronfactors.

  10. Root cause of incomplete control rod insertions at Westinghouse reactors

    SciTech Connect

    Ray, S.

    1997-01-01

    Within the past year, incomplete RCCA insertions have been observed on high burnup fuel assemblies at two Westinghouse PWRs. Initial tests at the Wolf Creek site indicated that the direct cause of the incomplete insertions observed at Wolf Creek was excessive fuel assembly thimble tube distortion. Westinghouse committed to the NRC to perform a root cause analysis by the end of August, 1996. The root cause analysis process used by Westinghouse included testing at ten sites to obtain drag, growth and other characteristics of high burnup fuel assemblies. It also included testing at the Westinghouse hot cell of two of the Wolf Creek incomplete insertion assemblies. A mechanical model was developed to calculate the response of fuel assemblies when subjected to compressive loads. Detailed manufacturing reviews were conducted to determine if this was a manufacturing related issue. In addition, a review of available worldwide experience was performed. Based on the above, it was concluded that the thimble tube distortion observed on the Wolf Creek incomplete insertion assemblies was caused by unusual fuel assembly growth over and above what would typically be expected as a result of irradiation exposure. It was determined that the unusual growth component is a combination of growth due to oxide accumulation and accelerated growth, and would only be expected in high temperature plants on fuel assemblies that see long residence times and high power duties.

  11. Temperature induced compositional redistribution in blended insertion electrodes

    NASA Astrophysics Data System (ADS)

    Heubner, C.; Lämmel, C.; Schneider, M.; Michaelis, A.

    2017-03-01

    Blending insertion compounds is a novel and promising approach to design advanced electrodes for future lithium-ion batteries. In spite of the considerable improvements regarding safety issues and power density, the understanding of basic interactions between the constituents of the blend and differences towards common single compound insertion electrodes is still ongoing. Herein we explore and verify the effect of temperature induced compositional redistribution of lithium-ions between the constituents of a blended insertion electrode for the first time. A model-like blend electrode and a special experimental setup is used to measure the compositional redistribution current between the constituents when subjected to a temperature change. The amount of lithium exchanged between the constituents of the blend is also derived theoretically based on the thermodynamic properties of the pure constituents, showing excellent agreement to the experimental results. Theoretical and experimental results proof that significant amounts of lithium are exchanged between the constituents without any cycling of the battery, suggesting that this effect may intrinsically reduce the cycle life of batteries with blended insertion electrodes.

  12. Astronaut Scott Carpenter inserted into Aurora 7 spacecraft

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Astronaut M. Scott Carpenter, pilot of the Mercury-Atlas 7 space flight, is inserted into Aurora 7 spacecraft during the prelaunch countdown. Carpenter is assisted into the spacecraft by Astronaut John Glenn and Gunter Vendt, McDonnell Douglas pad capsule test conducter.

  13. Computer Modeling of Protocellular Functions: Peptide Insertion in Membranes

    NASA Technical Reports Server (NTRS)

    Rodriquez-Gomez, D.; Darve, E.; Pohorille, A.

    2006-01-01

    Lipid vesicles became the precursors to protocells by acquiring the capabilities needed to survive and reproduce. These include transport of ions, nutrients and waste products across cell walls and capture of energy and its conversion into a chemically usable form. In modem organisms these functions are carried out by membrane-bound proteins (about 30% of the genome codes for this kind of proteins). A number of properties of alpha-helical peptides suggest that their associations are excellent candidates for protobiological precursors of proteins. In particular, some simple a-helical peptides can aggregate spontaneously and form functional channels. This process can be described conceptually by a three-step thermodynamic cycle: 1 - folding of helices at the water-membrane interface, 2 - helix insertion into the lipid bilayer and 3 - specific interactions of these helices that result in functional tertiary structures. Although a crucial step, helix insertion has not been adequately studied because of the insolubility and aggregation of hydrophobic peptides. In this work, we use computer simulation methods (Molecular Dynamics) to characterize the energetics of helix insertion and we discuss its importance in an evolutionary context. Specifically, helices could self-assemble only if their interactions were sufficiently strong to compensate the unfavorable Free Energy of insertion of individual helices into membranes, providing a selection mechanism for protobiological evolution.

  14. Insertion Success of the Laryngeal Tube in Emergency Airway Management

    PubMed Central

    Gries, André; Ramshorn-Zimmer, Alexandra; Wenzel, Volker

    2016-01-01

    Background. Emergency airway management (AM) is a priority when resuscitating critically ill or severely injured patients. The goal of this study was to determine the success rates of LT insertion during AM. Methods. Studies that included LT first-pass insertion (FPI) and overall-pass insertion (OPI) success by emergency medical services and in-hospital providers performing AM for emergency situations as well as for scheduled surgery published until July 2014 were searched systematically in Medline. Results. Data of 36 studies (n = 1,897) reported a LT FPI success by physicians of 82.5% with an OPI success of 93.6% (p < 0.001). A cumulative analysis of all 53 studies (n = 3,600) led to FPI and OPI success of 80.1% and 92.6% (p < 0.001), respectively. The results of 26 studies (n = 2,159) comparing the LT with the laryngeal mask airway (LMA) demonstrated a FPI success of 77.0 versus 78.7% (p = 0.36) and an OPI success of 92.2 versus 97.7% (p < 0.001). Conclusion. LT insertion failed in the first attempt in one out of five patients, with an overall failure rate in one out of 14 patients. When compared with the LT, the LMA had a cumulative 5.5% better OPI success rate. PMID:27642595

  15. Prediction tool for thrombi associated with peripherally inserted central catheters.

    PubMed

    Seeley, Maria A; Santiago, Mary; Shott, Susan

    2007-01-01

    The upper extremity deep vein thrombosis rate is increasing at the same time that the rate for insertions of peripherally inserted central catheters is on the rise. There is little information on whether the established risk factors for lower extremity deep vein thromboses are effective to predict the occurrence of upper extremity deep vein thrombosis. The purpose of this study was to identify patients at highest risk for upper extremity deep vein thrombosis in order to initiate effective prophylaxis. A retrospective review was undertaken of medical records of all patients with peripherally inserted central catheters inserted in a 6-month period at a Midwestern US hospital. Of the 233 charts reviewed, 17 (7.3%) recorded an upper extremity deep vein thrombosis during the patient's hospital stay. Of the multiple factors identified with deep vein thrombosis in the literature, a weighted risk factor measure, the upper extremity deep vein thrombosis prediction tool, was developed. Sensitivity of the instrument for upper extremity deep vein thrombosis is high (88%), as are its specificity (82%) and negative predictive value (99%), whereas the positive predictive value is low (28%). The total percentage of cases correctly classified is 82%. Further testing is indicated on a larger sample to extend the validity of this instrument.

  16. Folding and insertion thermodynamics of the transmembrane WALP peptide.

    PubMed

    Bereau, Tristan; Bennett, W F Drew; Pfaendtner, Jim; Deserno, Markus; Karttunen, Mikko

    2015-12-28

    The anchor of most integral membrane proteins consists of one or several helices spanning the lipid bilayer. The WALP peptide, GWW(LA)n (L)WWA, is a common model helix to study the fundamentals of protein insertion and folding, as well as helix-helix association in the membrane. Its structural properties have been illuminated in a large number of experimental and simulation studies. In this combined coarse-grained and atomistic simulation study, we probe the thermodynamics of a single WALP peptide, focusing on both the insertion across the water-membrane interface, as well as folding in both water and a membrane. The potential of mean force characterizing the peptide's insertion into the membrane shows qualitatively similar behavior across peptides and three force fields. However, the Martini force field exhibits a pronounced secondary minimum for an adsorbed interfacial state, which may even become the global minimum-in contrast to both atomistic simulations and the alternative PLUM force field. Even though the two coarse-grained models reproduce the free energy of insertion of individual amino acids side chains, they both underestimate its corresponding value for the full peptide (as compared with atomistic simulations), hinting at cooperative physics beyond the residue level. Folding of WALP in the two environments indicates the helix as the most stable structure, though with different relative stabilities and chain-length dependence.

  17. Effects of Rotational Motion in Robotic Needle Insertion

    PubMed Central

    Ramezanpour, H.; Yousefi, H.; Rezaei, M.; Rostami, M.

    2015-01-01

    Background Robotic needle insertion in biological tissues has been known as one the most applicable procedures in sampling, robotic injection and different medical therapies and operations. Objective In this paper, we would like to investigate the effects of angular velocity in soft tissue insertion procedure by considering force-displacement diagram. Non-homogenous camel liver can be exploited as a tissue sample under standard compression test with Zwick/Roell device employing 1-D axial load-cell. Methods Effects of rotational motion were studied by running needle insertion experiments in 5, 50 and 200 mm/min in two types of with or without rotational velocity of 50, 150 and 300 rpm. On further steps with deeper penetrations, friction force of the insertion procedure in needle shaft was acquired by a definite thickness of the tissue. Results Designed mechanism of fixture for providing different frequencies of rotational motion is available in this work. Results for comparison of different force graphs were also provided. Conclusion Derived force-displacement graphs showed a significant difference between two procedures; however, tissue bleeding and disorganized micro-structure would be among unavoidable results. PMID:26688800

  18. Analysis of HIV-2 Vpx by modeling and insertional mutagenesis

    SciTech Connect

    Mahnke, Lisa A. . E-mail: lmahnke@im.wustl.edu; Belshan, Michael; Ratner, Lee . E-mail: lratner@im.wustl.edu

    2006-04-25

    Vpx facilitates HIV-2 nuclear localization by a poorly understood mechanism. We have compared Vpx to an NMR structure HIV-1 Vpr in a central helical domain and probed regions of Vpx by insertional mutagenesis. A predicted loop between helices two and three appears to be unique, overlapping with a known novel nuclear localization signal. Overall, Vpx was found to be surprisingly flexible, tolerating a series of large insertions. We found that insertion within the polyproline-containing C-terminus destabilizes nuclear localization, whereas mutating a second helix in the central domain disrupts viral packaging. Other insertional mutants in the predicted loop and in a linker region between the central domain and the C-terminus may be useful as sites of intramolecular tags as they could be packaged adequately and retained preintegration complex associated integration activity in a serum starvation assay. An unexpected result was found within a previously defined nuclear localization motif near aa 71. This mutant retained robust nuclear localization in a GFP fusion assay and was competent for preintegration complex associated nuclear import. In summary, we have modeled helical content in Vpx and assessed potential sites of intramolecular tags which may prove useful for protein-protein interactions studies.

  19. Discovery of unfixed endogenous retrovirus insertions in diverse human populations

    PubMed Central

    Wildschutte, Julia Halo; Williams, Zachary H.; Montesion, Meagan; Subramanian, Ravi P.; Kidd, Jeffrey M.; Coffin, John M.

    2016-01-01

    Endogenous retroviruses (ERVs) have contributed to more than 8% of the human genome. The majority of these elements lack function due to accumulated mutations or internal recombination resulting in a solitary (solo) LTR, although members of one group of human ERVs (HERVs), HERV-K, were recently active with members that remain nearly intact, a subset of which is present as insertionally polymorphic loci that include approximately full-length (2-LTR) and solo-LTR alleles in addition to the unoccupied site. Several 2-LTR insertions have intact reading frames in some or all genes that are expressed as functional proteins. These properties reflect the activity of HERV-K and suggest the existence of additional unique loci within humans. We sought to determine the extent to which other polymorphic insertions are present in humans, using sequenced genomes from the 1000 Genomes Project and a subset of the Human Genome Diversity Project panel. We report analysis of a total of 36 nonreference polymorphic HERV-K proviruses, including 19 newly reported loci, with insertion frequencies ranging from <0.0005 to >0.75 that varied by population. Targeted screening of individual loci identified three new unfixed 2-LTR proviruses within our set, including an intact provirus present at Xq21.33 in some individuals, with the potential for retained infectivity. PMID:27001843

  20. Quench Module Insert Capabilities and Development Test Results

    NASA Technical Reports Server (NTRS)

    Carswell, B.; Crouch, M.; Farmer, J.; Breeding, S.; Rose, F.; Whitaker, Ann F. (Technical Monitor)

    2001-01-01

    Quench Module Insert is a directional solidification furnace, which will fly in the Materials Science Research Facility. The QMI provides high thermal gradient and quench capabilities for processing metals and alloys in microgravity. This paper will describe the capabilities and present of on-going analysis and development testing.

  1. Effects of lithium insertion on thermal conductivity of silicon nanowires

    SciTech Connect

    Xu, Wen; Zhang, Gang; Li, Baowen

    2015-04-27

    Recently, silicon nanowires (SiNWs) have been applied as high-performance Li battery anodes, since they can overcome the pulverization and mechanical fracture during lithiation. Although thermal stability is one of the most important parameters that determine safety of Li batteries, thermal conductivity of SiNWs with Li insertion remains unclear. In this letter, using molecular dynamics simulations, we study room temperature thermal conductivity of SiNWs with Li insertion. It is found that compared with the pristine SiNW, there is as much as 60% reduction in thermal conductivity with 10% concentration of inserted Li atoms, while under the same impurity concentration the reduction in thermal conductivity of the mass-disordered SiNW is only 30%. With lattice dynamics calculations and normal mode decomposition, it is revealed that the phonon lifetimes in SiNWs decrease greatly due to strong scattering of phonons by vibrational modes of Li atoms, especially for those high frequency phonons. The observed strong phonon scattering phenomenon in Li-inserted SiNWs is similar to the phonon rattling effect. Our study serves as an exploration of thermal properties of SiNWs as Li battery anodes or weakly coupled with impurity atoms.

  2. Cochlear implant insertion forces in microdissected human cochlea to evaluate a prototype array.

    PubMed

    Nguyen, Yann; Miroir, Mathieu; Kazmitcheff, Guillaume; Sutter, Jasmine; Bensidhoum, Morad; Ferrary, Evelyne; Sterkers, Olivier; Bozorg Grayeli, Alexis

    2012-01-01

    Cochlear implant array insertion forces are potentially related to cochlear trauma. We compared these forces between a standard (Digisonic SP; Neurelec, Vallauris, France) and an array prototype (Neurelec) with a smaller diameter. The arrays were inserted by a mechatronic tool in 23 dissected human cochlea specimens exposing the basilar membrane. The array progression under the basilar membrane was filmed together with dynamic force measurements. Insertion force profiles and depth of insertion were compared. The recordings showed lower insertion forces beyond 270° of insertion and deeper insertions with the thin prototype array. This will potentially allow larger cochlear coverage with less trauma.

  3. [Discussion on different methods of painless needle insertion based on positions].

    PubMed

    Chen, Chao; Shi, Xuemin

    2016-03-01

    Based on multiple painless needle insertion skills in clinic combined with own experience, the authors propose the opinion that different painless needle insertion methods need to be used according to the positions where acupoints are. The appropriate needle insertion mean must be selected in accord with the anatomical characteristic of acupoint position, namely, tapping insertion is applied in scalp part, skin-pinching up needle insertion is used in forehead and superficial nerve trunk, fingernail-pressure needle insertion in ocular region and positions where large blood vessels distribute, flying acupuncture insertion in face and finger tip, vibrating needle in occipitonuchal region, quivering insertion in abdomen, quick pricking by single hand in the four limbs and back, and chop acupuncture in palm and pelma. The insertion methods are systematically discussed in terms of preparation, acupoint position, manipulation and attention, etc, so that to provide the reference for painless needle insertion.

  4. Efficient simulation of chemical potentials and phase equilibria in associating fluids: monomer/dimer insertion versus gradual particle insertion in primitive water models

    NASA Astrophysics Data System (ADS)

    Vörtler, Horst L.; Kettler, Matthias

    Widom's test particle method for simulation of chemical potentials fails for associating (network forming) fluids. Therefore we study more sophisticated insertion methods, such as gradual particle insertion and the recent unbonded particle insertion of Tripathi and Chapman. We model strongly associating fluids by short-ranged primitive models (PMs) due to Nezbeda and co-workers. Recently, we showed that gradual insertion, in principle, is applicable to PM water. Here, we study systematically subcritical chemical potential isotherms, determine vapour-liquid coexistence densities and estimate the critical point density and temperature. For comparison we implement two variants of the Tripathi-Chapman algorithm. In the first case we use as unbonded test particles only particles which remain single molecules (monomers), when inserted and in the second case such test particles which remain as single molecules or form a two-particle cluster with an existing free molecule (monomer/dimer), when inserted. While monomer insertion improves Widom's method slightly, monomer/dimer insertion extends substantially the range of application to the subcritical temperatures studied by gradual insertion. But, for low temperatures monomer/dimer insertion requires extremely long Markov chains and significantly more computer time than gradual insertion. The Tripathi-Chapman algorithm - a direct extension of Widom's method - is much simpler than gradual insertion and therefore should be preferred at supercritical and critical conditions.

  5. The evolution of small insertions and deletions in the coding genes of Drosophila melanogaster.

    PubMed

    Chong, Zechen; Zhai, Weiwei; Li, Chunyan; Gao, Min; Gong, Qiang; Ruan, Jue; Li, Juan; Jiang, Lan; Lv, Xuemei; Hungate, Eric; Wu, Chung-I

    2013-12-01

    Studies of protein evolution have focused on amino acid substitutions with much less systematic analysis on insertion and deletions (indels) in protein coding genes. We hence surveyed 7,500 genes between Drosophila melanogaster and D. simulans, using D. yakuba as an outgroup for this purpose. The evolutionary rate of coding indels is indeed low, at only 3% of that of nonsynonymous substitutions. As coding indels follow a geometric distribution in size and tend to fall in low-complexity regions of proteins, it is unclear whether selection or mutation underlies this low rate. To resolve the issue, we collected genomic sequences from an isogenic African line of D. melanogaster (ZS30) at a high coverage of 70× and analyzed indel polymorphism between ZS30 and the reference genome. In comparing polymorphism and divergence, we found that the divergence to polymorphism ratio (i.e., fixation index) for smaller indels (size ≤ 10 bp) is very similar to that for synonymous changes, suggesting that most of the within-species polymorphism and between-species divergence for indels are selectively neutral. Interestingly, deletions of larger sizes (size ≥ 11 bp and ≤ 30 bp) have a much higher fixation index than synonymous mutations and 44.4% of fixed middle-sized deletions are estimated to be adaptive. To our surprise, this pattern is not found for insertions. Protein indel evolution appear to be in a dynamic flux of neutrally driven expansion (insertions) together with adaptive-driven contraction (deletions), and these observations provide important insights for understanding the fitness of new mutations as well as the evolutionary driving forces for genomic evolution in Drosophila species.

  6. Choice of Tevatron tune with matched low-. beta. insertions

    SciTech Connect

    Month, M.

    1984-06-20

    The Tevatron lattice, following closely the Main Ring lattice, is made up of approximately 90 FODO cells with six straight sections equally spaced. The straight sections are identical (neglecting the special high-..beta.. straight section used for resonant extraction) providing six-fold symmetry. Each straight section is comprised mainly of a pair of doublets, one at each end, in an antisymmetric configuration. The ..beta..-functions are matched to the building block cells but the dispersion function is not. The ..beta..-function matching implies that the structure is not sensitive to the tune of the sextant (i.e., structure resonances); however, the dispersion mismatch excites off-momentum dipole resonances and the dispersion structure of the ring is therefore sensitive to sextant tunes near an integer. The introduction of low-..beta.. insertions can be accomplished by replacing a standard insertion (expanded somewhat from the doublet pair straight section) with a low-..beta.. insertion. By matching the ends of the replacement units with respect to ..beta..-functions and dispersion function, the full lattice is made insensitive to linear structure resonances. Procedures are described here for introducing into the idealized Tevatron lattice considered here a single matched low-..beta.. insertion at BO, and two matched low-..beta.. insertions at BO and DO. The main consequence is the need for a significant number of independently powered quadrupoles. Another conclusion is related to the choice of tune, where it is found that the arbitrary rule of maintaining a fixed tune for all configurations should be modified.

  7. Effects of rotated square inserts on the longitudinal vibration band gaps in thin phononic crystal plates

    NASA Astrophysics Data System (ADS)

    Zhao, Haojiang; Liu, Rongqiang; Shi, Chuang; Guo, Hongwei; Deng, Zongquan

    2015-07-01

    Longitudinal vibration of thin phononic crystal plates with a hybrid square-like array of square inserts is investigated. The plane wave expansion method is used to calculate the vibration band structure of the plate. Numerical results show that rotated square inserts can open several vibration gaps, and the band structures are twisted because of the rotation of inserts. Filling fraction and material of the insert affect the change law of the gap width versus the rotation angles of square inserts.

  8. Effects on Subtalar Joint Stress Distribution After Cannulated Screw Insertion at Different Positions and Directions.

    PubMed

    Yuan, Cheng-song; Chen, Wan; Chen, Chen; Yang, Guang-hua; Hu, Chao; Tang, Kang-lai

    2015-01-01

    We investigated the effects on subtalar joint stress distribution after cannulated screw insertion at different positions and directions. After establishing a 3-dimensional geometric model of a normal subtalar joint, we analyzed the most ideal cannulated screw insertion position and approach for subtalar joint stress distribution and compared the differences in loading stress, antirotary strength, and anti-inversion/eversion strength among lateral-medial antiparallel screw insertion, traditional screw insertion, and ideal cannulated screw insertion. The screw insertion approach allowing the most uniform subtalar joint loading stress distribution was lateral screw insertion near the border of the talar neck plus medial screw insertion close to the ankle joint. For stress distribution uniformity, antirotary strength, and anti-inversion/eversion strength, lateral-medial antiparallel screw insertion was superior to traditional double-screw insertion. Compared with ideal cannulated screw insertion, slightly poorer stress distribution uniformity and better antirotary strength and anti-inversion/eversion strength were observed for lateral-medial antiparallel screw insertion. Traditional single-screw insertion was better than double-screw insertion for stress distribution uniformity but worse for anti-rotary strength and anti-inversion/eversion strength. Lateral-medial antiparallel screw insertion was slightly worse for stress distribution uniformity than was ideal cannulated screw insertion but superior to traditional screw insertion. It was better than both ideal cannulated screw insertion and traditional screw insertion for anti-rotary strength and anti-inversion/eversion strength. Lateral-medial antiparallel screw insertion is an approach with simple localization, convenient operation, and good safety.

  9. Cassini observations of Saturn's inner plasmasphere: Saturn orbit insertion results

    NASA Astrophysics Data System (ADS)

    Sittler, E. C.; Thomsen, M.; Johnson, R. E.; Hartle, R. E.; Burger, M.; Chornay, D.; Shappirio, M. D.; Simpson, D.; Smith, H. T.; Coates, A. J.; Rymer, A. M.; McComas, D. J.; Young, D. T.; Reisenfeld, D.; Dougherty, M.; Andre, N.

    2006-10-01

    We present new and definitive results of Cassini plasma spectrometer (CAPS) data acquired during passage through Saturn's inner plasmasphere by the Cassini spacecraft during the approach phase of the Saturn orbit insertion period. This analysis extends the original analysis of Sittler et al. [2005. Preliminary results on Saturn's inner plasmasphere as observed by Cassini: comparison with Voyager. Geophys. Res. Lett. 32, L14S07, doi:10.1029/2005GL022653] to L˜10 along with also providing a more comprehensive study of the interrelationship of the various fluid parameters. Coincidence data are sub-divided into protons and water group ions. Our revised analysis uses an improved convergence algorithm which provides a more definitive and independent estimate of the spacecraft potential ΦSC for which we enforce the protons and water group ions to co-move with each other. This has allowed us to include spacecraft charging corrections to our fluid parameter estimations and allow accurate estimations of fluctuations in the fluid parameters for future correlative studies. In the appendix we describe the ion moments algorithm, and minor corrections introduced by not weighting the moments with sin θ term in Sittler et al. [2005] (Correction offset by revisions to instruments geometric factor). Estimates of the spacecraft potential and revised proton densities are presented. Our total ion densities are in close agreement with the electron densities reported by Moncuquet et al. [2005. Quasi-thermal noise spectroscopy in the inner magnetosphere of Saturn with Cassini/RPWS: electron temperatures and density. Geophys. Res. Lett. 32, L20S02, doi:10.1029/2005GL022508] who used upper hybrid resonance (UHR) emission lines observed by the radio and plasma wave science (RPWS) instrument. We show a positive correlation between proton temperature and water group ion temperature. The proton and thermal electron temperatures track each with both having a positive radial gradient. These

  10. NSCLC harboring EGFR exon-20 insertions after the regulatory C-helix of kinase domain responds poorly to known EGFR inhibitors.

    PubMed

    Yang, Mengmeng; Xu, Xiaoxi; Cai, Jie; Ning, Jinying; Wery, Jean Pierre; Li, Qi-Xiang

    2016-07-01

    Anecdote clinical observations hint that non-small cell lung cancer (NSCLC) with exon-20 insertions might respond poorly to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), contrasting to those with classic mutations. Lack of patient-derived experimental models has been a major hurdle for the discovery of new treatment for the diseases. We established two NSCLC-PDXs harboring two different exon-20 insertions, LU0387-adenocarcinoma (ADC) with a nine-base insertion at 2319 (H773-V774insNPH) and LU3075-squamous cell carcinoma (SCC) with a nine-base insertion at 2316 (P772-H773insDNP). Both insertions immediately follow the regulatory C-helix of the kinase domain. Contrary to the generally good responses to EGFR inhibitors observed in PDXs with classic mutations, both exon-20 insertions are largely resistant to cetuximab and TKIs in vivo, suggesting fundamental difference from the classic EGFR mutations, consistent with the poor response rate to TKI seen in anecdotal clinic reports. It is worth noting that although responses are generally poor, they differ between the two exon-20 mutants depending on the type of TKI. In vitro drug sensitivity assays using established primary cell lines from our two PDXs largely confirmed the in vivo data. Our data from patient-derived experimental models confirmed that exon-20 insertions in domain immediately following the C-helix confer poor response to all known EGFR inhibitors, and suggested that these models can be utilized to facilitate the discovery of new therapies targeting NSCLC harboring exon-20 insertions.

  11. Ka-band full-360° analog phase shifter with low insertion loss

    NASA Astrophysics Data System (ADS)

    Mengyi, Cao; Yang, Lu; Jiaxing, Wei; Jiaxin, Zheng; Xiaohua, Ma; Yue, Hao

    2014-10-01

    A new reflection-type wideband 360° monolithic-microwave integrated-circuit (MMIC) analog phase shifter at the Ka-band is proposed. The phase shifter is designed based on the principle of vector synthesis. Three Lange couplers are employed in the phase shifter, which is fabricated by the standard 0.25-μm GaAs process. We use four 4 × 40 μm GaAs HEMTs as the reflection loads. A microstrip line in parallel with the device is used as an inductance to counteract the parasitic capacitance of the device so that the reflection load performs like a pure resistance and the insertion loss can be decreased. In this phase shifter, a folded Lange coupler is utilized to reduce the size of the chip. The size of the proposed MMIC phase shifter is only 2.0 × 1.2 mm2. The measurement results show that the insertion loss is 5.0 ± 0.8 dB and a 360° continuously tunable range across 27-32 GHz is obtained with miniscule DC power consumption.

  12. HACRE1, a recently inserted copia-like retrotransposon of sunflower (Helianthus annuus L.).

    PubMed

    Buti, M; Giordani, T; Vukich, M; Gentzbittel, L; Pistelli, L; Cattonaro, F; Morgante, M; Cavallini, A; Natali, L

    2009-11-01

    In this paper we report on the isolation and characterization, for the first time, of a complete 6511 bp retrotransposon of sunflower. Considering its protein domain order and sequence similarity to other copia elements of dicotyledons, this retrotransposon was assigned to the copia retrotransposon superfamily and named HACRE1 (Helianthus annuus copia-like retroelement 1). HACRE1 carries 5' and 3' long terminal repeats (LTRs) flanking an internal region of 4661 bp. The LTRs are identical in their sequence except for two deletions of 7 and 5 nucleotides in the 5' LTR. Based on the sequence identity of the LTRs, HACRE1 was estimated to have inserted within the last approximately 84 000 years. The isolated sequence contains a complete open reading frame with only one complete reading frame. The absence of nonsense mutations agrees with the very high sequence identity between LTRs, confirming that HACRE1 insertion is recent. The haploid genome of sunflower (inbred line HCM) contains about 160 copies of HACRE1. This retrotransposon is expressed in leaflets from 7-day-old plantlets under different light conditions, probably in relation to the occurrence of many putative light-related regulatory cis-elements in the LTRs. However, sequenced cDNAs show less variability than HACRE1 genomic sequences, indicating that only a subset of this family is expressed under these conditions.

  13. Functional gains in energy and cell metabolism after TSPO gene insertion.

    PubMed

    Liu, Guo-Jun; Middleton, Ryan J; Kam, Winnie Wai-Ying; Chin, David Y; Hatty, Claire R; Chan, Ronald H Y; Banati, Richard B

    2017-03-04

    Recent loss-of-function studies in tissue-specific as well as global Tspo (Translocator Protein 18 kDa) knockout mice have not confirmed its long assumed indispensability for the translocation of cholesterol across the mitochondrial inter-membrane space, a rate-limiting step in steroid biosynthesis. Instead, recent studies in global Tspo knockout mice indicate that TSPO may play a more fundamental role in cellular bioenergetics, which may include the indirect down-stream regulation of transport or metabolic functions. To examine whether overexpression of the TSPO protein alters the cellular bioenergetic profile, Jurkat cells with low to absent endogenous expression were transfected with a TSPO construct to create a stable cell line with de novo expression of exogenous TSPO protein. Expression of TSPO was confirmed by RT-qPCR, radioligand binding with [3H]PK11195 and immunocytochemistry with a TSPO antibody. We demonstrate that TSPO gene insertion causes increased transcription of genes involved in the mitochondrial electron transport chain. Furthermore, TSPO insertion increased mitochondrial ATP production as well as cell excitability, reflected in a decrease in patch clamp recorded rectified K channel currents. These functional changes were accompanied by an increase in cell proliferation and motility, which were inhibited by PK11195, a selective ligand for TSPO. We suggest that TSPO may serve a range of functions that can be viewed as downstream regulatory effects of its primary, evolutionary conserved role in cell metabolism and energy production.

  14. Sequencing human-gibbon breakpoints of synteny reveals mosaic new insertions at rearrangement sites.

    PubMed

    Girirajan, Santhosh; Chen, Lin; Graves, Tina; Marques-Bonet, Tomas; Ventura, Mario; Fronick, Catrina; Fulton, Lucinda; Rocchi, Mariano; Fulton, Robert S; Wilson, Richard K; Mardis, Elaine R; Eichler, Evan E

    2009-02-01

    The gibbon genome exhibits extensive karyotypic diversity with an increased rate of chromosomal rearrangements during evolution. In an effort to understand the mechanistic origin and implications of these rearrangement events, we sequenced 24 synteny breakpoint regions in the white-cheeked gibbon (Nomascus leucogenys, NLE) in the form of high-quality BAC insert sequences (4.2 Mbp). While there is a significant deficit of breakpoints in genes, we identified seven human gene structures involved in signaling pathways (DEPDC4, GNG10), phospholipid metabolism (ENPP5, PLSCR2), beta-oxidation (ECH1), cellular structure and transport (HEATR4), and transcription (ZNF461), that have been disrupted in the NLE gibbon lineage. Notably, only three of these genes show the expected evolutionary signatures of pseudogenization. Sequence analysis of the breakpoints suggested both nonclassical nonhomologous end-joining (NHEJ) and replication-based mechanisms of rearrangement. A substantial number (11/24) of human-NLE gibbon breakpoints showed new insertions of gibbon-specific repeats and mosaic structures formed from disparate sequences including segmental duplications, LINE, SINE, and LTR elements. Analysis of these sites provides a model for a replication-dependent repair mechanism for double-strand breaks (DSBs) at rearrangement sites and insights into the structure and formation of primate segmental duplications at sites of genomic rearrangements during evolution.

  15. Transgenic mouse model of hemifacial microsomia: Cloning and characterization of insertional mutation region on chromosome 10

    SciTech Connect

    Naora, Hiroyuki; Otani, Hiroki; Tanaka, Osamu

    1994-10-01

    The 643 transgenic mouse line carries an autosomal dominant insertional mutation that results in hemifacial microsomia (HFM), including microtia and/or abnormal biting. In this paper, we characterize the transgene integration site in transgenic mice and preintegration site of wildtype mice. The locus, designated Hfm (hemifacial microsomia-associated locus), was mapped to chromosome 10, B1-3, by chromosome in situ hybridization. We cloned the transgene insertion site from the transgenic DNA library. By using the 5{prime} and 3{prime} flanking sequences, the preintegration region was isolated. The analysis of these regions showed that a deletion of at least 23 kb DNA occurred in association with the transgene integration. Evolutionarily conserved regions were detected within and beside the deleted region. The result of mating between hemizygotes suggests that the phenotype of the homozygote is lethality in the prenatal period. These results suggests that the Hfm locus is necessary for prenatal development and that this strain is a useful animal model for investigating the genetic predisposition to HFM in humans.

  16. Functional gains in energy and cell metabolism after TSPO gene insertion

    PubMed Central

    Chin, David Y.; Hatty, Claire R.; Chan, Ronald H. Y.

    2017-01-01

    ABSTRACT Recent loss-of-function studies in tissue-specific as well as global Tspo (Translocator Protein 18 kDa) knockout mice have not confirmed its long assumed indispensability for the translocation of cholesterol across the mitochondrial inter-membrane space, a rate-limiting step in steroid biosynthesis. Instead, recent studies in global Tspo knockout mice indicate that TSPO may play a more fundamental role in cellular bioenergetics, which may include the indirect down-stream regulation of transport or metabolic functions. To examine whether overexpression of the TSPO protein alters the cellular bioenergetic profile, Jurkat cells with low to absent endogenous expression were transfected with a TSPO construct to create a stable cell line with de novo expression of exogenous TSPO protein. Expression of TSPO was confirmed by RT-qPCR, radioligand binding with [3H]PK11195 and immunocytochemistry with a TSPO antibody. We demonstrate that TSPO gene insertion causes increased transcription of genes involved in the mitochondrial electron transport chain. Furthermore, TSPO insertion increased mitochondrial ATP production as well as cell excitability, reflected in a decrease in patch clamp recorded rectified K channel currents. These functional changes were accompanied by an increase in cell proliferation and motility, which were inhibited by PK11195, a selective ligand for TSPO. We suggest that TSPO may serve a range of functions that can be viewed as downstream regulatory effects of its primary, evolutionary conserved role in cell metabolism and energy production. PMID:28103132

  17. Lesion insertion in the projection domain: Methods and initial results

    SciTech Connect

    Chen, Baiyu; Leng, Shuai; Yu, Lifeng; Yu, Zhicong; Ma, Chi; McCollough, Cynthia

    2015-12-15

    Purpose: To perform task-based image quality assessment in CT, it is desirable to have a large number of realistic patient images with known diagnostic truth. One effective way of achieving this objective is to create hybrid images that combine patient images with inserted lesions. Because conventional hybrid images generated in the image domain fails to reflect the impact of scan and reconstruction parameters on lesion appearance, this study explored a projection-domain approach. Methods: Lesions were segmented from patient images and forward projected to acquire lesion projections. The forward-projection geometry was designed according to a commercial CT scanner and accommodated both axial and helical modes with various focal spot movement patterns. The energy employed by the commercial CT scanner for beam hardening correction was measured and used for the forward projection. The lesion projections were inserted into patient projections decoded from commercial CT projection data. The combined projections were formatted to match those of commercial CT raw data, loaded onto a commercial CT scanner, and reconstructed to create the hybrid images. Two validations were performed. First, to validate the accuracy of the forward-projection geometry, images were reconstructed from the forward projections of a virtual ACR phantom and compared to physically acquired ACR phantom images in terms of CT number accuracy and high-contrast resolution. Second, to validate the realism of the lesion in hybrid images, liver lesions were segmented from patient images and inserted back into the same patients, each at a new location specified by a radiologist. The inserted lesions were compared to the original lesions and visually assessed for realism by two experienced radiologists in a blinded fashion. Results: For the validation of the forward-projection geometry, the images reconstructed from the forward projections of the virtual ACR phantom were consistent with the images physically

  18. Aberrant Splicing and Transcription Termination Caused by P Element Insertion into the Intron of a Drosophila Gene

    PubMed Central

    Horowitz, H.; Berg, C. A.

    1995-01-01

    Insertional mutagenesis screens using the P[lacZ, rosy(+)] (PZ) transposable element have provided thousands of mutant lines for analyzing genes of varied function in the fruitfly, Drosophila melanogaster. As has been observed with other P elements, many of the PZ-induced mutations result from insertion of the P element into the promoter or 5' untranslated regions of the affected gene. We document here a novel mechanism for mutagenesis by this element. We show that sequences present within the element direct aberrant splicing and termination events that produce a mRNA composed of 5' sequences from the mutated gene (in this case, pipsqueak) and 3' sequences from within the P[lacZ, rosy(+)] element. These truncated RNAs could yield proteins with dominant mutant effects. PMID:7705633

  19. Aberrant splicing and transcription termination caused by P element insertion into the intron of a Drosophila gene

    SciTech Connect

    Horowitz, H.; Berg, C.A.

    1995-01-01

    Insertional mutagenesis screens using the P[lacZ, rosy{sup +}] (PZ) transposable element have provided thousands of mutant lines for analyzing genes of varied function in the fruitfly, Drosophila melanogaster. As has been observed with other P elements, many of the PZ-induced mutations result from insertion of the P element into the promoter or 5{prime} untranslated regions of the affected gene. We document here a novel mechanism for mutagenesis by this element. We show that sequences present within the element direct aberrant splicing and termination events that produce an mRNA composed of 5{prime} sequences from the mutated gene (in this case, pipsqueak) and 3{prime} sequences from within the P[lacZ, rosy{sup +}] element. These truncated RNAs could yield proteins with dominant mutant effects. 43 refs., 4 figs.

  20. A Generalized Lossy Transmission-Line Model for Tunable Graphene-Based Transmission Lines with Attenuation Phenomenon

    PubMed Central

    Wu, Yongle; Qu, Meijun; Liu, Yuanan

    2016-01-01

    To investigate the frequency shift phenomenon by inserting graphene, a generalized lossy transmission-line model and the related electrical parameter-extraction theory are proposed in this paper. Three kinds of graphene-based transmission lines with attenuation phenomenon including microstrip line, double-side parallel strip line, and uniplanar coplanar waveguide are analyzed under the common conditions where different chemical potentials are loaded on graphene. The values of attenuation constant and phase constant, and the real and imaginary parts of the characteristic impedance of transmission lines are extracted to analyze in details. When the attenuation constant and the reactance part of the characteristic impedance are approximately equal to zero, this kind of transmission line has low or zero insertion loss. On the contrary, the transmission line is under the radiation mode with obvious insertion loss. The phase constant changes linearly under the transmission mode and can be varied with changing of chemical potentials which attributes to the property of frequency tunability. Furthermore, a bandwidth reconfigurable uniplanar coplanar waveguide power divider is simulated to demonstrate that this theory can be applied to the design of three-port devices. In summary, this work provides a strong potential approach and design theory to help design other kinds of terahertz and mid-infrared reconfigurable devices. PMID:27553759

  1. A Generalized Lossy Transmission-Line Model for Tunable Graphene-Based Transmission Lines with Attenuation Phenomenon.

    PubMed

    Wu, Yongle; Qu, Meijun; Liu, Yuanan

    2016-08-24

    To investigate the frequency shift phenomenon by inserting graphene, a generalized lossy transmission-line model and the related electrical parameter-extraction theory are proposed in this paper. Three kinds of graphene-based transmission lines with attenuation phenomenon including microstrip line, double-side parallel strip line, and uniplanar coplanar waveguide are analyzed under the common conditions where different chemical potentials are loaded on graphene. The values of attenuation constant and phase constant, and the real and imaginary parts of the characteristic impedance of transmission lines are extracted to analyze in details. When the attenuation constant and the reactance part of the characteristic impedance are approximately equal to zero, this kind of transmission line has low or zero insertion loss. On the contrary, the transmission line is under the radiation mode with obvious insertion loss. The phase constant changes linearly under the transmission mode and can be varied with changing of chemical potentials which attributes to the property of frequency tunability. Furthermore, a bandwidth reconfigurable uniplanar coplanar waveguide power divider is simulated to demonstrate that this theory can be applied to the design of three-port devices. In summary, this work provides a strong potential approach and design theory to help design other kinds of terahertz and mid-infrared reconfigurable devices.

  2. Generation of an Enhancer-Trapping Vector for Insertional Mutagenesis in Zebrafish

    PubMed Central

    Liu, Chunyan; Song, Guili; Mao, Lin; Long, Yong; Li, Qing; Cui, Zongbin

    2015-01-01

    Enhancer trapping (ET) is a powerful approach to establish tissue- or cell-specific reporters and identify expression patterns of uncharacterized genes. Although a number of enhancer-trapping vectors have been developed and a large library of fish lines with distinct tissue- or cell-specific expression of reporter genes have been generated, the specificity and efficiency of trapping vectors need to be improved because of the bias interaction of minimal promoters with genomic enhancers. Accordingly, we generated an enhancer-trapping vector pTME that contains a minimal mouse metallothionein gene (mMTI) promoter upstream of EGFP reporter. In the first round of screening, twelve zebrafish lines that carry a single copy of ET cassettes were characterized to have tissue- or cell-specific EGFP expression. One of the highly conserved noncoding elements near an insertion site of trapping cassettes was characterized as an enhancer that can specifically regulate the expression of EGFP in cells of the central nervous system. In addition, the pTME vector contains a mutation-cassette that is able to effectively block the transcription of an endogenous gene in an ET line with ubiquitous EGFP expression. Thus, the pTME vector can be used as an alternative tool for both enhancer trapping and mutagenesis across a target genome. PMID:26436547

  3. A transposon insertion in FLOWERING LOCUS T is associated with delayed flowering in Brassica rapa.

    PubMed

    Zhang, Xueming; Meng, Lin; Liu, Bo; Hu, Yunyan; Cheng, Feng; Liang, Jianli; Aarts, Mark G M; Wang, Xiaowu; Wu, Jian

    2015-12-01

    Long days and vernalization accelerate the transition from vegetative growth to reproductive growth in Brassica rapa. Bolting before plants reach the harvesting stage is a serious problem in B. rapa vegetable crop cultivation. The genetic dissection of flowering time is important for breeding of premature bolting-resistant B. rapa crops. Using a recombinant inbred line (RIL) population, we twice detected two major quantitative trait loci (QTLs) for flowering time in two different growing seasons that were located on chromosomes A02 and A07, respectively. We hypothesized that an orthologue of the Arabidopsis thaliana FLOWERING LOCUS T (FT) gene, named as BrFT2, was the candidate gene underlying the QTL localized to A07. A transposon insertion in the second intron of BrFT2 was detected in one of the parental lines, which was predicted to generate a loss-of-function allele. Transcription analysis revealed that the BrFT2 transcript was not present in the parental line that harbored the mutated allele. RILs carrying only the mutated BrFT2 allele showed delayed flowering regardless of growing seasons when compared to RILs carrying the wild-type BrFT2 allele. These data suggest that BrFT2 is involved in flowering time regulation in controlling flowering time in B. rapa.

  4. Disulfide bond bridge insertion turns hydrophobic anticancer prodrugs into self-assembled nanomedicines.

    PubMed

    Wang, Yongjun; Liu, Dan; Zheng, Qingchuan; Zhao, Qiang; Zhang, Hongjuan; Ma, Yan; Fallon, John K; Fu, Qiang; Haynes, Matthew T; Lin, Guimei; Zhang, Rong; Wang, Dun; Yang, Xinggang; Zhao, Linxiang; He, Zhonggui; Liu, Feng

    2014-10-08

    It is commonly observed that hydrophobic molecules alone cannot self-assemble into stable nanoparticles, requiring amphiphilic or ionic materials to support nanoparticle stability and function in vivo. We report herein newly self-assembled nanomedicines through entirely different mechanisms. We present proof-of-concept methodology and results in support of our hypothesis that disulfide-induced nanomedicines (DSINMs) are promoted and stabilized by the insertion of a single disulfide bond into hydrophobic molecules, in order to balance the competition between intermolecular forces involved in the self-assembly of nanomedicines. This hypothesis has been explored through diverse synthetic compounds, which include four first-line chemotherapy drugs (paclitaxel, doxorubicin, fluorouracil, and gemcitabine), two small-molecule natural products and their derivatives, as well as a fluorescent probe. Such an unprecedented and highly reproducible system has the potential to serve as a synthetic platform for a wide array of safe and effective therapeutic and diagnostic nanomedicine strategies.

  5. Disulfide Bond Bridge Insertion Turns Hydrophobic Anticancer Prodrugs into Self-Assembled Nanomedicines

    PubMed Central

    2015-01-01

    It is commonly observed that hydrophobic molecules alone cannot self-assemble into stable nanoparticles, requiring amphiphilic or ionic materials to support nanoparticle stability and function in vivo. We report herein newly self-assembled nanomedicines through entirely different mechanisms. We present proof-of-concept methodology and results in support of our hypothesis that disulfide-induced nanomedicines (DSINMs) are promoted and stabilized by the insertion of a single disulfide bond into hydrophobic molecules, in order to balance the competition between intermolecular forces involved in the self-assembly of nanomedicines. This hypothesis has been explored through diverse synthetic compounds, which include four first-line chemotherapy drugs (paclitaxel, doxorubicin, fluorouracil, and gemcitabine), two small-molecule natural products and their derivatives, as well as a fluorescent probe. Such an unprecedented and highly reproducible system has the potential to serve as a synthetic platform for a wide array of safe and effective therapeutic and diagnostic nanomedicine strategies. PMID:25188744

  6. Update on Insertion and Complications of Central Venous Catheters for Hemodialysis

    PubMed Central

    Bream, Peter R.

    2016-01-01

    Central venous catheters are a popular choice for the initiation of hemodialysis or for bridging between different types of access. Despite this, they have many drawbacks including a high morbidity from thrombosis and infection. Advances in technology have allowed placement of these lines relatively safely, and national guidelines have been established to help prevent complications. There is an established algorithm for location and technique for placement that minimizes harm to the patient; however, there are significant short- and long-term complications that proceduralists who place catheters should be able to recognize and manage. This review covers insertion and complications of central venous catheters for hemodialysis, and the social and economic impact of the use of catheters for initiating dialysis is reviewed. PMID:27011425

  7. Friend leukaemia insertion (Fli)-1 is a prediction marker candidate for radiotherapy resistant oral squamous cell carcinoma.

    PubMed

    Shintani, S; Hamakawa, H; Nakashiro, K; Shirota, T; Hatori, M; Tanaka, M; Kuroshita, Y; Kurokawa, Y

    2010-11-01

    Radiotherapy is commonly used to treat oral squamous cell carcinoma (OSCC), but its therapeutic effects are unpredictable. To determine which genes correlate with radiation resistance in oral cancer, the authors evaluated radiation sensitivity using a standard colony formation assay with a gene microarray system for seven OSCC cell lines. They found significant associations between dozens of gene-expression levels and radiation resistance of OSCC cell lines. Following analysis of the different radiosensitive cancer cell lines, the friend leukaemia insertion (Fli)-1 gene was selected as a prediction marker gene for OSCC radiotherapy resistance. Fli-1 expression was associated with radiation resistance in OSCC patients. These data help to predict the effects radiation therapy has on OSCC, in turn contributing to the development of alternative radiation therapies.

  8. Method of installing a metallic threaded insert in a composite/rubber panel

    NASA Astrophysics Data System (ADS)

    Izdebski, Stanley

    1994-02-01

    A method of installing a metallic threaded insert in a composite/rubber panel consists of threading a first insert section into a preformed hole in a fiberglass composite panel and bonding a rubber panel over the fiberglass composite panel. The rubber panel includes an opening which is positioned over the first insert section. A second insert section is threaded onto an elongated positioning bolt and then the positioning bolt is threaded into the first insert section. The second insert section is threaded downwardly on the positioning so as to be in adjacent relation to the first insert section, and the opening in the rubber panel is filled with a rubber compound. The rubber compound is cured, preferably at room temperature, to set the second insert section in position and the positioning bolt is threaded out of the first and second insert sections.

  9. Needle insertion with duty-cycled rotation into multiple media.

    PubMed

    Lehocky, Craig A; Riviere, Cameron N

    2012-01-01

    Thin, flexible needles can be steered along nonlinear paths to reach deep anatomical structures within the human body. This study builds upon previous work involving steering of bevel-tipped needles by inserting while rotating in a duty-cycled fashion. Here we investigate how needle material and radius, duty cycle, and tissue stiffness affect needle curvature. Needles were inserted into media while rotated at a specified duty cycle and the curvature was measured. A linear relationship between duty cycle and curvature was observed across all needle materials and radii, and tissue stiffnesses. Following these observations, we developed a model that encapsulates needle and tissue parameters in order to predict the duty cycle needed to achieve a desired curvature.

  10. Phase transitions in insertion electrodes for lithium batteries

    SciTech Connect

    Thackeray, M. M.

    2000-02-02

    Phase transitions that occur during lithium insertion into layered and framework structures are discussed in the context of their application as positive and negative electrodes in lithium-ion batteries. The discussion is focused on the two-dimensional structures of graphite, LiNi{sub 1{minus}x}M{sub x}O{sub 2} (M = Co, Ti and Mg), and Li{sub 1.2}V{sub 3}O{sub 8}; examples of framework structures with a three-dimensional interstitial space for Li{sup +}-ion transport include the spinel oxides and intermetallic compounds with zinc-blende-type structures. The phase transitions are discussed in terms of their tolerance to lithium insertion and extraction and to the chemical stability of the electrodes in the cell environment.

  11. [Neonatology nurses' knowledge about Peripherally Inserted Central Venous Catheter].

    PubMed

    Belo, Marcela Patricia Macêdo; Silva, Roberta Albuquerque Mello de Castro; Nogueira, Isis Larissa Maia; Mizoguti, Daniele Pereira; Ventura, Claudiane Maria Urbano

    2012-01-01

    The Peripherally Inserted Central Catheter (PICC) has been used as a safe venous access for infants at risk. The aim of this study was to describe the knowledge and practice of nurses from the five public Neonatal Intensive Care Units, of Recife-PE, Brazil, about the use of the PICC. The sample was comprised by 52 nurses; data were collected from January to February/2010. It was found that 64,8% of nurses did not have license for insertion of the PICC. Only two units routinely used the PICC. About the indication of the access, the accuracy was above 70%. In unit B only 8,3% of nurses reported adequate initial location of the catheter tip. It was concluded that is necessary greater incentives to train nurses to use the PICC.

  12. Metallocene Catalytic Insertion Polymerization of 1-Silene to Polycarbosilanes

    PubMed Central

    Tian, Yuelong; Ge, Min; Zhang, Weigang; Lv, Xiaoxu; Yu, Shouquan

    2015-01-01

    Metallocene of zirconium were used as a catalyst for an insertion polymerization of 1-methylsilene directly into pre-ceramic precursor polyzirconocenecarbosilane (PZCS) during dechlorination of dichlorodimethylesilane by sodium, which exhibits high catalytic effectiveness with the maximum conversion ratio of polycarbosilane up to 91%. The average molecular weights of polymers synthesized are less than 1400, all with very narrow polymolecularities. The mechanism of catalytic polymerization was assumed to be similar to a coordination insertion polymerization of 1-olefins by metallocenes. The obtained PZCS show high ceramic yields with formation of composite ceramics of ZrC-SiC, which are novel polymeric precursors of ultra-high temperature ceramic (UHTC) fiber and composite. PMID:26541636

  13. Insertion devices for the Advanced Light Source at LBL

    SciTech Connect

    Hassenzahl, W.; Chin, J.; Halbach, K.; Hoyer, E.; Humphries, D.; Kincaid, B.; Savoy, R.

    1989-03-01

    The Advanced Light Source (ALS) at the Lawrence Berkeley Laboratory will be the first of the new generation of dedicated synchrotron light sources to be put into operation. Specially designed insertion devices will be required to realize the high brightness photon beams made possible by the low emittance of the electron beam. The complement of insertion devices on the ALS will include undulators with periods as short as 3.9 cm and one or more high field wigglers. The first device to be designed is a 5 m long, 5 cm period, hybrid undulator. The goal of very high brightness and high harmonic output imposes unusually tight tolerances on the magnetic field quality and thus on the mechanical structure. The design process, using a generic structure for all undulators, is described. 5 refs., 4 figs., 1 tab.

  14. Convective heat transfer enhancement inside tubes using inserted helical coils

    NASA Astrophysics Data System (ADS)

    Ali, R. K.; Sharafeldeen, M. A.; Berbish, N. S.; Moawed, M. A.

    2016-01-01

    Convective heat transfer was experimentally investigated in tubes with helical coils inserts in turbulent flow regime within Reynolds number range of 14400 ≤ Re ≤ 42900. The present work aims to extend the experimental data available on wire coil inserts to cover wire diameter ratio from 0.044 to 0.133 and coil pitch ratio from 1 to 5. Uniform heat flux was applied to the external surface of the tube and air was selected as fluid. The effects of Reynolds number and wire diameter and coil pitch ratios on the Nusselt number and friction factor were studied. The enhancement efficiency and performance criteria ranges are of (46.9-82.6%) and (100.1-128%) within the investigated range of the different parameters, respectively. Correlations are obtained for the average Nusselt number and friction factor utilizing the present measurements within the investigated range of geometrical parameters and Re.

  15. Calculation of magnetic error fields in hybrid insertion devices

    NASA Astrophysics Data System (ADS)

    Savoy, R.; Halbach, K.; Hassenzahl, W.; Hoyer, E.; Humphries, D.; Kincaid, B.

    1990-05-01

    The Advanced Light Source (ALS) at the Lawrence Berkeley Laboratory requires insertion devices with fields sufficiently accurate to take advantage of the small emittance of the ALS electron beam. To maintain the spectral performance of the synchrotron radiation and to limit steering effects on the electron beam these errors must be smaller than 0.25%. This paper develops a procedure for calculating the steering error due to misalignment of the easy axis of the permanent-magnet material. The procedure is based on a three-dimensional theory of the design of hybrid insertion devices developed by one of us. The acceptable tolerance for easy axis misalignment is found for a 5-cm-period undulator proposed for the ALS.

  16. Massive reflux and aspiration after radiographically inserted gastrostomy tube placement.

    PubMed

    Chesoni, Sandra A; Bach, John R; Okamura, Erica Mia

    2015-01-01

    To the authors' knowledge, fatal postgastrostomy aspiration within 2 days of enteral nutrition has not been reported. The authors report consecutive cases of severe postgastrotomy aspiration with one being fatal for a 26-yr-old with Duchenne muscular dystrophy 2 days after initiation of gastrostomy feedings. Previous to these consecutive radiographically inserted gastrostomies, all gastrotomies at the institution were percutaneous endoscopic gastrostomies or open gastrostomies. Radiographically inserted gastrostomy tubes have an increased likelihood of being oriented toward the esophagus as opposed to the duodenum, which may increase the risk for reflux. Elimination of invasive airway tubes should be delayed until after gastrostomy feedings are documented to be well tolerated. Oximetry and repeated measurements of vital capacity can suggest changes in the status of airway clearance.

  17. The Ziegler—Natta olefin insertion reaction for cationic metals

    NASA Astrophysics Data System (ADS)

    Jensen, Vidar R.; Siegban, Per E. M.

    1993-09-01

    The catalytic Ziegler—Natta polymerization reaction has been studied for a set of metal cations, in order to identify the role of the positive charge on this process. Geometry optimizations have been performed for the reactant metal—methyl systems, the π-coordinated olefin systems, the transition states for the olefin insertion and finally for the product metal—propyl systems. All valence electrons are correlated. The cations selected for this study are the transition metals Zr + and Ti +, the non-transition metals Be +, Mg +, Al + and finally also Si +. The transition metal cations are found to have very low barriers for the insertion, but the lowest barrier is actually found for Be +. The results are discussed in terms of the ionization energies and the accessibility to valence p and d orbitals. Comparisons are made to previous theoretical work on cationic model systems.

  18. Flexibility in targeting and insertion during bacterial membrane protein biogenesis

    SciTech Connect

    Bloois, Edwin van; Hagen-Jongman, Corinne M. ten; Luirink, Joen

    2007-10-26

    The biogenesis of Escherichia coli inner membrane proteins (IMPs) is assisted by targeting and insertion factors such as the signal recognition particle (SRP), the Sec-translocon and YidC with translocation of (large) periplasmic domains energized by SecA and the proton motive force (pmf). The use of these factors and forces is probably primarily determined by specific structural features of an IMP. To analyze these features we have engineered a set of model IMPs based on endogenous E. coli IMPs known to follow distinct targeting and insertion pathways. The modified model IMPs were analyzed for altered routing using an in vivo protease mapping approach. The data suggest a facultative use of different combinations of factors.

  19. Wear mechanisms of milling inserts: Dry and wet cutting

    SciTech Connect

    Gu, J.; Tung, S.C.; Barber, G.C.

    1998-07-01

    There is less literature on wear of milling tools than on wear of turning tools because milling is one of the most complicated machining operations. The intermittent milling action creates mechanical and thermal surges that distinguish milling from single-point machining. A systematic tool life study for face milling inserts was conducted with and without coolant. Workpieces made of 4140 steel were cut by C5 grade carbide inserts under various cutting conditions. The comparison between dry and wet cutting shows that caution should be taken when applying a coolant for milling operations. Special tests should be carried out in evaluating potential coolant candidates. It is not always true that coolant enhances tool life for milling. Wear mechanisms are presented by means of war maps. Identified wear mechanisms are: micro-attrition, micro-abrasion, mechanical fatigue, thermal fatigue, thermal pitting, and edge chipping.

  20. Chromaticity and low-β insertions in particle colliders

    NASA Astrophysics Data System (ADS)

    Autin, Bruno

    1990-12-01

    Besides its energy range, the main characteristic of a particle collider is its luminosity. A very important piece of equipment to achieve high luminosities is a low-β insertion, a device which focuses the beam down to a tiny spot size. The focusing limits are not only technological, they are also associated with the aberrations of such systems. This paper deals with the chromatic aberrations. In a first part, the treatment of the focusing errors is reviewed and a formalism using symbolic computation is set up so that high-order perturbations can be evaluated, although results are limited to the second-order betatron tune shift and to its interpretation. In a second part, a completely analytical theory of the final doublet of a low-β insertion is given for a special model of flat-beam focusing.

  1. Insertion sequences enrichment in extreme Red sea brine pool vent.

    PubMed

    Elbehery, Ali H A; Aziz, Ramy K; Siam, Rania

    2017-03-01

    Mobile genetic elements are major agents of genome diversification and evolution. Limited studies addressed their characteristics, including abundance, and role in extreme habitats. One of the rare natural habitats exposed to multiple-extreme conditions, including high temperature, salinity and concentration of heavy metals, are the Red Sea brine pools. We assessed the abundance and distribution of different mobile genetic elements in four Red Sea brine pools including the world's largest known multiple-extreme deep-sea environment, the Red Sea Atlantis II Deep. We report a gradient in the abundance of mobile genetic elements, dramatically increasing in the harshest environment of the pool. Additionally, we identified a strong association between the abundance of insertion sequences and extreme conditions, being highest in the harshest and deepest layer of the Red Sea Atlantis II Deep. Our comparative analyses of mobile genetic elements in secluded, extreme and relatively non-extreme environments, suggest that insertion sequences predominantly contribute to polyextremophiles genome plasticity.

  2. Needle Insertion with Duty-Cycled Rotation into Multiple Media

    PubMed Central

    Lehocky, Craig A.; Riviere, Cameron N.

    2012-01-01

    Thin, flexible needles can be steered along nonlinear paths to reach deep anatomical structures within the human body. This study builds upon previous work involving steering of bevel-tipped needles by inserting while rotating in a duty-cycled fashion. Here we investigate how needle material and radius, duty cycle, and tissue stiffness affect needle curvature. Needles were inserted into media while rotated at a specified duty cycle and the curvature was measured. A linear relationship between duty cycle and curvature was observed across all needle materials and radii, and tissue stiffnesses. Following these observations, we developed a model that encapsulates needle and tissue parameters in order to predict the duty cycle needed to achieve a desired curvature. PMID:23366042

  3. Insertional hypermutation in mineral oil-induced plasmacytomas.

    PubMed

    Knittel, Gero; Metzner, Mirjam; Beck-Engeser, Gabriele; Kan, Ada; Ahrends, Tomasz; Eilat, Dan; Huppi, Konrad; Wabl, Matthias

    2014-09-01

    Unless stimulated by a chronic inflammatory agent, such as mineral oil, plasma cell tumors are rare in young BALB/c mice. This raises the questions: What do inflammatory tissues provide to promote mutagenesis? And what is the nature of mutagenesis? We determined that mineral oil-induced plasmacytomas produce large amounts of endogenous retroelements--ecotropic and polytropic murine leukemia virus and intracisternal A particles. Therefore, plasmacytoma formation might occur, in part, by de novo insertion of these retroelements, induced or helped by the inflammation. We recovered up to ten de novo insertions in a single plasmacytoma, mostly in genes with common retroviral integration sites. Additional integrations accompany tumor evolution from a solid tumor through several generations in cell culture. The high frequency of de novo integrations into cancer genes suggests that endogenous retroelements are coresponsible for plasmacytoma formation and progression in BALB/c mice.

  4. Challenges for Insertion of Structural Nanomaterials in Aerospace Applications

    NASA Technical Reports Server (NTRS)

    Sochi, Emilie J.

    2012-01-01

    In the two decades since Iijima's report on carbon nanotubes (CNT), there has been great interest in realizing the benefits of mechanical properties observed at the nanoscale in large-scale structures. The weight savings possible due to dramatic improvements in mechanical properties relative to state-of-the-art material systems can be game changing for applications like aerospace vehicles. While there has been significant progress in commercial production of CNTs, major aerospace applications that take advantage of properties offered by this material have yet to be realized. This paper provides a perspective on the technical challenges and barriers for insertion of CNTs as an emerging material technology in aerospace applications and proposes approaches that may reduce the typical timeframe for technology maturation and insertion into aerospace structures.

  5. A Medicago truncatula Tobacco Retrotransposon Insertion Mutant Collection with Defects in Nodule Development and Symbiotic Nitrogen Fixation1[W][OA

    PubMed Central

    Pislariu, Catalina I.; D. Murray, Jeremy; Wen, JiangQi; Cosson, Viviane; Muni, RajaSekhara Reddy Duvvuru; Wang, Mingyi; A. Benedito, Vagner; Andriankaja, Andry; Cheng, Xiaofei; Jerez, Ivone Torres; Mondy, Samuel; Zhang, Shulan; Taylor, Mark E.; Tadege, Million; Ratet, Pascal; Mysore, Kirankumar S.; Chen, Rujin; Udvardi, Michael K.

    2012-01-01

    A Tnt1-insertion mutant population of Medicago truncatula ecotype R108 was screened for defects in nodulation and symbiotic nitrogen fixation. Primary screening of 9,300 mutant lines yielded 317 lines with putative defects in nodule development and/or nitrogen fixation. Of these, 230 lines were rescreened, and 156 lines were confirmed with defective symbiotic nitrogen fixation. Mutants were sorted into six distinct phenotypic categories: 72 nonnodulating mutants (Nod−), 51 mutants with totally ineffective nodules (Nod+ Fix−), 17 mutants with partially ineffective nodules (Nod+ Fix+/−), 27 mutants defective in nodule emergence, elongation, and nitrogen fixation (Nod+/− Fix−), one mutant with delayed and reduced nodulation but effective in nitrogen fixation (dNod+/− Fix+), and 11 supernodulating mutants (Nod++Fix+/−). A total of 2,801 flanking sequence tags were generated from the 156 symbiotic mutant lines. Analysis of flanking sequence tags revealed 14 insertion alleles of the following known symbiotic genes: NODULE INCEPTION (NIN), DOESN’T MAKE INFECTIONS3 (DMI3/CCaMK), ERF REQUIRED FOR NODULATION, and SUPERNUMERARY NODULES (SUNN). In parallel, a polymerase chain reaction-based strategy was used to identify Tnt1 insertions in known symbiotic genes, which revealed 25 additional insertion alleles in the following genes: DMI1, DMI2, DMI3, NIN, NODULATION SIGNALING PATHWAY1 (NSP1), NSP2, SUNN, and SICKLE. Thirty-nine Nod− lines were also screened for arbuscular mycorrhizal symbiosis phenotypes, and 30 mutants exhibited defects in arbuscular mycorrhizal symbiosis. Morphological and developmental features of several new symbiotic mutants are reported. The collection of mutants described here is a source of novel alleles of known symbiotic genes and a resource for cloning novel symbiotic genes via Tnt1 tagging. PMID:22679222

  6. Design and optimization of a HTS insert for solenoid magnets

    NASA Astrophysics Data System (ADS)

    Tomassetti, Giordano; de Marzi, Gianluca; Muzzi, Luigi; Celentano, Giuseppe; della Corte, Antonio

    2016-12-01

    With the availability of High-Temperature Superconducting (HTS) prototype cables, based on high-performance REBCO Coated Conductor (CC) tapes, new designs can now be made for large bore high-field inserts in superconducting solenoids, thus extending the magnet operating point to higher magnetic fields. In this work, as an alternative approach to the standard trial-and-error design process, an optimization procedure for a HTS grading section design is proposed, including parametric electro-magnetic and structural analyses, using the ANSYS software coupled with a numerically-efficient optimization algorithm. This HTS grading section is designed to be inserted into a 12 T large bore Low-Temperature Superconducting (LTS) solenoid (diameter about 1 m) to increase the field up to a maximum value of at least 17 T. The optimization variables taken into consideration are the number of turns and layers and the circle-in-square jacket inner diameter in order to minimize the total needed conductor length to achieve a peak field of at least 17 T, while guaranteeing the structural integrity and manufacturing constraints. By means of the optimization, an optimal 360 m total conductor length was found, achieving 17.2 T with an operating current of 22.4 kA and a coil comprised of 18 × 12 turns, shortened of about 20% with respect to the best initial candidate architectural design. The optimal HTS insert has a bore compatible with manufacturing constraints (inner bore radius larger than 30 cm). A scaled HTS insert for validation purposes, with a reduced conductor length, to be tested in an advanced experimental facility currently under construction, is also mentioned.

  7. Crystallographic snapshots of sulfur insertion by lipoyl synthase

    PubMed Central

    McLaughlin, Martin I.; Lanz, Nicholas D.; Goldman, Peter J.; Lee, Kyung-Hoon; Booker, Squire J.; Drennan, Catherine L.

    2016-01-01

    Lipoyl synthase (LipA) catalyzes the insertion of two sulfur atoms at the unactivated C6 and C8 positions of a protein-bound octanoyl chain to produce the lipoyl cofactor. To activate its substrate for sulfur insertion, LipA uses a [4Fe-4S] cluster and S-adenosylmethionine (AdoMet) radical chemistry; the remainder of the reaction mechanism, especially the source of the sulfur, has been less clear. One controversial proposal involves the removal of sulfur from a second (auxiliary) [4Fe-4S] cluster on the enzyme, resulting in destruction of the cluster during each round of catalysis. Here, we present two high-resolution crystal structures of LipA from Mycobacterium tuberculosis: one in its resting state and one at an intermediate state during turnover. In the resting state, an auxiliary [4Fe-4S] cluster has an unusual serine ligation to one of the irons. After reaction with an octanoyllysine-containing 8-mer peptide substrate and 1 eq AdoMet, conditions that allow for the first sulfur insertion but not the second insertion, the serine ligand dissociates from the cluster, the iron ion is lost, and a sulfur atom that is still part of the cluster becomes covalently attached to C6 of the octanoyl substrate. This intermediate structure provides a clear picture of iron–sulfur cluster destruction in action, supporting the role of the auxiliary cluster as the sulfur source in the LipA reaction and describing a radical strategy for sulfur incorporation into completely unactivated substrates. PMID:27506792

  8. Evolution of implantable and insertable drug delivery systems.

    PubMed

    Kleiner, Lothar W; Wright, Jeremy C; Wang, Yunbing

    2014-05-10

    The paper describes the development of implantable and insertable drug delivery systems (IDDS) from their early stage in the 1960s until the current stage in the 2010s. It gives a detailed summary of non-degradable and biodegradable systems and their applications in different areas such as vascular disease treatment, birth control, cancer treatment, and eye disease treatment. It also describes the development of various implantable pump systems and some other atypical IDDS, the challenges and the future of IDDS.

  9. Protease-sensitive prions with 144-bp insertion mutations

    PubMed Central

    Puoti, Gianfranco; Yuan, Jue; Qing, Liuting; Wang, Heming; Kong, Qingzhong; Gambetti, Pierluigi; Zou, Wen-Quan

    2013-01-01

    Insertion of 144-base pair (bp) containing six extra octapeptide repeats between residues 51 and 91 of prion protein (PrP) gene is associated with inherited prion diseases. Most cases linked to this insertion examined by Western blotting showed detectable proteinase K-resistant PrPSc (rPrPSc) resembling PrPSc type 1 and type 2 in sporadic Creutzfeldt-Jakob disease (sCJD), or PrP7-8 in Gerstmann-Sträussler-Scheinker disease. However, cases lacking detectable rPrPSc also have been reported. Which PrP conformer is associated with neuropathological changes in the cases without detectable rPrPSc remains to be determined. Here we report that while all six but one subjects with the 144-bp insertion mutations examined display the pathognomonic PrP patches in the cerebellum, one of them exhibits no detectable typical rPrPSc even in PrPSc-enriched preparations. Instead, a large amount of abnormal PrP is captured from this case by gene 5 protein and sodium phosphotungstate, reagents that have been proved to specifically capture abnormal PrP. All captured abnormal PrP from the cerebellum and other brain regions is virtually sensitive to PK-digestion (termed sPrPSc). The presence of the predominant sPrPSc but absence of rPrPSc in this 144-bp insertion-linked inherited CJD case suggests that mutant sPrPSc is the main component of the PrP deposit patches and sPrPSc is sufficient to cause neurotoxicity and prion disease. PMID:23515139

  10. Accurate Insertion Loss Measurements of the Juno Patch Array Antennas

    NASA Technical Reports Server (NTRS)

    Chamberlain, Neil; Chen, Jacqueline; Hodges, Richard; Demas, John

    2010-01-01

    This paper describes two independent methods for estimating the insertion loss of patch array antennas that were developed for the Juno Microwave Radiometer instrument. One method is based principally on pattern measurements while the other method is based solely on network analyzer measurements. The methods are accurate to within 0.1 dB for the measured antennas and show good agreement (to within 0.1dB) of separate radiometric measurements.

  11. Theoretical model for electrophilic oxygen atom insertion into hydrocarbons

    SciTech Connect

    Bach, R.D.; Su, M.D. ); Andres, J.L. Wayne State Univ., Detroit, MI ); McDouall, J.J.W. )

    1993-06-30

    A theoretical model suggesting the mechanistic pathway for the oxidation of saturated-alkanes to their corresponding alcohols and ketones is described. Water oxide (H[sub 2]O-O) is employed as a model singlet oxygen atom donor. Molecular orbital calculations with the 6-31G basis set at the MP2, QCISD, QCISD(T), CASSCF, and MRCI levels of theory suggest that oxygen insertion by water oxide occurs by the interaction of an electrophilic oxygen atom with a doubly occupied hydrocarbon fragment orbital. The electrophilic oxygen approaches the hydrocarbon along the axis of the atomic carbon p orbital comprising a [pi]-[sub CH(2)] or [pi]-[sub CHCH(3)] fragment orbital to form a carbon-oxygen [sigma] bond. A concerted hydrogen migration to an adjacent oxygen lone pair of electrons affords the alcohol insertion product in a stereoselective fashion with predictable stereochemistry. Subsequent oxidation of the alcohol to a ketone (or aldehyde) occurs in a similar fashion and has a lower activation barrier. The calculated (MP4/6-31G*//MP2/6-31G*) activation barriers for oxygen atom insertion into the C-H bonds of methane, ethane, propane, butane, isobutane, and methanol are 10.7, 8.2, 3.9, 4.8, 4.5, and 3.3 kcal/mol, respectively. We use ab initio molecular orbital calculations in support of a frontier MO theory that provides a unique rationale for both the stereospecificity and the stereoselectivity of insertion of electrophilic oxygen and related electrophiles into the carbon-hydrogen bond. 13 refs., 7 figs., 2 tabs.

  12. Osteometry at muscle origin and insertion in sex determination.

    PubMed

    France, D L

    1988-08-01

    Multivariate statistical analyses were performed on 22 size measurements of the humerus from five sample populations: Sudanese Nubians, Arikara, Pecos Pueblos, American blacks, and American whites. The effects of muscle use ("occupational" differences) on identification of sex in this long bone are examined, particularly at points of muscle origins and insertions. The ramifications of the results of this research on sexual dimorphism studies of other bones are discussed.

  13. Non-harmful insertion of data mimicking computer network attacks

    DOEpatents

    Neil, Joshua Charles; Kent, Alexander; Hash, Jr, Curtis Lee

    2016-06-21

    Non-harmful data mimicking computer network attacks may be inserted in a computer network. Anomalous real network connections may be generated between a plurality of computing systems in the network. Data mimicking an attack may also be generated. The generated data may be transmitted between the plurality of computing systems using the real network connections and measured to determine whether an attack is detected.

  14. The first insertion devices at SSRL - some personal recollections

    SciTech Connect

    Winick, H.

    1995-02-01

    The author recounts his experiences with insertion devices at the Stanford Synchrotron Radiation Laboratory. His first experiences with wigglers occured at the Cambridge Electron Accelerator, and was carried over to SSRL with the proposal for a six pole electromagnetic wiggler. Most modern undulators, and many wigglers are now designed around permanent magnets, and the origin of this transition at SSRL was rather fortuitous and humorous. It reflects some of the personality characteristics of Klaus Halbach.

  15. Development of an ultrasound-guided PICC insertion service.

    PubMed

    Nicholson, Jackie

    Use of the peripherally inserted central catheter (PICC) has gained popularity over the past 30 years in the USA and over the past 15 years in the UK. The PICC was initially used for specific purposes, such as the delivery of parenteral nutrition and chemotherapy, but its use has expanded both within the acute hospital and community settings. This article describes an ultrasound-guided PICC insertion service that has been set up in a district general hospital to expand the service within the oncology department and to begin a service for non-oncology patients. As expected, patients who have benefited from the development of this service include those requiring parenteral nutrition, chemotherapy and intravenous therapy in the community. However, retrospective audit has shown that the largest group of patients to benefit from the ultrasound-guided PICC insertion service was those with difficult venous access for a variety of reasons. Some of these reasons are discussed, along with an overview of the benefits the service has provided to other groups of patients, and associated benefits to the hospital trust.

  16. Examining Prebiotic Chemistry Using O(^1D) Insertion Reactions

    NASA Astrophysics Data System (ADS)

    Hays, Brian M.; Laas, Jacob C.; Weaver, Susanna L. Widicus

    2013-06-01

    Aminomethanol, methanediol, and methoxymethanol are all prebiotic molecules expected to form via photo-driven grain surface chemistry in the interstellar medium (ISM). These molecules are expected to be precursors for larger, biologically-relevant molecules in the ISM such as sugars and amino acids. These three molecules have not yet been detected in the ISM because of the lack of available rotational spectra. A high resolution (sub)millimeter spectrometer coupled to a molecular source is being used to study these molecules using O(^1D) insertion reactions. The O(^1D) chemistry is initiated using an excimer laser, and the products of the insertion reactions are adiabatically cooled using a supersonic expansion. Experimental parameters are being optimized by examination of methanol formed from O(^1D) insertion into methane. Theoretical studies of the structure and reaction energies for aminomethanol, methanediol, and methoxymethanol have been conducted to guide the laboratory studies once the methanol experiment has been optimized. The results of the calculations and initial experimental results will be presented.

  17. Levator Ani Muscle Anatomy Evaluated by Origin-Insertion Pairs

    PubMed Central

    Kearney, Rohna; Sawhney, Raja; DeLancey, John O. L.

    2005-01-01

    OBJECTIVE To examine the published literature and suggest a resolution to the confusion that exists in levator ani muscle descriptions and terminology. DATA SOURCES A MEDLINE search was performed using the keyword “levator ani,” limited to human studies in women. References found in these articles were reviewed to identify research reported before 1966 and articles not included in the search. STUDY SELECTION Studies were accepted if they contained direct observations of female specimens. Only those that contained specific descriptions or illustrations of the muscle origins and insertions in more than 5 female specimens were included. Review of 265 human studies yielded 9 qualifying articles, and reference tracing disclosed 3 additional reports. TABULATION, INTEGRATION, AND RESULTS The literature review identified 5 origin-insertion pairs consistently described in studies directly examining the levator ani muscle in women, but 16 terms were used by authors for these 5 components of the muscle. Labeled illustrations often provided more precise information than was provided in the text. Terms were reviewed for inconsistencies of usage and appropriateness of term choice. The terms puboperineal, pubovaginal, and puboanal (for components of the pubovisceral [“pubococcygeal”] muscle), along with puborectal and iliococcygeal, are sufficient to describe the divisions of the levator ani muscle. CONCLUSION Although there was great diversity and conflict in terms chosen among the original articles, the number of origin and insertion pairs was relatively consistent among authors and confusion can be avoided by standardizing terminology. PMID:15229017

  18. Effects of Tools Inserted through Snake-like Surgical Manipulators

    PubMed Central

    Murphy, Ryan J.; Otake, Yoshito; Wolfe, Kevin C.; Taylor, Russell H.; Armand, Mehran

    2015-01-01

    Snake-like manipulators with a large, open lumen can offer improved treatment alternatives for minimally- and less-invasive surgeries. In these procedures, surgeons use the manipulator to introduce and control flexible tools in the surgical environment. This paper describes a predictive algorithm for estimating manipulator configuration given tip position for nonconstant curvature, cable-driven manipulators using energy minimization. During experimental bending of the manipulator with and without a tool inserted in its lumen, images were recorded from an overhead camera in conjunction with actuation cable tension and length. To investigate the accuracy, the estimated manipulator configuration from the model and the ground-truth configuration measured from the image were compared. Additional analysis focused on the response differences for the manipulator with and without a tool inserted through the lumen. Results indicate that the energy minimization model predicts manipulator configuration with an error of 0.24 ± 0.22mm without tools in the lumen and 0.24 ± 0.19mm with tools in the lumen (no significant difference, p = 0.81). Moreover, tools did not introduce noticeable perturbations in the manipulator trajectory; however, there was an increase in requisite force required to reach a configuration. These results support the use of the proposed estimation method for calculating the shape of the manipulator with an tool inserted in its lumen when an accuracy range of at least 1mm is required. PMID:25571571

  19. Internal carotid artery rupture caused by carotid shunt insertion

    PubMed Central

    Illuminati, Giulio; Caliò, Francesco G.; Pizzardi, Giulia; Vietri, Francesco

    2015-01-01

    Introduction Shunting is a well-accepted method of maintaining cerebral perfusion during carotid endarterectomy (CEA). Nonetheless, shunt insertion may lead to complications including arterial dissection, embolization, and thrombosis. We present a complication of shunt insertion consisting of arterial wall rupture, not reported previously. Presentation of case A 78-year-old woman underwent CEA combined with coronary artery bypass grafting (CABG). At the time of shunt insertion an arterial rupture at the distal tip of the shunt was detected and was repaired via a small saphenous vein patch. Eversion CEA and subsequent CABG completed the procedure whose postoperative course was uneventful. Discussion Shunting during combined CEA-CABG may be advisable to assure cerebral protection from possible hypoperfusion due to potential hemodynamic instability of patients with severe coronary artery disease. Awareness and prompt management of possible shunt-related complications, including the newly reported one, may contribute to limiting their harmful effect. Conclusion Arterial wall rupture is a possible, previously not reported, shunt-related complication to be aware of when performing CEA. PMID:26255001

  20. Retroviral gene insertion in breast milk mediated lymphomagenesis.

    PubMed

    Chakraborty, Joana; Okonta, Henry; Bagalb, Hussein; Lee, Soon Jin; Fink, Brian; Changanamkandat, Rajesh; Duggan, Joan

    2008-07-20

    We have demonstrated breast milk transmitted MoMuLV-ts1 retrovirus infection and subsequent lymphoma development in offspring of uninfected mothers suckled by infected surrogate mothers. Additionally, we have shown that the lymphoma development occurs as a result of viral gene integration into host genome. A total of 146 pups from Balb/C mice were divided into 5 groups; one control and 4 experimental. All offspring suckled from surrogate infected or control mothers, except one group of infected pups left with their biological mothers. Thirteen of 91 infected pups developed lymphoma. Inverse-PCR, DNA cloning, and quantitative real-time PCR (qRT-PCR) were used to study the virus integration sites (VIS) and alterations in gene expression. VIS were randomly distributed throughout the genome. The majority of insertion sites were found in chromosomes 10, 12 and 13. A total of 209 proviral genomic insertion sites were located with 52 intragenic and 157 intergenic sites. We have identified 29 target genes. Four genes including Tacc3, Aurka, Gfi1 and Ahi1 showed the maximum upregulation of mRNA expression. These four genes can be considered as candidate genes based on their association with cancer. Upregulation of these genes may be involved in this type of lymphoma development. This model provides an important opportunity to gain insight into the relationship of viral gene insertion into host genome and development of lymphoma via natural transmission route such as breast milk.

  1. ADC's Insertion Devices and Magnetic Measurement Systems Capabilities

    NASA Astrophysics Data System (ADS)

    Deyhim, A.; Kulesza, J.

    2013-03-01

    In this paper Advance Design Consulting USA, Inc. (ADC) will discuss ADC's major improved capabilities for building Wiggler Insertion Devices, Undulator Planar Devices, Elliptical Polarizing Undulators (EPU), In-Vacuum Undulators (IVU), Cryogenically Cooled in-vacuum Undulators (CPMU), Super Conductive Undulator, and Insertion Device Magnetic Measurement Systems. ADC has designed, built and delivered Insertion Devices and Magnetic Measurement Systems to such facilities as MAX-lab (two EPUs, a Planar, and Measurement System), ALBA and ASP (Wigglers), BNL (CPMU), SSRF (two IVUs and a Measurement System), PAL (one IVU and Measurement System), NSRRC (one 4m EPU), and SRC (Planar and EPU). ADC's magnetic field measurement system is a sophisticated and sensitive machine for the measurement of magnetic fields in undulators (Planar and EPU), wigglers and in-vacuum ID units. The magnetic fields are measured using 3 axis hall-effect probes, mounted orthogonally, to a thin wand. The wand is mounted to a carriage that rides on vacuum air bearings. The base is granite. A flip coil is provided on two vertical towers with X, Y and Theta axes. Special software is provided to assist in homing, movement, and data collection and analysis.

  2. Optimising daytime deliveries when inducing labour using prostaglandin vaginal inserts

    PubMed Central

    Miller, Hugh; Goetzl, Laura; Wing, Deborah A.; Powers, Barbara; Rugarn, Olof

    2016-01-01

    Abstract Objective: To determine induction start time(s) that would maximise daytime deliveries when using prostaglandin vaginal inserts. Methods: Women enrolled into the Phase III trial, EXPEDITE (clinical trial registration: NCT01127581), had labour induced with either a misoprostol or dinoprostone vaginal insert (MVI or DVI). A secondary analysis was conducted to determine the optimal start times for induction by identifying the 12-h period with the highest proportion of deliveries by parity and treatment. Results: Optimal start times for achieving daytime deliveries when using MVI appear to be 19:00 in nulliparae and 23:00 in multiparae. Applying these start times, the median time of onset of active labour would be approximately 08:30 for both parities and the median time of delivery would be the following day at approximately 16:30 for nulliparae and 12:00 (midday) for multiparae. Optimal start times when using DVI appear to be 07:00 for nulliparae and 23:00 for multiparae. Using these start times, the median time of onset of active labour would be the following day at approximately 04:00 and 11:50, and the median time of delivery would be approximately 13:40 and 16:10, respectively. Conclusions: When optimising daytime deliveries, different times to initiate induction of labour may be appropriate depending on parity and the type of retrievable prostaglandin vaginal insert used. PMID:25758619

  3. Development of Needle Insertion Manipulator for Central Venous Catheterization

    NASA Astrophysics Data System (ADS)

    Kobayashi, Yo; Hong, Jaesung; Hamano, Ryutaro; Hashizume, Makoto; Okada, Kaoru; Fujie, Masakatsu G.

    Central venous catheterization is a procedure, which a doctor insert a catheter into the patient’s vein for transfusion. Since there are risks of bleeding from arterial puncture or pneumothorax from pleural puncture. Physicians are strictly required to make needle reach up into the vein and to stop the needle in the middle of vein. We proposed a robot system for assisting the venous puncture, which can relieve the difficulties in conventional procedure, and the risks of complication. This paper reports the design structuring and experimental results of needle insertion manipulator. First, we investigated the relationship between insertion force and angle into the vein. The results indicated that the judgment of perforation using the reaction force is possible in case where the needling angle is from 10 to 20 degree. The experiment to evaluate accuracy of the robot also revealed that it has beyond 0.5 mm accuracy. We also evaluated the positioning accuracy in the ultrasound images. The results displays that the accuracy is beyond 1.0 mm and it has enough for venous puncture. We also carried out the venous puncture experiment to the phantom and confirm our manipulator realized to make needle reach up into the vein.

  4. Coaxial needle insertion assistant with enhanced force feedback.

    PubMed

    De Lorenzo, Danilo; Koseki, Yoshihiko; De Momi, Elena; Chinzei, Kiyoyuki; Okamura, Allison M

    2013-02-01

    Many medical procedures involving needle insertion into soft tissues, such as anesthesia, biopsy, brachytherapy, and placement of electrodes, are performed without image guidance. In such procedures, haptic detection of changing tissue properties at different depths during needle insertion is important for needle localization and detection of subsurface structures. However, changes in tissue mechanical properties deep inside the tissue are difficult for human operators to sense, because the relatively large friction force between the needle shaft and the surrounding tissue masks the smaller tip forces. A novel robotic coaxial needle insertion assistant, which enhances operator force perception, is presented. This one-degree-of-freedom cable-driven robot provides to the operator a scaled version of the force applied by the needle tip to the tissue, using a novel design and sensors that separate the needle tip force from the shaft friction force. The ability of human operators to use the robot to detect membranes embedded in artificial soft tissue was tested under the conditions of 1) tip force and shaft force feedback, and 2) tip force only feedback. The ratio of successful to unsuccessful membrane detections was significantly higher (up to 50%) when only the needle tip force was provided to the user.

  5. Genome annotation in the presence of insertional RNA editing

    PubMed Central

    Beargie, Christina; Liu, Tsunglin; Corriveau, Mark; Lee, Ha Youn; Gott, Jonatha; Bundschuh, Ralf

    2008-01-01

    Motivation: Insertional RNA editing renders gene prediction very difficult compared to organisms without such RNA editing. A case in point is the mitochondrial genome of Physarum polycephalum in which only about one-third of the number of genes that are to be expected given its length are annotated. Thus, gene prediction methods that explicitly take into account insertional editing are needed for successful annotation of such genomes. Results: We annotate the mitochondrial genome of P.polycephalum using several different approaches for gene prediction in organisms with insertional RNA editing. We computationally validate our annotations by comparing the results from different methods against each other and as proof of concept experimentally validate two of the newly predicted genes. We more than double the number of annotated putative genes in this organism and find several intriguing candidate genes that are not expected in a mitochondrial genome. Availability: The C source code of the programs described here are available upon request from the corresponding author. Contact: bundschuh@mps.ohio-state.edu PMID:18819938

  6. Local Dispersion Insert: The {gamma}T knob for accelerators

    SciTech Connect

    Visnjic, V.

    1994-05-01

    The Local Dispersion Insert is a novel three-quadrupole cell which produces local dispersion and beta waves while leaving all other beam parameters unchanged. The obtained dispersion bump can be effectively used to control the transition energy and the so-called {gamma}T jump of an accelerator. Since the induced beta wave is local and the tune shift is zero, the Local Dispersion Insert is an extremely efficient cell, typically requiring only a small number of magnets in order to achieve a {gamma}T jump. Since the desired {gamma}T jump can be obtained by locally decreasing the dispersion, the well known problems which accompany the conventional {gamma}T jump schemes, like large maximal dispersion, transverse emittance growth etc. are absent. The paper contains three applications of the Local Dispersion Insert: the {gamma}T jump in the Main Injector, the {eta} adjustment in the Antiproton Debuncher (both at Fermi National Accelerator Laboratory), and the {gamma}T jump in the Relativistic Heavy Ion Collider at Brookhaven National Laboratory.

  7. Comparing Intravenous Insertion Instructional Methods with Haptic Simulators

    PubMed Central

    Malecha, Ann

    2017-01-01

    Objective. The objective of this review was to compare traditional intravenous (IV) insertion instructional methods with the use of haptic IV simulators. Design. An integrative research design was used to analyze the current literature. Data Sources. A search was conducted using key words intravenous (IV) insertion or cannulation or venipuncture and simulation from 2000 to 2015 in the English language. The databases included Academic Search Complete, CINAHL Complete, Education Resource Information Center, and Medline. Review Methods. Whittemore and Knafl's (2005) strategies were used to critique the articles for themes and similarities. Results. Comparisons of outcomes between traditional IV instructional methods and the use of haptic IV simulators continue to show various results. Positive results indicate that the use of the haptic IV simulator decreases both band constriction and total procedure time. While students are satisfied with practicing on the haptic simulators, they still desire faculty involvement. Conclusion. Combining the haptic IV simulator with practical experience on the IV arm may be the best practice for learning IV insertion. Research employing active learning strategies while using a haptic IV simulator during the learning process may reduce cost and faculty time. PMID:28250987

  8. A comprehensive map of mobile element insertion polymorphisms in humans.

    PubMed

    Stewart, Chip; Kural, Deniz; Strömberg, Michael P; Walker, Jerilyn A; Konkel, Miriam K; Stütz, Adrian M; Urban, Alexander E; Grubert, Fabian; Lam, Hugo Y K; Lee, Wan-Ping; Busby, Michele; Indap, Amit R; Garrison, Erik; Huff, Chad; Xing, Jinchuan; Snyder, Michael P; Jorde, Lynn B; Batzer, Mark A; Korbel, Jan O; Marth, Gabor T

    2011-08-01

    As a consequence of the accumulation of insertion events over evolutionary time, mobile elements now comprise nearly half of the human genome. The Alu, L1, and SVA mobile element families are still duplicating, generating variation between individual genomes. Mobile element insertions (MEI) have been identified as causes for genetic diseases, including hemophilia, neurofibromatosis, and various cancers. Here we present a comprehensive map of 7,380 MEI polymorphisms from the 1000 Genomes Project whole-genome sequencing data of 185 samples in three major populations detected with two detection methods. This catalog enables us to systematically study mutation rates, population segregation, genomic distribution, and functional properties of MEI polymorphisms and to compare MEI to SNP variation from the same individuals. Population allele frequencies of MEI and SNPs are described, broadly, by the same neutral ancestral processes despite vastly different mutation mechanisms and rates, except in coding regions where MEI are virtually absent, presumably due to strong negative selection. A direct comparison of MEI and SNP diversity levels suggests a differential mobile element insertion rate among populations.

  9. Comparing Intravenous Insertion Instructional Methods with Haptic Simulators.

    PubMed

    McWilliams, Lenora A; Malecha, Ann

    2017-01-01

    Objective. The objective of this review was to compare traditional intravenous (IV) insertion instructional methods with the use of haptic IV simulators. Design. An integrative research design was used to analyze the current literature. Data Sources. A search was conducted using key words intravenous (IV) insertion or cannulation or venipuncture and simulation from 2000 to 2015 in the English language. The databases included Academic Search Complete, CINAHL Complete, Education Resource Information Center, and Medline. Review Methods. Whittemore and Knafl's (2005) strategies were used to critique the articles for themes and similarities. Results. Comparisons of outcomes between traditional IV instructional methods and the use of haptic IV simulators continue to show various results. Positive results indicate that the use of the haptic IV simulator decreases both band constriction and total procedure time. While students are satisfied with practicing on the haptic simulators, they still desire faculty involvement. Conclusion. Combining the haptic IV simulator with practical experience on the IV arm may be the best practice for learning IV insertion. Research employing active learning strategies while using a haptic IV simulator during the learning process may reduce cost and faculty time.

  10. Central line infections - hospitals

    MedlinePlus

    ... infection; CVC - infection; Central venous device - infection; Infection control - central line infection; Nosocomial infection - central line infection; Hospital acquired infection - central line infection; Patient safety - central ...

  11. Role of the import motor in insertion of transmembrane segments by the mitochondrial TIM23 complex.

    PubMed

    Popov-Čeleketić, Dušan; Waegemann, Karin; Mapa, Koyeli; Neupert, Walter; Mokranjac, Dejana

    2011-06-01

    The TIM23 complex mediates translocation of proteins across, and their lateral insertion into, the mitochondrial inner membrane. Translocation of proteins requires both the membrane-embedded core of the complex and its ATP-dependent import motor. Insertion of some proteins, however, occurs in the absence of ATP, questioning the need for the import motor during lateral insertion. We show here that the import motor associates with laterally inserted proteins even when its ATPase activity is not required. Furthermore, our results suggest a role for the import motor in lateral insertion. Thus, the import motor is involved in ATP-dependent translocation and ATP-independent lateral insertion.

  12. Insertion of Foreign DNA into an Established Mammalian Genome Can Alter the Methylation of Cellular DNA Sequences†

    PubMed Central

    Remus, Ralph; Kämmer, Christina; Heller, Hilde; Schmitz, Birgit; Schell, Gudrun; Doerfler, Walter

    1999-01-01

    The insertion of adenovirus type 12 (Ad12) DNA into the hamster genome and the transformation of these cells by Ad12 can lead to marked alterations in the levels of DNA methylation in several cellular genes and DNA segments. Since such alterations in DNA methylation patterns are likely to affect the transcription patterns of cellular genes, it is conceivable that these changes have played a role in the generation or the maintenance of the Ad12-transformed phenotype. We have now isolated clonal BHK21 hamster cell lines that carry in their genomes bacteriophage λ and plasmid pSV2neo DNAs in an integrated state. Most of these cell lines contain one or multiple copies of integrated λ DNA, which often colocalize with the pSV2neo DNA, usually in a single chromosomal site as determined by the fluorescent in situ hybridization technique. In different cell lines, the loci of foreign DNA insertion are different. The inserted bacteriophage λ DNA frequently becomes de novo methylated. In some of the thus-generated hamster cell lines, the levels of DNA methylation in the retrotransposon genomes of the endogenous intracisternal A particles (IAP) are increased in comparison to those in the non-λ-DNA-transgenic BHK21 cell lines. These changes in the methylation patterns of the IAP subclone I (IAPI) segment have been documented by restriction analyses with methylation-sensitive restriction endonucleases followed by Southern transfer hybridization and phosphorimager quantitation. The results of genomic sequencing experiments using the bisulfite protocol yielded additional evidence for alterations in the patterns of DNA methylation in selected segments of the IAPI sequences. In these experiments, the nucleotide sequences in >330 PCR-generated cloned DNA molecules were determined. Upon prolonged cultivation of cell lines with altered cellular methylation patterns, these differences became less apparent, perhaps due to counterselection of the transgenic cells. The possibility

  13. Modified mariner transposons for random inducible-expression insertions and transcriptional reporter fusion insertions in Bacillus subtilis.

    PubMed

    Pozsgai, Eric R; Blair, Kris M; Kearns, Daniel B

    2012-02-01

    Transposons are mobile genetic elements bounded by insertion sequences that are recognized by a specific mobilizing transposase enzyme. The transposase may mobilize not only the insertion sequences but also intervening DNA. mariner is a particularly efficient transposon for the random chromosomal integration of genes and insertional mutagenesis. Here, we modify an existing mariner transposon, TnYLB, such that it can easily be genetically manipulated and introduced into Bacillus subtilis. We generate a series of three new mariner derivatives that mobilize spectinomycin, chloramphenicol, and kanamycin antibiotic resistance cassettes. Furthermore, we generate a series of transposons with a strong, outward-oriented, optionally isopropyl-β-D-thiogalactopyranoside (IPTG)-inducible promoter for the random overexpression of neighboring genes and a series of transposons with a promoterless lacZ gene for the random generation of transcriptional reporter fusions. We note that the modification of the base transposon is not restricted to B. subtilis and should be applicable to any mariner-compatible host organism, provided that in vitro mutagenesis or an in vivo species-specific delivery vector is employed.

  14. The nature of the transition mismatches with Watson-Crick architecture: the G*·T or G·T* DNA base mispair or both? A QM/QTAIM perspective for the biological problem.

    PubMed

    Brovarets', Ol'ha O; Hovorun, Dmytro M

    2015-01-01

    This study provides the first accurate investigation of the tautomerization of the biologically important guanine*·thymine (G*·T) DNA base mispair with Watson-Crick geometry, involving the enol mutagenic tautomer of the G and the keto tautomer of the T, into the G·T* mispair (∆G = .99 kcal mol(-1), population = 15.8% obtained at the MP2 level of quantum-mechanical theory in the continuum with ε = 4), formed by the keto tautomer of the G and the enol mutagenic tautomer of the T base, using DFT and MP2 methods in vacuum and in the weakly polar medium (ε = 4), characteristic for the hydrophobic interfaces of specific protein-nucleic acid interactions. We were first able to show that the G*·T↔G·T* tautomerization occurs through the asynchronous concerted double proton transfer along two antiparallel O6H···O4 and N1···HN3 H-bonds and is assisted by the third N2H···O2 H-bond, that exists along the entire reaction pathway. The obtained results indicate that the G·T* base mispair is stable from the thermodynamic point of view complex, while it is dynamically unstable structure in vacuum and dynamically stable structure in the continuum with ε = 4 with lifetime of 6.4·10(-12) s, that, on the one side, makes it possible to develop all six low-frequency intermolecular vibrations, but, on the other side, it is by three orders less than the time (several ns) required for the replication machinery to forcibly dissociate a base pair into the monomers during DNA replication. One of the more significant findings to emerge from this study is that the short-lived G·T* base mispair, which electronic interaction energy between the bases (-23.76 kcal mol(-1)) exceeds the analogical value for the G·C Watson-Crick nucleobase pair (-20.38 kcal mol(-1)), "escapes from the hands" of the DNA replication machinery by fast transforming into the G*·T mismatch playing an indirect role of its supplier during the DNA replication. So

  15. Excitation of Toroidicity-Induced Alfven Eigenmodes by the Electrodes Inserted in a Heliotron/Torsatron Plasma

    SciTech Connect

    Matsunaga, G.; Kawada, S.; Kotani, J.; Toi, K.; Suzuki, C.; Matsuoka, K.

    2005-06-10

    A novel method of exciting shear Alfven waves using electrodes inserted in a plasma was developed for basic study of Alfven eigenmodes in a heliotron/torsatron plasma. The electrodes can induce excitation current along the confinement field line, and generate magnetic perturbations perpendicular to the confinement field. By sweeping the frequency of the current in a cold plasma, the toroidicity-induced Alfven eigenmode was resonantly excited at the predicted frequency and radial location. Plasma response to the applied magnetic perturbations indicates a fairly large damping rate caused by continuum damping.0.

  16. Twenty-one-base-pair insertion polymorphism creates an enhancer element and potentiates SLC6A1 GABA transporter promoter activity

    PubMed Central

    Hirunsatit, Rungnapa; George, Elizabeth D.; Lipska, Barbara K.; Elwafi, Hani M.; Sander, Lisa; Yrigollen, Carolyn M.; Gelernter, Joel; Grigorenko, Elena L.; Lappalainen, Jaakko; Mane, Shrikant; Nairn, Angus C.; Kleinman, Joel E.; Simen, Arthur A.

    2009-01-01

    Objective Sodium-dependent and chloride-dependent γ-aminobutyric acid (GABA) transporter 1 (SLC6A1) is the target of a number of drugs of clinical importance and is a major determinant of synaptic GABA concentrations. We resequenced the human SLC6A1 gene previously and discovered a novel 21 bp insertion in the predicted promoter region that creates a second tandem copy of the sequence. Here we sought to determine the functional relevance of this variation. Methods We used reporter assays, mobility shift assays, quantitative PCR, and proteomics methods as well as postmortem expression analysis for this work. Results Reporter assays showed that the insertion allele significantly increases promoter activity in multiple cell lines. The zinc finger transcription factor ZNF148 was found to significantly transactivate the promoter and increase expression when overexpressed but could not account for the differences in activity between the two alleles of the promoter. Copy number of the insertion sequence was associated with exponentially increasing activity of a downstream promoter, suggesting that the insertion sequence has enhancer activity when present in multiple copies. SLC6A1 promoter genotype was found to predict SLC6A1 RNA expression in human postmortem hippocampal samples. These results suggest that the insertion polymorphism leads to increased SLC6A1 promoter activity because, in part, of creation of an enhancer element when present as multiple copies. Genotyping individuals from Tanzania in this study suggested that the insertion allele has its origin in Africa. Conclusion On account of the effect of the insertion on promoter activity, this relatively common polymorphism may prove useful in predicting clinical response to pharmacological modulators of SLC6A1 as well as GABAergic function in individuals of African descent. PMID:19077666

  17. A novel eight amino acid insertion contributes to the hemagglutinin cleavability and the virulence of a highly pathogenic avian influenza A (H7N3) virus in mice

    SciTech Connect

    Sun, Xiangjie; Belser, Jessica A.; Tumpey, Terrence M.

    2016-01-15

    In 2012, an avian influenza A H7N3 (A/Mexico/InDRE7218/2012; Mx/7218) virus was responsible for two confirmed cases of human infection and led to the death or culling of more than 22 million chickens in Jalisco, Mexico. Interestingly, this virus acquired an 8-amino acid (aa)-insertion (..PENPK-DRKSRHRR-TR/GLF) near the hemagglutinin (HA) cleavage site by nonhomologous recombination with host rRNA. It remains unclear which specific residues at the cleavage site contribute to the virulence of H7N3 viruses in mammals. Using loss-of-function approaches, we generated a series of cleavage site mutant viruses by reverse genetics and characterized the viruses in vitro and in vivo. We found that the 8-aa insertion and the arginine at position P4 of the Mx/7218 HA cleavage site are essential for intracellular HA cleavage in 293T cells, but have no effect on the pH of membrane fusion. However, we identified a role for the histidine residue at P5 position in viral fusion pH. In mice, the 8-aa insertion is required for Mx/7218 virus virulence; however, the basic residues upstream of the P4 position are dispensable for virulence. Overall, our study provides the first line of evidence that the insertion in the Mx/7218 virus HA cleavage site confers its intracellular cleavability, and consequently contributes to enhanced virulence in mice. - Highlights: • An avian influenza H7N3 virus acquired a unique 8-amino acid (aa) insertion. • The role of specific basic residues in the HA insertion in viral pathogenesis was determined. • In mice, the 8-aa insertion is required for H7N3 virus virulence. • The R residue at position P4 is essential for HA intracellular cleavage and virus virulence.

  18. Characterization and event specific-detection by quantitative real-time PCR of T25 maize insert.

    PubMed

    Collonnier, Cécile; Schattner, Alexandra; Berthier, Georges; Boyer, Francine; Coué-Philippe, Géraldine; Diolez, Annick; Duplan, Marie-Noëlle; Fernandez, Sophie; Kebdani, Naïma; Kobilinsky, André; Romaniuk, Marcel; de Beuckeleer, Marc; de Loose, Marc; Windels, Pieter; Bertheau, Yves

    2005-01-01

    T25 is one of the 4 maize transformation events from which commercial lines have so far been authorized in Europe. It was created by polyethylene glycol-mediated transformation using a construct bearing one copy of the synthetic pat gene associated with both promoter and terminator of the 35S ribosomal gene from cauliflower mosaic virus. In this article, we report the sequencing of the whole T25 insert and the characterization of its integration site by using a genome walking strategy. Our results confirmed that one intact copy of the initial construct had been integrated in the plant genome. They also revealed, at the 5' junction of the insert, the presence of a second truncated 35S promoter, probably resulting from rearrangements which may have occurred before or during integration of the plasmid DNA. The analysis of the junction fragments showed that the integration site of the insert presented high homologies with the Huck retrotransposon family. By using one primer annealing in the maize genome and the other in the 5' end of the integrated DNA, we developed a reliable event-specific detection system for T25 maize. To provide means to comply with the European regulation, a real-time PCR test was designed for specific quantitation of T25 event by using Taqman chemistry.

  19. Insertion mechanism and stability of boron nitride nanotubes in lipid bilayers.

    PubMed

    Thomas, Michael; Enciso, Marta; Hilder, Tamsyn A

    2015-04-16

    We provide insight into the interaction of boron nitride nanotubes (BNNTs) with cell membranes to better understand their improved biocompatibility compared to carbon nanotubes (CNTs). Contrary to CNTs, no computational studies exist investigating the insertion mechanism and stability of BNNTs in membranes. Our molecular dynamics simulations demonstrate that BNNTs are spontaneously attracted to lipid bilayers and are stable once inserted. They insert via a lipid-mediated, passive insertion mechanism. BNNTs demonstrate similar characteristics to more biocompatible functionalized CNTs.

  20. Efficient Merge and Insert Operations for Binary Heaps and Trees

    NASA Technical Reports Server (NTRS)

    Kuszmaul, Christopher Lee; Woo, Alex C. (Technical Monitor)

    2000-01-01

    Binary heaps and binary search trees merge efficiently. We introduce a new amortized analysis that allows us to prove the cost of merging either binary heaps or balanced binary trees is O(l), in the amortized sense. The standard set of other operations (create, insert, delete, extract minimum, in the case of binary heaps, and balanced binary trees, as well as a search operation for balanced binary trees) remain with a cost of O(log n). For binary heaps implemented as arrays, we show a new merge algorithm that has a single operation cost for merging two heaps, a and b, of O(absolute value of a + min(log absolute value of b log log absolute value of b. log absolute value of a log absolute value of b). This is an improvement over O(absolute value of a + log absolute value of a log absolute value of b). The cost of the new merge is so low that it can be used in a new structure which we call shadow heaps. to implement the insert operation to a tunable efficiency. Shadow heaps support the insert operation for simple priority queues in an amortized time of O(f(n)) and other operations in time O((log n log log n)/f (n)), where 1 less than or equal to f (n) less than or equal to log log n. More generally, the results here show that any data structure with operations that change its size by at most one, with the exception of a merge (aka meld) operation, can efficiently amortize the cost of the merge under conditions that are true for most implementations of binary heaps and search trees.

  1. Flexor Hallucis Longus Tendon Transfer for Calcific Insertional Achilles Tendinopathy.

    PubMed

    Howell, Michael A; Catanzariti, Alan R

    2016-01-01

    Calcific insertional Achilles tendinopathy can result in significant pain and disability. Although some patients respond to nonoperative therapy, many patients are at risk for long-term morbidity and unpredictable clinical outcomes. There is no evidence-based data to support the timing of operative invention, choice of procedures, or whether equinus requires treatment. This article suggests the need for a classification system based on physical examination and imaging to help guide treatment. There is an obvious need for evidence-based studies evaluating outcomes and for properly conducted scientific research to establish appropriate treatment protocols.

  2. Re-visiting the trans insertion model for complexin clamping

    PubMed Central

    Krishnakumar, Shyam S; Li, Feng; Coleman, Jeff; Schauder, Curtis M; Kümmel, Daniel; Pincet, Frederic; Rothman, James E; Reinisch, Karin M

    2015-01-01

    We have previously proposed that complexin cross-links multiple pre-fusion SNARE complexes via a trans interaction to function as a clamp on SNARE-mediated neurotransmitter release. A recent NMR study was unable to detect the trans clamping interaction of complexin and therefore questioned the previous interpretation of the fluorescence resonance energy transfer and isothermal titration calorimetry data on which the trans clamping model was originally based. Here we present new biochemical data that underscore the validity of our previous interpretation and the continued relevancy of the trans insertion model for complexin clamping. DOI: http://dx.doi.org/10.7554/eLife.04463.001 PMID:25831964

  3. Modification of commercial force feedback hardware for needle insertion simulation.

    PubMed

    Coles, Timothy R; John, Nigel W; Sofia, Giuseppe; Gould, Derek A; Caldwell, Darwin G

    2011-01-01

    A SensAble Omni force feedback device has been modified to increase the face validity of a needle insertion simulation. The new end effector uses a real needle hub and shortened needle shaft in place of the Omni's pre-fitted pen shaped end effector. This modification facilitates correct procedural training through the simulation of co-located visual and haptic cues in an augmented reality approach to simulation. The development of the new end effector is described and a pictorial guide to its manufacture and the fitting process is provided. Initial results from face validation studies bode well for the fidelity of this low cost device.

  4. Development and testing of the rack insertion device

    NASA Technical Reports Server (NTRS)

    Strickland, G. Scott

    1995-01-01

    Installing and removing experiment racks in a Space Station Logistics Module will become a repetitive operation at Kennedy Space Center (KSC) in the near future. A Rack Insertion Device (RID) consisting of an Extendible Boom, End Effector, and Positioning Base is being developed for the task. This paper discusses the key elements of the RlD's function and design. Prototype test results for the RlD's Extendible Boom and End Effector are presented. Also discussed are future end effectors that will further enhance the RlD's Space Station processing capability.

  5. The ATLAS Insertable B-Layer: from construction to operation

    NASA Astrophysics Data System (ADS)

    La Rosa, A.

    2016-12-01

    The ATLAS Insertable B-Layer (IBL) is the innermost layer of pixel detectors, and was installed in May 2014 at a radius of 3.3 cm from the beam axis, between the existing Pixel detector and a new smaller radius beam-pipe. The new detector, built to cope with high radiation and occupancy, is the first large scale application of 3D sensors and CMOS 130 nm technology. The IBL detector construction was completed within about two years (2012 - 2014), and the key features and challenges met during the IBL project are presented, as well as its commissioning and operational experience at the LHC.

  6. Crossing professional boundaries: nurse-led catheter insertion.

    PubMed

    Linda, Kelly

    2009-10-01

    The steady expansion of nursing roles is enabling staff to perform tasks traditionally undertaken by doctors. This change is perceived as being in the interests of patients, particularly because it can reduce waiting times, but it can also increase job satisfaction and enhance opportunities for the recruitment and retention of staff. This article describes the establishment of a nurse-led service for tunnelled central venous catheter insertion for renal dialysis. The result of a retrospective audit describes what impact this service has had.

  7. RHIC INSERTION REGION, SHUNT POWER SUPPLY CURRENT ERRORS.

    SciTech Connect

    BRUNO,D.; GANETIS,G.; LAMBIASE,R.F.; SANDBERG,J.

    2001-06-18

    The Relativistic Heavy Ion Collider (RHIC) was commissioned in 1999 and 2000. RHIC requires power supplies to supply currents to highly inductive superconducting magnets. The RHIC Insertion Region contain's many shunt power supplies to trim the current of different magnet elements in a large superconducting magnet circuit. Power Supply current error measurements were performed during the commissioning of RHIC. Models of these power supply systems were produced to predict and improve these power supply current errors using the circuit analysis program MicroCap V by Spectrum Software (TM). Results of the power supply current errors are presented from the models and from the measurements performed during the commissioning of RHIC.

  8. Pediatric Enteric Feeding Techniques: Insertion, Maintenance, and Management of Problems

    SciTech Connect

    Nijs, Els L. F.; Cahill, Anne Marie

    2010-12-15

    Enteral feeding is considered a widespread, well-accepted means of delivering nutrition to adults and children who are unable to consume food by mouth or who need support in maintaining adequate nutrition for a variety of reasons, including acute and chronic disease states. Delivery of enteral feeding to nutritionally deprived patients may be achieved by several means. In this article, the indications and insertion of enteral access in children will be reviewed. In addition, common complications and management of problems will be discussed.

  9. Postpartum and postabortal insertion of intrauterine contraceptive devices.

    PubMed

    Martorella, L A; Esposito, J M

    1975-04-01

    One hundred forty-four patients are studied over an 18 month period. Postpartum and postabortal insertions of intrauterine contraceptive devices using the Lippes Loop and the Dalkon Shield are compared. The age, parity, previous methods of contraception, and marital status of the study group indicated a poorly motivated patient population. Although accidental pregnancy rates and expulsion rates are higher, the technique is considered worth-while in the population studied. Future research should be directed toward improved devices in order to decrease side effects and to increase appeal of the technique to those who will benefit most.

  10. Reliability Technology to Achieve Insertion of Advanced Packaging (RELTECH) program

    NASA Technical Reports Server (NTRS)

    Fayette, Daniel F.; Speicher, Patricia; Stoklosa, Mark J.; Evans, Jillian V.; Evans, John W.; Gentile, Mike; Pagel, Chuck A.; Hakim, Edward

    1993-01-01

    A joint military-commercial effort to evaluate multichip module (MCM) structures is discussed. The program, Reliability Technology to Achieve Insertion of Advanced Packaging (RELTECH), has been designed to identify the failure mechanisms that are possible in MCM structures. The RELTECH test vehicles, technical assessment task, product evaluation plan, reliability modeling task, accelerated and environmental testing, and post-test physical analysis and failure analysis are described. The information obtained through RELTECH can be used to address standardization issues, through development of cost effective qualification and appropriate screening criteria, for inclusion into a commercial specification and the MIL-H-38534 general specification for hybrid microcircuits.

  11. ARTEMIS Lunar Orbit Insertion and Science Orbit Design Through 2013

    NASA Technical Reports Server (NTRS)

    Broschart, Stephen B.; Sweetser, Theodore H.; Angelopoulos, Vassilis; Folta, David; Woodard, Mark

    2015-01-01

    As of late-July 2011, the ARTEMIS mission is transferring two spacecraft from Lissajous orbits around Earth-Moon Lagrange Point #1 into highly-eccentric lunar science orbits. This paper presents the trajectory design for the transfer from Lissajous orbit to lunar orbit insertion, the period reduction maneuvers, and the science orbits through 2013. The design accommodates large perturbations from Earth's gravity and restrictive spacecraft capabilities to enable opportunities for a range of heliophysics and planetary science measurements. The process used to design the highly-eccentric ARTEMIS science orbits is outlined. The approach may inform the design of future planetary moon missions.

  12. The Design and Analysis of Transposon-Insertion Sequencing Experiments

    PubMed Central

    Chao, Michael C.; Abel, Sören; Davis, Brigid M.; Waldor, Matthew K.

    2016-01-01

    Preface Transposon-insertion sequencing (TIS) is a powerful approach that can be widely applied to genome-wide definition of loci that are required for growth in diverse conditions. However, experimental design choices and stochastic biological processes can heavily influence the results of TIS experiments and affect downstream statistical analysis. Here, we discuss TIS experimental parameters and how these factors relate to the benefits and limitations of the various statistical frameworks that can be applied to computational analysis of TIS data. PMID:26775926

  13. Infrasonic and low-frequency insert earphone hearing threshold.

    PubMed

    Kuehler, Robert; Fedtke, Thomas; Hensel, Johannes

    2015-04-01

    Low-frequency and infrasonic pure-tone monaural hearing threshold data down to 2.5 Hz are presented. These measurements were made by means of a newly developed insert-earphone source. The source is able to generate pure-tone sound pressure levels up to 130 dB between 2 and 250 Hz with very low harmonic distortions. Behavioral hearing thresholds were determined in the frequency range from 2.5 to 125 Hz for 18 otologically normal test persons. The median hearing thresholds are comparable to values given in the literature. They are intended for stimulus calibration in subsequent brain imaging investigations.

  14. Systemic Air Embolism Associated with Pleural Pigtail Chest Tube Insertion

    PubMed Central

    Alkhankan, Emad; Nusair, Ahmad; Mazagri, Rida

    2016-01-01

    Pleural pigtail catheter placement is associated with many complications including pneumothorax, hemorrhage, and chest pain. Air embolism is a known but rare complication of pleural pigtail catheter insertion and has a high risk of occurrence with positive pressure ventilation. In this case report, we present a 50-year-old male with bilateral pneumonia who developed a pneumothorax while on mechanical ventilation with continuous positive airway pressure mode. During the placement of the pleural pigtail catheter to correct the pneumothorax, the patient developed a sudden left sided body weakness and became unresponsive. An air embolism was identified in the right main cerebral artery, which was fatal. PMID:27630781

  15. Improving Peripherally Inserted Central Catheter (PICC) care on a Trauma and Orthopaedics ward.

    PubMed

    Piorkowska, Marta; Al-Raweshidy, Zahra; Yeong, Keefai

    2013-01-01

    Peripherally Inserted Central Catheter (PICC) blockage rate was audited over a two month period on the Trauma & Orthopaedics ward at our District General Hospital. A 70% (five out of seven) PICC blockage rate was observed. High blockage rates lead to potential treatment complications, delays in delivery of treatment, increase in costs, and reduction in patient satisfaction. The factors contributing to the significant blockage rate include, long and contradictory PICC care guidelines, no information sheets in the patient notes, lack of training and awareness about care of, and flushing of, PICC lines, and lack of accountability for PICC flushing. Our project aimed to achieve a greater rate of PICC patency. We produced one succinct and comprehensive PICC care guideline, carried out staff training sessions, introduced a sticker reminding staff to flush the PICC line after use, and introduced a prescription of weekly heparin saline and PRN saline flushes (for monitoring and accountability). We used questionnaires to assess competency of hospital staff pre-teaching (doctors 6%, nurses 0%), and post-teaching (doctors 70%, nurses 38%). Blockage rate data post-intervention is pending. Education improved awareness of guidelines amongst staff and we anticipate that the proposed interventions will translate into reduced blockage rates, improving patient outcomes and reducing costs.

  16. Insertable system for fast turnaround time microwave experiments in a dilution refrigerator.

    PubMed

    Ong, Florian R; Orgiazzi, Jean-Luc; de Waard, Arlette; Frossati, Giorgio; Lupascu, Adrian

    2012-09-01

    Microwave experiments in dilution refrigerators are a central tool in the field of superconducting quantum circuits and other research areas. This type of experiments relied so far on attaching a device to the mixing chamber of a dilution refrigerator. The minimum turnaround time in this case is a few days as required by cooling down and warming up the entire refrigerator. We developed a new approach, in which a suitable sample holder is attached to a cold-insertable probe and brought in contact with transmission lines permanently mounted inside the cryostat. The total turnaround time is 8 h if the target temperature is 80 mK. The lowest attainable temperature is 30 mK. Our system can accommodate up to six transmission lines, with a measurement bandwidth tested from zero frequency to 12 GHz. This bandwidth is limited by low-pass components in the setup; we expect the intrinsic bandwidth to be at least 18 GHz. We present our setup, discuss the experimental procedure, and give examples of experiments enabled by this system. This new measurement method will have a major impact on systematic ultra-low temperature studies using microwave signals, including those requiring quantum coherence.

  17. Insertable system for fast turnaround time microwave experiments in a dilution refrigerator

    NASA Astrophysics Data System (ADS)

    Ong, Florian R.; Orgiazzi, Jean-Luc; de Waard, Arlette; Frossati, Giorgio; Lupascu, Adrian

    2012-09-01

    Microwave experiments in dilution refrigerators are a central tool in the field of superconducting quantum circuits and other research areas. This type of experiments relied so far on attaching a device to the mixing chamber of a dilution refrigerator. The minimum turnaround time in this case is a few days as required by cooling down and warming up the entire refrigerator. We developed a new approach, in which a suitable sample holder is attached to a cold-insertable probe and brought in contact with transmission lines permanently mounted inside the cryostat. The total turnaround time is 8 h if the target temperature is 80 mK. The lowest attainable temperature is 30 mK. Our system can accommodate up to six transmission lines, with a measurement bandwidth tested from zero frequency to 12 GHz. This bandwidth is limited by low-pass components in the setup; we expect the intrinsic bandwidth to be at least 18 GHz. We present our setup, discuss the experimental procedure, and give examples of experiments enabled by this system. This new measurement method will have a major impact on systematic ultra-low temperature studies using microwave signals, including those requiring quantum coherence.

  18. 21 CFR 876.5830 - Hemodialyzer with disposable insert (Kiil type).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Hemodialyzer with disposable insert (Kiil type... Hemodialyzer with disposable insert (Kiil type). (a) Identification. A hemodialyzer with disposable inserts (Kiil type) is a device that is used as a part of an artificial kidney system for the treatment...

  19. 21 CFR 876.5830 - Hemodialyzer with disposable insert (Kiil type).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Hemodialyzer with disposable insert (Kiil type... Hemodialyzer with disposable insert (Kiil type). (a) Identification. A hemodialyzer with disposable inserts (Kiil type) is a device that is used as a part of an artificial kidney system for the treatment...

  20. 21 CFR 876.5830 - Hemodialyzer with disposable insert (Kiil type).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Hemodialyzer with disposable insert (Kiil type... Hemodialyzer with disposable insert (Kiil type). (a) Identification. A hemodialyzer with disposable inserts (Kiil type) is a device that is used as a part of an artificial kidney system for the treatment...