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Sample records for t-dna insertion line

  1. Phenotypic analyses of Arabidopsis T-DNA insertion lines and expression profiling reveal that multiple L-type lectin receptor kinases are involved in plant immunity.

    PubMed

    Wang, Yan; Bouwmeester, Klaas; Beseh, Patrick; Shan, Weixing; Govers, Francine

    2014-12-01

    L-type lectin receptor kinases (LecRK) are membrane-spanning receptor-like kinases with putative roles in biotic and abiotic stress responses and in plant development. In Arabidopsis, 45 LecRK were identified but their functions are largely unknown. Here, a systematic functional analysis was carried out by evaluating phenotypic changes of Arabidopsis LecRK T-DNA insertion lines in plant development and upon exposure to various external stimuli. None of the LecRK T-DNA insertion lines showed clear developmental changes, either under normal conditions or upon abiotic stress treatment. However, many of the T-DNA insertion lines showed altered resistance to Phytophthora brassicae, Phytophthora capsici, Pseudomonas syringae, or Alternaria brassicicola. One mutant defective in LecRK-V.5 expression was compromised in resistance to two Phytophthora spp. but showed enhanced resistance to Pseudomonas syringae. LecRK-V.5 overexpression confirmed its dual role in resistance and susceptibility depending on the pathogen. Combined analysis of these phenotypic data and LecRK expression profiles retrieved from public datasets revealed that LecRK which are hardly induced upon infection or even suppressed are also involved in pathogen resistance. Computed coexpression analysis revealed that LecRK with similar function displayed diverse expression patterns. Because LecRK are widespread in plants, the results presented here provide invaluable information for exploring the potential of LecRK as novel sources of resistance in crops. PMID:25083911

  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. PMID:20444194

  3. Spectrum of T-DNA integrations for insertional mutagenesis of Histoplasma capsulatum

    PubMed Central

    Kemski, Megan M.; Stevens, Bryan; Rappleye, Chad A.

    2012-01-01

    Agrobacterium-mediated transformation is being increasingly used for insertional mutagenesis of fungi. To better evaluate its effectiveness as a mutagen for the fungal pathogen Histoplasma capsulatum, we analyzed a collection of randomly selected T-DNA insertion mutants. Testing of different T-DNA element vectors engineered for transformation of fungi showed that pBHt2 provides the highest transformation efficiency and the lowest rate of vector backbone carryover. Sixty-eight individual T-DNA integrations were characterized by recovery of T-DNA ends and flanking genomic sequences. The right border end of the T-DNA is largely preserved whereas the left border end is frequently truncated. Analysis of T-DNA insertion sites confirms the lack of any integration hotspots in the Histoplasma genome. Relative to genes, T-DNA integrations show significant bias towards promoter regions at the expense of coding sequences. With consideration for potential promoter interruption and the demonstrated efficacy of intronic insertions, 61% of mapped T-DNA insertions should impair gene expression or function. Mapping of T-DNA flanking sequences demonstrates 67% of T-DNA integrations are integrations at a single chromosomal site and 31% of T-DNA integrations are associated with large-scale chromosomal rearrangements. This characterization of T-DNA insertions in mutants selected without regard to phenotype supports application of Agrobacterium-mediated transformation as an insertional mutagen for genome-based screens and functional discovery of genes in Histoplasma. PMID:23332832

  4. T-DNA insertion alters the terpenoid content composition and bioactivity of transgenic Artemisia annua.

    PubMed

    Karaket, Netiya; Wiyakrutta, Suthep; Lacaille-Dubois, Marie-Aleth; Supaibulwatana, Kanyaratt

    2014-03-01

    In this study, the interference of T-DNA insertion upon Agrobacterium-mediated transformation on the biochemical expression of the host genome is discussed. Plant extracts of transgenic Artemisia annua L. with or without an overexpressed famesyl pyrophosphate synthase gene have been investigated for their bioactivity and metabolic profile in comparison with wild type A. annua. The highest antimicrobial activity against Staphylococcus aureus, Bacillus subtilis and Candida albicans was observed in the T253 transgenic lines. Moreover, the crude extract from T253 showed higher antimalarial activity against the Plasmodium faciparum K1 strain than those of the others. The terpenoid constituents and antimicrobial properties of the plant samples were grouped by hierarchical clustering analysis. The clustering showed that squalene is a putative compound that might be involved in increasing the bioactivity of the transgenic line. In addition, T253 had a triterpene content that was about twice as great as that of the T253-2 line, which had a higher content of sesquiterpenes. However, both lines were transformed by the same FPS gene. These results suggested that the different bioactive properties observed in each transgenic line may be caused by variations in their terpenoid composition, which is affected by T-DNA insertion at different positions in the host plant. PMID:24689216

  5. Decreased Expression of a Gene Caused by a T-DNA Insertion in an Adjacent Gene in Arabidopsis

    PubMed Central

    Tamura, Kentaro; Kawabayashi, Takenori; Shikanai, Toshiharu; Hara-Nishimura, Ikuko

    2016-01-01

    ALADIN is a component of the nuclear pore complex in higher eukaryotes. An Arabidopsis knockout line that had a T-DNA insertion in the ALADIN gene was defective in plant growth and thylakoid development and had reduced photosynthetic activity resulting from lower chlorophyll accumulation. The mutation appeared to decrease the level of chloroplast RuBisCO subunits and PSBA and PGL35 proteins. Unexpectedly, the T-DNA insertion in the ALADIN gene decreased the expression of the neighboring gene PSRP5, which functions in translation in chloroplasts. The mutant phenotype was rescued by expressing PSRP5, but not by expressing ALADIN. The abnormal phenotypes were also detected in an artificial microRNA (amiRNA)-mediated PSRPS5 knockdown, but not in an amiRNA-mediated ALADIN knockdown line. Thus, users of T-DNA insertions should be aware that a T-DNA insertion in one gene can have effects on the expression of neighboring genes. PMID:26828726

  6. GABI-Kat SimpleSearch: an Arabidopsis thaliana T-DNA mutant database with detailed information for confirmed insertions

    PubMed Central

    Li, Yong; Rosso, Mario G.; Viehoever, Prisca; Weisshaar, Bernd

    2007-01-01

    Insertional mutagenesis approaches, especially by T-DNA, play important roles in gene function studies of the model plant Arabidopsis thaliana. GABI-Kat SimpleSearch () is a Flanking Sequence Tag (FST)-based database for T-DNA insertion mutants generated by the GABI-Kat project. Currently, the database contains >108 000 mapped FSTs from ?64 000 lines which cover 64% of all annotated A.thaliana protein-coding genes. The web interface allows searching for relevant insertions by gene code, keyword, line identifier, GenBank accession number of the FST, and also by BLAST. A graphic display of the genome region around the gene or the FST assists users to select insertion lines of their interests. About 3500 insertions were confirmed in the offspring of the plant from which the original FST was generated, and the seeds of these lines are available from the Nottingham Arabidopsis Stock Centre. The database now also contains additional information such as segregation data, gene-specific primers and confirmation sequences. This information not only helps users to evaluate the usefulness of the mutant lines, but also covers a big part of the molecular characterization of the insertion alleles. PMID:17062622

  7. Studies on rice seed quality through analysis of a large-scale T-DNA insertion population.

    PubMed

    Fu, Fang-Fang; Ye, Rui; Xu, Shu-Ping; Xue, Hong-Wei

    2009-03-01

    A rice (Oryza sativa) T-DNA insertion population, which included more than 63 000 independent transgenic lines and 8 840 identified flanking sequence tags (FSTs) that were mapped onto the rice genome, was developed to systemically study the rice seed quality control. Genome-wide analysis of the FST distribution showed that T-DNA insertions were positively correlated with expressed genes, but negatively with transposable elements and small RNAs. In addition, the recovered T-DNAs were preferentially located at the untranslated region of the expressed genes. More than 11 000 putative homozygous lines were obtained through multi-generations of planting and resistance screening, and measurement of seed quality of around half of them, including the contents of starch, amylose, protein and fat, with a nondestructive near-infrared spectroscopy method, identified 551 mutants with unique or multiple altered parameters of seed quality. Analysis of the corresponding FSTs showed that genes participating in diverse functions, including metabolic processes and transcriptional regulation, were involved, indicating that seed quality is regulated by a complex network. PMID:19223856

  8. 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 promoter-trapping Ti vector, combined with induction medium that contains root exudates, can be useful for identification of pathogenicity-related and functional genes in V. dahliae. PMID:24993736

  9. T?DNA insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus Laccaria bicolor

    PubMed Central

    Kemppainen, Minna; Duplessis, Sebastien; Martin, Francis; Pardo, Alejandro G.

    2008-01-01

    Summary Ectomycorrhiza is a mutualistic symbiosis formed between fine roots of trees and the mycelium of soil fungi. This symbiosis plays a key role in forest ecosystems for the mineral nutrition of trees and the biology of the fungal communities associated. The characterization of genes involved in developmental and metabolic processes is important to understand the complex interactions that control the ectomycorrhizal symbiosis. Agrobacterium?mediated gene transfer (AMT) in fungi is currently opening a new era for fungal research. As whole genome sequences of several fungi are being released studies about T?DNA integration patterns are needed in order to understand the integration mechanisms involved and to evaluate the AMT as an insertional mutagenesis tool for different fungal species. The first genome sequence of a mycorrhizal fungus, the basidiomycete Laccaria bicolor, became public in July 2006. Release of Laccaria genome sequence and the availability of AMT makes this fungus an excellent model for functional genomic studies in ectomycorrhizal research. No data on the integration pattern in Laccaria genome were available, thus we optimized a plasmid rescue approach for this fungus. To this end the transformation vector (pHg/pBSk) was constructed allowing the rescue of the T?DNA right border (RB)–genomic DNA junctions in Escherichia coli. Fifty?one Agrobacterium?transformed fungal strains, picked up at random from a larger collection of T?DNA tagged strains (about 500), were analysed. Sixty?nine per cent were successfully rescued for the RB of which 87% were resolved for genomic integration sequences. Our results demonstrate that the plasmid rescue approach can be used for resolving T?DNA integration sites in Laccaria. The RB was well conserved during transformation of this fungus and the integration analysis showed no clear sequence homology between different genomic sites. Neither obvious sequence similarities were found between these sites and the T?DNA borders indicating non?homologous integration of the transgenes. Majority (75%) of the integrations were located in predicted genes. Agrobacterium?mediated gene transfer is a powerful tool that can be used for functional gene studies in Laccaria and will be helpful along with plasmid rescue in searching for relevant fungal genes involved in the symbiotic process. PMID:21261845

  10. [Genetic analysis of a heading-delayed mutant by T-DNA insertion in rice (Oryza sativa L.)].

    PubMed

    Chen, Zhao-Gui; Wang, Jiang; Zhang, Ze-Min; Liu, Fang; Zhu, Hai-Tao; Wan, Xin-Shan; Zhang, Jing-Liu; Zhang, Gui-Quan

    2004-02-01

    Transposon tagging was used to isolate genes in higher plant. In this study, a delayed heading mutant caused by T-DNA insertion in rice was identified. Genetic analysis of the mutant showed that the three types of phenotype, normal heading, delayed heading and overly delayed heading in the segregating populations derived from the T-DNA heterozygotes fit the ratio of 1:2:1. Test for Basta resistance showed the delayed heading plants were all resistant while the normal heading plants were susceptible, and the ratio of resistant and susceptible plants was 3:1, which indicated that the delayed heading mutant was co-segregated with Basta resistance. The delayed heading mutant caused by T-DNA insertion was confirmed by T-DNA detection using PCR method. This delayed heading mutant will be used for isolation of the tagged gene in rice. PMID:15583412

  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

    2015-11-01

    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. PMID:26343971

  12. Analysis of the early-flowering mechanisms and generation of T-DNA tagging lines in Kitaake, a model rice cultivar

    PubMed Central

    An, Gynheung

    2013-01-01

    As an extremely early flowering cultivar, rice cultivar Kitaake is a suitable model system for molecular studies. Expression analyses revealed that transcript levels of the flowering repressor Ghd7 were decreased while those of its downstream genes, Ehd1, Hd3a, and RFT1, were increased. Sequencing the known flowering-regulator genes revealed mutations in Ghd7 and OsPRR37 that cause early translation termination and amino acid substitutions, respectively. Genetic analysis of F2 progeny from a cross between cv. Kitaake and cv. Dongjin indicated that those mutations additively contribute to the early-flowering phenotype in cv. Kitaake. Because the short life cycle facilitates genetics research, this study generated 10 000 T-DNA tagging lines and deduced 6758 flanking sequence tags (FSTs), in which 3122 were genic and 3636 were intergenic. Among the genic lines, 367 (11.8%) were inserted into new genes that were not previously tagged. Because the lines were generated by T-DNA that contained the promoterless GUS reporter gene, which had an intron with triple splicing donors/acceptors in the right border region, a high efficiency of GUS expression was shown in various organs. Sequencing of the GUS-positive lines demonstrated that the third splicing donor and the first splicing acceptor of the vector were extensively used. The FST data have now been released into the public domain for seed distribution and facilitation of rice research. PMID:23966593

  13. Reduced tillering in Basmati rice T-DNA insertional mutant OsTEF1 associates with differential expression of stress related genes and transcription factors.

    PubMed

    Paul, Priyanka; Awasthi, Anjali; Rai, Amit Kumar; Gupta, Santosh Kumar; Prasad, R; Sharma, T R; Dhaliwal, H S

    2012-06-01

    A T-DNA insertional mutant OsTEF1 of rice gives 60-80% reduced tillering, retarded growth of seminal roots, and sensitivity to salt stress compared to wild type Basmati 370. The insertion occurred in a gene encoding a transcription elongation factor homologous to yeast elf1, on chromosome 2 of rice. Detailed transcriptomic profiling of OsTEF1 revealed that mutation in the transcription elongation factor differentially regulates the expression of more than 100 genes with known function and finely regulates tillering process in rice by inducing the expression of cytochrome P450. Along with different transcription factors, several stress associated genes were also affected due to a single insertion. In silico analysis of the TEF1 protein showed high conservation among different organisms. This transcription elongation factor predicted to interact with other proteins that directly or indirectly positively regulate tillering in rice. PMID:22367482

  14. Oryza Tag Line, a phenotypic mutant database for the Génoplante rice insertion line library

    PubMed Central

    Larmande, Pierre; Gay, Céline; Lorieux, Mathias; Périn, Christophe; Bouniol, Matthieu; Droc, Gaëtan; Sallaud, Christophe; Perez, Pascual; Barnola, Isabelle; Biderre-Petit, Corinne; Martin, Jérôme; Morel, Jean Benoît; Johnson, Alexander A. T.; Bourgis, Fabienne; Ghesquière, Alain; Ruiz, Manuel; Courtois, Brigitte; Guiderdoni, Emmanuel

    2008-01-01

    To organize data resulting from the phenotypic characterization of a library of 30 000 T-DNA enhancer trap (ET) insertion lines of rice (Oryza sativa L cv. Nipponbare), we developed the Oryza Tag Line (OTL) database (http://urgi.versailles.inra.fr/OryzaTagLine/). OTL structure facilitates forward genetic search for specific phenotypes, putatively resulting from gene disruption, and/or for GUSA or GFP reporter gene expression patterns, reflecting ET-mediated endogenous gene detection. In the latest version, OTL gathers the detailed morpho-physiological alterations observed during field evaluation and specific screens in a first set of 13 928 lines. Detection of GUS or GFP activity in specific organ/tissues in a subset of the library is also provided. Search in OTL can be achieved through trait ontology category, organ and/or developmental stage, keywords, expression of reporter gene in specific organ/tissue as well as line identification number. OTL now contains the description of 9721 mutant phenotypic traits observed in 2636 lines and 1234 GUS or GFP expression patterns. Each insertion line is documented through a generic passport data including production records, seed stocks and FST information. 8004 and 6101 of the 13 928 lines are characterized by at least one T-DNA and one Tos17 FST, respectively that OTL links to the rice genome browser OryGenesDB. PMID:17947330

  15. T-DNA-associated duplication/translocations in Arabidopsis. Implications for mutant analysis and functional genomics.

    PubMed

    Tax, F E; Vernon, D M

    2001-08-01

    T-DNA insertion mutants have become a valuable resource for studies of gene function in Arabidopsis. In the course of both forward and reverse genetic projects, we have identified novel interchromosomal rearrangements in two Arabidopsis T-DNA insertion lines. Both rearrangements were unilateral translocations associated with the left borders of T-DNA inserts that exhibited normal Mendelian segregation. In one study, we characterized the embryo-defective88 mutation. Although emb88 had been mapped to chromosome I, molecular analysis of DNA adjacent to the T-DNA left border revealed sequence from chromosome V. Simple sequence length polymorphism mapping of the T-DNA insertion demonstrated that a >40-kbp region of chromosome V had inserted with the T-DNA into the emb88 locus on chromosome I. A similar scenario was observed with a prospective T-DNA knockout allele of the LIGHT-REGULATED RECEPTOR PROTEIN KINASE (LRRPK) gene. Whereas wild-type LRRPK is on lower chromosome IV, mapping of the T-DNA localized the disrupted LRRPK allele to chromosome V. In both these cases, the sequence of a single T-DNA-flanking region did not provide an accurate picture of DNA disruption because flanking sequences had duplicated and inserted, with the T-DNA, into other chromosomal locations. Our results indicate that T-DNA insertion lines--even those that exhibit straightforward genetic behavior--may contain an unexpectedly high frequency of rearrangements. Such duplication/translocations can interfere with reverse genetic analyses and provide misleading information about the molecular basis of mutant phenotypes. Simple mapping and polymerase chain reaction methods for detecting such rearrangements should be included as a standard step in T-DNA mutant analysis. PMID:11500551

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

    DOE PAGESBeta

    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,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

  17. 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.

  18. High-Throughput Analysis of T-DNA Location and Structure Using Sequence Capture

    PubMed Central

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

    2015-01-01

    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. Our method greatly facilitates the first step of this characterization of transgenic plants by providing an efficient screen for the selection of promising lines. PMID:26445462

  19. An Intron Sense Suppression Construct Targeting Soybean FAD2-1 Requires a Double-Stranded RNA-Producing Inverted Repeat T-DNA Insert[OA

    PubMed Central

    Mroczka, Andrew; Roberts, Peter D.; Fillatti, JoAnne J.; Wiggins, B. Elizabeth; Ulmasov, Tim; Voelker, Toni

    2010-01-01

    We demonstrate that the transformation of soybean (Glycine max) with sense suppression constructs using intron sequences from the fatty acid oleyl ?12 desaturase gene FAD2-1A leads to efficient and specific reduction of FAD2-1 transcripts in developing seeds, increased oleic acid, and decreased polyunsaturated fatty acids. The related FAD2-2 transcripts are only marginally affected. Despite screening a large number of independent transformants, no single-copy efficacious transformants could be found. Invariably, all the least complex transgenic loci have two T-DNA copies in an inverted repeat configuration, centered at the right borders. We show that this T-DNA configuration produces an inverted repeat transcript and that small interfering RNAs accumulate against the target sequence. PMID:20424004

  20. 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. PMID:25392478

  1. Genetics, structure, and prevalence of FP967 (CDC Triffid) T-DNA in flax.

    PubMed

    Young, Lester; Hammerlindl, Joseph; Babic, Vivijan; McLeod, Jamille; Sharpe, Andrew; Matsalla, Chad; Bekkaoui, Faouzi; Marquess, Leigh; Booker, Helen M

    2015-01-01

    The detection of T-DNA from a genetically modified flaxseed line (FP967, formally CDC Triffid) in a shipment of Canadian flaxseed exported to Europe resulted in a large decrease in the amount of flax planted in Canada. The Canadian flaxseed industry undertook major changes to ensure the removal of FP967 from the supply chain. This study aimed to resolve the genetics and structure of the FP967 transfer DNA (T-DNA). The FP967 T-DNA is thought to be inserted in at single genomic locus. The junction between the T-DNA and genomic DNA consisted of two inverted Right Borders with no Left Border (LB) flanking genomic DNA sequences recovered. This information was used to develop an event-specific quantitative PCR (qPCR) assay. This assay and an existing assay specific to the T-DNA construct were used to determine the genetics and prevalence of the FP967 T-DNA. These data supported the hypothesis that the T-DNA is present at a single location in the genome. The FP967 T-DNA is present at a low level (between 0.01 and 0.1%) in breeder seed lots from 2009 and 2010. None of the 11,000 and 16,000 lines selected for advancement through the Flax Breeding Program in 2010 and 2011, respectively, tested positive for the FP967 T-DNA, however. Most of the FP967 T-DNA sequence was resolved via PCR cloning and next generation sequencing. A 3,720 bp duplication of an internal portion of the T-DNA (including a Right Border) was discovered between the flanking genomic DNA and the LB. An event-specific assay, SAT2-LB, was developed for the junction between this repeat and the LB. PMID:25883881

  2. Forward genetics screening of Medicago truncatula Tnt1 insertion lines.

    PubMed

    Yarce, Juan Carlos Serrani; Lee, Hee-Kyung; Tadege, Million; Ratet, Pascal; Mysore, Kirankumar S

    2013-01-01

    A large population of Medicago truncatula insertion lines has been generated using the Tnt1 retrotransposon. More than 21,000 insertion lines have been generated, representing more than 500,000 insertion events. This mutant population is being used by the legume research community to screen for various different mutants using a forward genetics approach. Some of the phenotypes that have been screened using this population include developmentally abnormal phenotypes in leaves, stem, flowers, and roots. In addition to these, mutants with defects in symbiosis with Rhizobium and mycorrhiza, mutants with altered nonhost resistance against Asian Soybean Rust and switch grass rust pathogens, mutants with altered lignin content, mutants with altered cell wall structure, etc. have been identified. Here, we describe the high throughput methodology that is being used to identify these M. truncatula mutants. PMID:23996311

  3. GABI-Kat SimpleSearch: new features of the Arabidopsis thaliana T-DNA mutant database

    PubMed Central

    Kleinboelting, Nils; Huep, Gunnar; Kloetgen, Andreas; Viehoever, Prisca; Weisshaar, Bernd

    2012-01-01

    T-DNA insertion mutants are very valuable for reverse genetics in Arabidopsis thaliana. Several projects have generated large sequence-indexed collections of T-DNA insertion lines, of which GABI-Kat is the second largest resource worldwide. User access to the collection and its Flanking Sequence Tags (FSTs) is provided by the front end SimpleSearch (http://www.GABI-Kat.de). Several significant improvements have been implemented recently. The database now relies on the TAIRv10 genome sequence and annotation dataset. All FSTs have been newly mapped using an optimized procedure that leads to improved accuracy of insertion site predictions. A fraction of the collection with weak FST yield was re-analysed by generating new FSTs. Along with newly found predictions for older sequences about 20?000 new FSTs were included in the database. Information about groups of FSTs pointing to the same insertion site that is found in several lines but is real only in a single line are included, and many problematic FST-to-line links have been corrected using new wet-lab data. SimpleSearch currently contains data from ?71?000 lines with predicted insertions covering 62.5% of the 27?206 nuclear protein coding genes, and offers insertion allele-specific data from 9545 confirmed lines that are available from the Nottingham Arabidopsis Stock Centre. PMID:22080561

  4. 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. PMID:11437280

  5. Marked reduction in compliance with central line insertion practices (CLIP) when accounting for missing CLIP data and incomplete line capture.

    PubMed

    Scott, Shruti K; Gohil, Shruti K; Quan, Kathleen; Huang, Susan S

    2016-02-01

    Adherence to central line insertion practices can significantly reduce infections and is used as a hospital benchmark for quality. However, current national standards for central line insertion practices (CLIP) compliance calculation do not include missing CLIP forms. We found adherence rates significantly decreased when accounting for all lines at an academic medical center. PMID:26518499

  6. Prepackaged central line kits reduce procedural mistakes during central line insertion: a randomized controlled prospective trial

    PubMed Central

    2013-01-01

    Background Central line catheter insertion is a complex procedure with a high cognitive load for novices. Providing a prepackaged all-inclusive kit is a simple measure that may reduce the cognitive load. We assessed whether the use of prepackaged all-inclusive central line insertion kits reduces procedural mistakes during central line catheter insertion by novices. Methods Thirty final year medical students and recently qualified physicians were randomized into two equal groups. One group used a prepackaged all-inclusive kit and the other used a standard kit containing only the central vein catheter and all other separately packaged components provided in a materials cart. The procedure was videotaped and analyzed by two blinded raters using a checklist. Both groups performed central line catheter insertion on a manikin, assisted by nursing students. Results The prepackaged kit group outperformed the standard kit group in four of the five quality indicators: procedure duration (26:26?±?3:50 min vs. 31:27?±?5:57 min, p?=?.01); major technical mistakes (3.1?±?1.4 vs. 4.8?±?2.6, p?=?.03); minor technical mistakes (5.2?±?1.7 vs. 8.0?±?3.2, p?=?.01); and correct steps (83?±?5% vs. 75?±?11%, p?=?.02). The difference for breaches of aseptic technique (1.2?±?0.8 vs. 3.0?±?3.6, p?=?.06) was not statistically significant. Conclusions Prepackaged all-inclusive kits for novices improved the procedure quality and saved staff time resources in a controlled simulation environment. Future studies are needed to address whether central line kits also improve patient safety in hospital settings. PMID:23631396

  7. 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

  8. 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 ...

  9. Gene identification with sequenced T-DNA tags generated by transformation of Arabidopsis cell suspension.

    PubMed

    Mathur, J; Szabados, L; Schaefer, S; Grunenberg, B; Lossow, A; Jonas-Straube, E; Schell, J; Koncz, C; Koncz-Kálmán, Z

    1998-03-01

    A protocol for establishment and high-frequency Agrobacterium-mediated transformation of morphogenic Arabidopsis cell suspensions was developed to facilitate saturation mutagenesis and identification of plant genes by sequenced T-DNA tags. Thirty-two self-circularized T-DNA tagged chromosomal loci were isolated from 21 transgenic plants by plasmid rescue and long-range inverse polymerase chain reaction (LR-iPCR). By bidirectional sequencing of the ends of T-DNA-linked plant DNA segments, nine T-DNA inserts were thus localized in genes coding for the Arabidopsis ASK1 kinase, cyclin 3b, J-domain protein, farnesyl diphosphate synthase, ORF02, an unknown EST, and homologues of a copper amine oxidase, a peripheral Golgi protein and a maize pollen-specific transcript. In addition, 16 genes were identified in the vicinity of sequenced T-DNA tags illustrating the efficiency of genome analysis by insertional mutagenesis. PMID:9681013

  10. Genome-wide analysis of T-DNA integration into the chromosomes of Magnaporthe oryzae

    PubMed Central

    Choi, Jaehyuk; Park, Jongsun; Jeon, Junhyun; Chi, Myoung-Hwan; Goh, Jaeduk; Yoo, Sung-Yong; Park, Jaejin; Jung, Kyongyong; Kim, Hyojeong; Park, Sook-Young; Rho, Hee-Sool; Kim, Soonok; Kim, Byeong Ryun; Han, Seong-Sook; Kang, Seogchan; Lee, Yong-Hwan

    2007-01-01

    Agrobacterium tumefaciens-mediated transformation (ATMT) has become a prevalent tool for functional genomics of fungi, but our understanding of T-DNA integration into the fungal genome remains limited relative to that in plants. Using a model plant-pathogenic fungus, Magnaporthe oryzae, here we report the most comprehensive analysis of T-DNA integration events in fungi and the development of an informatics infrastructure, termed a T-DNA analysis platform (TAP). We identified a total of 1110 T-DNA-tagged locations (TTLs) and processed the resulting data via TAP. Analysis of the TTLs showed that T-DNA integration was biased among chromosomes and preferred the promoter region of genes. In addition, irregular patterns of T-DNA integration, such as chromosomal rearrangement and readthrough of plasmid vectors, were also observed, showing that T-DNA integration patterns into the fungal genome are as diverse as those of their plant counterparts. However, overall the observed junction structures between T-DNA borders and flanking genomic DNA sequences revealed that T-DNA integration into the fungal genome was more canonical than those observed in plants. Our results support the potential of ATMT as a tool for functional genomics of fungi and show that the TAP is an effective informatics platform for handling data from large-scale insertional mutagenesis. PMID:17850257

  11. Integrated mechanism for the generation of the 5? junctions of LINE inserts

    PubMed Central

    Yamaguchi, Katsumi; Kajikawa, Masaki; Okada, Norihiro

    2014-01-01

    To elucidate the molecular mechanism of the integration of long interspersed elements (LINEs), we characterized the 5? ends of more than 200 LINE de novo retrotransposition events into chicken DT40 or human HeLa cells. Human L1 inserts produced 15-bp target-site duplications (TSDs) and zebrafish ZfL2-1 inserts produced 5-bp TSDs in DT40 cells, suggesting that TSD length depends on the LINE species. Further analysis of 5? junctions revealed that the 5?-end-joining pathways of LINEs can be divided into two fundamental types—annealing or direct. We also found that the generation of 5? inversions depends on host and LINE species. These results led us to propose a new model for 5?-end joining, the type of which is determined by the extent of exposure of 3? overhangs generated after the second-strand cleavage and by the involvement of host factors. PMID:25378331

  12. 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

  13. [Doctor-nurse delegation of the insertion of central venous lines].

    PubMed

    Cellupica, Mary

    2015-01-01

    The Léon Bérard Cancer Centre treats the disease in all its complexity with numerous disciplines such as surgery, medicine, radiotherapy, palliative care, home care, etc. The insertion of a central venous line is an essential part of cancer care and the nursing profession. It enables patients to have their treatment administered in the best possible conditions and without risk. PMID:26144516

  14. 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. PMID:26670088

  15. 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 ?. Images PMID:16593241

  16. Paramutation-Like Interaction of T-DNA Loci in Arabidopsis

    PubMed Central

    Xue, Weiya; Ruprecht, Colin; Street, Nathaniel; Hematy, Kian; Chang, Christine; Frommer, Wolf B.; Persson, Staffan; Niittylä, Totte

    2012-01-01

    In paramutation, epigenetic information is transferred from one allele to another to create a gene expression state which is stably inherited over generations. Typically, paramutation describes a phenomenon where one allele of a gene down-regulates the expression of another allele. Paramutation has been described in several eukaryotes and is best understood in plants. Here we describe an unexpected paramutation-like trans SALK T-DNA interaction in Arabidopsis. Unlike most of the previously described paramutations, which led to gene silencing, the trans SALK T-DNA interaction caused an increase in the transcript levels of the endogenous gene (COBRA) where the T-DNA was inserted. This increased COBRA expression state was stably inherited for several generations and led to the partial suppression of the cobra phenotype. DNA methylation was implicated in this trans SALK T-DNA interaction since mutation of the DNA methyltransferase 1 in the suppressed cobra caused a reversal of the suppression. In addition, null mutants of the DNA demethylase ROS1 caused a similar COBRA transcript increase in the cobra SALK T-DNA mutant as the trans T-DNA interaction. Our results provide a new example of a paramutation-like trans T-DNA interaction in Arabidopsis, and establish a convenient hypocotyl elongation assay to study this phenomenon. The results also alert to the possibility of unexpected endogenous transcript increase when two T-DNAs are combined in the same genetic background. PMID:23272131

  17. 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 disease. This study is still recruiting participants (expected recruitment: up to 165 participants) and is due to be completed in December 2017. There were no completed studies. There were no studies that compared no platelet transfusions to a platelet transfusion threshold. Authors’ conclusions There is no evidence from RCTs to determine whether platelet transfusions are required prior to central line insertion in patients with thrombocytopenia, and, if a platelet transfusion is required, what is the correct platelet transfusion threshold. Further randomised trials with robust methodology are required to develop the optimal transfusion strategy for such patients. The one ongoing RCT involving people with cirrhosis will not be able to answer this review’s questions, because it is a small study that assesses one patient group and does not address all of the comparisons included in this review. To detect an increase in the proportion of participants who had major bleeding from 1 in 100 to 2 in 100 would require a study containing at least 4634 participants (80% power, 5% significance). PMID:26627708

  18. Formation of Complex Extrachromosomal T-DNA Structures in Agrobacterium tumefaciens-Infected Plants1[C][W][OA

    PubMed Central

    Singer, Kamy; Shiboleth, Yoel M.; Li, Jianming; Tzfira, Tzvi

    2012-01-01

    Agrobacterium tumefaciens is a unique plant pathogenic bacterium renowned for its ability to transform plants. The integration of transferred DNA (T-DNA) and the formation of complex insertions in the genome of transgenic plants during A. tumefaciens-mediated transformation are still poorly understood. Here, we show that complex extrachromosomal T-DNA structures form in A. tumefaciens-infected plants immediately after infection. Furthermore, these extrachromosomal complex DNA molecules can circularize in planta. We recovered circular T-DNA molecules (T-circles) using a novel plasmid-rescue method. Sequencing analysis of the T-circles revealed patterns similar to the insertion patterns commonly found in transgenic plants. The patterns include illegitimate DNA end joining, T-DNA truncations, T-DNA repeats, binary vector sequences, and other unknown “filler” sequences. Our data suggest that prior to T-DNA integration, a transferred single-stranded T-DNA is converted into a double-stranded form. We propose that termini of linear double-stranded T-DNAs are recognized and repaired by the plant’s DNA double-strand break-repair machinery. This can lead to circularization, integration, or the formation of extrachromosomal complex T-DNA structures that subsequently may integrate. PMID:22797657

  19. Rescuing Alu: Recovery of New Inserts Shows LINE-1 Preserves Alu Activity through A-Tail Expansion

    PubMed Central

    Wagstaff, Bradley J.; Hedges, Dale J.; Derbes, Rebecca S.; Campos Sanchez, Rebeca; Chiaromonte, Francesca; Makova, Kateryna D.; Roy-Engel, Astrid M.

    2012-01-01

    Alu elements are trans-mobilized by the autonomous non-LTR retroelement, LINE-1 (L1). Alu-induced insertion mutagenesis contributes to about 0.1% human genetic disease and is responsible for the majority of the documented instances of human retroelement insertion-induced disease. Here we introduce a SINE recovery method that provides a complementary approach for comprehensive analysis of the impact and biological mechanisms of Alu retrotransposition. Using this approach, we recovered 226 de novo tagged Alu inserts in HeLa cells. Our analysis reveals that in human cells marked Alu inserts driven by either exogenously supplied full length L1 or ORF2 protein are indistinguishable. Four percent of de novo Alu inserts were associated with genomic deletions and rearrangements and lacked the hallmarks of retrotransposition. In contrast to L1 inserts, 5′ truncations of Alu inserts are rare, as most of the recovered inserts (96.5%) are full length. De novo Alus show a random pattern of insertion across chromosomes, but further characterization revealed an Alu insertion bias exists favoring insertion near other SINEs, highly conserved elements, with almost 60% landing within genes. De novo Alu inserts show no evidence of RNA editing. Priming for reverse transcription rarely occurred within the first 20 bp (most 5′) of the A-tail. The A-tails of recovered inserts show significant expansion, with many at least doubling in length. Sequence manipulation of the construct led to the demonstration that the A-tail expansion likely occurs during insertion due to slippage by the L1 ORF2 protein. We postulate that the A-tail expansion directly impacts Alu evolution by reintroducing new active source elements to counteract the natural loss of active Alus and minimizing Alu extinction. PMID:22912586

  20. 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. PMID:15744539

  1. Increased Polyamine Biosynthesis Enhances Stress Tolerance by Preventing the Accumulation of Reactive Oxygen Species: T-DNA Mutational Analysis of Oryza sativa Lysine Decarboxylase-like Protein 1

    PubMed Central

    Jang, Su Jin; Wi, Soo Jin; Choi, Yoo Jin; An, Gynheung; Park, Ky Young

    2012-01-01

    A highly oxidative stress-tolerant japonica rice line was isolated by T-DNA insertion mutation followed by screening in the presence of 50 mM H2O2. The T-DNA insertion was mapped to locus Os09g0547500, the gene product of which was annotated as lysine decarboxylase-like protein (GenBank accession No. AK062595). We termed this gene OsLDC-like 1, for Oryza sativa lysine decarboxylase-like 1. The insertion site was in the second exon and resulted in a 27 amino acid N-terminal deletion. Despite this defect in OsLDC-like 1, the mutant line exhibited enhanced accumulation of the polyamines (PAs) putrescine, spermidine, and spermine under conditions of oxidative stress. The generation of reactive oxygen species (ROS) in the mutant line was assessed by qRT-PCR analysis of NADPH oxidase (RbohD and RbohF), and by DCFH-DA staining. Cellular levels of ROS in osldc-like 1 leaves were significantly lower than those in the wild-type (WT) rice after exposure to oxidative, high salt and acid stresses. Exogenously-applied PAs such as spermidine and spermine significantly inhibited the stress-induced accumulation of ROS and cell damage in WT leaves. Additionally, the activities of ROS-detoxifying enzymes were increased in the homozygous mutant line in the presence or absence of H2O2. Thus, mutation of OsLDC-like 1 conferred an oxidative stress-tolerant phenotype. These results suggest that increased cellular PA levels have a physiological role in preventing stress-induced ROS and ethylene accumulation and the resultant cell damage. PMID:22965749

  2. An Entamoeba histolytica LINE/SINE Pair Inserts at Common Target Sites Cleaved by the Restriction Enzyme-Like LINE-Encoded Endonuclease

    PubMed Central

    Mandal, Prabhat K.; Bagchi, Anindya; Bhattacharya, Alok; Bhattacharya, Sudha

    2004-01-01

    The non-long-terminal-repeat (non-LTR) retrotransposons (also called long interspersed repetitive elements [LINEs]) are among the oldest retroelements. Here we describe the properties of such an element from a primitive protozoan parasite, Entamoeba histolytica, that infects the human gut. This 4.8-kb element, called EhLINE1, is present in about 140 copies dispersed throughout the genome. The element belongs to the R4 clade of non-LTR elements. It has a centrally located reverse transcriptase domain and a restriction enzyme-like endonuclease (EN) domain at the carboxy terminus. We have cloned and expressed a 794-bp fragment containing the EN domain in Escherichia coli. The purified protein could nick supercoiled pBluescript DNA to yield open circular and linear DNAs. The conserved PDX12-14D motif was required for activity. Genomic sequences flanking the sites of insertion of EhLINE1 and the putative partner short interspersed repetitive element (SINE), EhSINE1, were analyzed. Both elements resulted in short target site duplications (TSD) upon insertion. A common feature was the presence of a short T-rich stretch just upstream of the TSD in most insertion sites. By sequence analysis an empty target site in the E. histolytica genome, known to be occupied by EhSINE1, was identified. When a 176-bp fragment containing the empty site was used as a substrate for EN, it was prominently nicked on the bottom strand at the precise point of insertion of EhSINE1, showing that this SINE could use the LINE-encoded endonuclease for its insertion. The nick on the bottom strand was toward the right of the TSD, which is uncommon. The lack of strict target site-specificity of the restriction enzyme-like EN encoded by EhLINE1 is also exceptional. A model for retrotransposition of EhLINE1/SINE1 is presented. PMID:14871947

  3. Characterization and isolation of a T-DNA tagged banana promoter active during in vitro culture and low temperature stress

    PubMed Central

    Santos, Efrén; Remy, Serge; Thiry, Els; Windelinckx, Saskia; Swennen, Rony; Sági, László

    2009-01-01

    Background Next-generation transgenic plants will require a more precise regulation of transgene expression, preferably under the control of native promoters. A genome-wide T-DNA tagging strategy was therefore performed for the identification and characterization of novel banana promoters. Embryogenic cell suspensions of a plantain-type banana were transformed with a promoterless, codon-optimized luciferase (luc+) gene and low temperature-responsive luciferase activation was monitored in real time. Results Around 16,000 transgenic cell colonies were screened for baseline luciferase activity at room temperature 2 months after transformation. After discarding positive colonies, cultures were re-screened in real-time at 26°C followed by a gradual decrease to 8°C. The baseline activation frequency was 0.98%, while the frequency of low temperature-responsive luciferase activity was 0.61% in the same population of cell cultures. Transgenic colonies with luciferase activity responsive to low temperature were regenerated to plantlets and luciferase expression patterns monitored during different regeneration stages. Twenty four banana DNA sequences flanking the right T-DNA borders in seven independent lines were cloned via PCR walking. RT-PCR analysis in one line containing five inserts allowed the identification of the sequence that had activated luciferase expression under low temperature stress in a developmentally regulated manner. This activating sequence was fused to the uidA reporter gene and back-transformed into a commercial dessert banana cultivar, in which its original expression pattern was confirmed. Conclusion This promoter tagging and real-time screening platform proved valuable for the identification of novel promoters and genes in banana and for monitoring expression patterns throughout in vitro development and low temperature treatment. Combination of PCR walking techniques was efficient for the isolation of candidate promoters even in a multicopy T-DNA line. Qualitative and quantitative GUS expression analyses of one tagged promoter in a commercial cultivar demonstrated a reproducible promoter activity pattern during in vitro culture. Thus, this promoter could be used during in vitro selection and generation of commercial transgenic plants. PMID:19552803

  4. Incidence of Genome Structure, DNA Asymmetry, and Cell Physiology on T-DNA Integration in Chromosomes of the Phytopathogenic Fungus Leptosphaeria maculans

    PubMed Central

    Bourras, Salim; Meyer, Michel; Grandaubert, Jonathan; Lapalu, Nicolas; Fudal, Isabelle; Linglin, Juliette; Ollivier, Benedicte; Blaise, Françoise; Balesdent, Marie-Hélène; Rouxel, Thierry

    2012-01-01

    The ever-increasing generation of sequence data is accompanied by unsatisfactory functional annotation, and complex genomes, such as those of plants and filamentous fungi, show a large number of genes with no predicted or known function. For functional annotation of unknown or hypothetical genes, the production of collections of mutants using Agrobacterium tumefaciens–mediated transformation (ATMT) associated with genotyping and phenotyping has gained wide acceptance. ATMT is also widely used to identify pathogenicity determinants in pathogenic fungi. A systematic analysis of T-DNA borders was performed in an ATMT-mutagenized collection of the phytopathogenic fungus Leptosphaeria maculans to evaluate the features of T-DNA integration in its particular transposable element-rich compartmentalized genome. A total of 318 T-DNA tags were recovered and analyzed for biases in chromosome and genic compartments, existence of CG/AT skews at the insertion site, and occurrence of microhomologies between the T-DNA left border (LB) and the target sequence. Functional annotation of targeted genes was done using the Gene Ontology annotation. The T-DNA integration mainly targeted gene-rich, transcriptionally active regions, and it favored biological processes consistent with the physiological status of a germinating spore. T-DNA integration was strongly biased toward regulatory regions, and mainly promoters. Consistent with the T-DNA intranuclear-targeting model, the density of T-DNA insertion correlated with CG skew near the transcription initiation site. The existence of microhomologies between promoter sequences and the T-DNA LB flanking sequence was also consistent with T-DNA integration to host DNA mediated by homologous recombination based on the microhomology-mediated end-joining pathway. PMID:22908038

  5. 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.

  6. Automatic computation of wing-fuselage intersection lines and fillet inserts with fixed-area constraint

    NASA Technical Reports Server (NTRS)

    Barger, Raymond L.; Adams, Mary S.

    1993-01-01

    Procedures for automatic computation of wing-fuselage juncture geometry are described. These procedures begin with a geometry in wave-drag format. First, an intersection line is computed by extrapolating the wing to the fuselage. Then two types of filleting procedures are described, both of which utilize a combination of analytical and numerical techniques appropriate for automatic calculation. An analytical technique for estimating the added volume due to the fillet is derived, and an iterative procedure for revising the fuselage to compensate for this additional volume is given. Sample results are included in graphical form.

  7. Power electron beam front shortening in magnetically insulated transmission line with inner coaxial dielectric insert

    SciTech Connect

    Galstjan, E.A.; Kazanskiy, L.N.

    1995-12-31

    By now the technology of high-power high-current relativistic electron beams with microsecond duration has been developed. The same technology is used in high-power electric pulse generation. However at present the high-power beams of subnanosecond duration are necessary for some investigations and applications. The maximum power parameter achieved by means of the usual technology is limited by a value in the range of 100-300 MW. For this reason a search for new ways to generate high-power REB (relativistic electron beam) is a topical problem. It is obvious that this problem may be reduced to the generation of pulses with a subnanosecond front duration. There are many methods to shorten a pulse duration to its front duration for instance, by using parts of short-circuited transmission lines.

  8. Identification of Lactobacillus species using tDNA-PCR.

    PubMed

    Baele, Margo; Vaneechoutte, Mario; Verhelst, Rita; Vancanneyt, Marc; Devriese, Luc A; Haesebrouck, Freddy

    2002-08-01

    tDNA intergenic spacer PCR (tDNA-PCR) using consensus primers complementary to the conserved edges of the tRNA genes can amplify the intergenic spacers. Separation of the PCR products with capillary electrophoresis enables discrimination between fragments differing only one basepair in length. This method was applied to a collection of 82 Lactobacillus strains belonging to 37 species in order to evaluate the discriminatory power of this technique within this genus. Twenty-one species could be distinguished to species level on the basis of a unique tDNA fingerprint pattern. The other species grouped by two (e.g. L. fermentum and L. cellobiosus) or three (L. acidophilus, L. gallinarum and L. helveticus). Inclusion of the resulting fingerprints in a numerical database containing fingerprints of numerous other Gram-positive and Gram-negative species makes the identification of unknown strains possible. PMID:12031576

  9. Site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana mediated by Cre recombinase.

    PubMed

    Vergunst, A C; Jansen, L E; Hooykaas, P J

    1998-06-01

    In this study Agrobacterium tumefaciens transferred DNA (T-DNA) was targeted to a chromosomally introduced lox site in Arabidopsis thaliana by employing the Cre recombinase system. To this end, Arabidopsis target lines were constructed which harboured an active chimeric promoter-lox-cre gene stably integrated in the plant genome. A T-DNA vector with a promoterless lox -neomycin phosphotransferase (nptII) fusion was targeted to this genomic lox site with an efficiency of 1.2-2.3% of the number of random events. Cre-catalyzed site-specific recombination resulted in restoration of nptII expression by translational fusion of the lox-nptII sequence in the integration vector with the transcription and translation initiation sequences present at the target site, allowing selective enrichment on medium containing kanamycin. Simultaneously, the coding sequence of the Cre recombinase was disconnected from these same transcription and translation initiation signals by displacement, aimed at preventing the efficient reversible excision reaction. Of the site-specific recombinants, 89% were the result of precise integration. Furthermore, approximately 50% of these integrants were single copy transformants, based on PCR analysis. Agrobacterium T-DNA, which is transferred to plant cells as a single-stranded linear DNA structure, is in principle incompatible with Cre-mediated integration. Nevertheless, the results presented here clearly demonstrate the feasibility of the Agrobacterium -mediated transformation system, which is generally used for transformation of plants, to obtain site-specific integration. PMID:9592161

  10. Site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana mediated by Cre recombinase.

    PubMed Central

    Vergunst, A C; Jansen, L E; Hooykaas, P J

    1998-01-01

    In this study Agrobacterium tumefaciens transferred DNA (T-DNA) was targeted to a chromosomally introduced lox site in Arabidopsis thaliana by employing the Cre recombinase system. To this end, Arabidopsis target lines were constructed which harboured an active chimeric promoter-lox-cre gene stably integrated in the plant genome. A T-DNA vector with a promoterless lox -neomycin phosphotransferase (nptII) fusion was targeted to this genomic lox site with an efficiency of 1.2-2.3% of the number of random events. Cre-catalyzed site-specific recombination resulted in restoration of nptII expression by translational fusion of the lox-nptII sequence in the integration vector with the transcription and translation initiation sequences present at the target site, allowing selective enrichment on medium containing kanamycin. Simultaneously, the coding sequence of the Cre recombinase was disconnected from these same transcription and translation initiation signals by displacement, aimed at preventing the efficient reversible excision reaction. Of the site-specific recombinants, 89% were the result of precise integration. Furthermore, approximately 50% of these integrants were single copy transformants, based on PCR analysis. Agrobacterium T-DNA, which is transferred to plant cells as a single-stranded linear DNA structure, is in principle incompatible with Cre-mediated integration. Nevertheless, the results presented here clearly demonstrate the feasibility of the Agrobacterium -mediated transformation system, which is generally used for transformation of plants, to obtain site-specific integration. PMID:9592161

  11. 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

  12. 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. PMID:26360633

  13. On-line air-tightness and insertion loss simultaneous detection method of high air-tightness fiber optic penetration connector

    NASA Astrophysics Data System (ADS)

    Zhang, Jingchuan; Yang, Xiaoning; Wang, Jing; Jiang, Junfeng

    2015-08-01

    The high air-tightness multicore fiber optic penetration connector is a core component for the optical fiber sensing and communication technologies applied in the space environment simulator under the vacuum thermal environment. High air-tightness and insertion loss are the two key indexes of the fiber optic penetration connector. The air-tightness and insertion loss on-line synchronous detection method was proposed. First, established hardware-in-the-loop testing platform by using the vacuum pumping system, the vacuum vessel, the helium mass spectrometer leak detector and optical time-domain reflectmeter, then, described the air tightness and insertion loss on-line detection principle, finally, designed a detection test scheme and air-tightness and insertion loss were tested. Experimental results indicate that the leakage rate is lower than 1.0×10-7Pa•L/S, the minimum of which is1.0×10-10Pa•L/S and the insertion loss at wave length window 1550 nm is +/-0.07db, which is less than +/-0.1db. It can lay the data basis for the design of opto-mechanical combination and later period fine processing.

  14. 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...

  15. Frequent problems and their resolutions by using thermal asymmetric interlaced PCR (TAIL-PCR) to clone genes in Arabidopsis T-DNA tagged mutants

    PubMed Central

    Wu, Lei; Di, Dong-Wei; Zhang, Dan; Song, Bin; Luo, Pan; Guo, Guang-Qin

    2015-01-01

    T-DNA insertional mutagenesis is a powerful tool in Arabidopsis functional genomics research. Previous studies have developed thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) as an efficient strategy in isolation of DNA sequences adjacent to known sequences in T-DNA tagged mutants. However, a number of problems are encountered when attempts are made to clone flanking sequences in T-DNA tagged mutants. Therefore, it is necessary to improve the efficiency of cloning mutagenesis. Here, we present the most frequent problems and provide an improved method to increase TAIL-PCR efficiency. Even then, it is not always possible to successfully obtain flanking sequences; in such cases, we recommend using high-throughput sequencing to determine the mutations. PMID:26019639

  16. 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.

  17. 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

  18. 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. PMID:25922687

  19. Neonatal late-onset sepsis following peripherally inserted central catheter removal: association with antibiotic use and adverse line events.

    PubMed

    Hoffman, Michelle A; Snowden, Jessica N; Simonsen, Kari A; Nenninger, Tabitha M; Lyden, Elizabeth R; Anderson-Berry, Ann L

    2015-01-01

    The objective of the study was to evaluate incidence of and risk factors for sepsis following peripherally inserted central catheter (PICC) removal. The retrospective cohort study looked at neonatal intensive care unit patients with PICC placement between February 2003 and June 2010 at a single medical center in the United States. Results showed that 14/216 patients (6.5%) had sepsis within 5 days of PICC removal. PICC removal because of adverse events was significantly associated with sepsis (P = .017). Antibiotic use before PICC removal did not have a significant impact on sepsis. The conclusions of the study are that removal of PICCs because of adverse events is significantly associated with late-onset neonatal sepsis and that antibiotic use at the time of PICC removal is not associated with a decline in sepsis rate. PMID:25723835

  20. Retrieval of Embolized Intracardiac Peripherally Inserted Central Catheter Line: Novel Percutaneous Technique by Utilizing a Flexible Biopsy Forceps

    PubMed Central

    Pande, Arindam; Sarkar, Achyut; Ahmed, Imran; Patil, Shailesh K.

    2015-01-01

    Peripheral catheter embolization to the heart is common but infrequently reported. In view of the hazardous complications of thrombosis, embolism, infection, arrhythmia and even death, percutaneous retrieval of such foreign bodies is usually attempted. Previously reported percutaneous technique of retrieval mainly involved the snaring technique. Herein, we report a novel nonsurgical retrieval technique for successful removal of a 46 cm long embolized intracardiac peripherally inserted central catheter by utilizing a flexible biopsy forceps. To the best of our knowledge, the use of flexible biopsy forceps for retrieval has hitherto been unreported and this case report therefore adds to the repertoire of percutaneous retrieval techniques for safe and easy removal of embolized catheters to the heart. PMID:26900421

  1. A LINE-1 Insertion in DLX6 Is Responsible for Cleft Palate and Mandibular Abnormalities in a Canine Model of Pierre Robin Sequence

    PubMed Central

    Wolf, Zena T.; Leslie, Elizabeth J.; Arzi, Boaz; Jayashankar, Kartika; Karmi, Nili; Jia, Zhonglin; Rowland, Douglas J.; Young, Amy; Safra, Noa; Sliskovic, Saundra; Murray, Jeffrey C.; Wade, Claire M.; Bannasch, Danika L.

    2014-01-01

    Cleft palate (CP) is one of the most commonly occurring craniofacial birth defects in humans. In order to study cleft palate in a naturally occurring model system, we utilized the Nova Scotia Duck Tolling Retriever (NSDTR) dog breed. Micro-computed tomography analysis of CP NSDTR craniofacial structures revealed that these dogs exhibit defects similar to those observed in a recognizable subgroup of humans with CP: Pierre Robin Sequence (PRS). We refer to this phenotype in NSDTRs as CP1. Individuals with PRS have a triad of birth defects: shortened mandible, posteriorly placed tongue, and cleft palate. A genome-wide association study in 14 CP NSDTRs and 72 unaffected NSDTRs identified a significantly associated region on canine chromosome 14 (24.2 Mb–29.3 Mb; praw?=?4.64×10?15). Sequencing of two regional candidate homeobox genes in NSDTRs, distal-less homeobox 5 (DLX5) and distal-less homeobox 6 (DLX6), identified a 2.1 kb LINE-1 insertion within DLX6 in CP1 NSDTRs. The LINE-1 insertion is predicted to insert a premature stop codon within the homeodomain of DLX6. This prompted the sequencing of DLX5 and DLX6 in a human cohort with CP, where a missense mutation within the highly conserved DLX5 homeobox of a patient with PRS was identified. This suggests the involvement of DLX5 in the development of PRS. These results demonstrate the power of the canine animal model as a genetically tractable approach to understanding naturally occurring craniofacial birth defects in humans. PMID:24699068

  2. A LINE-1 insertion in DLX6 is responsible for cleft palate and mandibular abnormalities in a canine model of Pierre Robin sequence.

    PubMed

    Wolf, Zena T; Leslie, Elizabeth J; Arzi, Boaz; Jayashankar, Kartika; Karmi, Nili; Jia, Zhonglin; Rowland, Douglas J; Young, Amy; Safra, Noa; Sliskovic, Saundra; Murray, Jeffrey C; Wade, Claire M; Bannasch, Danika L

    2014-04-01

    Cleft palate (CP) is one of the most commonly occurring craniofacial birth defects in humans. In order to study cleft palate in a naturally occurring model system, we utilized the Nova Scotia Duck Tolling Retriever (NSDTR) dog breed. Micro-computed tomography analysis of CP NSDTR craniofacial structures revealed that these dogs exhibit defects similar to those observed in a recognizable subgroup of humans with CP: Pierre Robin Sequence (PRS). We refer to this phenotype in NSDTRs as CP1. Individuals with PRS have a triad of birth defects: shortened mandible, posteriorly placed tongue, and cleft palate. A genome-wide association study in 14 CP NSDTRs and 72 unaffected NSDTRs identified a significantly associated region on canine chromosome 14 (24.2 Mb-29.3 Mb; p(raw?)=?4.64 × 10(-15)). Sequencing of two regional candidate homeobox genes in NSDTRs, distal-less homeobox 5 (DLX5) and distal-less homeobox 6 (DLX6), identified a 2.1 kb LINE-1 insertion within DLX6 in CP1 NSDTRs. The LINE-1 insertion is predicted to insert a premature stop codon within the homeodomain of DLX6. This prompted the sequencing of DLX5 and DLX6 in a human cohort with CP, where a missense mutation within the highly conserved DLX5 homeobox of a patient with PRS was identified. This suggests the involvement of DLX5 in the development of PRS. These results demonstrate the power of the canine animal model as a genetically tractable approach to understanding naturally occurring craniofacial birth defects in humans. PMID:24699068

  3. Generation and identification of DNA sequence flanking T-DNA integration site of Trichoderma atroviride mutants with high dichlorvos-degrading capacity.

    PubMed

    Sun, Wenliang; Liu, Lixing; Hu, Xiaolu; Tang, Jun; Liu, Peng; Chen, Jie; Chen, Yunpeng

    2009-12-01

    A protocol for efficient Agrobacterium tumefaciens-mediated transformation (ATMT) of biocontrol fungus Trichoderma atroviride strain T23 was developed to construct mutants with improved dichlorvos-degradation ability. A transformation frequency of 5x10(-6) was achieved. Among 110 genetically stable T-DNA transformants of T. atroviride T23, two transformants, AMT-12 and AMT-28, confirmed by Southern blot analysis to have single-copy inserts of T-DNA, showed an increase in dichlorvos-degradation ability of more than 10% compared to that of the wild type, exhibited similar tolerance to the pesticide, but lower spore formation ability. Five transformants exhibited a reduction in degradation of more than 70%, exhibited wild-type spore formation, and tolerated up to 800 microg/mL of dichlorvos. The left-flanking sequence of the insertion site in AMT-12 was cloned as a 1845-bp fragment and shown to have 89% identity to the DNA from T. atroviride IMI 206040; however, the involvement of this DNA in dichlorvos degradation remains still to be determined. This study can promote both a more efficient isolation of DNA sequence flanking T-DNA integration site in T. atroviride mutants and a more rational utilization of these transformants in dichlorvos degradation. PMID:19577921

  4. Insertional mutagenesis using Tnt1 retrotransposon in potato

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Potato is the third most important food crop in the world. However, genetics and genomics research of potato has lagged behind many major crop species due to its autotetraploidy and a highly heterogeneous genome. Insertional mutagenesis using T-DNA or transposable elements, which is available in sev...

  5. ?-Thalassemia due to intronic LINE-1 insertion in the ?-globin gene (HBB): molecular mechanisms underlying reduced transcript levels of the ?-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 ?(+)-thalassemia that is caused by the insertion of the full-length transposable element LINE-1 (L1) into the intron-2 of the ?-globin gene (HBB). The transcript level of the affected ?-globin gene was severely reduced. The remaining transcripts consisted of full-length, correctly processed ?-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 ?-globin(L1) allele also resulted from a reduced rate of transcription and decreased production of full-length ?-globin primary transcripts. The promoter and enhancer sequences of the ?-globin(L1) allele were hypermethylated; however, treatment with a demethylating agent did not restore the impaired transcription. A histone deacetylase inhibitor partially reactivated the ?-globin(L1) transcription despite permanent ?-globin(L1) promoter CpG methylation. This result indicates that the decreased rate of transcription from the ?-globin(L1) allele is associated with an altered chromatin structure. Therefore, the molecular defect caused by intronic L1 insertion in the ?-globin gene represents a novel etiology of ?-thalassemia. PMID:23878091

  6. Electrically heatable catalytic converter insert

    SciTech Connect

    Whittenberger, W.A.

    1992-09-15

    This patent describes an electrically heatable catalytic converter insert for insertion in an exhaust line from an internal combustion engine, it comprises: a catalyst bearing corrugated thin metal polycellular honeycomb monolith dimensioned and having a circular configuration for placement within an exhaust line.

  7. Agrobacterium tumefaciens transfers single-stranded transferred DNA (T-DNA) into the plant cell nucleus.

    PubMed Central

    Tinland, B; Hohn, B; Puchta, H

    1994-01-01

    Transferred DNA (T-DNA) is transferred as a single-stranded derivative from Agrobacterium to the plant cell nucleus. This conclusion is drawn from experiments exploiting the different properties of single- and double-stranded DNA to perform extrachromosomal homologous recombination in plant cells. After transfer from Agrobacterium to plant cells, T-DNA molecules recombined much more efficiently if the homologous sequences were of opposite polarity than if they were of the same polarity. This observation reflects the properties of single-stranded DNA; single-stranded DNA molecules of opposite polarity can anneal directly, whereas single-stranded DNA molecules of the same polarity first have to become double stranded to anneal. Judging from the relative amounts of single- to double-stranded T-DNA derivatives undergoing recombination, we infer that the T-DNA derivatives enter the plant nucleus in their single-stranded form. Images PMID:11607492

  8. 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 ...

  9. Evidence of frequent integration of non-T-DNA vector backbone sequences in transgenic strawberry plant.

    PubMed

    Abdal-Aziz, Samia Abdallah; Pliego-Alfaro, Fernando; Quesada, Miguel A; Mercado, José A

    2006-06-01

    We have studied the occurrence of the integration of non-T-DNA sequences in transgenic strawberry plants obtained through Agrobacterium inoculation. DNA from these plants was subjected to PCR amplification of the sequence of the gene trfA, which is located outside the T-DNA. The percentage of trfA-positive plants varied from 40% to 90%, with a mean of 65.7%. Backbone sequences were confirmed by Southern blot analysis. PMID:16935253

  10. 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

  11. An intronic LINE-1 element insertion in the dystrophin gene aborts dystrophin expression and results in Duchenne-like muscular dystrophy in the corgi breed

    PubMed Central

    Smith, Bruce F.; Yue, Yongping; Woods, Philip R.; Kornegay, Joe N.; Shin, Jin-Hong; Williams, Regina R.; Duan, Dongsheng

    2010-01-01

    Duchenne muscular dystrophy (DMD) is a dystrophin-deficient lethal muscle disease. To date, the catastrophic muscle wasting phenotype has only been seen in dystrophin-deficient humans and dogs. While Duchenne-like symptoms have been observed in more than a dozen dog breeds, the mutation is often not known and research colonies are rarely established. Here we report an independent canine DMD model originally derived from the Pembroke Welsh corgi breed. The affected dogs presented clinical signs of muscular dystrophy. Immunostaining revealed the absence of dystrophin and up-regulation of utrophin. Histopathologic examination showed variable fiber size, central nucleation, calcification, fibrosis, neutrophil and macrophage infiltration and cardiac focal vacuolar degeneration. Carrier dogs also displayed mild myopathy. The mutation was identified as a long interspersed repetitive element-1 (LINE-1) insertion in intron 13 which introduced a new exon containing an in-frame stop codon. Similar mutations have been seen in human patients. A colony was generated by crossing carrier females with normal males. Affected puppies had a normal birth weight but they experienced a striking growth delay in the first 5 days. In summary, the new corgi DMD model offers an excellent opportunity to study DMD pathogenesis and to develop novel therapies. PMID:20714321

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

    PubMed

    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

    2015-10-01

    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

  13. An intronic LINE-1 element insertion in the dystrophin gene aborts dystrophin expression and results in Duchenne-like muscular dystrophy in the corgi breed.

    PubMed

    Smith, Bruce F; Yue, Yongping; Woods, Philip R; Kornegay, Joe N; Shin, Jin-Hong; Williams, Regina R; Duan, Dongsheng

    2011-02-01

    Duchenne muscular dystrophy (DMD) is a dystrophin-deficient lethal muscle disease. To date, the catastrophic muscle wasting phenotype has only been seen in dystrophin-deficient humans and dogs. Although Duchenne-like symptoms have been observed in more than a dozen dog breeds, the mutation is often not known and research colonies are rarely established. Here, we report an independent canine DMD model originally derived from the Pembroke Welsh corgi breed. The affected dogs presented clinical signs of muscular dystrophy. Immunostaining revealed the absence of dystrophin and upregulation of utrophin. Histopathologic examination showed variable fiber size, central nucleation, calcification, fibrosis, neutrophil and macrophage infiltration and cardiac focal vacuolar degeneration. Carrier dogs also displayed mild myopathy. The mutation was identified as a long interspersed repetitive element-1 (LINE-1) insertion in intron 13, which introduced a new exon containing an in-frame stop codon. Similar mutations have been seen in human patients. A colony was generated by crossing carrier females with normal males. Affected puppies had a normal birth weight but they experienced a striking growth delay in the first 5 days. In summary, the new corgi DMD model offers an excellent opportunity to study DMD pathogenesis and to develop novel therapies. PMID:20714321

  14. 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 Central

    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 evidence for the link between melanocyte development and the epidermal microenvironment. PMID:26930598

  15. 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-collection/) and through the Phytozome genome browser (http://phytozome.jgi.doe.gov/pz/portal.html). The collection has been heavily utilized by the research community and, as of October 23, 2015, 223 orders for 12,069 seeds packets have been filled. In addition to creating this resource, we also optimized methods for transformation and sequencing DNA flanking insertion sites.

  16. 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.

  17. Arabidopsis genes essential for seedling viability: isolation of insertional mutants and molecular cloning.

    PubMed Central

    Budziszewski, G J; Lewis, S P; Glover, L W; Reineke, J; Jones, G; Ziemnik, L S; Lonowski, J; Nyfeler, B; Aux, G; Zhou, Q; McElver, J; Patton, D A; Martienssen, R; Grossniklaus, U; Ma, H; Law, M; Levin, J Z

    2001-01-01

    We have undertaken a large-scale genetic screen to identify genes with a seedling-lethal mutant phenotype. From screening approximately 38,000 insertional mutant lines, we identified >500 seedling-lethal mutants, completed cosegregation analysis of the insertion and the lethal phenotype for >200 mutants, molecularly characterized 54 mutants, and provided a detailed description for 22 of them. Most of the seedling-lethal mutants seem to affect chloroplast function because they display altered pigmentation and affect genes encoding proteins predicted to have chloroplast localization. Although a high level of functional redundancy in Arabidopsis might be expected because 65% of genes are members of gene families, we found that 41% of the essential genes found in this study are members of Arabidopsis gene families. In addition, we isolated several interesting classes of mutants and genes. We found three mutants in the recently discovered nonmevalonate isoprenoid biosynthetic pathway and mutants disrupting genes similar to Tic40 and tatC, which are likely to be involved in chloroplast protein translocation. Finally, we directly compared T-DNA and Ac/Ds transposon mutagenesis methods in Arabidopsis on a genome scale. In each population, we found only about one-third of the insertion mutations cosegregated with a mutant phenotype. PMID:11779813

  18. The Genomic Pattern of tDNA Operon Expression in E. coli

    PubMed Central

    Ardell, David H; Kirsebom, Leif A

    2005-01-01

    In fast-growing microorganisms, a tRNA concentration profile enriched in major isoacceptors selects for the biased usage of cognate codons. This optimizes translational rate for the least mass invested in the translational apparatus. Such translational streamlining is thought to be growth-regulated, but its genetic basis is poorly understood. First, we found in reanalysis of the E. coli tRNA profile that the degree to which it is translationally streamlined is nearly invariant with growth rate. Then, using least squares multiple regression, we partitioned tRNA isoacceptor pools to predicted tDNA operons from the E. coli K12 genome. Co-expression of tDNAs in operons explains the tRNA profile significantly better than tDNA gene dosage alone. Also, operon expression increases significantly with proximity to the origin of replication, oriC, at all growth rates. Genome location explains about 15% of expression variation in a form, at a given growth rate, that is consistent with replication-dependent gene concentration effects. Yet the change in the tRNA profile with growth rate is less than would be expected from such effects. We estimated per-copy expression rates for all tDNA operons that were consistent with independent estimates for rDNA operons. We also found that tDNA operon location, and the location dependence of expression, were significantly different in the leading and lagging strands. The operonic organization and genomic location of tDNA operons are significant factors influencing their expression. Nonrandom patterns of location and strandedness shown by tDNA operons in E. coli suggest that their genomic architecture may be under selection to satisfy physiological demand for tRNA expression at high growth rates. PMID:16103901

  19. 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

  20. A Resource of Mapped Dissociation Launch Pads for Targeted Insertional Mutagenesis in the Arabidopsis Genome1

    PubMed Central

    Muskett, Paul R.; Clissold, Leah; Marocco, Adriano; Springer, Patricia S.; Martienssen, Robert; Dean, Caroline

    2003-01-01

    We describe a new resource for targeted insertional mutagenesis in Arabidopsis using a maize (Zea mays) Activator/Dissociation (Ds) two-element system. The two components of the system, T-DNA vectors carrying a Ds launch pad and a stable Activator transposase source, were designed to simplify selection of transposition events and maximize their usefulness. Because Ds elements preferentially transpose to nearby genomic sites, they can be used in targeted mutagenesis of linked genes. To efficiently target all genes throughout the genome, we generated a large population of transgenic Arabidopsis plants containing the Ds launch pad construct, identified lines containing single Ds launch pad inserts, and mapped the positions of Ds launch pads in 89 lines. The integration sites of the Ds launch pads were relatively evenly distributed on all five chromosomes, except for a region of chromosomes 2 and 4 and the centromeric regions. This resource therefore provides access to the majority of the Arabidopsis genome for targeted tagging. PMID:12805583

  1. Pacemaker insertion

    PubMed Central

    Kotsakou, Maria; Kioumis, Ioannis; Lazaridis, George; Pitsiou, Georgia; Lampaki, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Tsakiridis, Kosmas; Katsikogiannis, Nikolaos; Karapantzos, Ilias; Karapantzou, Chrysanthi; Baka, Sofia; Mpoukovinas, Ioannis; Karavasilis, Vasilis; Rapti, Aggeliki; Trakada, Georgia; Zissimopoulos, Athanasios; Zarogoulidis, Konstantinos

    2015-01-01

    A pacemaker (PM) (or artificial PM, so as not to be confused with the heart’s natural PM) is a medical device that uses electrical impulses, delivered by electrodes contracting the heart muscles, to regulate the beating of the heart. The primary purpose of this device is to maintain an adequate heart rate, either because the heart’s natural PM is not fast enough, or there is a block in the heart’s electrical conduction system. Modern PMs are externally programmable and allow the cardiologist to select the optimum pacing modes for individual patients. Some combine a PM and defibrillator in a single implantable device. PMs can be temporary or permanent. Temporary PMs are used to treat short-term heart problems, such as a slow heartbeat that’s caused by a heart attack, heart surgery, or an overdose of medicine. Permanent PMs are used to control long-term heart rhythm problems. A PM can relieve some arrhythmia symptoms, such as fatigue and fainting. A PM also can help a person who has abnormal HRs resume a more active lifestyle. In the current mini review we will focus on the insertion of a PM and the possible pneumothorax that can be caused. PMID:25815303

  2. Genetic analysis of the virD operon of Agrobacterium tumefaciens: a search for functions involved in transport of T-DNA into the plant cell nucleus and in T-DNA integration.

    PubMed Central

    Koukolíková-Nicola, Z; Raineri, D; Stephens, K; Ramos, C; Tinland, B; Nester, E W; Hohn, B

    1993-01-01

    The transferred DNA (T-DNA) is transported from Agrobacterium tumefaciens to the nucleus and is stably integrated into the genome of many plant species. It has been proposed that the VirD2 protein, tightly attached to the T-DNA, pilots the T-DNA into the plant cell nucleus and that it is involved in integration. Using agroinfection and beta-glucuronidase expression as two different very sensitive transient assays for T-DNA transfer, together with assays for stable integration, we have shown that the C-terminal half of the VirD2 protein and the VirD3 protein are not involved in T-DNA integration. However, the bipartite nuclear localization signal, which is located within the C terminus of the VirD2 protein and which has previously been shown to be able to target a foreign protein into the plant cell nucleus, was shown to be required for efficient T-DNA transfer. virD4 mutants were shown by agroinfection to be completely inactive in T-DNA transfer. Images PMID:8380800

  3. 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)

  4. 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. PMID:16827753

  5. Traversing the Cell: Agrobacterium T-DNA’s Journey to the Host Genome

    PubMed Central

    Gelvin, Stanton B.

    2012-01-01

    The genus Agrobacterium is unique in its ability to conduct interkingdom genetic exchange. Virulent Agrobacterium strains transfer single-strand forms of T-DNA (T-strands) and several Virulence effector proteins through a bacterial type IV secretion system into plant host cells. T-strands must traverse the plant wall and plasma membrane, traffic through the cytoplasm, enter the nucleus, and ultimately target host chromatin for stable integration. Because any DNA sequence placed between T-DNA “borders” can be transferred to plants and integrated into the plant genome, the transfer and intracellular trafficking processes must be mediated by bacterial and host proteins that form complexes with T-strands. This review summarizes current knowledge of proteins that interact with T-strands in the plant cell, and discusses several models of T-complex (T-strand and associated proteins) trafficking. A detailed understanding of how these macromolecular complexes enter the host cell and traverse the plant cytoplasm will require development of novel technologies to follow molecules from their bacterial site of synthesis into the plant cell, and how these transferred molecules interact with host proteins and sub-cellular structures within the host cytoplasm and nucleus. PMID:22645590

  6. Analysis of the T-DNA structure in a large number of transgenic petunias generated by Agrobacterium-mediated transformation.

    PubMed

    Deroles, S C; Gardner, R C

    1988-05-01

    Southern hybridisation was performed on ninety-six transgenic petunias that had been selected for resistance to kanamycin. Just over half of the plants contained intact copies of the T-DNA. The most common rearrangements (at least 24 plants out of 96) were simple deleted derivatives that had lost one or both ends of the T-DNA. T-DNAs lacking the left border occurred at a frequency of 20%, and estimates of the frequency of T-DNAs lacking the right border were at least this high. Three plants contained grossly rearranged T-DNAs, of which all expressed the kanamycin resistance gene but only one transmitted the gene to progeny. Two plants lacked T-DNA homology altogether and did not express kanamycin resistance in their leaves or their progeny. Circumstantial evidence suggests that plants containing a chimaeric kanamycin resistance gene driven by the ocs promoter do not root efficiently in the presence of kanamycin. There was no correlation between intactness of the T-DNA and Mendelian inheritance of the kanamycin-resistance phenotype. However, a disproportionate number of plants showing non-Mendelian inheritance had a high copy number of their T-DNA. PMID:24272349

  7. Identification of virulence genes in the crucifer anthracnose fungus Colletotrichum higginsianum by insertional mutagenesis.

    PubMed

    Liu, Liping; Zhao, Dian; Zheng, Lu; Hsiang, Tom; Wei, Yangdou; Fu, Yanping; Huang, Junbin

    2013-11-01

    To investigate the molecular and genetic mechanisms underlying virulence of Colletotrichum higginsianum on Arabidopsis thaliana, a T-DNA insertion mutant library of C. higginsianum, the causal agent of crucifer anthracnose, was established using Agrobacterium tumefaciens-mediated transformation. Among 875 transformants tested for virulence on Arabidopsis, six mutants with altered virulence, including an appressorial melanin-deficient mutant T734, two mutants defective in penetration, T45 and B30, and three mutants, T679, T732 and T801, that cause hypersensitive reactions on host Arabidopsis, were obtained. Southern blot analysis indicated that the mutants T732 and T734 harbored single-site T-DNA integrations, while B30 harbored two T-DNA insertions. Border flanking sequences of T-DNAs from these mutants were recovered by inverse polymerase chain reaction (PCR) and thermal asymmetric interlaced PCR. Sequence analyses revealed that single T-DNA insertions in mutant T734 targeted the coding region of a gene with unknown function, and in mutant T732 targeted a gene encoding a copper amine oxidase. The two T-DNA insertion sites in mutant B30 were found in the coding region of a gene encoding an exosome component and in the upstream region of a DUF221-domain gene. None of these genes have previously been implicated in virulence of the phytopathogenic fungi. Among these avirulent mutants, T734 showed altered color in colony growth and produced melanin-deficient, albino appressoria. The T-DNA insert in T734 was detected in the coding region of a gene named C. higginsianum melanin-deficiency gene (Ch-MEL1), which is highly similar to a gene encoding a hypothetical protein in Colletotrichum gloeosporioides (GenBank ELA33048). To validate whether the Ch-MEL1 gene was associated with virulence of the mutant T734, a targeted gene disruption and complementation approach was used. The appressoria of ?Ch-mel1 null mutants were defective in melanization and failed to penetrate the host epidermal cells. When inoculated onto the wounded leaf tissues, the ?Ch-mel1 mutants grew on host tissues but failed to cause lesions beyond the wound site. In contrast, both the complement C?Ch-mel1-2 and the wild type produced melanized appressoria and caused necrosis on leaves of Arabidopsis. Ch-MEL1 is required for both appressorial melanin production in C. higginsianum and post-invasive growth in host tissues. Together with identification of other avirulent mutants and their associated genes, this study provides novel insights into molecular mechanisms underlying virulence of the hemibiotroph, C. higginsianum. PMID:23806215

  8. The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals.

    PubMed Central

    Tinland, B; Koukolíková-Nicola, Z; Hall, M N; Hohn, B

    1992-01-01

    Agrobacterium tumefaciens causes neoplastic growth in plants by transferring a piece of DNA, called T-DNA, into the nucleus of the plant cell. The virulence protein VirD2 of A. tumefaciens is tightly linked to the T-DNA and is thought to direct it to the plant genome. Here we show that the VirD2 protein contains two nuclear localization signals that are functional both in yeast and in plant cells. One signal is located in the N-terminal part of the protein and resembles a single-cluster-type nuclear localization signal. The second signal is near the C terminus and is a bipartite-type nuclear localization signal. The involvement of these sequences in the entry of the T-DNA into the nucleus is discussed. Images PMID:1502156

  9. 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…

  10. 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.

  11. Cloning quantitative trait loci by insertional mutagenesis.

    PubMed

    Soller, M; Beckmann, J S

    1987-07-01

    This study explores the theoretical potential of "insertional mutagenesis" (i.e., mutagenesis as a result of integration of novel DNA sequences into the germ line), as a means of cloning quantitative trait loci (QTL). The approach presented is based on a direct search for mutagenic effects of a quantitative nature, and makes no assumptions as to the nature of the loci affecting quantitative trait value. Since there are a very large number of potential insertion sites in the genome but only a limited number of target sites that can affect any particular trait, large numbers of inserts will have to be generated and screened. The effects of allelic variants at any single QTL on phenotype value are expected to be small relative to sampling variation. Thus two of three stages of replicate testing will be required for each insert in order to bring overall Type I error down to negligible proportions and yet maintain good statistical power for inserts with true effects on the quantitative traits under consideration. The overall effort involved will depend on the spectrum of mutagenic effects produced by insertional mutagenesis. This spectrum is presently unknown, but using reasonable estimates, about 10,000 inserts would have to be tested, at reasonable replicate numbers (n ? 30) and Type I error (?=0.01) in the first testing stage, to provide a high likelihood of detecting at least one insert with a true effect on a given quantitative trait of interest. Total offspring numbers required per true quantitative mutagenic effect detected decrease strongly with increased number of traits scored and increased number of inserts per initial transformed parent. In fact, it would appear that successful implementation of experiments of this sort will require the introduction of multiple independent inserts in the original parent individuals, by means of repeated transformation, or use of transposable elements as inserts. When biologically feasible, selfing would appear to be the method of choice for insert replication, and in all cases the experiments must be carried out in inbred lines to reduce error variation due to genetic segregation, and avoid confounding mutational effects of the insert with effects due to linkage with nearby segregating QTL. The special qualifications of Arabidopsis thaliana for studies of this sort are emphasized, and problems raised by somaclonal variation are discussed. PMID:24241675

  12. [Inserts for foot deformities].

    PubMed

    Stinus, H; Weber, F

    2005-08-01

    Inserts are orthopedic aids in the treatment of foot disorders that result from changes of the static or dynamic situation. Provision of appropriate orthopedic devices can relieve the pain caused by forefoot deformities either in lieu of surgical intervention or in rare cases also following surgical treatment to improve the symptoms of residual pain.Available materials provide support, padding, and cushioning. Inserts are custom-made to measure and/or based on a plaster impression. Determining the indication, prescribing the inlay, and checking the orthosis are the tasks of the physician. One treatment option for relieving the pain of forefoot deformities consists in conservative therapy with an insert combining features of padding and support as well as adjusting a ready-made shoe. The shoe and inlay should constitute a functional unit since often the optimal effect is only achieved with a combination of insert and orthopedic adjustment of the ready-made shoe. PMID:15991037

  13. 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...

  14. 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...

  15. 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.

  16. 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 cytokinin and auxin levels for further cell proliferation. PMID:25615824

  17. Regulation of Oncogene Expression in T-DNA-Transformed Host Plant Cells

    PubMed Central

    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 cytokinin and auxin levels for further cell proliferation. PMID:25615824

  18. ALS insertion devices

    SciTech Connect

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

    1990-11-01

    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. 18 refs., 9 figs., 5 tabs.

  19. Insertion in Persian

    ERIC Educational Resources Information Center

    Kambuziya, Aliyeh Kord-e Zafaranlu; Dehghan, Masoud

    2011-01-01

    This paper investigates epenthesis process in Persian to catch some results in relating to vowel and consonant insertion in Persian lexicon. This survey has a close relationship to the description of epenthetic consonants and the conditions in which these consonants are used. Since no word in Persian may begin with a vowel, so that hiatus can't be…

  20. Interfacial insert for electrical connectors

    NASA Technical Reports Server (NTRS)

    Macavay, D.

    1975-01-01

    The development of interfacial inserts for improved electric connectors is discussed. The inserts were manufactured from epoxy resins. The design features of the inserts and the manufacturing equipment are described. The reliability test program is reported. Drawings of the interfacial inserts are provided.

  1. Hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA

    SciTech Connect

    Hood, E.E.; Helmer, G.L.; Fraley, R.T.; Chilton, M.D.

    1986-12-01

    A binary-vectory strategy was used to study the hypervirulence of Agrobacterium tumefaciens A281, an L,L-succinamopine strain. Strain A281 is hypervirulent on several solanaceous plants. Plasmids were constructed (pCS65 and pCS277) carrying either the transferred DNA (T-DNA) or the remainder of the tumor-inducing (Ti) plasmid (pEHA101) from this strain and tested each of these constructs were tested in trans with complementary each of regions from heterologous Ti plasmids. Hypervirulence on tobacco could be reconstructed in a bipartite strain with the L,L-succinamopine T-DNA and the vir region on separate plasmids. pEHA101 was able to complement octopine T-DNA to hypervirulence on tobacco and tomato plants. Nopaline T-DNA was complemented better on tomato plants by pEHA101 than it was by its own nopaline vir region, but not to hypervirulence. L,L-Succinamopine T-DNA could not be complemented to hypervirulence on tobacco and tomato plants with either heterologous vir region. From these results the authors suggest that the hypervirulence of strain A281 is due to non-T-DNA sequences on the Ti plasmid.

  2. Ear tube insertion - series (image)

    MedlinePLUS

    Ear tube insertion is recommended for: fluid in the middle ear (present for more than 12 weeks) recurrent ear ... the accumulated fluid is suctioned out. A small tube is inserted through the incised eardrum in order ...

  3. Effects of P Element Insertions on Quantitative Traits in Drosophila Melanogaster

    PubMed Central

    Mackay, TFC.; Lyman, R. F.; Jackson, M. S.

    1992-01-01

    P element mutagenesis was used to construct 94 third chromosome lines of Drosophila melanogaster which contained on average 3.1 stable P element inserts, in an inbred host strain background previously free of P elements. The homozygous and heterozygous effects of the inserts on viability and abdominal and sternopleural bristle number were ascertained by comparing the chromosome lines with inserts to insert-free control lines of the inbred host strain. P elements reduced average homozygous viability by 12.2% per insert and average heterozygous viability by 5.5% per insert, and induced recessive lethal mutations at a rate of 3.8% per insert. Mutational variation for the bristle traits averaged over both sexes was 0.03V(e) per homozygous P insert and 0.003V(e) per heterozygous P insert, where V(e) is the environmental variance. Mutational variation was greater for the sexes considered separately because inserts had large pleiotropic effects on sex dimorphism of bristle characters. The distributions of homozygous effects of inserts on the bristle traits were asymmetrical, with the largest effects in the direction of reducing bristle number; and highly leptokurtic, with most of the increase in variance contributed by a few lines with large effects. The inserts had partially recessive effects on the bristle traits. Insert lines with extreme bristle effects had on average greatly reduced viability. PMID:1311697

  4. Insight into the molecular requirements for pathogenicity of Fusarium oxysporum f. sp. lycopersici through large-scale insertional mutagenesis

    PubMed Central

    Michielse, Caroline B; van Wijk, Ringo; Reijnen, Linda; Cornelissen, Ben JC; Rep, Martijn

    2009-01-01

    Background Fusarium oxysporum f. sp. lycopersici is the causal agent of vascular wilt disease in tomato. In order to gain more insight into the molecular processes in F. oxysporum necessary for pathogenesis and to uncover the genes involved, we used Agrobacterium-mediated insertional mutagenesis to generate 10,290 transformants and screened the transformants for loss or reduction of pathogenicity. Results This led to the identification of 106 pathogenicity mutants. Southern analysis revealed that the average T-DNA insertion is 1.4 and that 66% of the mutants carry a single T-DNA. Using TAIL-PCR, chromosomal T-DNA flanking regions were isolated and 111 potential pathogenicity genes were identified. Conclusions Functional categorization of the potential pathogenicity genes indicates that certain cellular processes, such as amino acid and lipid metabolism, cell wall remodeling, protein translocation and protein degradation, seem to be important for full pathogenicity of F. oxysporum. Several known pathogenicity genes were identified, such as those encoding chitin synthase V, developmental regulator FlbA and phosphomannose isomerase. In addition, complementation and gene knock-out experiments confirmed that a glycosylphosphatidylinositol-anchored protein, thought to be involved in cell wall integrity, a transcriptional regulator, a protein with unknown function and peroxisome biogenesis are required for full pathogenicity of F. oxysporum. PMID:19134172

  5. Cre/lox-mediated site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana by transient expression of cre.

    PubMed

    Vergunst, A C; Hooykaas, P J

    1998-10-01

    The Cre/lox system was used to obtain targeted integration of an Agrobacterium T-DNA at a lox site in the genome of Arabidopsis thaliana. Site-specific recombinants, and not random events, were preferentially selected by activation of a silent lox-neomycin phosphotransferase (nptII) target gene. To analyse the effectiveness of Agrobacterium-mediated transfer we used T-DNA vectors harbouring a single lox sequence (this vector had to circularize at the T-DNA left- and right-border sequences prior to site-specific integration) or two lox sequences (this vector allowed circularization at the lox sequences within the T-DNA either prior to or after random integration, followed by targeting of the circularized vector), respectively. Furthermore, to control the reversibility of the integration reaction, Cre recombinase was provided transiently by using a cotransformation approach. One precise stable integrant was found amongst the recombinant calli obtained after transformation with a double-lox T-DNA vector. The results indicate that Agrobacterium-mediated transformation can be used as a tool to obtain site-specific integration. PMID:9747847

  6. Application and Evaluation of the Interlaboratory Reproducibility of tRNA Intergenic Length Polymorphism Analysis (tDNA-PCR) for Identification of Streptococcus Species

    PubMed Central

    Baele, Margo; Storms, Virginie; Haesebrouck, Freddy; Devriese, Luc A.; Gillis, Monique; Verschraegen, Gerda; de Baere, Thierry; Vaneechoutte, Mario

    2001-01-01

    The discriminatory power, speed, and interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) combined with capillary electrophoresis was evaluated for the identification of streptococci. This method was carried out in three different laboratories under highly standardized conditions for 54 strains belonging to 18 different species. It was concluded that interlaboratory reproducibility of tDNA fingerprints produced by means of capillary electrophoresis was sufficiently high to permit the exchange between different laboratories and the construction of common libraries which can be consulted for comparison with fingerprints obtained independently in separate laboratories. In a second step, 17 other species were included in the study and examined in one of the participating laboratories. All Streptococcus species studied, except S. mitis, S. oralis, S. parasanguinis, S. pneumoniae, S. thermophilus, and S. vestibularis, showed distinguishable tDNA fingerprints. A database of well-characterized strains was constructed to enable computer-aided identification of unknown streptococcal isolates. PMID:11283068

  7. Networking Senescence-Regulating Pathways by Using Arabidopsis Enhancer Trap Lines1

    PubMed Central

    He, Yuehui; Tang, Weining; Swain, Johnnie D.; Green, Anthony L.; Jack, Thomas P.; Gan, Susheng

    2001-01-01

    The last phase of leaf development, generally referred to as leaf senescence, is an integral part of plant development that involves massive programmed cell death. Due to a sharp decline of photosynthetic capacity in a leaf, senescence limits crop yield and forest plant biomass production. However, the biochemical components and regulatory mechanisms underlying leaf senescence are poorly characterized. Although several approaches such as differential cDNA screening, differential display, and cDNA subtraction have been employed to isolate senescence-associated genes (SAGs), only a limited number of SAGs have been identified, and information regarding the regulation of these genes is fragmentary. Here we report on the utilization of enhancer trap approach toward the identification and analysis of SAGs. We have developed a sensitive large-scale screening method and have screened 1,300 Arabidopsis enhancer trap lines and have identified 147 lines in which the reporter gene GUS (?-glucuronidase) is expressed in senescing leaves but not in non-senescing ones. We have systematically analyzed the regulation of ?-glucuronidase expression in 125 lines (genetically, each contains single T-DNA insertion) by six senescence-promoting factors, namely abscisic acid, ethylene, jasmonic acid, brassinosteroid, darkness, and dehydration. This analysis not only reveals the complexity of the regulatory circuitry but also allows us to postulate the existence of a network of senescence-promoting pathways. We have also cloned three SAGs from randomly selected enhancer trap lines, demonstrating that reporter expression pattern reflects the expression pattern of the endogenous gene. PMID:11402199

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

    PubMed

    Rodi?, 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. PMID:26259033

  9. 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 the putative gene had 79% homology with the aspartate aminotransferase gene in Magnaporthe oryzae (XP_003719674.1). PMID:25361073

  10. 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 of the putative gene had 79% homology with the aspartate aminotransferase gene in Magnaporthe oryzae (XP_003719674.1). PMID:25361073

  11. Agrobacterium tumefaciens-mediated transformation of Aspergillus aculeatus for insertional mutagenesis

    PubMed Central

    2011-01-01

    Agrobacterium tumefaciens-mediated transformation (AMT) was applied to Aspergillus aculeatus. Transformants carrying the T-DNA from a binary vector pBIG2RHPH2 were sufficiently mitotically stable to allow functional genomic analyses. The AMT technique was optimized by altering the concentration of acetosyringone, the ratio and concentration of A. tumefaciens and A. aculeatus cells, the duration of co-cultivation, and the status of A. aculeatus cells when using conidia, protoplasts, or germlings. On average, 30 transformants per 104 conidia or 217 transformants per 107 conidia were obtained under the optimized conditions when A. tumefaciens co-cultured with fungi using solid or liquid induction media (IM). Although the transformation frequency in liquid IM was 100-fold lower than that on solid IM, the AMT method using liquid IM is better suited for high-throughput insertional mutagenesis because the transformants can be isolated on fewer selection media plates by concentrating the transformed germlings. The production of two albino A. aculeatus mutants by AMT confirmed that the inserted T-DNA disrupted the polyketide synthase gene AapksP, which is involved in pigment production. Considering the efficiency of AMT and the correlation between the phenotypes and genotypes of the transformants, the established AMT technique offers a highly efficient means for characterizing the gene function in A. aculeatus. PMID:22166586

  12. Characterization of pre-insertion loci of de novo L1 insertions

    PubMed Central

    Gasior, Stephen L.; Preston, Graeme; Hedges, Dale J.; Gilbert, Nicolas; Moran, John V.; Deininger, Prescott L.

    2007-01-01

    The human Long Interspersed Element-1 (LINE-1) and the Short Interspersed Element (SINE) Alu comprise 28% of the human genome. They share the same L1-encoded endonuclease for insertion, which recognizes an A+T-rich sequence. Under a simple model of insertion distribution, this nucleotide preference would lead to the prediction that the populations of both elements would be biased towards A+T-rich regions. Genomic L1 elements do show an A+T-rich bias. In contrast, Alu is biased towards G+C-rich regions when compared to the genome average. Several analyses have demonstrated that relatively recent insertions of both elements show less G+C content bias relative to older elements. We have analyzed the repetitive element and G+C composition of more than 100 pre-insertion loci derived from de novo L1 insertions in cultured human cancer cells, which should represent an evolutionarily unbiased set of insertions. An A+T-rich bias is observed in the 50 bp flanking the endonuclease target site, consistent with the known target site for the L1 endonuclease. The L1, Alu, and G+C content of 20 kb of the de novo pre-insertion loci show a different set of biases than those observed for fixed L1s in the human genome. In contrast to the insertion sites of genomic L1s, the de novo L1 pre-insertion loci are relatively L1-poor, Alu-rich and G+C-neutral. Finally, a statistically significant cluster of de novo L1 insertions was localized in the vicinity of the c-myc gene. These results suggest that the initial insertion preference of L1, while A+T-rich in the initial vicinity of the break site, can be influenced by the broader content of the flanking genomic region and have implications for understanding the dynamics of L1 and Alu distributions in the human genome. PMID:17067767

  13. Mars Observer Orbit Insertion Briefing

    NASA Technical Reports Server (NTRS)

    1993-01-01

    For the first part of this briefing, see NONP-NASA-VT-2000081556. Marvin Traxler continues his discussion on signal tracking from the Mars Observer. Julie Webster, Lead Engineer, Telecommunications Subsystem, is introduced. She explains how signals coming back from Mars are detected. Dr. Pasquale Esposito talks about flyby orbits and capture orbits. He says that frequencies coming from the spacecraft can determine if the spacecraft has flown by Mars, or if a capture orbit has occurred. Charles Whetsel, System Engineer Spacecraft Team, presents a computer program. He shows where the signal will appear on the computer from the Spacecraft. Suzanne Dodd presents orbit insertion geometry. Dr. Arden Albee, Project Scientist Mars Observer Project, Cal Tech tech, says that Mars is studied to get more data to confirm their hypotheses derived from previous Mars Missions such as the Viking Mars Program and the Mariner Program. Dr. Albee also describes instrumentation on the Mars Observer such as the Ultra Stable Oscillator, Mars Orbiter Laser Altimeter, and Magnetometer. The camera on the spacecraft is similar to a fax machine because it scans one line at a time as the spacecraft orbits Mars. Dr. Michael Malin, Principle Investigator Mars Observer Camera, Malin Space Science Systems, Inc., describe this process.

  14. 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.

  15. 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...

  16. 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.

  17. Sequence and temperature effect on electrical properties of modified poly(dA)-poly(dT) DNA wire

    NASA Astrophysics Data System (ADS)

    Suhendro, Daniel K.; Yudiarsah, Efta; Saleh, Rosari

    2016-03-01

    We studied the effect of thermal fluctuations on charge transport in two types of modified poly(dA)-poly(dT) DNA by calculating the transmission probability and I-V characteristics. The modification is carried out by replacing some of the AT base pairs with CG or GC base pairs placed randomly along the chain. The DNA model is placed in between two metallic electrodes. We used tight binding Hamiltonian to model the DNA molecule and electrodes. The thermal fluctuation is modeled by varying the twist angles of each base in the DNA. The transfer matrix and scattering matrix methods were employed in calculating the transmission probability. Transmission probability is used as an input in calculating I-V characteristics using Landauer-Büttiker formalism. The twist angle disorder is influenced by the temperature and frequency. In order to prevent spurious results, we take the average value transmission probability of 1000 realizations of twist angles. Our results indicate that the DNA modified by CG base pairs has its electrical properties impeded by increasing thermal fluctuation (increasing temperature and decreasing frequency). However, different behavior is observed on the DNA modified by GC base pairs, at low temperature its electrical properties are enhanced by decreasing frequency. Moreover, electrical properties of this DNA at high frequency are also enhanced as temperature increases.

  18. SPONTANEOUS TRANSMEMBRANE HELIX INSERTION THERMODYNAMICALLY MIMICS TRANSLOCON-GUIDED INSERTION

    PubMed Central

    Ulmschneider, Martin B.; Ulmschneider, Jakob P.; Schiller, Nina; Wallace, B. A.; von Heijne, Gunnar; White, Stephen H.

    2014-01-01

    The favorable transfer free energy for a transmembrane (TM) α-helix between the aqueous phase and lipid bilayer underlies the stability of membrane proteins. However, the connection between the energetics and process of membrane protein assembly by the Sec61/SecY translocon complex in vivo is not clear. Here, we directly determine the partitioning free energies of a family of designed peptides using three independent approaches: an experimental microsomal Sec61 translocon assay, a biophysical (spectroscopic) characterization of peptide insertion into hydrated planar lipid bilayer arrays, and an unbiased atomic-detail equilibrium folding-partitioning molecular dynamics simulation. Remarkably, the measured free energies of insertion are quantitatively similar for all three approaches. The molecular dynamics simulations show that TM helix insertion involves equilibrium with the membrane interface, suggesting that the interface may play a role in translocon-guided insertion. PMID:25204588

  19. Immediate Postpartum Intrauterine Contraception Insertion.

    PubMed

    Prager, Sarah W; McCoy, Erin E

    2015-12-01

    The immediate postpartum period is a favorable time for initiating contraception because women who have recently given birth are often highly motivated to use contraception, pregnancy is excluded, and the hospital setting offers convenience for patients and providers. This article addresses immediate postpartum intrauterine contraception (IUC) insertion for copper and levonorgestrel IUC. Immediate postpartum IUC is safe and effective, with a majority of IUC devices retained at 6 and 12 months. There are increased rates of expulsion, compared with delayed postpartum insertion and interval insertion, which need to be weighed against the risk of patients not returning for postpartum follow-up. PMID:26598300

  20. Chest tube insertion - series (image)

    MedlinePLUS

    ... lung can collapse, preventing adequate air exchange. Chest tubes are used to treat conditions that can cause ... Chest tubes are inserted to drain blood, fluid, or air and allow full expansion of the lungs. The tube ...

  1. 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 motivate 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.

  2. Live insertion method used for main renewal

    SciTech Connect

    Solkowitz, M. )

    1992-04-01

    Baltimore Gas and Electric's pilot project using the live insertion method to replace a cast iron main provided excellent results. Its use on Eastern Avenue, a major state highway, was cost effective, provided gas service to customers during the work, required relatively short construction time and resulted in only minor traffic disruptions. Gas service transfers to the new main were done at customer convenience and resulted in outages of only a few hours per customer. This paper reports that the project involved inserting a 6-in. plastic line inside an existing 10-in. cast iron main. Miller Pipeline Corp., Indianapolis, supplier of the Insertec {l angle}R{r angle} live insertion method was contracted for the job. Miller technicians assisted BG and E forces by providing a load analysis of the main, a pushing machine and related supplies, foaming equipment and pipe cutting tools. Company forces were responsible for all preparatory work, including opening all excavations, installing bypasses, and fusing and testing the plastic pipe. Service transfers and renewals were also completed by company employees.

  3. Inducible Expression of Agrobacterium Virulence Gene VirE2 for Stringent Regulation of T-DNA Transfer in Plant Transient Expression Systems.

    PubMed

    Denkovskienė, Erna; Paškevičius, Šarūnas; Werner, Stefan; Gleba, Yuri; Ražanskienė, Aušra

    2015-11-01

    Agrotransfection with viral vectors is an effective solution for the transient production of valuable proteins in plants grown in contained facilities. Transfection methods suitable for field applications are desirable for the production of high-volume products and for the transient molecular reprogramming of plants. The use of genetically modified (GM) Agrobacterium strains for plant transfections faces substantial biosafety issues. The environmental biosafety of GM Agrobacterium strains could be improved by regulating their T-DNA transfer via chemically inducible expression of virE2, one of the essential Agrobacterium virulence genes. In order to identify strong and stringently regulated promoters in Agrobacterium strains, we evaluated isopropyl-β-d-thiogalactoside-inducible promoters Plac, Ptac, PT7/lacO, and PT5/lacOlacO and cumic acid-inducible promoters PlacUV5/CuO, Ptac/CuO, PT5/CuO, and PvirE/CuO. Nicotiana benthamiana plants were transfected with a virE2-deficient A. tumefaciens strain containing transient expression vectors harboring inducible virE2 expression cassettes and containing a marker green fluorescent protein (GFP) gene in their T-DNA region. Evaluation of T-DNA transfer was achieved by counting GFP expression foci on plant leaves. The virE2 expression from cumic acid-induced promoters resulted in 47 to 72% of wild-type T-DNA transfer. Here, we present efficient and tightly regulated promoters for gene expression in A. tumefaciens and a novel approach to address environmental biosafety concerns in agrobiotechnology. PMID:26292850

  4. [Factors influencing agrobacterium-mediated transformation of maize elite inbred lines].

    PubMed

    Huang, Xue Qing; Wei, Zhi Ming

    2004-10-01

    Using the maize elite inbred lines 9046, Qi319, 414, Mo17 as target genotypes, a highly efficient transformation system was developed based on the study of factors influencing the Agrobacterium-mediated maize transformation. The results showed that the immature embryos of 1.0-2.0 mm in length were optimal transformation explants. Inclusion of acetosyringone (200 micromol/L) and ascorbatic acid (50 mg/L) in both infection medium and co-cultivation medium led to a significantly increase in the transformation efficiency. However, high osmotic treatment on the explants before inoculation didn't improve transformation efficiency. Delaying selection was beneficial to the survival of resistant calli. Using the optimized transformation procedure, 42 PCR-positive transgenic plants were obtained from the 4 elite inbred lines and the frequency of PCR-positive plant ranged from 1.71%-4.09%. The integration of the transgenes into the maize nuclear genome was confirmed by PCR analysis using bar- and gus-specific primers and by Southern blot using gus- specific probe. Most of transgenic plants (71.4%) had one copy of T-DNA insert. The establishment of the transformation system in maize provides an efficient way for transferring useful foreign genes to maize plants. PMID:15636368

  5. Molecular analysis of high-copy insertion sites in maize

    PubMed Central

    Settles, A. Mark; Latshaw, Susan; McCarty, Donald R.

    2004-01-01

    High-copy transposon mutagenesis is an effective tool for creating gene disruptions in maize. In order to molecularly define transposon-induced disruptions on a genome-wide scale, we optimized TAIL-PCR to amplify genomic DNA flanking maize Robertson’s Mutator insertions. Sample sequencing from 43 Mutator stocks and the W22 inbred line identified 676 non-redundant insertions, and only a small fraction of the flanking sequences showed significant similarity to maize repetitive sequences. We further designed and tested 79 arbitrary primers to identify 12 primers that amplify all Mutator insertions within a DNA sample at 3.1-fold redundancy. Importantly, the products are of sufficient size to use as substrates or probes for hybridization-based identification of gene disruptions. Our adaptation simplifies previously published TAIL-PCR protocols and should be transferable to other high-copy insertional mutagens. PMID:15060129

  6. 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.

  7. Chemo-port insertion: A cause of vocal cord palsy.

    PubMed

    Alazzawi, Sarmad; Hindi, Khalid; Malik, Ausama; Wee, Chong Aun; Prepageran, Narayanan

    2015-11-01

    We describe extremely rare cases of vocal cord palsy following surgical insertion of a chemo port. Our cohort consisted of patients with cancer who developed hoarseness immediately after central venous line placement for the administration of chemotherapy, with vocal cord palsy confirmed with flexible laryngoscopy. Given the timing, central venous line placement appears to be the most likely cause. Laryngoscope, 125:2551-2552, 2015. PMID:26108861

  8. Precise-Conductance Valve Insert

    NASA Technical Reports Server (NTRS)

    Outlaw, R. A.; Hoyt, R. F.

    1986-01-01

    Valve modification provides two operating modes fully open and small, precise leak. Copper insert with radially oriented holes allows small, controllable, precise effusion rate when valve closed or nearly unobstructed flow when valve open. Numerous applications in surface physics, vacuum physics, materials science, gas kinetics, thin films, and other areas of research requiring measured flows of gas into or out of system.

  9. Biomechanical Analysis of Differing Pedicle Screw Insertion Angles

    PubMed Central

    Sterba, William; Kim, Do-Gyoon; Fyhrie, David P.; Yeni, Yener N.; Vaidya, Rahul

    2007-01-01

    Background Pedicle screw fixation to stabilize lumbar spinal fusion has become the gold standard for posterior stabilization. A significant percentage of surgical candidates are classified as obese or morbidly obese. For these patients, the depth of the incisions and soft tissue makes it extremely difficult to insert pedicle screws along the pedicle axis. As such, the pedicle screws could only be inserted in a much more sagittal axis. However, biomechanical stability of the angled screw insertion has been controversial. We hypothesized that the straight or parallel screw was a more stable construct compared to the angled or axially inserted screw when subjected to caudal cyclic loading. Methods We obtained 12 fresh frozen lumbar vertebrae from L3 to L5 from five cadavers. Schantz screws (6.0mm) were inserted into each pedicle, one angled and along the axis of the pedicle and the other parallel to the spinous process. Fluoroscopic imaging was used to guide insertion. Each screw was then subjected to caudal cyclic loads of 50N for 2000 cycles at 2Hz. Analysis of initial damage, initial rate, and total damage during cyclic loading was undertaken. Findings Average total fatigue damage for straight screws measured 0.398±0.38 mm, and 0.689±0.96 mm for angled screws. Statistical analysis for total fatigue damage ratio of angled to straight screws revealed that a significant stability was achieved in straight- screw construct (p<0.03). Interpretation This study showed that straight screw insertion results in a more stable pedicle-screw construct. The angled screw insertion technique resulted in more scattered values of damage indicating that the outcome from the angled screw fixation is less predictable. This validates the use of this technique to implant pedicle screws across the axis of the pedicle rather than along the axis, (parallel to the midline sagittal line), and has broad implications in instrumented posterior lumbar spinal surgery. PMID:17208340

  10. 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.

  11. 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

  12. 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. PMID:25815301

  13. Large-Scale Mapping of Transposable Element Insertion Sites Using Digital Encoding of Sample Identity

    PubMed Central

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

    2014-01-01

    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. PMID:24374352

  14. Insertional mutagenesis using Tnt1 retrotransposon in potato.

    PubMed

    Duangpan, Saowapa; Zhang, Wenli; Wu, Yufang; Jansky, Shelley H; Jiang, Jiming

    2013-09-01

    Insertional mutagenesis using transfer DNA or transposable elements, which is an important tool in functional genomics and is well established in several crops, has not been developed in potato (Solanum tuberosum). Here, we report the application of the tobacco (Nicotiana tabacum) Tnt1 retrotransposon as an insertional mutagen in potato. The Tnt1 retrotransposon was introduced into a highly homozygous and self-compatible clone, 523-3, of the diploid wild potato species Solanum chacoense. Transposition of the Tnt1 elements introduced into 523-3 can be efficiently induced by tissue culture. Tnt1 preferentially inserted into genic regions in the potato genome and the insertions were stable during sexual reproduction, making Tnt1 an ideal mutagen in potato. Several distinct phenotypes associated with plant stature and leaf morphology were discovered in mutation screening from a total of 38 families derived from Tnt1-containing lines. We demonstrate that the insertional mutagenesis system based on Tnt1 and the 523-3 clone can be expanded to the genome-wide level to potentially tag every gene in the potato genome. PMID:23898040

  15. Insertional Mutagenesis Using Tnt1 Retrotransposon in Potato1[OPEN

    PubMed Central

    Duangpan, Saowapa; Zhang, Wenli; Wu, Yufang; Jansky, Shelley H.; Jiang, Jiming

    2013-01-01

    Insertional mutagenesis using transfer DNA or transposable elements, which is an important tool in functional genomics and is well established in several crops, has not been developed in potato (Solanum tuberosum). Here, we report the application of the tobacco (Nicotiana tabacum) Tnt1 retrotransposon as an insertional mutagen in potato. The Tnt1 retrotransposon was introduced into a highly homozygous and self-compatible clone, 523-3, of the diploid wild potato species Solanum chacoense. Transposition of the Tnt1 elements introduced into 523-3 can be efficiently induced by tissue culture. Tnt1 preferentially inserted into genic regions in the potato genome and the insertions were stable during sexual reproduction, making Tnt1 an ideal mutagen in potato. Several distinct phenotypes associated with plant stature and leaf morphology were discovered in mutation screening from a total of 38 families derived from Tnt1-containing lines. We demonstrate that the insertional mutagenesis system based on Tnt1 and the 523-3 clone can be expanded to the genome-wide level to potentially tag every gene in the potato genome. PMID:23898040

  16. 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.

  17. 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.

  18. Retargeting Sleeping Beauty Transposon Insertions by Engineered Zinc Finger DNA-binding Domains

    PubMed Central

    Voigt, Katrin; Gogol-Döring, Andreas; Miskey, Csaba; Chen, Wei; Cathomen, Toni; Izsvák, Zsuzsanna; Ivics, Zoltán

    2012-01-01

    The Sleeping Beauty (SB) transposon is a nonviral, integrating vector system with proven efficacy in preclinical animal models, and thus holds promise for future clinical applications. However, SB has a close-to-random insertion profile that could lead to genotoxic effects, thereby presenting a potential safety issue. We evaluated zinc finger (ZF) DNA-binding domains (DBDs) for their abilities to introduce a bias into SB's insertion profile. E2C, that binds a unique site in the erbB-2 gene, mediated locus-specific transposon insertions at low frequencies. A novel ZF targeting LINE1 repeats, ZF-B, showed specific binding to an 18-bp site represented by ~12,000 copies in the human genome. We mapped SB insertions using linear-amplification (LAM)-PCR and Illumina sequencing. Targeted insertions with ZF-B peaked at approximately fourfold enrichment of transposition around ZF-B binding sites yielding ~45% overall frequency of insertion into LINE1. A decrease in the ZF-B dataset with respect to transposon insertions in genes was found, suggesting that LINE1 repeats act as a sponge that “soak up” a fraction of SB insertions and thereby redirect them away from genes. Improvements in ZF technology and a careful choice of targeted genomic regions may improve the safety profile of SB for future clinical applications. PMID:22776959

  19. 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.

  20. Analysis of inserts in prokaryote genomes

    NASA Astrophysics Data System (ADS)

    Cristea, Paul Dan; Tuduce, Rodica Aurora

    2008-02-01

    Nucleotide genomic signals satisfy regularities that reveal restrictions in the distribution of nucleotides and pairs of nucleotides along DNA sequences. Structurally, a chromosome appears to be more than a plain text, by satisfying symmetry constrains that evoke the rhythm and rhyme in poems. These regularities make it easy to identify exogenous inserts in the genomes of prokaryotes, because such inserts obey different regularities than the background sequence. The paper presents instances of inserts found in the genomes of Bacillus subtilis, Mycobacterium tuberculosis and other prokaryotes. Inserts of exogenous material are frequently accompanied by complementary inserts tending to restore the original constrains.

  1. 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.

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

    PubMed Central

    Cotogni, Paolo; Pittiruti, Mauro

    2014-01-01

    Venous access devices are of pivotal importance for an increasing number of critically ill patients in a variety of disease states and in a variety of clinical settings (emergency, intensive care, surgery) and for different purposes (fluids or drugs infusions, parenteral nutrition, antibiotic therapy, hemodynamic monitoring, procedures of dialysis/apheresis). However, healthcare professionals are commonly worried about the possible consequences that may result using a central venous access device (CVAD) (mainly, bloodstream infections and thrombosis), both peripherally inserted central catheters (PICCs) and centrally inserted central catheters (CICCs). This review aims to discuss indications, insertion techniques, and care of PICCs in critically ill patients. PICCs have many advantages over standard CICCs. First of all, their insertion is easy and safe -due to their placement into peripheral veins of the arm- and the advantage of a central location of catheter tip suitable for all osmolarity and pH solutions. Using the ultrasound-guidance for the PICC insertion, the risk of hemothorax and pneumothorax can be avoided, as well as the possibility of primary malposition is very low. PICC placement is also appropriate to avoid post-procedural hemorrhage in patients with an abnormal coagulative state who need a CVAD. Some limits previously ascribed to PICCs (i.e., low flow rates, difficult central venous pressure monitoring, lack of safety for radio-diagnostic procedures, single-lumen) have delayed their start up in the intensive care units as common practice. Though, the recent development of power-injectable PICCs overcomes these technical limitations and PICCs have started to spread in critical care settings. Two important take-home messages may be drawn from this review. First, the incidence of complications varies depending on venous accesses and healthcare professionals should be aware of the different clinical performance as well as of the different risks 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

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

    PubMed

    Cotogni, Paolo; Pittiruti, Mauro

    2014-11-01

    Venous access devices are of pivotal importance for an increasing number of critically ill patients in a variety of disease states and in a variety of clinical settings (emergency, intensive care, surgery) and for different purposes (fluids or drugs infusions, parenteral nutrition, antibiotic therapy, hemodynamic monitoring, procedures of dialysis/apheresis). However, healthcare professionals are commonly worried about the possible consequences that may result using a central venous access device (CVAD) (mainly, bloodstream infections and thrombosis), both peripherally inserted central catheters (PICCs) and centrally inserted central catheters (CICCs). This review aims to discuss indications, insertion techniques, and care of PICCs in critically ill patients. PICCs have many advantages over standard CICCs. First of all, their insertion is easy and safe -due to their placement into peripheral veins of the arm- and the advantage of a central location of catheter tip suitable for all osmolarity and pH solutions. Using the ultrasound-guidance for the PICC insertion, the risk of hemothorax and pneumothorax can be avoided, as well as the possibility of primary malposition is very low. PICC placement is also appropriate to avoid post-procedural hemorrhage in patients with an abnormal coagulative state who need a CVAD. Some limits previously ascribed to PICCs (i.e., low flow rates, difficult central venous pressure monitoring, lack of safety for radio-diagnostic procedures, single-lumen) have delayed their start up in the intensive care units as common practice. Though, the recent development of power-injectable PICCs overcomes these technical limitations and PICCs have started to spread in critical care settings. Two important take-home messages may be drawn from this review. First, the incidence of complications varies depending on venous accesses and healthcare professionals should be aware of the different clinical performance as well as of the different risks 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

  4. 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; Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-26 Naka-cho, Koganei, Tokyo 184-8588

    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.

  5. Preoperative measurement of ACL insertion sites.

    PubMed

    Schwartzberg, Randy; Snyder, Kevin; Reuss, Bryan

    2015-02-01

    This study was designed to evaluate preoperative predictions of the femoral and tibial ACL insertion lengths. Intraoperative measurements were made of the femoral and tibial ACL insertion lengths with an arthroscopic ruler. A musculoskeletal radiologist and a sports medicine orthopedic surgeon measured the ACL insertions from preoperative magnetic resonance imagings (MRIs) in a blinded fashion. Correlations between height, weight, body mass index, and gender with intraoperative ACL insertion length measures were also evaluated. Patient height and male gender demonstrated strong correlations with intraoperative ACL insertion measures. A moderate correlation was seen between patient weight and intraoperative ACL insertion measures. When evaluating the frequency with which the raters' respective MRI measures were exactly the same as the intraoperative measures, moderate correlations were seen. The radiologist and orthopedic surgeons' MRI measures were within 3 mm of the intraoperative measures 98 and 75% of the time, respectively, for the femur and 92 and 83% of the time, respectively, for the tibia. This study concludes that preoperative MRI measures of ACL insertion lengths can be used to predict intraoperative ACL insertion lengths. Also, it is expected that taller patients and male patients should have greater ACL insertion lengths seen arthroscopically. PMID:24622911

  6. Nozzle insert for mixed mode fuel injector

    DOEpatents

    Lawrence, Keith E. (Peoria, IL)

    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.

  7. Repeated Transapical Transcatheter Aortic Valve Insertion.

    PubMed

    Khullar, Vishal; Greason, Kevin L; Sandhu, Gurpreet S; Pislaru, Sorin V

    2016-02-01

    Transcatheter aortic valves may develop structural valve deterioration. With that development the issue arises of repeated transcatheter aortic valve insertion. There are, unfortunately, limited data about repeated transapical valve insertion, with only a single case report in the literature. We report an additional successful case. PMID:26777928

  8. Lipid-insertion enables targeting functionalization of erythrocyte membrane-cloaked nanoparticles

    NASA Astrophysics Data System (ADS)

    Fang, Ronnie H.; Hu, Che-Ming J.; Chen, Kevin N. H.; Luk, Brian T.; Carpenter, Cody W.; Gao, Weiwei; Li, Shulin; Zhang, Dong-Er; Lu, Weiyue; Zhang, Liangfang

    2013-09-01

    RBC membrane-cloaked polymeric nanoparticles represent an emerging nanocarrier platform with extended circulation in vivo. A lipid-insertion method is employed to functionalize these nanoparticles without the need for direct chemical conjugation. Insertion of both folate and the nucleolin-targeting aptamer AS1411 shows receptor-specific targeting against model cancer cell lines.RBC membrane-cloaked polymeric nanoparticles represent an emerging nanocarrier platform with extended circulation in vivo. A lipid-insertion method is employed to functionalize these nanoparticles without the need for direct chemical conjugation. Insertion of both folate and the nucleolin-targeting aptamer AS1411 shows receptor-specific targeting against model cancer cell lines. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr03064d

  9. 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 the Company and ITER. The Fabrication Specifications may reflect some national requirements and regulations that are not fully provided here. This document presents the ITER CSI specifications.

  10. Localization properties of electronic states in a polaron model of poly(dG)-poly(dC) and poly(dA)-poly(dT) DNA polymers.

    PubMed

    Yamada, H; Starikov, E B; Hennig, D; Archilla, J F R

    2005-06-01

    We numerically investigate localization properties of electronic states in a static model of poly(dG)-poly(dC) and poly(dA)-poly(dT) DNA polymers with realistic parameters obtained by quantum-chemical calculation. The randomness in the on-site energies caused by the electron-phonon coupling is completely correlated to the off-diagonal parts. In the single electron model, the effect of the hydrogen-bond stretchings, the twist angles between the base pairs and the finite system size effects on the energy dependence of the localization length and on the Lyapunov exponent are given. The localization length is reduced by the influence of the fluctuations in the hydrogen bond stretchings. It is also shown that the helical twist angle affects the localization length in the poly(dG)-poly(dC) DNA polymer more strongly than in the poly(dA)-poly(dT) one. Furthermore, we show resonance structures in the energy dependence of the localization length when the system size is relatively small. PMID:15906117

  11. Elliptically polarizing adjustable phase insertion device

    DOEpatents

    Carr, Roger (Redwood City, CA)

    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. Sequential cooling insert for turbine stator vane

    DOEpatents

    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.

  13. 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.

  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. Insertion Homo- and Copolymerization of Diallyl Ether.

    PubMed

    Jian, Zhongbao; Mecking, Stefan

    2015-12-21

    The previously unresolved issue of polymerization of allyl monomers CH2 ?CHCH2 X is overcome by a palladium-catalyzed insertion polymerization of diallyl ether as a monomer. An enhanced 2,1-insertion of diallyl ether as compared to mono-allyl ether retards the formation of an unreactive five-membered cyclic O-chelate (after 1,2-insertion) that otherwise hinders further polymerization, and also enhances incorporation in ethylene polymers (20.4?mol?%). Cyclic ether repeat units are formed selectively (96?%-99?%) by an intramolecular insertion of the second allyl moiety of the monomer. These features even enable a homopolymerization to yield polymers (poly-diallyl ether) with degrees of polymerization of DPn ?44. PMID:26564774

  16. Shoe Inserts and Prescription Custom Orthotics

    MedlinePLUS

    ... from the wide variety of devices on the market can be overwhelming. Here are some podiatrist-tested tips to help you find the insert that best meets your needs: Consider your health. Do you have diabetes? Problems ...

  17. Insertable fluid flow passage bridgepiece and method

    DOEpatents

    Jones, Daniel O. (Glenville, NV)

    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.

  18. 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.

  19. The Basques according to polymorphic Alu insertions.

    PubMed

    de Pancorbo, M M; López-Martínez, M; Martínez-Bouzas, C; Castro, A; Fernández-Fernández, I; de Mayolo, G A; de Mayolo, A A; de Mayolo, P A; Rowold, D J; Herrera, R J

    2001-08-01

    Polymorphic Alu insertions provide a set of DNA markers of interest in human population genetics. Approximately 1000-2000 of these insertions have not reached fixation within the human genome. Each one of these polymorphic loci most probably resulted from a unique insertional event, and therefore all individuals possessing the insertion are related by descent not just state. In addition, the direction of mutational change is toward the gain of the Alu element at a particular locus. Therefore, the improved knowledge of both the ancestral state and the direction of mutational change greatly facilitates the analysis of population relationships. As a result, Alu insertion polymorphisms represent a significant tool for population genetic studies. In this study, polymorphic Alu insertions have been employed to ascertain phylogenetic relationships among Basque groups and worldwide populations. The Basques are considered to be a geographic isolate with a unique language and customs. They may be direct descendants of Cro-Magnon enclaves from the upper Paleolithic (38,000 to 10,000 years). The Basques are distributed among narrow valleys in northeastern Spain with little migration between them until recently. This characteristic may have had an effect on allelic frequency distributions. With the aim of studying this possible effect, we have analyzed six autosomal polymorphic Alu loci from four different sites within the Spanish Basque region in order to ascertain any genetic heterogeneity among the Basques. The results are consistent with a lack of homogeneity among these four autochthonous Basque groups. PMID:11511929

  20. 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.

  1. 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. PMID:1681170

  2. [Evaluation of peripherally inserted central catheters in a pediatric population].

    PubMed

    Baudin, G; Occelli, A; Boyer, C; Geoffray, A; Chevallier, P

    2013-10-01

    A peripherally inserted central catheter (PICC) is a central venous access mostly used in France in the adult population, whereas it is only rarely used in the pediatric population. The main objective of this study was to analyze a cohort of children treated with PICCs inserted under radiological guidance. We conducted a single-center study in the Radiology department of Nice University Hospital and the Lenval Foundation Children's Hospital. During a 43-month period between November 2008 and June 2012, a total of 67 catheter placement attempts were performed in 57 pediatric patients aged from 7 months to 18 years. We achieved 95.5% technical success with a median procedure duration of 17min. Only 6% of the PICC placements required light intravenous sedation; all the others were performed using a combination of local anesthesia, EMLA cream, and equimolar mixture of oxygen and nitrous oxide (EMONO). Subjective scale analysis of pain during catheter insertion showed a median score of 2.1. Catheter life ranged from 1 to 210 days (median, 38.3 days) with the treatment fully completed in 75% of the cases. The overall complication rate was 18.7% (4.9 per 1000 catheter-days), largely dominated by mechanical complications (9.4%) such as accidental removal (6.2%) or catheter obstruction (3.1%). Infectious complications occurred in 7.8% of the patients. The duration of catheterization and the use of tape to secure the catheter significantly affected the occurrence of complications. Peripheral insertion of central catheters was highly feasible in infants and children. It is a simple, safe, and effective alternative to intravenous central devices in the pediatric population. The occurrence of complications, typically mechanical, must be reduced and prevented by strict management of this type of central line by the nursing team. PMID:23953872

  3. Mechanisms for sodium insertion in carbon materials

    NASA Astrophysics Data System (ADS)

    Stevens, David Andrew

    2000-12-01

    This thesis details the mechanisms for sodium insertion into different carbons using both electrochemical and vapour techniques. Room temperature electrochemical measurements were completed to examine the insertion and removal of sodium from soft (graphitizable) and nanoporous hard (non-graphitizable) carbons prepared by the heat treatment of organic precursors to a range of temperatures. The mechanisms identified from these studies were further investigated through a series of in situ x-ray scattering studies on operating electrochemical cells. The results obtained were then compared with x-ray scattering measurements on carbons after exposure to sodium vapour at 890C. This work is primarily driven by the aluminium industry's need to understand how sodium insertion causes carbon cathode blocks in aluminium reduction cells to swell. The results obtained are also of relevance to the lithium-ion battery field as they help to verify mechanisms proposed in the literature for lithium insertion into carbon hosts. Some carbons were also identified that could accommodate large amounts of sodium, making them attractive candidates for anodes in rechargeable sodium ion batteries. For soft carbons, the results showed that both sodium and lithium insert between approximately parallel carbon layers along the sloping voltage region of the electrochemical curves, increasing the average interlayer spacing. The sodium and lithium capacities decreased with increasing carbon heat treatment temperature. For the soft carbons studied, the sodium capacity was found to be consistently lower than the lithium capacity, implying that some lithium-accessible sites were unavailable for sodium insertion. The electrochemical profiles for the hard carbons also contained capacity along a sloping voltage region and, as with the soft carbons, this was shown to result from the insertion of sodium and lithium between approximately parallel carbon layers. In contrast to the soft carbons, however, the electrochemical profiles for the hard carbons also exhibited significant capacity along low voltage plateaus at chemical potentials close to the chemical potential of the metal. An in situ small angle x-ray scattering experimental method was therefore developed to examine changes in scattering from the nanopores during sodium and lithium insertion/removal. The results from this technique clearly showed that the low voltage plateau capacity resulted from the insertion of sodium and lithium into nanopores in the carbon.

  4. Nozzle cavity impingement/area reduction insert

    DOEpatents

    Yu, Yufeng Phillip (Simpsonville, SC); Itzel, Gary Michael (Simpsonville, SC); Osgood, Sarah Jane (East Thetford, VT)

    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.

  5. Cancer gene discovery: exploiting insertional mutagenesis

    PubMed Central

    Ranzani, Marco; Annunziato, Stefano; Adams, David J.; Montini, Eugenio

    2013-01-01

    Insertional mutagenesis has been utilized as a functional forward genetics screen for the identification of novel genes involved in the pathogenesis of human cancers. Different insertional mutagens have been successfully used to reveal new cancer genes. For example, retroviruses (RVs) are integrating viruses with the capacity to induce the deregulation of genes in the neighborhood of the insertion site. RVs have been employed for more than 30 years to identify cancer genes in the hematopoietic system and mammary gland. Similarly, another tool that has revolutionized cancer gene discovery is the cut-and-paste transposons. These DNA elements have been engineered to contain strong promoters and stop cassettes that may function to perturb gene expression upon integration proximal to genes. In addition, complex mouse models characterized by tissue-restricted activity of transposons have been developed to identify oncogenes and tumor suppressor genes that control the development of a wide range of solid tumor types, extending beyond those tissues accessible using RV-based approaches. Most recently, lentiviral vectors (LVs) have appeared on the scene for use in cancer gene screens. LVs are replication defective integrating vectors that have the advantage of being able to infect non-dividing cells, in a wide range of cell types and tissues. In this review, we describe the various insertional mutagens focusing on their advantages/limitations and we discuss the new and promising tools that will improve the insertional mutagenesis screens of the future. PMID:23928056

  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. Insertional mutagenesis enables cleistothecial formation in a non-mating strain of Histoplasma capsulatum

    PubMed Central

    2010-01-01

    Background Histoplasma capsulatum is a pathogenic ascomycete fungus that rapidly loses mating ability in culture. Loss of mating ability, as well as the organism's low rate of targeted gene replacement, limits techniques available for genetic studies in H. capsulatum. Understanding molecular mechanisms regulating mating in this organism may allow us to reverse or prevent loss of mating in H. capsulatum strains, introducing a variety of classical genetics techniques to the field. We generated a strain, UC1, by insertional mutagenesis of the laboratory strain G217B, and found that UC1 acquired the ability to form mating structures called cleistothecia. The aim of this study was to determine the mechanism by which UC1 gained the ability to form cleistothecia. We also present initial studies demonstrating that UC1 can be used as a tool to determine molecular correlates of mating in H. capsulatum. Results The strain UC1 was found to have increased RNA levels of the mating locus transcription factor (MAT1-1-1), and the putative alpha pheromone (PPG1) compared to G217B. Agrobacterium-mediated transformation and integration of T-DNA from the vector pCB301-GFP-HYG were found to be partially responsible for the increased RNA levels of these genes; however, the site of integration appeared to play the largest role in the strain's ability to form cleistothecia. Silencing HMK1, a putative FUS3/KSS1 homolog, had no effect on cleistothecial production by UC1. Protein kinase C (PKC1) RNA and protein levels were increased in UC1 compared to G217B, and pheromone production was found to be linked with Pkc1 activity in H. capsulatum. Conclusions The site of the T-DNA integration event appears to play the largest role in UC1's ability to form cleistothecia. We show that the UC1 strain can be used as a tool to study cleistothecia production in H. capsulatum by manipulating the strain, or by identifying differences between UC1 and G217B. Using these approaches, we were able to link Pkc1 activity with pheromone production in H. capsulatum; however, further studies are required to determine molecular mechanisms behind this. These studies may reveal regulatory mechanisms that can be manipulated to restore mating ability in H. capsulatum laboratory strains. PMID:20158914

  8. 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.

  9. 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.

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

    PubMed Central

    Franklin, Iain; Gilmore, Christopher

    2015-01-01

    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. PMID:26229681

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

    PubMed

    Franklin, Iain; Gilmore, Christopher

    2015-06-01

    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. PMID:26229681

  12. 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

  13. Mosaic retroposon insertion patterns in placental mammals.

    TOXLINE Toxicology Bibliographic Information

    Churakov G; Kriegs JO; Baertsch R; Zemann A; Brosius J; Schmitz J

    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.

  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. PMID:19261842

  15. 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.

  16. Circular permutant GFP insertion folding reporters

    SciTech Connect

    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.

  17. Circular permutant GFP insertion folding reporters

    SciTech Connect

    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.

  18. RERTR-12 Insertion 2 Irradiation Summary Report

    SciTech Connect

    D. M. Perez; G. S. Chang; D. M. Wachs; G. A. Roth; N. E. Woolstenhulme

    2012-09-01

    The Reduced Enrichment for Research and Test Reactor (RERTR) experiment RERTR-12 was designed to provide comprehensive information on the performance of uranium-molybdenum (U-Mo) based monolithic fuels for research reactor applications.1 RERTR-12 insertion 2 includes the capsules irradiated during the last three irradiation cycles. These capsules include Z, Y1, Y2 and Y3 type capsules. The following report summarizes the life of the RERTR-12 insertion 2 experiment through end of irradiation, including as-run neutronic analysis results, thermal analysis results and hydraulic testing results.

  19. 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.

  20. ALS insertion device block measurement and inspection

    SciTech Connect

    Marks, S.; Carrieri, J.; Cook, C.; Hassenzahl, W.V.; Hoyer, E.; Plate, D.

    1991-05-01

    The performance specifications for ALS insertion devices require detailed knowledge and strict control of the Nd-Fe-B permanent magnet blocks incorporated in these devices. This paper describes the measurement and inspection apparatus and the procedures designed to qualify and characterize these blocks. A detailed description of a new, automated Helmholtz coil facility for measurement of the three components of magnetic moment is included. Physical block inspection and magnetic moment measurement procedures are described. Together they provide a basis for qualifying blocks and for specifying placement of blocks within an insertion devices' magnetic structures. 1 ref., 4 figs.

  1. Matching Sequences under Deletion/Insertion Constraints

    PubMed Central

    Sankoff, David

    1972-01-01

    Given two finite sequences, we wish to find the longest common subsequences satisfying certain deletion/insertion constraints. Consider two successive terms in the desired subsequence. The distance between their positions must be the same in the two original sequences for all but a limited number of such pairs of successive terms. Needleman and Wunsch gave an algorithm for finding longest common subsequences without constraints. This is improved from the viewpoint of computational economy. An economical algorithm is then elaborated for finding subsequences satisfying deletion/insertion constraints. This result is useful in the study of genetic homology based on nucleotide or amino-acid sequences. PMID:4500555

  2. Active interlock for storage ring insertion devices

    SciTech Connect

    Rochman, J.L.; Nawrocky, R.J.

    1989-01-01

    The insertion devices in the NSLS x-ray ring produce such intense photon beams that passive measures alone are not adequate to protect the vacuum chamber. An active interlock is used to prevent thermal damage from improperly steered electron beams. The interlock system measures the position of the electron beam on both sides of the insertion devices using pick-up electrodes and interrupts the rf if the beam moves outside a safe window. The interlock features redundant circuitry as well as an automatic testing procedure. 1 ref., 4 figs.

  3. 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.

  4. 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

  5. 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

  6. 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.

  7. 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.

  8. 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.'

  9. 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.

  10. 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.

  11. 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...

  12. 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...

  13. Gene expression variation in Drosophila melanogaster due to rare transposable element insertion alleles of large effect.

    PubMed

    Cridland, Julie M; Thornton, Kevin R; Long, Anthony D

    2015-01-01

    Transposable elements are a common source of genetic variation that may play a substantial role in contributing to gene expression variation. However, the contribution of transposable elements to expression variation thus far consists of a handful of examples. We used previously published gene expression data from 37 inbred Drosophila melanogaster lines from the Drosophila Genetic Reference Panel to perform a genome-wide assessment of the effects of transposable elements on gene expression. We found thousands of transcripts with transposable element insertions in or near the transcript and that the presence of a transposable element in or near a transcript is significantly associated with reductions in expression. We estimate that within this example population, ∼2.2% of transcripts have a transposable element insertion, which significantly reduces expression in the line containing the transposable element. We also find that transcripts with insertions within 500 bp of the transcript show on average a 0.67 standard deviation decrease in expression level. These large decreases in expression level are most pronounced for transposable element insertions close to transcripts and the effect diminishes for more distant insertions. This work represents the first genome-wide analysis of gene expression variation due to transposable elements and suggests that transposable elements are an important class of mutation underlying expression variation in Drosophila and likely in other systems, given the ubiquity of these mobile elements in eukaryotic genomes. PMID:25335504

  14. The Genomic Distribution of L1 Elements: The Role of Insertion Bias and Natural Selection

    PubMed Central

    Graham, Todd; Boissinot, Stephane

    2006-01-01

    LINE-1 (L1) retrotransposons constitute the most successful family of retroelements in mammals and account for as much as 20% of mammalian DNA. L1 elements can be found in all genomic regions but they are far more abundant in AT-rich, gene-poor, and low-recombining regions of the genome. In addition, the sex chromosomes and some genes seem disproportionately enriched in L1 elements. Insertion bias and selective processes can both account for this biased distribution of L1 elements. L1 elements do not appear to insert randomly in the genome and this insertion bias can at least partially explain the genomic distribution of L1. The contrasted distribution of L1 and Alu elements suggests that postinsertional processes play a major role in shaping L1 distribution. The most likely mechanism is the loss of recently integrated L1 elements that are deleterious (negative selection) either because of disruption of gene function or their ability to mediate ectopic recombination. By comparison, the retention of L1 elements because of some positive effect is limited to a small fraction of the genome. Understanding the respective importance of insertion bias and selection will require a better knowledge of insertion mechanisms and the dynamics of L1 inserts in populations. PMID:16877820

  15. Functional Genomics of Eukaryotic Photosynthesis Using Insertional Mutagenesis of Chlamydomonas reinhardtii1

    PubMed Central

    Dent, Rachel M.; Haglund, Cat M.; Chin, Brian L.; Kobayashi, Marilyn C.; Niyogi, Krishna K.

    2005-01-01

    The unicellular green alga Chlamydomonas reinhardtii is a widely used model organism for studies of oxygenic photosynthesis in eukaryotes. Here we describe the development of a resource for functional genomics of photosynthesis using insertional mutagenesis of the Chlamydomonas nuclear genome. Chlamydomonas cells were transformed with either of two plasmids conferring zeocin resistance, and insertional mutants were selected in the dark on acetate-containing medium to recover light-sensitive and nonphotosynthetic mutants. The population of insertional mutants was subjected to a battery of primary and secondary phenotypic screens to identify photosynthesis-related mutants that were pigment deficient, light sensitive, nonphotosynthetic, or hypersensitive to reactive oxygen species. Approximately 9% of the insertional mutants exhibited 1 or more of these phenotypes. Molecular analysis showed that each mutant line contains an average of 1.4 insertions, and genetic analysis indicated that approximately 50% of the mutations are tagged by the transforming DNA. Flanking DNA was isolated from the mutants, and sequence data for the insertion sites in 50 mutants are presented and discussed. PMID:15653810

  16. Screw-Thread Inserts As Temporary Flow Restrictors

    NASA Technical Reports Server (NTRS)

    Trimarchi, Paul

    1992-01-01

    Coil-spring screw-thread inserts found useful as temporary flow restrictors. Inserts placed in holes through which flow restricted, effectively reducing cross sections available for flow. Friction alone holds inserts against moderate upstream pressures. Use of coil-spring thread inserts as flow restrictors conceived as inexpensive solution to problem of adjusting flow of oxygen through orifices in faceplate into hydrogen/oxygen combustion chamber. Installation and removal of threaded inserts gentle enough not to deform orifice tubes.

  17. 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 styloid, and the center of the PBF was 8.96 mm (8.2-9.7) from the tip of the fibular styloid. Conclusion: A tibial footprint, distal fibular footprint, medial fibular footprint, and proximal fibular footprint were all consistent components of the insertion of the biceps femoris. Consistent relationships existed between the biceps femoris and insertions of the ALL and FCL. Clinical Relevance: The size of these footprints and distances from pertinent surgical landmarks will guide repairs of biceps femoris avulsion injuries. PMID:26535398

  18. 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. PMID:25931804

  19. PET Performance Evaluation of an MR-Compatible PET Insert.

    PubMed

    Wu, Yibao; Catana, Ciprian; Farrell, Richard; Dokhale, Purushottam A; Shah, Kanai S; Qi, Jinyi; Cherry, Simon R

    2009-06-16

    A magnetic resonance (MR) compatible positron emission tomography (PET) insert has been developed in our laboratory for simultaneous small animal PET/MR imaging. This system is based on lutetium oxyorthosilicate (LSO) scintillator arrays with position-sensitive avalanche photodiode (PSAPD) photodetectors. The PET performance of this insert has been measured. The average reconstructed image spatial resolution was 1.51 mm. The sensitivity at the center of the field of view (CFOV) was 0.35%, which is comparable to the simulation predictions of 0.40%. The average photopeak energy resolution was 25%. The scatter fraction inside the MRI scanner with a line source was 12% (with a mouse-sized phantom and standard 35 mm Bruker 1H RF coil), 7% (with RF coil only) and 5% (without phantom or RF coil) for an energy window of 350-650 keV. The front-end electronics had a dead time of 390 ns, and a trigger extension dead time of 7.32 ?s that degraded counting rate performance for injected doses above ~0.75 mCi (28 MBq). The peak noise-equivalent count rate (NECR) of 1.27 kcps was achieved at 290 ?Ci (10.7 MBq). The system showed good imaging performance inside a 7-T animal MRI system; however improvements in data acquisition electronics and reduction of the coincidence timing window are needed to realize improved NECR performance. PMID:21072320

  20. Insertion and Management of Temporary Pacemakers.

    PubMed

    Sullivan, Breandan Lawrence; Bartels, Karsten; Hamilton, Natalie

    2016-03-01

    Temporary pacemakers are used in a variety of critical care settings. These life-saving devices are reviewed in 2 major categories in this review: first, the insertion and management of epicardial pacemakers after and during cardiac surgery; and second, the insertion of transvenous temporary pacemakers for the emergent treatment of bradyarrhythmias. Temporary epicardial pacemakers are used routinely in patients recovering from cardiac surgery. Borrowing from advances in cardiac resynchronization therapy there are many theoretical and untested benefits to pacing the postoperative cardiac surgery patient. Temporary transvenous pacing is traditionally an emergency procedure to stabilize patients suffering from hemodynamically unstable bradyarrhythmia. We review the traditional and expanding use of transvenous pacemakers inside and outside the operating room. PMID:26134176

  1. 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.

  2. Electrochemical insertion of sodium into carbon

    NASA Astrophysics Data System (ADS)

    Doeff, Marca M.; Ma, Yanping; Visco, Steven J.; Dejonghe, Lutgard C.

    1993-12-01

    Electrochemical insertion of sodium ions into carbon using solid polymer electrolytes or organic liquid electrolytes is described. Cells with the configuration Na P(EO)(sub *)NaCF3SO3 CP(EO) = polyethylene oxide or Na/liquid electrolyte/C were galvanostatically discharged, charged, and cycled. The extent of insertion into C (i.e., x in Na(+)C(sub x -)) was found to be a strong function of the type and particle size of the carbon used, and the reversibility of the process was highly dependent upon the type of electrolyte used. The possibility of designing sodium ion rocking chair cells is discussed, and a first-generation example, using a petroleum coke anode, polymer electrolyte, and sodium cobalt bronze cathode is described.

  3. 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

  4. A small, insertable oven for boronization

    NASA Astrophysics Data System (ADS)

    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.

  5. Symplectic models for general insertion devices

    SciTech Connect

    Wu, Y.; Forest, E.; Robin, D. S.; Nishimura, H.; Wolski, A.; Litvinenko, V. N.

    2001-06-01

    A variety of insertion devices (IDs), wigglers and undulators, linearly or elliptically polarized,are widely used as high brightness radiation sources at the modern light source rings. Long and high-field wigglers have also been proposed as the main source of radiation damping at next generation damping rings. As a result, it becomes increasingly important to understand the impact of IDs on the charged particle dynamics in the storage ring. In this paper, we report our recent development of a general explicit symplectic model for IDs with the paraxial ray approximation. High-order explicit symplectic integrators are developed to study real-world insertion devices with a number of wiggler harmonics and arbitrary polarizations.

  6. Complex central venous catheter insertion for hemodialysis.

    PubMed

    Powell, Steven; Belfield, Jane

    2014-01-01

    Despite the introduction of payment by results in the UK, there has been no decrease in central venous catheter (CVC) use. In part, this may relate to a requirement to dialyse through a CVC while autogenous access matures. Mortality data have improved in parallel and patients on hemodialysis live longer, which may lead to an increased exposure to CVCs.Exposure to CVCs carries a significant risk of infection and occlusion requiring their repositioning or exchange. The mid to long-term sequelae of CVC use is central venous occlusion leaving clinical teams with an ever increasing challenge to find adequate venous access.In this article, we will discuss the challenges faced by operators inserting CVCs into the hemodialysis-dependent patient who has exhausted more tradition insertion sites. These include translumbar caval catheters, transocclusion and transcollateral catheters, transjugular Inferior Vena Cava catheter positioning, and transhepatic catheters. We will demonstrate the techniques employed, complications, and anticipated longevity of function. PMID:24817471

  7. 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}

  8. Electrochemical insertion of sodium into carbon

    SciTech Connect

    Doeff, M.M.; Yanping Ma; Visco, S.J.; De Jonghe, L.C.

    1993-12-01

    Electrochemical insertion of sodium ions into carbon using solid polymer electrolytes or organic liquid electrolytes is described. Cells with the configuration Na/P(EO){sub 8}NaCF{sub 3}SO{sub 3}/C where P(EO) = polyethylene oxide or Na/liquid electrolyte/C were galvanostatically discharged, charged, and cycled. The extent of insertion into C (i.e., x in Na{sup +}C{sub x{minus}}) was found to be a strong function of the type and particle size of the carbon used, and the reversibility of the process was highly dependent upon the type of electrolyte used. The possibility of designing a sodium ion rocking chair cell is discussed, and a first-generation example, using a petroleum coke anode, polymer electrolyte, and sodium cobalt bronze cathode is described.

  9. A Monte Carlo study of peptide insertion into lipid bilayers: equilibrium conformations and insertion mechanisms.

    PubMed Central

    Maddox, Michael W; Longo, Marjorie L

    2002-01-01

    The membrane insertion behavior of two peptides, Magainin2 and M2 delta, was investigated by applying the Monte Carlo simulation technique to a theoretical model. The model included many novel aspects, such as a new semi-empirical lipid bilayer model and a new set of semi-empirical transfer energies, which reproduced the experimental insertion behavior of Magainin2 and M2 delta without parameter fitting. Additionally, we have taken into account diminished internal (intramolecular) hydrogen bonding at the N- and C-termini of helical peptides. All simulations were carried out at 305 K, above the membrane thermal phase transition temperature, and at pH 7.0. The peptide equilibrium conformations are discussed for a range of bilayers with tail polarities varying from octanol-like to alkane-like. Probability distributions of the individual amino-acid-residue positions show the dynamic nature of these equilibrium conformations. Two different insertion mechanisms for M2 delta, and a translocation mechanism for Magainin2, are described. A study of the effect of bilayer thickness on M2 delta insertion suggests a critical thickness above which insertion is unfavorable. Additionally, we did not need to use an orientational potential or array of hard cylinders to persuade M2 delta to insert perpendicular to the membrane surface. Instead, we found that diminished internal hydrogen bonding in the helical conformation anchored the termini in the headgroups and resulted in a nearly perpendicular orientation. PMID:11751313

  10. 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.

  11. Comparison of Insertional RNA Editing in Myxomycetes

    PubMed Central

    Chen, Cai; Frankhouser, David; Bundschuh, Ralf

    2012-01-01

    RNA editing describes the process in which individual or short stretches of nucleotides in a messenger or structural RNA are inserted, deleted, or substituted. A high level of RNA editing has been observed in the mitochondrial genome of Physarum polycephalum. The most frequent editing type in Physarum is the insertion of individual Cs. RNA editing is extremely accurate in Physarum; however, little is known about its mechanism. Here, we demonstrate how analyzing two organisms from the Myxomycetes, namely Physarum polycephalum and Didymium iridis, allows us to test hypotheses about the editing mechanism that can not be tested from a single organism alone. First, we show that using the recently determined full transcriptome information of Physarum dramatically improves the accuracy of computational editing site prediction in Didymium. We use this approach to predict genes in the mitochondrial genome of Didymium and identify six new edited genes as well as one new gene that appears unedited. Next we investigate sequence conservation in the vicinity of editing sites between the two organisms in order to identify sites that harbor the information for the location of editing sites based on increased conservation. Our results imply that the information contained within only nine or ten nucleotides on either side of the editing site (a distance previously suggested through experiments) is not enough to locate the editing sites. Finally, we show that the codon position bias in C insertional RNA editing of these two organisms is correlated with the selection pressure on the respective genes thereby directly testing an evolutionary theory on the origin of this codon bias. Beyond revealing interesting properties of insertional RNA editing in Myxomycetes, our work suggests possible approaches to be used when finding sequence motifs for any biological process fails. PMID:22383871

  12. Comparison of insertional RNA editing in Myxomycetes.

    PubMed

    Chen, Cai; Frankhouser, David; Bundschuh, Ralf

    2012-01-01

    RNA editing describes the process in which individual or short stretches of nucleotides in a messenger or structural RNA are inserted, deleted, or substituted. A high level of RNA editing has been observed in the mitochondrial genome of Physarum polycephalum. The most frequent editing type in Physarum is the insertion of individual Cs. RNA editing is extremely accurate in Physarum; however, little is known about its mechanism. Here, we demonstrate how analyzing two organisms from the Myxomycetes, namely Physarum polycephalum and Didymium iridis, allows us to test hypotheses about the editing mechanism that can not be tested from a single organism alone. First, we show that using the recently determined full transcriptome information of Physarum dramatically improves the accuracy of computational editing site prediction in Didymium. We use this approach to predict genes in the mitochondrial genome of Didymium and identify six new edited genes as well as one new gene that appears unedited. Next we investigate sequence conservation in the vicinity of editing sites between the two organisms in order to identify sites that harbor the information for the location of editing sites based on increased conservation. Our results imply that the information contained within only nine or ten nucleotides on either side of the editing site (a distance previously suggested through experiments) is not enough to locate the editing sites. Finally, we show that the codon position bias in C insertional RNA editing of these two organisms is correlated with the selection pressure on the respective genes thereby directly testing an evolutionary theory on the origin of this codon bias. Beyond revealing interesting properties of insertional RNA editing in Myxomycetes, our work suggests possible approaches to be used when finding sequence motifs for any biological process fails. PMID:22383871

  13. Insertional Achilles tendinosis: pathogenesis and treatment.

    PubMed

    Den Hartog, Bryan D

    2009-12-01

    Insertional Achilles tendinopathy can be a painful debilitating condition that should initially be treated non-operatively. If pain becomes chronic and debilitating, despite appropriate conservative treatment, debridement of the diseased portion of the Achilles tendon and removal of the impinging calcaneal prominence and transfer of the flexor hallucis longus through a single incision can be a reliable pain relieving procedure with relatively high patient satisfaction. PMID:19857838

  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. Investigation of flexible hose insertion forces and selected factors.

    PubMed

    Drinkaus, P; Armstrong, T; Foulke, J

    2009-01-01

    Flexible hoses are widely used in automobiles. Understanding the force required to insert a hose, independent of worker variation, is important for engineers, designers and health professionals. Various parameters affect the insertion forces including the interference between the hose and flange. Also, it has been observed that workers often add motions during the insertion process, including rotation and oscillation. This paper describes the effects of these parameters on axial insertion forces. A programmable mill was used to isolate the effects of four variables on hose insertion force: fit, insertion speed, rotational velocity and oscillation magnitude. The results show that the magnitude of the interference of the 19 mm inside diameters (ID) heater hose and the flange, and the insertion technique affect the insertion forces; resulting in a range of insertion forces from 11.1+/-0.2 to 128.4+/-11.3N. PMID:18339355

  16. Hollow polymer microneedles array resistance and insertion tests.

    PubMed

    Lhernould, Marion Sausse; Deleers, Michel; Delchambre, Alain

    2015-03-01

    Microneedles are developed in order to become the transdermal administration method of the future. They however still face numerous challenges. This paper addresses the challenge to effectively insert the microneedle arrays into membranes. A recently proposed model membrane and test method for microneedles insertion, published in International Journal of Pharmaceutics, is used in this aim. A moulded 4 by 4 hollow polymer microneedle array developed at the Université Libre de Bruxelles is tested for insertion using this model. Results show that the array is extremely resistant to insertion, it can withstand very high forces and even multiple insertions without blunting. Different insertion tests were performed on a folded in eight Parafilm® film because it exhibits excellent similarity to porcine skin. The insertion force, the insertion speed and the holding time of the array against membranes must be optimised in order to get efficient reliable insertions at, at least, 500?m depth. PMID:25595569

  17. Rack Insertion End Effector (RIEE) guidance

    NASA Astrophysics Data System (ADS)

    Malladi, Narasimha S.

    1994-10-01

    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.

  18. 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.

  19. Contemporary practice of testicular prosthesis insertion

    PubMed Central

    Mohammed, Aza; Yassin, Musaab; Hendry, David; Walker, Gregory

    2015-01-01

    Objectives To assess the practice of testicular prosthesis insertion (TPI) related to orchidectomy in one geographical region and to identify the difference in the rates of insertion among different age groups. Patients and methods Males who underwent orchidectomy between 1989 and 2009 were identified from data collected from Scottish Morbidity Records. Patients were classified into six age groups. The TPI rate and relation to original orchidectomy were analysed according to different age groups. Results In all, 3364 patients underwent orchidectomy in the 20-year period of the study. The most common indications for orchidectomy were atrophy, undescended testes, torsion, and tumour. In the same period, 530 patients had a TPI, with 59.4% of them (316 patients) having TPI at initial surgery, 17.3% (92) as a second surgical procedure, and 22.8% (122) having the TPI without prior history of orchidectomy. Among patients who underwent TPI, postpubertal males were more likely to have simultaneous insertion at the time of orchidectomy than prepubertal males (83% vs 32%; odds ratio 10.44, 95% confidence interval 5.23–20.82; P < 0.01). Conclusion Younger males are more likely to have TPI at a later date. Paediatric urologists should be mindful of the possibility of concurrent TPI at the time of initial scrotal/groin exploration. PMID:26609448

  20. Software-implemented fault insertion: An FTMP example

    NASA Technical Reports Server (NTRS)

    Czeck, Edward W.; Siewiorek, Daniel P.; Segall, Zary Z.

    1987-01-01

    This report presents a model for fault insertion through software; describes its implementation on a fault-tolerant computer, FTMP; presents a summary of fault detection, identification, and reconfiguration data collected with software-implemented fault insertion; and compares the results to hardware fault insertion data. Experimental results show detection time to be a function of time of insertion and system workload. For the fault detection time, there is no correlation between software-inserted faults and hardware-inserted faults; this is because hardware-inserted faults must manifest as errors before detection, whereas software-inserted faults immediately exercise the error detection mechanisms. In summary, the software-implemented fault insertion is able to be used as an evaluation technique for the fault-handling capabilities of a system in fault detection, identification and recovery. Although the software-inserted faults do not map directly to hardware-inserted faults, experiments show software-implemented fault insertion is capable of emulating hardware fault insertion, with greater ease and automation.

  1. Autodirected insertion: preinserted VDAC channels greatly shorten the delay to the insertion of new channels.

    PubMed Central

    Xu, X; Colombini, M

    1997-01-01

    VDAC, a mitochondrial outer membrane channel, has the ability to catalyze and direct the insertion of other VDAC channels into planar phospholipid membranes. The spontaneous rate of insertion of detergent-solubilized VDAC channels into phospholipid membranes is estimated to be 1.5 x 10(-5) channels min-1 micron-2. VDAC channels already in the membrane can increase this rate by a factor of 10(9). The presence of 5 M urea on the opposite side of the membrane increases this 10-fold to 4.5 x 10(5) channels min-1 microns-2. Similar but weaker effects are observed with Triton X100 addition (10(-3)% (v/v)). These agents are not acting on uninserted channels because they do not affect the delay from sample addition to first insertion. Under the chosen conditions, this delay is long (240 s) without preinserted channels. However, the presence of a few VDAC channels in the membrane reduces this delay to 14 s, close to the diffusion limit. Therefore, urea and Triton, added to the side of the membrane opposite that to which the VDAC sample was added, likely increase the flexibility of the VDAC channels in the membrane, allowing them to be more efficient catalysts for VDAC insertion. There are obvious implications for membrane protein insertion and targeting. PMID:9129814

  2. 38. INTERIOR VIEW, DENISON MULTIPRESS FOR INSERTION OF WEDGES ONTO ...

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

    38. INTERIOR VIEW, DENISON MULTI-PRESS FOR INSERTION OF WEDGES ONTO HANDLES AND CUTTING OFF SCRAP END OF HANDLE FOLLOWING WEDGE INSERTION, BRIAN KIMBLE, OPERATOR - Warwood Tool Company, Foot of Nineteenth Street, Wheeling, Ohio County, WV

  3. Transposon Insertion Finder (TIF): a novel program for detection of de novo transpositions of transposable elements

    PubMed Central

    2014-01-01

    Background Transposition event detection of transposable element (TE) in the genome using short reads from the next-generation sequence (NGS) was difficult, because the nucleotide sequence of TE itself is repetitive, making it difficult to identify locations of its insertions by alignment programs for NGS. We have developed a program with a new algorithm to detect the transpositions from NGS data. Results In the process of tool development, we used next-generation sequence (NGS) data of derivative lines (ttm2 and ttm5) of japonica rice cv. Nipponbare, regenerated through cell culture. The new program, called a transposon insertion finder (TIF), was applied to detect the de novo transpositions of Tos17 in the regenerated lines. TIF searched 300 million reads of a line within 20 min, identifying 4 and 12 de novo transposition in ttm2 and ttm5 lines, respectively. All of the transpositions were confirmed by PCR/electrophoresis and sequencing. Using the program, we also detected new transposon insertions of P-element from NGS data of Drosophila melanogaster. Conclusion TIF operates to find the transposition of any elements provided that target site duplications (TSDs) are generated by their transpositions. PMID:24629057

  4. Analytic closed orbit analysis for RHIC insertion

    SciTech Connect

    Lee, S.Y. . Dept. of Physics); Tepikian, S. )

    1991-01-01

    Analytic closed orbit analysis is performed to evaluate the tolerance of quadrupole misalignment and dipole errors (b{sub 0},a{sub 0}) in the RHIC insertion. Sensitivity coefficients of these errors are tabulated for different {beta}{sup 0} values. Using these sensitivity tables, we found that the power supplies ripple of 10{sup {minus}4} can cause closed orbit motion of 0.05 mm at the IP in comparison with the rms beam size of 0.3 mm. It is desirable to have the power supply ripple less than 10{sup {minus}5}. 2 refs., 1 fig., 2 tabs.

  5. Transcatheter aortic valve insertion (TAVI): a review

    PubMed Central

    Morgan-Hughes, G; Roobottom, C

    2014-01-01

    The introduction of transcatheter aortic valve insertion (TAVI) has transformed the care provided for patients with severe aortic stenosis. The uptake of this procedure is increasing rapidly, and clinicians from all disciplines are likely to increasingly encounter patients being assessed for or having undergone this intervention. Successful TAVI heavily relies on careful and comprehensive imaging assessment, before, during and after the procedure, using a range of modalities. This review outlines the background and development of TAVI, describes the nature of the procedure and considers the contribution of imaging techniques, both to successful intervention and to potential complications. PMID:24258463

  6. Shoe inserts for small deformed feet.

    PubMed

    Platts, R G; Knight, S; Jakins, I

    1982-08-01

    Modern materials and a better understanding of the biomechanical requirements enable adaptations to shoes to be make quickly and easily in cases where the deformed foot is small enough to fit satisfactorily into standard shop-bought or standard deep footwear. A flexible self-generating polyurethane foam is used inside the shoe. It expands to the internal shape of the shoe and the external shape of the foot. It can be used either against the patient's own foot or against a positive cast of the foot. The technique has been used for 75 patients and has proved successful. The insert so made is durable and economical. PMID:7110918

  7. 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.

  8. One-ended insertion of IS911.

    PubMed Central

    Polard, P; Seroude, L; Fayet, O; Prère, M F; Chandler, M

    1994-01-01

    An apparently nonreplicative integration reaction mediated by the insertion sequence IS911 has been analyzed. It is shown to involve the right-end inverted repeat (IRR) of the element and sequences in the flanking vector DNA. The flanking sequences appear to behave as a surrogate IS911 end, since integration is greatly reduced when limited similarities with IRR are eliminated by site-directed mutagenesis. Data are presented which suggest that the activity of the IRR junction results from the proximity of the transposase gene and may therefore reflect preferential transposase recognition of IRR in cis. Images PMID:8106332

  9. Self-Inserted Needles in the Heart.

    PubMed

    Di Paolo, Marco; Guidi, Benedetta; Vergaro, Giuseppe; Emdin, Michele

    2015-10-15

    Cardiac injuries due to penetration by sharp foreign bodies usually have a clear clinical presentation. A case of a 38-year-old man with self-inserted cardiac lesions and a misleading presentation is reported. The patient was admitted to the emergency room because of chest pain, with increase in biomarkers of myocardial necrosis, and electrocardiographic and echocardiographic abnormalities inducing initial diagnostic and therapeutic workup of acute coronary syndrome. Once clinical destabilization followed by death had occurred, the suspicion of an alternative diagnosis came from delayed chest radiography, confirmed by autopsy, revealing the presence of multiple metallic objects within the thorax. PMID:26277296

  10. 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.

  11. 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.

  12. Forceps insertion with incisional anesthesia for Sensar intraocular lenses.

    PubMed

    Dewey, Steven H

    2002-07-01

    This technique to prevent self-adherence of the Sensar acrylic intraocular lens (IOL) optic during forceps insertion with wound stretch includes modifications to the 3-and-9 insertion technique to prevent difficulties that may be encountered with this phenomenon. To decrease patient sensation during IOL insertion and thus improve surgeon control, the corneal tunnel incision is directly anesthetized. PMID:12106716

  13. 21 CFR 310.515 - Patient package inserts for estrogens.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 5 2014-04-01 2014-04-01 false Patient package inserts for estrogens. 310.515... package inserts for estrogens. (a) Requirement for a patient package insert. FDA concludes that the safe and effective use of drug products containing estrogens requires that patients be fully informed...

  14. 21 CFR 310.515 - Patient package inserts for estrogens.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 5 2011-04-01 2011-04-01 false Patient package inserts for estrogens. 310.515... package inserts for estrogens. (a) Requirement for a patient package insert. FDA concludes that the safe and effective use of drug products containing estrogens requires that patients be fully informed...

  15. 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... package inserts for estrogens. (a) Requirement for a patient package insert. FDA concludes that the safe and effective use of drug products containing estrogens requires that patients be fully informed...

  16. 21 CFR 310.515 - Patient package inserts for estrogens.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 5 2013-04-01 2013-04-01 false Patient package inserts for estrogens. 310.515... package inserts for estrogens. (a) Requirement for a patient package insert. FDA concludes that the safe and effective use of drug products containing estrogens requires that patients be fully informed...

  17. 21 CFR 310.515 - Patient package inserts for estrogens.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 5 2012-04-01 2012-04-01 false Patient package inserts for estrogens. 310.515... package inserts for estrogens. (a) Requirement for a patient package insert. FDA concludes that the safe and effective use of drug products containing estrogens requires that patients be fully informed...

  18. 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

  19. Transposon insertion mapping with PIMMS – Pragmatic Insertional Mutation Mapping System

    PubMed Central

    Blanchard, Adam M.; Leigh, James A.; Egan, Sharon A.; Emes, Richard D.

    2015-01-01

    The PIMMS (Pragmatic Insertional Mutation Mapping System) pipeline has been developed for simple conditionally essential genome discovery experiments in bacteria. Capable of using raw sequence data files alongside a FASTA sequence of the reference genome and GFF file, PIMMS will generate a tabulated output of each coding sequence with corresponding mapped insertions accompanied with normalized results enabling streamlined analysis. This allows for a quick assay of the genome to identify conditionally essential genes on a standard desktop computer prioritizing results for further investigation. Availability: The PIMMS script, manual and accompanying test data is freely available at https://github.com/ADAC-UoN/PIMMS PMID:25914720

  20. 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.

  1. 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.

  2. Mammalian NUMT insertion is non-random.

    PubMed

    Tsuji, Junko; Frith, Martin C; Tomii, Kentaro; Horton, Paul

    2012-10-01

    It is well known that remnants of partial or whole copies of mitochondrial DNA, known as Nuclear MiTochondrial sequences (NUMTs), are found in nuclear genomes. Since whole genome sequences have become available, many bioinformatics studies have identified putative NUMTs and from those attempted to infer the factors involved in NUMT creation. These studies conclude that NUMTs represent randomly chosen regions of the mitochondrial genome. There is less consensus regarding the nuclear insertion sites of NUMTs - previous studies have discussed the possible role of retrotransposons, but some recent ones have reported no correlation or even anti-correlation between NUMT sites and retrotransposons. These studies have generally defined NUMT sites using BLAST with default parameters. We analyze a redefined set of human NUMTs, computed with a carefully considered protocol. We discover that the inferred insertion points of NUMTs have a strong tendency to have high-predicted DNA curvature, occur in experimentally defined open chromatin regions and often occur immediately adjacent to A?+?T oligomers. We also show clear evidence that their flanking regions are indeed rich in retrotransposons. Finally we show that parts of the mitochondrial genome D-loop are under-represented as a source of NUMTs in primate evolution. PMID:22761406

  3. 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.

  4. Evidence for insertional RNA editing in humans.

    PubMed

    Zougman, Alexandre; Zió?kowski, Piotr; Mann, Matthias; Wi?niewski, Jacek R

    2008-11-25

    Large-scale analysis directly at the protein level holds the promise of uncovering features not apparent or present at the gene level [1-3]. Although mass spectrometry (MS)-based proteomics can now identify and quantify thousands of cellular proteins in large-scale proteomics experiments, much of the peptide information contained in these experiments remains unassigned [4]. Here, we use such information to discover a previously unreported mechanism creating altered protein forms. Linker histones H1 and high-mobility group (HMG) proteins are abundant nuclear proteins that regulate gene expression through modulation of chromatin structure [5-8]. In the high-resolution MS analysis of histone H1 and HMG protein fractions isolated from human cells, we discovered peptides that mapped upstream of the known translation start sites of these genes. No alternative upstream start site exists in the genome, but analysis of Expressed Sequence Tag (EST) databases revealed that these N-terminally extended (ET) proteins are due to in-frame translation of the 5' untranslated region (5'UTR) sequences of the transcripts. The new translation start sites are created by a single uridine insertion between AG, reflecting a previously unreported RNA-editing mechanism. To our knowledge, this is the first report of RNA-insertion editing in humans and may be an example of the type of discoveries possible with modern proteomics methods. PMID:18993075

  5. Phoenix Conductivity Probe Inserted in Martian Soil

    NASA Technical Reports Server (NTRS)

    2008-01-01

    This series of six images from the Robotic Arm Camera on NASA's Phoenix Mars Lander records the first time that the four spikes of the lander's thermal and electrical conductivity probe were inserted into Martian soil.

    The images were taken on July 8, 2008, during the Phoenix mission's 43rd Martian day, or sol, since landing. The insertion visible from the shadows cast on the ground on that sol was a validation test of the procedure. The spikes on the probe are about 1.5 centimeters or half an inch long.

    The science team will use the probe tool to assess how easily heat and electricity move through the soil from one spike to another. Such measurements can provide information about frozen or unfrozen water in the soil. The probe is mounted on the 'knuckle' of Phoenix's Robotic Arm. It has already been used for assessing water vapor in the atmosphere when it is held above the ground.

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

  6. Recessed impingement insert metering plate for gas turbine nozzles

    DOEpatents

    Itzel, Gary Michael (218 Quail Ridge Dr., Greenville, SC 29680); Burdgick, Steven Sebastian (7006 Kevin La., Schenectady, NY 12303)

    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.

  7. Insertion of coherence requests for debugging a multiprocessor

    DOEpatents

    Blumrich, Matthias A. (Ridgefield, CT); Salapura, Valentina (Chappaqua, NY)

    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.

  8. Cell Lines

    PubMed Central

    Cherbas, Lucy; Gong, Lei

    2014-01-01

    We review the properties and uses of cell lines in Drosophila research, emphasizing the variety of lines, the large body of genomic and transcriptional data available for many of the lines, and the variety of ways the lines have been used to provide tools for and insights into the developmental, molecular, and cell biology of Drosophila and mammals. PMID:24434506

  9. 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. PMID:19387483

  10. Migration of tendinous insertions. I. Cause and mechanism.

    PubMed Central

    Dörfl, J

    1980-01-01

    The cause and mechanisms of the migration of tendons and ligaments were studied in young rabbits. Three techniques were used: (1) Marking of insertions, the neighbouring periosteum and the diaphysis with metallic markers. (2) Marking of insertion sites by tetracycline as an indicator of osteogenesis. (3) Histological examination. The insertions used in the study were of three different characters: (1) Insertions subject to muscular traction (patellar ligament, quadratus femoris muscle, tibialis anterior muscle). (2) The distal insertions of the medial collateral ligament of the knee, stretched by the activity of the proximal epiphyseal cartilage of the tibia. (3) The proximal and distal insertions of the anterior annular ligament of the tibia, inserted solely in bone and periosteum. The cause of migration is the growth of periosteum dragging the insertions during its stretching, caused itself by the activity of the epiphyseal plates. The local mechanism governing migration while ensuring a continuous connexion with the bone is not the same in all sites. It depends upon the character of the bony surface at the insertion and of the function of the insertion zone, which can be osteogenic, resorptive or both. A plexus of precollagenous fibres is present at all resorptive insertion sites, and at some of the osteogenic sites. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 PMID:7440401

  11. A piezoelectric vibration-based syringe for reducing insertion force

    NASA Astrophysics Data System (ADS)

    Huang, Y. C.; Tsai, M. C.; Lin, C. H.

    2012-12-01

    Puncturing of the human skin with a needle is perhaps the most common invasive medical procedure. Clinical studies have revealed that tissue deformation and needle deflection are the primary problem for needle misplacement in percutaneous procedures. To avoid this, various techniques for reducing insertion forces during needle insertion have been considered. This paper presents a piezoelectric vibration-based syringe to reduce insertion force. AC power was applied to the piezoelectric elements to vibrate the needle with high frequency and thereby reduce the friction and cutting forces between the needle and tissue. Vibration mode shapes of the needle were observed by finite element analysis and verified by experimental results. Effects of reducing insertion force via the vibrating needle were also confirmed by inserting the needle into the porcine tissues. The proposed syringe, which minimizes the insertion force and overcomes limitations of needle materials, can be widely utilized in robot-assisted needle insertion systems.

  12. Method Of Wire Insertion For Electric Machine Stators

    DOEpatents

    Brown, David L (Westfield, IN); Stabel, Gerald R (Swartz Creek, MI); Lawrence, Robert Anthony (Kokomo, IN)

    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.

  13. 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.

  14. Radiographic assessment of the femoral insertion of the posterior cruciate ligament

    PubMed Central

    Gali, Julio Cesar; Soares, André Schmidt; de Lima, Bruno Spagnuolo; Vianna, Filippo Santos Zozoloto; da Silva, Phelipe Augusto Cintra; Caetano, Edie Benedito

    2013-01-01

    Objective To establish the radiographic images of the femoral insertion of the posterior cruciate ligament (PCL), in order to assist the creation of anatomical femoral tunnels during surgeries, and to be used as parameters in postoperative evaluation of the location of these tunnels. Methods Thirty adult cadaver knees were used. The PCL anterolateral (AL) and posteromedial (PM) bundles' center was marked with a metallic marker. Radiographs were taken and a grid system was established to locate the position of bundles insertion. The percentile in which the projection of each bundle's center was in relation to the Blumensaat line was also determined. Results In the anteroposterior view, AL and PM bundles' centers were on average, on the 42.5% and 38.18% percentiles of Blumensaat's line, respectively. In lateral views, the AL and PM bundles' centers corresponded to the 72.94% and 55.56% percentiles of the line, respectively. In 73.33% of the knees the AL bundle center was on the 3D quadrant and in 70% of samples the PM bundle center was in quadrant 2D. Conclusions We established an X-ray pattern of femoral insertion of PCL that may be of interest for intraoperative control, before tunnel drilling, and also for post-operative evaluation of tunnel location. Controlled Laboratory Study. PMID:24453690

  15. 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. PMID:17151812

  16. Stabilization of insertion electrodes for lithium batteries.

    SciTech Connect

    Thackeray, M. M.

    1998-09-03

    This paper discusses the techniques that are being employed to stabilize LiMn{sub 2}O{sub 4} spinel and composite Li{sub x}MnO{sub 2} positive electrodes. The critical role that spinel domains play in stabilizing these electrodes for operation at both 4 V and 3 V is highlighted. The concept of using an intermetallic electrode MM{prime} where M is an active alloying element and M{prime} is an inactive element (or elements) is proposed as an alternative negative electrode (to carbon) for lithium-ion cells. An analogy to metal oxide insertion electrodes, such as MnO{sub 2}, in which Mn is the electrochemically active ion and O is the inactive ion, is made. Performance data are given for the copper-tin electrode system, which includes the intermetallic phases eta-Cu{sub 6}Sn{sub 5} and Li{sub 2}CuSn.

  17. Increasing Prion Propensity by Hydrophobic Insertion

    PubMed Central

    Petri, Michelina; Flores, Noe; Rogge, Ryan A.; Cascarina, Sean M.; Ross, Eric D.

    2014-01-01

    Prion formation involves the conversion of proteins from a soluble form into an infectious amyloid form. Most yeast prion proteins contain glutamine/asparagine-rich regions that are responsible for prion aggregation. Prion formation by these domains is driven primarily by amino acid composition, not primary sequence, yet there is a surprising disconnect between the amino acids thought to have the highest aggregation propensity and those that are actually found in yeast prion domains. Specifically, a recent mutagenic screen suggested that both aromatic and non-aromatic hydrophobic residues strongly promote prion formation. However, while aromatic residues are common in yeast prion domains, non-aromatic hydrophobic residues are strongly under-represented. Here, we directly test the effects of hydrophobic and aromatic residues on prion formation. Remarkably, we found that insertion of as few as two hydrophobic residues resulted in a multiple orders-of-magnitude increase in prion formation, and significant acceleration of in vitro amyloid formation. Thus, insertion or deletion of hydrophobic residues provides a simple tool to control the prion activity of a protein. These data, combined with bioinformatics analysis, suggest a limit on the number of strongly prion-promoting residues tolerated in glutamine/asparagine-rich domains. This limit may explain the under-representation of non-aromatic hydrophobic residues in yeast prion domains. Prion activity requires not only that a protein be able to form prion fibers, but also that these fibers be cleaved to generate new independently-segregating aggregates to offset dilution by cell division. Recent studies suggest that aromatic residues, but not non-aromatic hydrophobic residues, support the fiber cleavage step. Therefore, we propose that while both aromatic and non-aromatic hydrophobic residues promote prion formation, aromatic residues are favored in yeast prion domains because they serve a dual function, promoting both prion formation and chaperone-dependent prion propagation. PMID:24586661

  18. 48 CFR 252.204-7011 - Alternative Line Item Structure.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Structure. 252.204-7011 Section 252.204-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION... of Provisions And Clauses 252.204-7011 Alternative Line Item Structure. As prescribed in 204.7109(b), insert the following provision: Alternative Line-Item Structure (SEP 2011) (a) Line items are the...

  19. 48 CFR 252.204-7011 - Alternative Line Item Structure.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Structure. 252.204-7011 Section 252.204-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION... of Provisions And Clauses 252.204-7011 Alternative Line Item Structure. As prescribed in 204.7109(b), insert the following provision: ALTERNATIVE LINE-ITEM STRUCTURE (SEP 2011) (a) Line items are the...

  20. 48 CFR 252.204-7011 - Alternative Line Item Structure.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Structure. 252.204-7011 Section 252.204-7011 Federal Acquisition Regulations System DEFENSE ACQUISITION... of Provisions And Clauses 252.204-7011 Alternative Line Item Structure. As prescribed in 204.7109(b), insert the following provision: Alternative Line-Item Structure (SEP 2011) (a) Line items are the...

  1. Peripherally inserted central catheter: compliance with evidence-based indications for insertion in an inpatient setting.

    PubMed

    Wojnar, Danuta G; Beaman, Margaret L

    2013-01-01

    A randomized, retrospective chart review was conducted at a medium-sized community hospital to verify appropriateness of peripherally inserted central catheter (PICC) use in 49 inpatient patients. Study results support the Infusion Nurses Society recommendation to use PICCs to facilitate either short- or long-term intravenous therapy of vesicants, irritants, and any medications with a pH less than 5 or greater than 9 and osmolarity greater than 600 mOsm/L. All PICC insertion criteria recommended by the Centers for Disease Control and Prevention were met except one--the intended duration of intravenous therapy of more than 6 days. Identical PICC selection criteria are needed to standardize clinical practice. PMID:23823005

  2. Assessment of infants with peripherally inserted central catheters: Part 1. Detecting the most frequently occurring complications.

    PubMed

    Pettit, Janet

    2002-12-01

    Inserting, maintaining, and monitoring vascular access are integral components of neonatal care. Advances in vascular access technology have led to the insertion of peripherally inserted central catheters (PICC) to provide stable venous access for early and aggressive parenteral nutrition. Medications that are irritating or damaging, or those with a high osmolality or a nonphysiologic pH, can also be safely administered into the central venous system. The need for repeated peripheral intravenous attempts, as well as the associated pain and physiologic instability, are virtually eliminated once a PICC line is placed. Complications related to PICCs may occur at any phase of therapy: during insertion, while indwelling, or after discontinuing the line. The risk factors associated with PICCs are distinctly different from peripheral intravenous lines because of their long dwell time, central placement, and potential to migrate. Part 1 of Focus on the Physical offers a review of the relevant anatomy of the vascular system and a discussion of the appropriate sites for catheter tip placement. Guidelines for a systematic physical assessment, along with recommendations for standardized PICC documentation, are provided. A review of the signs and symptoms of more frequently occurring complications, such as catheter occlusion and bloodstream infections, is presented to enhance awareness of PICC-specific complications in the neonate and to expedite early detection and treatment. Part 2 of this series will focus on systematic assessment for less common complications such as catheter migration, dislodgement, breakage, and thrombosis, as well as the life-threatening complications of pleural and pericardial effusion. PMID:12881943

  3. Measure Lines

    ERIC Educational Resources Information Center

    Crissman, Sally

    2011-01-01

    One tool for enhancing students' work with data in the science classroom is the measure line. As a coteacher and curriculum developer for The Inquiry Project, the author has seen how measure lines--a number line in which the numbers refer to units of measure--help students not only represent data but also analyze it in ways that generate…

  4. Large insert environmental genomic library production.

    PubMed

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

    2009-01-01

    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 in execution to improve overall quality and efficiency of library production. The whole process of fosmid library production and automated colony picking takes at least 7-10 days as there are many incubation steps included. However, there are several stopping points possible which are mentioned within the protocol. PMID:19776717

  5. An evaluation of consumable inserts for gas tungsten arc welding

    SciTech Connect

    Chavez, A.E.; Marburger, S.J.

    1987-03-01

    This study presents information on the use of consumable inserts to make 100% penetration welds in 304 and modified A286 (designated JBK-75) stainless steels. Current and voltage ranges which produced acceptable welds were found for different combinations of joint preparations and inserts. Three third member insert designs were examined: Type A (AWS Class 1), Type G (AWS Class 3), and Type K (AWS Class 5). One joint design incorporated the configuration of the Type G insert into the root face of the joint (termed an ''integral insert''). Joints welded with and without filler wire addition were made for comparison. Metallurgical and radiographic tests were done on all welds. It was found that the consumable insert root pass was comparable to the wire-fed root pass in weld shape and size using the same parameters; it also had additional advantages in ease of use, cleanliness, inspectability and reduced procedure complexity.

  6. Effect of insertion torque on bone screw pullout strength.

    PubMed

    Lawson, K J; Brems, J

    2001-05-01

    The effect of insertion torque on the holding strength of 4.5-mm ASIF/AO cortical bone screws was studied in vitro. Screw holding strength was determined using an Instron materials testing machine (Bristol, United Kingdom) on 55 lamb femora and 30 human tibiocortical bone sections. Holding strength was defined as tensile stress at pullout with rapid loading to construct failure. Different insertion torques were tested, normalizing to the thickness of cortical bone specimen engaged. These represented low, intermediate, high, and thread-damaging insertion torque. All screws inserted with thread-damaging torque and single cortex engaging screws inserted to high torque tightening moments showed diminished holding strength. This loss of strength amounted to 40%-50% less than screws inserted with less torque. PMID:11379993

  7. The Effect of Transposable Element Insertions on Gene Expression Evolution in Rodents

    PubMed Central

    Pereira, Vini; Enard, David; Eyre-Walker, Adam

    2009-01-01

    Background Many genomes contain a substantial number of transposable elements (TEs), a few of which are known to be involved in regulating gene expression. However, recent observations suggest that TEs may have played a very important role in the evolution of gene expression because many conserved non-genic sequences, some of which are know to be involved in gene regulation, resemble TEs. Results Here we investigate whether new TE insertions affect gene expression profiles by testing whether gene expression divergence between mouse and rat is correlated to the numbers of new transposable elements inserted near genes. We show that expression divergence is significantly correlated to the number of new LTR and SINE elements, but not to the numbers of LINEs. We also show that expression divergence is not significantly correlated to the numbers of ancestral TEs in most cases, which suggests that the correlations between expression divergence and the numbers of new TEs are causal in nature. We quantify the effect and estimate that TE insertion has accounted for ?20% (95% confidence interval: 12% to 26%) of all expression profile divergence in rodents. Conclusions We conclude that TE insertions may have had a major impact on the evolution of gene expression levels in rodents. PMID:19183808

  8. Critical monitoring of the Cassini Saturn Orbit Insertion maneuver

    NASA Technical Reports Server (NTRS)

    Asmar, S. W.; Johnston, D. V.; Maize, E.; Mitchell, R. T.

    2004-01-01

    The Cassini-Huygens spacecraft will have spent seven years since launch by the time it reaches Saturn and its satellites. The primary mission will begin after the Saturn Orbit Insertion (SOI) scheduled for early July 2004. the insertion will require a critical navigational maneuver that involves firing the orbiter's thrusters leading to the capture in the gravity well of the planet. This paper will describe the challenges and plans by the Cassini Program to monitor the Saturn Orbit Insertion Maneuver.

  9. 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.

  10. 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.

  11. AKATSUKI status after the Venus orbit insertion

    NASA Astrophysics Data System (ADS)

    Nakamura, M.; Ishii, N.; Imamura, T.; Ueno, M.; Yamazaki, A.; Satoh, T.; Suzuki, M.; Iwagami, N.; Taguchi, M.; Watanabe, S.; Takahashi, Y.; Fukuhara, T.; Ohtsuki, S.; PLANET-C Project Team

    2010-12-01

    Japanese Venus Climate Orbiter 'AKATSUKI' (PLANET-C) was successfully launched at 06:58:22JST on May 21, by H-IIA F17. After the separation from H-IIA, the telemetry from AKATSUKI was normally detected by DSN Goldstone station (10:00JST) and the solar cell paddles' expansion was confirmed. AKATSUKI was put into the 3-axis stabilized mode in the initial operation from Uchinoura station and the critical operation was finished at 20:00JST on the same day. AKATSUKI will arrive at Venus on Dec 7, 2010. From the orbit around Venus, it will take pictures of the distribution of clouds and minor gas components at different wavelength by 4 cameras. These data will be analyzed to study the meteorology on Venus, especially the mechanism of the super-rotation of Venusian atmosphere which is still the top mystery of the Venus science. Simultaneous observation with ESA's Venus Express is highly expected to study the Venusian atmosphere. We will present the present status of AKATSUKI after the Venus orbit insertion with the latest results from the cameras.

  12. Does Feeding Tube Insertion and its Timing Improve Survival?

    PubMed Central

    Teno, Joan M.; Gozalo, Pedro L.; Mitchell, Susan L.; Kuo, Sylvia; Rhodes, Ramona L.; Bynum, Julie P.W.; Mor, Vincent

    2012-01-01

    Background/Objectives The objective of this study was to examine survival with and without a percutaneous endoscopic gastrostomy (PEG) feeding tube using rigorous methods to account for selection bias. A second objective was to examine whether the timing of feeding tube insertion affected survival. Design Prospective cohort study Setting All U.S. Nursing Homes Participants 36,492 nursing home residents with advanced cognitive impairment from dementia and new problems eating studied between 1999–2007. Measurements Survival following the development of the need for eating assistance and feeding tube insertion. Results Of the 36,492 nursing home (NH) residents (88.4% white, mean age 84.9, 87.4% with one feeding tube risk factor), 1,957 (5.4%) had a feeding tube inserted within 1 year of developing eating problems. After multivariate analysis correcting for selection bias with propensity score weights, no difference was found in the survival of the 2 groups (AHR 1.03, 95% CI 0.94–1.13). Among residents who were tube-fed, the timing of PEG tube insertion relative to the onset of eating problems was not associated with improved survival post feeding tube insertion (AHR 1.01, 95% CI 0.86–1.20) comparing those persons with a PEG inserted within a month of developing an eating problem compared to later (4 months) insertion. Conclusion This national study confirms that neither insertion of PEG feeding tubes nor the timing of the insertion improve survival. PMID:23002947

  13. Wind tunnel supplementary Mach number minimum section insert

    NASA Technical Reports Server (NTRS)

    Couch, L. M. (inventor)

    1981-01-01

    A device is described which changes the Mach number capability of a wind tunnel without permanently altering the existing nozzle of the tunnel. An insert is removably attached to the wall of the existing nozzle expansion area thereby creating a second minimum section upstream of the model test section. The added insert may be removed without complicated and expensive changes to the basic wind tunnel. In one embodiment, a removable insert is disposed within wind tunnel nozzle walls with a portion of the flow boundary layer being bled off from the tunnel via passageway and tunnel exit to reduce the extent of separated flow normally occuring upstream of the insert contraction section.

  14. 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.

  15. Vector-hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus.

    PubMed

    Herring, C D; Chevillard, C; Johnston, S L; Wettstein, P J; Riblet, R

    1998-06-01

    We have developed a simple PCR strategy, termed vector-hexamer PCR, that is unique in its ability to easily recover every insert end from large insert clones in YAC and BAC vectors. We used this method to amplify and isolate all insert ends from a YAC contig covering the mouse Igh locus. Seventy-seven ends were amplified and sequenced from 36 YAC clones from four libraries in the pYAC4 vector. Unexpectedly, 40% of the insert ends of these YACs were LINE1 repeats. Nonrepetitive ends were suitable for use as probes on Southern blots of digested YACs to identify overlaps and construct a contig. The same strategy was used successfully to amplify insert ends from YACs in the pRML vector from the Whitehead Institute/MIT-820 mouse YAC library and from BACs in pBeloBAC11. The simplicity of this technique and its ability to isolate every end from large insert clones are of great utility in genomic investigation. [The nucleotide sequence data reported in this paper are accessible in GenBank under accession nos. B07512-B07598.] PMID:9647641

  16. Vector–Hexamer PCR Isolation of All Insert Ends from a YAC Contig of the Mouse Igh?Locus

    PubMed Central

    Herring, Christopher D.; Chevillard, Christophe; Johnston, Sean L.; Wettstein, Peter J.; Riblet, Roy

    1998-01-01

    We have developed a simple PCR strategy, termed vector–hexamer PCR, that is unique in its ability to easily recover every insert end from large insert clones in YAC and BAC vectors. We used this method to amplify and isolate all insert ends from a YAC contig covering the mouse Igh locus. Seventy-seven ends were amplified and sequenced from 36 YAC clones from four libraries in the pYAC4 vector. Unexpectedly, 40% of the insert ends of these YACs were LINE1 repeats. Nonrepetitive ends were suitable for use as probes on Southern blots of digested YACs to identify overlaps and construct a contig. The same strategy was used successfully to amplify insert ends from YACs in the pRML vector from the Whitehead Institute/MIT-820 mouse YAC library and from BACs in pBeloBAC11. The simplicity of this technique and its ability to isolate every end from large insert clones are of great utility in genomic investigation. [The nucleotide sequence data reported in this paper are accessible in GenBank under accession nos. B07512–B07598.] PMID:9647641

  17. A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes.

    PubMed

    Wang, Weiquan; Wu, Yaorong; Li, Yin; Xie, Jiaying; Zhang, Zhonghui; Deng, Zhiyong; Zhang, Yiyue; Yang, Cuiping; Lai, Jianbin; Zhang, Huawei; Bao, Hongyan; Tang, Sanyuan; Yang, Chengwei; Gao, Peng; Xia, Guixian; Guo, Huishan; Xie, Qi

    2010-01-01

    Salt cress (Thellungiella halophila), a salt-tolerant relative of Arabidopsis, has turned to be an important model plant for studying abiotic stress tolerance. One binary bacterial artificial chromosome (BIBAC) library was constructed which represents the first plant-transformation-competent large-insert DNA library generated for Thellungiella halophila. The BIBAC library was constructed in BamHI site of binary vector pBIBAC2 by ligation of partial digested nuclear DNA of Thellungiella halophila. This library consists of 23,040 clones with an average insert size of 75 kb, and covers 4x Thellungiella halophila haploid genomes. BIBAC clones which contain inserts over 50 kb were selected and transformed into Arabidopsis for salt tolerant plant screening. One transgenic line was found to be more salt tolerant than wild type plants from the screen of 200 lines. It was demonstrated that the library contains candidates of stress tolerance genes and the approach is suitable for the transformation of stress susceptible plants for genetic improvement. PMID:19787433

  18. Identification of the lacZ insertion site and beta-galactosidase expression in transgenic chickens.

    PubMed

    Mozdziak, Paul E; Wu, Qian; Bradford, Jennifer M; Pardue, Samuel L; Borwornpinyo, Suparerk; Giamario, Carol; Petitte, James N

    2006-04-01

    The quail:chick chimera system is a classical research model in developmental biology. An improvement over the quail:chick chimera system would be a line of transgenic chickens expressing a reporter gene. Transgenic chickens carrying lacZ and expressing bacterial beta-galactosidase have been generated, but complete characterization of the insertion event and characterization of beta-galactosidase expression have not previously been available. The genomic sequences flanking the retroviral insertion site have now been identified by using inverse polymerase chain reaction (PCR), homozygous individuals have been identified by using PCR-based genotyping, and beta-galactosidase expression has been evaluated by using Western analysis and histochemistry. Based upon the current draft of the chicken genome, the viral insertion carrying the lacZ gene has been located on chromosome 11 within the predicted gene for neurotactin/fractalkine (CX3CL1); neurotactin mRNA expression appears to be missing from the brain of homozygous individuals. When Generation 2 (G2) lacZ-positive individuals were inter-mated, they generated 361 G3 progeny; 82 were homozyous for lacZ (22.7%), 97 were wild-type non-transgenic (26.9%), and 182 (50.4%) were hemizygous for lacZ. Western analysis revealed the highest expression in the muscle and liver. With the identification of homozygous birds, the line of chickens is now designated NCSU-Blue1. PMID:16408197

  19. Everyman's Guide to Bacterial Insertion Sequences.

    PubMed

    Siguier, Patricia; Gourbeyre, Edith; Varani, Alessandro; Ton-Hoang, Bao; Chandler, Mick

    2015-04-01

    The number and diversity of known prokaryotic insertion sequences (IS) have increased enormously since their discovery in the late 1960s. At present the sequences of more than 4000 different IS have been deposited in the specialized ISfinder database. Over time it has become increasingly apparent that they are important actors in the evolution of their host genomes and are involved in sequestering, transmitting, mutating and activating genes, and in the rearrangement of both plasmids and chromosomes. This review presents an overview of our current understanding of these transposable elements (TE), their organization and their transposition mechanism as well as their distribution and genomic impact. In spite of their diversity, they share only a very limited number of transposition mechanisms which we outline here. Prokaryotic IS are but one example of a variety of diverse TE which are being revealed due to the advent of extensive genome sequencing projects. A major conclusion from sequence comparisons of various TE is that frontiers between the different types are becoming less clear. We detail these receding frontiers between different IS-related TE. Several, more specialized chapters in this volume include additional detailed information concerning a number of these.In a second section of the review, we provide a detailed description of the expanding variety of IS, which we have divided into families for convenience. Our perception of these families continues to evolve and families emerge regularly as more IS are identified. This section is designed as an aid and a source of information for consultation by interested specialist readers. PMID:26104715

  20. Meta-analysis of gross insertions causing human genetic disease: novel mutational mechanisms and the role of replication slippage.

    PubMed

    Chen, Jian-Min; Chuzhanova, Nadia; Stenson, Peter D; Férec, Claude; Cooper, David N

    2005-02-01

    Although gross insertions (>20 bp) comprise <1% of disease-causing mutations, they nevertheless represent an important category of pathological lesion. In an attempt to study these insertions in a systematic way, 158 gross insertions ranging in size between 21 bp and approximately 10 kb were identified using the Human Gene Mutation Database (www.hgmd.org). A careful meta-analytical study revealed extensive diversity in terms of the nature of the inserted DNA sequence and has provided new insights into the underlying mutational mechanisms. Some 70% of gross insertions were found to represent sequence duplications of different types (tandem, partial tandem, or complex). Although most of the tandem duplications were explicable by simple replication slippage, the three complex duplications appear to result from multiple slippage events. Some 11% of gross insertions were attributable to nonpolyglutamine repeat expansions (including octapeptide repeat expansions in the prion protein gene [PRNP] and polyalanine tract expansions) and evidence is presented to support the contention that these mutations are also caused by replication slippage rather than by unequal crossing over. Some 17% of gross insertions, all >or=276 bp in length, were found to be due to LINE-1 (L1) retrotransposition involving different types of element (L1 trans-driven Alu, L1 direct, and L1 trans-driven SVA). A second example of pathological mitochondrial-nuclear sequence transfer was identified in the USH1C gene but appears to arise via a novel mechanism, trans-replication slippage. Finally, evidence for another novel mechanism of human genetic disease, involving the possible capture of DNA oligonucleotides, is presented in the context of a 26-bp insertion into the ERCC6 gene. PMID:15643617

  1. A standardised approach to the insertion of Hickman catheters.

    PubMed Central

    Ebbs, S. R.; Cameron, A. E.

    1988-01-01

    A standardised technique for the insertion of Hickman catheters under local anaesthetic is described. The external jugular vein is to be preferred, but if this proves too small the catheter may be introduced via the internal jugular vein using a Seldinger technique. A total of 44 Hickman catheters were inserted into 44 patients using this approach without major complication. Images fig. 1 PMID:3190127

  2. 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

  3. Cruciate retaining and cruciate substituting ultra-congruent insert.

    PubMed

    Mazzucchelli, Luca; 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. 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.

  5. Endoscopic Achilles tenodesis: a surgical alternative for chronic insertional tendinopathy.

    PubMed

    Maquirriain, Javier

    2007-07-01

    This report describes an endoscopic approach for visualization and repair of heel structures potentially involved in patients presenting chronic calcaneal tendon insertion pain. Tendon-bone junction separation allows enthesis debridement. Tenodesis with a knotless absorbable anchor completes this minimally invasive surgical procedure for chronic insertional Achilles tendinopathy. PMID:17053930

  6. 21 CFR 529.1940 - Progesterone intravaginal inserts.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...)(ii)(A) of this section. See § 522.690(c) of this chapter. (e) Conditions of use—(1) Cows—(i) Amount... Progesterone intravaginal inserts. (a) Specifications. Each insert contains: (1) 1.38 grams (g) progesterone in molded silicone over a nylon spine. (2) 0.3 g progesterone in molded silicone over a flexible nylon...

  7. 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.

  8. 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. PMID:26339941

  9. 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 summarizes testing that was performed to determine a k value for helicoil inserts in the Z2 suit and how the insert design was modified to resist a higher pull out tension.

  10. Method for improving the durability of ion insertion materials

    DOEpatents

    Lee, Se-Hee (Lakewood, CO); Tracy, C. Edwin (Golden, CO); Cheong, Hyeonsik M. (Seoul, KR)

    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. Gas turbine nozzle vane insert and methods of installation

    DOEpatents

    Miller, William John (Simpsonville, SC); Predmore, Daniel Ross (Clifton Park, NY); Placko, James Michael (West Chester, OH)

    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.

  12. 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.

  13. [Insertion and maintenance of peripheral venous catheters in neonates].

    PubMed

    Pastor Rodríguez, Jesús David; Serrano Matás, Encarnación; Muñoz Escolar, Dolores Angeles

    2008-01-01

    Insertion of peripheral venous catheters in premature and term newborns is a common practice in neonatology units and neonatal intensive care units. Nurses are responsible for the insertion and maintenance of peripheral venous catheters and for the prevention of complications. Although this technique is routine, a series of recommendations, supported by evidence-based practice, should be bourne in mind when inserting these catheters. Following these recommendations guarantees successful insertion, and the absence of risks and complications. To achieve this aim, the following steps should be carried out: preparing the material, selecting the vein, selecting the catheter, cleaning and disinfecting the area, inserting the catheter, fixing the catheter, and restoring intravenous therapy. In addition, attention must be paid to potential risks in order to resolve them as quickly as possible, thereby avoiding complications. PMID:18724919

  14. Functional Genomics of Rice Pollen and Seed Development by Genome-wide Transcript Profiling and Ds Insertion Mutagenesis

    PubMed Central

    Jiang, Shu-Ye; Ramachandran, Srinivasan

    2011-01-01

    Rice pollen and seed development are directly related to grain yield. To further improve rice yield, it is important for us to functionally annotate the genes controlling pollen/seed development and to use them for rice breeding. Here we first carried out a genome-wide expression analysis with an emphasis on genes being involved in rice pollen and seed development. Based on the transcript profiling, we have identified and functionally classified 82 highly expressed pollen-specific, 12 developing seed-specific and 19 germinating seed-specific genes. We then presented the utilization of the maize transposon Dissociation (Ds) insertion lines for functional genomics of rice pollen and seed development and as alternative germplasm resources for rice breeding. We have established a two-element Activator/Dissociation (Ac/Ds) gene trap tagging system and generated around 20,000 Ds insertion lines. We have subjected these lines for screens to obtain high and low yield Ds insertion lines. Some interesting lines have been obtained with higher yield or male sterility. Flanking Sequence Tags (FSTs) analyses showed that these Ds-tagged genes encoded various proteins including transcription factors, transport proteins, unknown functional proteins and so on. They exhibited diversified expression patterns. Our results suggested that rice could be improved not only by introducing foreign genes but also by knocking out its endogenous genes. This finding might provide a new way for rice breeder to further improve rice varieties. PMID:21209789

  15. Modulation of the pHLIP transmembrane helix insertion pathway.

    PubMed

    Karabadzhak, Alexander G; Weerakkody, Dhammika; Wijesinghe, Dayanjali; Thakur, Mak S; Engelman, Donald M; Andreev, Oleg A; Markin, Vladislav S; Reshetnyak, Yana K

    2012-04-18

    The membrane-associated folding/unfolding of pH (low) insertion peptide (pHLIP) provides an opportunity to study how sequence variations influence the kinetics and pathway of peptide insertion into bilayers. Here, we present the results of steady-state and kinetics investigations of several pHLIP variants with different numbers of charged residues, with attached polar cargoes at the peptide's membrane-inserting end, and with three single-Trp variants placed at the beginning, middle, and end of the transmembrane helix. Each pHLIP variant exhibits a pH-dependent interaction with a lipid bilayer. Although the number of protonatable residues at the inserting end does not affect the ultimate formation of helical structure across a membrane, it correlates with the time for peptide insertion, the number of intermediate states on the folding pathway, and the rates of unfolding and exit. The presence of polar cargoes at the peptide's inserting end leads to the appearance of intermediate states on the insertion pathway. Cargo polarity correlates with a decrease of the insertion rate. We conclude that the existence of intermediate states on the folding and unfolding pathways is not mandatory and, in the simple case of a polypeptide with a noncharged and nonpolar inserting end, the folding and unfolding appears as an all-or-none transition. We propose a model for membrane-associated insertion/folding and exit/unfolding and discuss the importance of these observations for the design of new delivery agents for direct translocation of polar therapeutic and diagnostic cargo molecules across cellular membranes. PMID:22768940

  16. Trocar insertion: the neglected task of VR simulation.

    PubMed

    Brümmer, Vera; Carnahan, Heather; Okrainec, Allan; Dubrowski, Adam

    2008-01-01

    We hypothesized that simulation based practice will results in skill improvement on trocar insertion task. Novices, junior trainees and surgeons inserted a trocar into a simulated abdomen. The depth of penetration (plunge) was the similar for novice and junior trainees (p=.98), and exceeded that observed for practicing surgeons (p<.05). Trocar insertion may be an important variable to consider for surgical skill training. To date, no virtual reality (VR) trainers allow for practice of this skill. We will present a prototype of a VR trainer designed. PMID:18391255

  17. 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.

  18. 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.

  19. Insertions of a Novel Class of Transposable Elements with a Strong Target Site Preference at the R Locus of Maize

    PubMed Central

    Walker, E. L.; Eggleston, W. B.; Demopulos, D.; Kermicle, J.; Dellaporta, S. L.

    1997-01-01

    The r locus of maize regulates anthocyanin synthesis in various tissues of maize through the production of helix-loop-helix DNA binding proteins capable of inducing expression of structural genes in the anthocyanin biosynthetic pathway. The complex r variant, R-r:standard (R-r), undergoes frequent mutation through a variety of mechanisms including displaced synapsis and crossing over, and intrachromosomal recombination. Here we report a new mechanism for mutation at the R-r complex: insertion of a novel family of transposable elements. Because the elements were first identified in the R-p gene of the R-r complex, they have been named P Instability Factor (PIF). Two different PIF elements were cloned and found to have identical sequences at their termini but divergent internal sequences. In addition, the PIF elements showed a marked specificity of insertion sites. Six out of seven PIF-containing derivatives examined had an element inserted at an identical location. Two different members of the PIF element family were identified at this position. The seventh PIF-containing derivative examined had the element inserted at a distinct position within r. Even at this location, however, the element inserted into a conserved target sequence. The timing of PIF excision is unusual. Germinal excision rates can range up to several percent of progeny. Yet somatic sectors are rare, even in lines exhibiting high germinal reversion rates. PMID:9178016

  20. 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. genesis 54:65-77, 2016. © 2016 Wiley Periodicals, Inc. PMID:26713866

  1. 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.

  2. Water exchange vs. water immersion during colonoscope insertion.

    PubMed

    Rex, Douglas K

    2014-09-01

    Water exchange (water infusion with water removal primarily during insertion) and water immersion (water infusion with water removal during withdrawal) reduce patient discomfort during colonoscope insertion compared with air insufflation, and represent a major achievement in colonoscopy. Hsieh et al. found that water exchange, relative to water immersion, resulted in more painless insertions to the cecum and improved adenoma detection in the right colon. However, water exchange is also associated with better bowel cleansing and longer insertion and procedure times. These factors are not specific to water exchange, but could account for all or part of the better results with water exchange. Additional controlled investigation is needed to define the benefits of water exchange compared with water immersion. PMID:25196871

  3. 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.

  4. Insertion device magnet measurements for the Advanced Light Source

    SciTech Connect

    Marks, S.; Cork, C.; Hoyer, E.; Humphries, D.; Kincaid, B.; Plate, D.; Robb, A.; Schlueter, R.; Wang, C.; Hassenzahl, W.

    1993-05-01

    Allowable magnetic field errors for the 4.6 m long insertion devices for the Advanced Light Source (ALS) are extremely small and are driven by electron beam and radiation requirements. Detailed measurements and adjustments of each insertion device are performed to qualify them for installation in the ALS. To accomplish this, a high speed, precision magnetic measurement facility has been designed and built. Hall probe mapping equipment, capable of completing a 2500 sample, 6 m scan with precision axial position monitoring using a laser interferometer in under one minute, is used to obtain both local and integrated field information. A 5.5 m long, 1 cm wide coil is used to measure the field integral through an entire insertion device. This paper describes magnetic measurement equipment, and results of measurements on IDA, the first of the ALS insertion devices.

  5. 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

  6. 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.

  7. [PERIPHERALLY INSERTED CENTRAL VENOUS CATHETER THROUGH DRUM SYSTEM].

    PubMed

    Salas Campos, Luis

    2015-11-01

    It described a one lumen central venous catheter peripherally inserted drum system whose insertion into the vein is performed through a plastic splittable cannula by braunula technique. The braunula technique is particularly important in certain urgent or emergency situations, where speed and aseptic conditions of which the procedure is performed represent a further improvement over other insertion techniques. The catheter mantains the advantages, in terms of easy placement, that all devices equipped with a drum system have but at the same time eliminates the complications presented by other similar catheters whose insertion to the venous circulation used to be performed through a needle. These catheters allow quick, simple and safe access to the central venous circulation and reduce risks associated with the procedure, improving patient safety. PMID:26749753

  8. Transmigrated intrauterine device discovered 17 years after its insertion.

    PubMed

    Bartalena, T; Pascali, E; Rinaldi, M F; Marasco, R; Bassi, F; Alboni, C; Gavelli, G

    2007-12-01

    We present the case of an intraperitoneal IUD incidentally noted on lumbar spine X-rays and confirmed by CT. This was secondary to asymptomatic uterine perforation occurred at the time of insertion 17 years before. PMID:17991086

  9. Insertional mutation of the motor endplate disease (med) locus on mouse chromosome 15

    SciTech Connect

    Kohrman, D.C.; Plummer, N.W.; Schuster, T.

    1995-03-20

    Homozygous transgenic mice from line A4 have an early-onset progressive neuromuscular disorder characterized by paralysis of the rear limbs, muscle atrophy, and lethality by 4 weeks of age. The transgene insertion site was mapped to distal chromosome 15 close to the locus motor endplate disease (med). The sequence of mouse DNA flanking the insertion site junctions was determined. A small (<20 kb) deletion was detected at the insertion site, with no evidence of additional rearrangement of the chromosomal DNA. Noncomplementation of the transgene-induced mutation and med was demonstrated in a cross with med{sup J}/ + mice. The new allele is designated med{sup TgNA4Bs}(med{sup tg}). The homologous human locus MED was assigned to chromosome 12. Synaptotagmin 1 and contactin 1 were eliminated as candidate genes for the med mutation. The transgene-induced allele provides molecular access to the med gene, whose function is required for synaptic transmission at the neuromuscular junction and long-term survival of cerebellar Purkinje cells. 49 refs., 5 figs., 1 tab.

  10. Construction of finite element model and stress analysis of anterior cruciate ligament tibial insertion

    PubMed Central

    Dai, Can; Yang, Liu; Guo, Lin; Wang, Fuyou; Gou, Jingyue; Deng, Zhilong

    2015-01-01

    Objective: The aim of the present study was to develop a more realistic finite element (FE) model of the human anterior cruciate ligament (ACL) tibial insertion and to analyze the stress distribution in the ACL internal fibers under load. Methods: The ACL tibial insertions were processed histologically. With Photoshop software, digital images taken from the histological slides were collaged, contour lines were drawn, and different gray values were filled based on the structure. The data were exported to Amira software and saved as “.hmascii” file. This document was imported into HyperMesh software. The solid mesh model generated using HyperMesh software was imported into Abaqus software. The material properties were introduced, boundary conditions were set, and load was added to carry out the FE analysis. Results: The stress distribution of the ACL internal fibers was uneven. The lowest stress could be observed in the ACL lateral fibers under tensile and shear load. Conclusion: The establishment of ACL tibial insertion FE model and mechanical analysis could reveal the stress distribution in the ACL internal fibers under load. There was greater load carrying capacity in the ACL lateral fibers which could sustain greater tensile and shear forces. PMID:26150858

  11. International Conference on Insertion Devices for Synchrotron Sources, Stanford, CA, October 27-30, 1985, Proceedings

    NASA Astrophysics Data System (ADS)

    Tatchyn, R.; Lindau, I.

    Papers are presented on a new formulation of synchrotron radiation optics using the Wigner distribution, undulators as a primary source of coherent X-rays, programs for the calculation of undulator radiation spectra, and random errors in undulators and their effects on the radiation spectrum. Also considered are REC and NdFe magnetic moment irreversibility from temperature cycling, the design potential of insertion device storage rings, FEL undulator technology and synchrotron radiation source requirements, and the Orsay undulators. Other topics include a high quality hybrid wiggler for infrared FEL and coherent harmonic generation, the development of a NdFe-steel hybrid wiggler for SSRL, the design of a 3.0 Tesla wiggler for the European Synchrotron Radiation Facility, and optics for insertion-device beam lines. Papers are also presented on mirror degradation and performance requirements in FELs, novel radiation sources for infrared to gamma rays, variational theory of insertion devices, and the evolution of the radiation-beam in long undulators.

  12. Shoe inserts and orthotics for sport and physical activities.

    PubMed

    Nigg, B M; Nurse, M A; Stefanyshyn, D J

    1999-07-01

    The purposes of this paper were to discuss the perceived benefits of inserts and orthotics for sport activities and to propose a new concept for inserts and orthotics. There is evidence that inserts or orthotics reduce or prevent movement-related injuries. However, there is limited knowledge about the specific functioning an orthotic or insert provides. The same orthotic or insert is often proposed for different problems. Changes in skeletal movement due to inserts or orthotics seem to be small and not systematic. Based on the results of a study using bone pins, one may question the idea that a major function of orthotics or inserts consists in aligning the skeleton. Impact cushioning with shoe inserts or orthotics is typically below 10%. Such small reductions might not be important for injury reduction. It has been suggested that changes in material properties might produce adjustments in the muscular response of the locomotor system. The foot has various sensors to detect input signals with subject specific thresholds. Subjects with similar sensitivity threshold levels seem to respond in their movement pattern in a similar way. Comfort is an important variable. From a biomechanical point of view, comfort may be related to fit, additional stabilizing muscle work, fatigue, and damping of soft tissue vibrations. Based on the presented evidence, the concept of minimizing muscle work is proposed when using orthotics or inserts. A force signal acts as an input variable on the shoe. The shoe sole acts as a first filter, the insert or orthotic as a second filter, the plantar surface of the foot as a third filter for the force input signal. The filtered information is transferred to the central nervous system that provides a subject specific dynamic response. The subject performs the movement for the task at hand. For a given movement task, the skeleton has a preferred path. If an intervention supports/counteracts the preferred movement path, muscle activity can/must be reduced/increased. Based on this concept, an optimal insert or orthotic would reduce muscle activity, feel comfortable, and should increase performance. PMID:10416543

  13. Production Recesses for Replaceable Cutting Inserts of Milling Tools

    NASA Astrophysics Data System (ADS)

    Kousal, Lukáš; Sadílek, Marek

    2014-12-01

    This article describes the production of recesses for replaceable cutting inserts of milling tools. The recess manufacture is exemplified on a D20 shanktype milling cutter with three replaceable cutting inserts. A new production technology and its operating cycle are demonstrated. A com-parison of the old and the new technologies shows the percentage saving of change cycles and used tools, including overall evaluation of the tech-nology.

  14. Metal-Matrix Composite Parts With Metal Inserts

    NASA Technical Reports Server (NTRS)

    Majkowski, T.; Kashalikar, U.

    1995-01-01

    Developmental fabrication process produces metal-matrix composite (MMC) parts with integral metal inserts. With inserts, MMC parts readily joined to similar parts by use of brazing, welding, or mechanical fasteners. Process conceived to make strong, lightweight components of structures erected in outer space. Also useful on Earth, in such automotive parts as rocker arms, cylinder liners, and pistons. Potential industrial applications include parts subjected to high stresses at high temperatures, as in power-generation, mining, and oil-drilling equipment.

  15. 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

  16. Deep electrode insertion and sound coding in cochlear implants.

    PubMed

    Hochmair, Ingeborg; Hochmair, Erwin; Nopp, Peter; Waller, Melissa; Jolly, Claude

    2015-04-01

    Present-day cochlear implants demonstrate remarkable speech understanding performance despite the use of non-optimized coding strategies concerning the transmission of tonal information. Most systems rely on place pitch information despite possibly large deviations from correct tonotopic placement of stimulation sites. Low frequency information is limited as well because of the constant pulse rate stimulation generally used and, being even more restrictive, of the limited insertion depth of the electrodes. This results in a compromised perception of music and tonal languages. Newly available flexible long straight electrodes permit deep insertion reaching the apical region with little or no insertion trauma. This article discusses the potential benefits of deep insertion which are obtained using pitch-locked temporal stimulation patterns. Besides the access to low frequency information, further advantages of deeply inserted long electrodes are the possibility to better approximate the correct tonotopic location of contacts, the coverage of a wider range of cochlear locations, and the somewhat reduced channel interaction due to the wider contact separation for a given number of channels. A newly developed set of strategies has been shown to improve speech understanding in noise and to enhance sound quality by providing a more "natural" impression, which especially becomes obvious when listening to music. The benefits of deep insertion should not, however, be compromised by structural damage during insertion. The small cross section and the high flexibility of the new electrodes can help to ensure less traumatic insertions as demonstrated by patients' hearing preservation rate. This article is part of a Special Issue entitled . PMID:25456089

  17. 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).

  18. Site-Specific Insertion of IS492 in Pseudoalteromonas atlantica?

    PubMed Central

    Higgins, Brian P.; Popkowski, Adam C.; Caruana, Peter R.; Karls, Anna C.

    2009-01-01

    Reversible insertion of IS492 at a site within epsG on the Pseudoalteromonas atlantica chromosome controls peripheral extracellular polysaccharide production and biofilm formation by P. atlantica. High-frequency precise excision of IS492 from epsG requires 5 and 7 bp of flanking DNA, suggesting that IS492 transposition involves a site-specific recombination mechanism. The site specificity of IS492 insertion was examined in P. atlantica and shown to be specific for a 7-bp target, 5?-CTTGTTA-3?. Characterization of numerous insertion events at the target site in epsG indicated that insertion is also orientation specific. The frequency of IS492 insertion at the epsG target site (2.7 × 10?7/cell/generation), determined by quantitative PCR, is 4 to 5 orders of magnitude lower than the frequency of IS492 precise excision from the same site. Comparison of insertion sites for IS492 and the highly related ISPtu2 from Pseudoalteromonas tunicata suggests DNA sequence and/or structural features that may contribute to site recognition and recombination by the transposase of IS492. PMID:19684137

  19. Seamless Insertion of Pulmonary Nodules in Chest CT Images.

    PubMed

    Pezeshk, Aria; Sahiner, Berkman; Zeng, Rongping; Wunderlich, Adam; Chen, Weijie; Petrick, Nicholas

    2015-12-01

    The availability of large medical image datasets is critical in many applications, such as training and testing of computer-aided diagnosis systems, evaluation of segmentation algorithms, and conducting perceptual studies. However, collection of data and establishment of ground truth for medical images are both costly and difficult. To address this problem, we are developing an image blending tool that allows users to modify or supplement existing datasets by seamlessly inserting a lesion extracted from a source image into a target image. In this study, we focus on the application of this tool to pulmonary nodules in chest CT exams. We minimize the impact of user skill on the perceived quality of the composite image by limiting user involvement to two simple steps: the user first draws a casual boundary around a nodule in the source, and, then, selects the center of desired insertion area in the target. We demonstrate the performance of our system on clinical samples, and report the results of a reader study evaluating the realism of inserted nodules compared to clinical nodules. We further evaluate our image blending techniques using phantoms simulated under different noise levels and reconstruction filters. Specifically, we compute the area under the ROC curve of the Hotelling observer (HO) and noise power spectrum of regions of interest enclosing native and inserted nodules, and compare the detectability, noise texture, and noise magnitude of inserted and native nodules. Our results indicate the viability of our approach for insertion of pulmonary nodules in clinical CT images. PMID:26080378

  20. Bits with diamond-coated inserts reduce gauge problems

    SciTech Connect

    Eckstrom, D. )

    1991-06-17

    In highly abrasive formations, failure of the gauge row cutters on tungsten carbide insert bits may occur rapidly, resulting in short bit runs, poor performance, and undergauge hole. In certain applications, polycrystalline diamond (PCD) enhanced insert bits have longer bit runs and maintain an in-gauge hole which reduces reaming time and wear on downhole equipment. These bits with PCD-coated inserts have reduced drilling costs in several areas of Canada. PCD has been applied to rock drilling tools for several years because of its high wear resistance. Polycrystalline diamond compact (PDC) bits use polycrystalline diamonds formed in flat wafers applied to the flat surfaces on carbide inserts. The flat PDC cutters drill by shearing the formation. Smith International Canada Ltd. developed a patented process to apply PCD to curved surfaces, which now allows PCD-enhanced inserts to be used for percussion and rotary cone applications. These diamond-enhanced inserts combine the wear resistance properties of diamond with the durability of tungsten carbide.

  1. Evaluation of postabortion IUD insertion in Egyptian women.

    PubMed

    Moussa, A

    2001-06-01

    This study was carried out at Alhussein University Hospital and Elmonera General Hospital to assess the safety and efficacy of intrauterine device (IUD) insertion immediately after spontaneous abortion compared with insertion 2 weeks after abortion. One hundred women between ages 18 and 40 years were recruited from those admitted via the emergency room with first trimester spontaneous abortion. All women were counseled about a method of contraception, particularly copper T-380, and divided into two groups: Group I, which included 69 women who preferred immediate IUD insertion, and Group II, which included 31 women who asked for late IUD insertion 2 weeks after an abortion. All women were followed at 2, 6, and 10 weeks after insertion of IUDs. Bleeding patterns were comparable in both groups. Mild bleeding occurred in 9.2% and 16% in Groups I and II, respectively; moderate bleeding occurred in 80% and 64%, respectively, and severe bleeding was observed in 10.8% and 20%, respectively. This was not significant. Expulsion rate was 4.5% and 3.4% in Groups I and II, respectively, which was also not significant. There were no cases of perforation or pelvic infections. This study showed that insertion of an IUD immediately after a spontaneous abortion is safe and could be offered to those who have had an abortion and who ask for a method of contraception. PMID:11672553

  2. 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

  3. Design of Revolute Joints for In-Mold Assembly Using Insert Molding.

    PubMed

    Ananthanarayanan, Arvind; Ehrlich, Leicester; Desai, Jaydev P; Gupta, Satyandra K

    2011-12-01

    Creating highly articulated miniature structures requires assembling a large number of small parts. This is a very challenging task and increases cost of mechanical assemblies. Insert molding presents the possibility of creating a highly articulated structure in a single molding step. This can be accomplished by placing multiple metallic bearings in the mold and injecting plastic on top of them. In theory, this idea can generate a multi degree of freedom structures in just one processing step without requiring any post molding assembly operations. However, the polymer material has a tendency to shrink on top of the metal bearings and hence jam the joints. Hence, until now insert molding has not been used to create articulated structures. This paper presents a theoretical model for estimating the extent of joint jamming that occurs due to the shrinkage of the polymer on top of the metal bearings. The level of joint jamming is seen as the effective torque needed to overcome the friction in the revolute joints formed by insert molding. We then use this model to select the optimum design parameters which can be used to fabricate functional, highly articulating assemblies while meeting manufacturing constraints. Our analysis shows that the strength of weld-lines formed during the in-mold assembly process play a significant role in determining the minimum joint dimensions necessary for fabricating functional revolute joints. We have used the models and methods described in this paper to successfully fabricate the structure for a minimally invasive medical robot prototype with potential applications in neurosurgery. To the best of our knowledge, this is the first demonstration of building an articulated structure with multiple degrees of freedom using insert molding. PMID:23904746

  4. Study of Insertion Characteristics of Si Neural Probe with Sharpened Tip for Minimally Invasive Insertion to Brain

    NASA Astrophysics Data System (ADS)

    Lee, Sanghoon; Kanno, Soichiro; Kino, Hisashi; Tanaka, Tetsu

    2013-04-01

    We have fabricated various types of Si neural probe for in vivo and in vitro neuronal signal recordings by combining standard photolithography with a bulk micromachining process. To place the probe tip at target areas in the brain precisely, the mechanical properties of the Si neural probes with various tip shapes were carefully investigated under different insertion conditions to brain phantoms. As results, the minimum length and penetration force of the Si neural probe were determined from conditions where the Si neural probe began to penetrate the surface of the brain phantom. To control buckling of the Si neural probe, it is necessary to optimize the insertion rate in accordance with the conditions of the Si neural probe. Although the insertion forces of the Si neural probe with a sharpened tip were smaller than those of the Si neural probe with a normal tip, the effect of the probe tip shape became small with increased insertion speeds.

  5. Effects of Delay Lines on Surface Acoustic Wave Resonator Filters and Their Applications

    NASA Astrophysics Data System (ADS)

    Chu, Sheng-Yuan; Water, Walter

    1998-11-01

    The surface acoustic wave (SAW) resonator filters consisting of a pair of interdigital-electrode transducers can generate and transmit acoustic waves. By inserting a grating, we can improve the frequency response and lower the insertion loss; therefore appropriate design of the delay line will be very important. In this paper, we used the coupling-of-mode (COM) theory to model the frequency response of the SAW device and discuss the influence of the delay line on the frequency response. We found that the optimum delay line distance and the corresponding insertion loss is very small, which will be very helpful in designing corresponding SAW-coupled resonator filters.

  6. Time-elapsed screw insertion with microCT imaging.

    PubMed

    Ryan, M K; Mohtar, A A; Cleek, T M; Reynolds, K J

    2016-01-25

    Time-elapsed analysis of bone is an innovative technique that uses sequential image data to analyze bone mechanics under a given loading regime. This paper presents the development of a novel device capable of performing step-wise screw insertion into excised bone specimens, within the microCT environment, whilst simultaneously recording insertion torque, compression under the screw head and rotation angle. The system is computer controlled and screw insertion is performed in incremental steps of insertion torque. A series of screw insertion tests to failure were performed (n=21) to establish a relationship between the torque at head contact and stripping torque (R(2)=0.89). The test-device was then used to perform step-wise screw insertion, stopping at intervals of 20%, 40%, 60% and 80% between screw head contact and screw stripping. Image data-sets were acquired at each of these time-points as well as at head contact and post-failure. Examination of the image data revealed the trabecular deformation as a result of increased insertion torque was restricted to within 1mm of the outer diameter of the screw thread. Minimal deformation occurred prior to the step between the 80% time-point and post-failure. The device presented has allowed, for the first time, visualization of the micro-mechanical response in the peri-implant bone with increased tightening torque. Further testing on more samples is expected to increase our understanding of the effects of increased tightening torque at the micro-structural level, and the failure mechanisms of trabeculae. PMID:26747514

  7. Gene Transfer and Genome-Wide Insertional Mutagenesis by Retroviral Transduction in Fish Stem Cells

    PubMed Central

    Liu, Qizhi; Wang, Yunzhi; Lin, Fan; Zhang, Lei; Li, Yan; Ge, Ruowen; Hong, Yunhan

    2015-01-01

    Retrovirus (RV) is efficient for gene transfer and integration in dividing cells of diverse organisms. RV provides a powerful tool for insertional mutagenesis (IM) to identify and functionally analyze genes essential for normal and pathological processes. Here we report RV-mediated gene transfer and genome-wide IM in fish stem cells from medaka and zebrafish. Three RVs were produced for fish cell transduction: rvLegfp and rvLcherry produce green fluorescent protein (GFP) and mCherry fluorescent protein respectively under control of human cytomegalovirus immediate early promoter upon any chromosomal integration, whereas rvGTgfp contains a splicing acceptor and expresses GFP only upon gene trapping (GT) via intronic in-frame integration and spliced to endogenous active genes. We show that rvLegfp and rvLcherry produce a transduction efficiency of 11~23% in medaka and zebrafish stem cell lines, which is as 30~67% efficient as the positive control in NIH/3T3. Upon co-infection with rvGTgfp and rvLcherry, GFP-positive cells were much fewer than Cherry-positive cells, consistent with rareness of productive gene trapping events versus random integration. Importantly, rvGTgfp infection in the medaka haploid embryonic stem (ES) cell line HX1 generated GTgfp insertion on all 24 chromosomes of the haploid genome. Similar to the mammalian haploid cells, these insertion events were presented predominantly in intergenic regions and introns but rarely in exons. RV-transduced HX1 retained the ES cell properties such as stable growth, embryoid body formation and pluripotency gene expression. Therefore, RV is proficient for gene transfer and IM in fish stem cells. Our results open new avenue for genome-wide IM in medaka haploid ES cells in culture. PMID:26029933

  8. Possible Application of NbTi Artificial Pinning Centers Wire for Insertion Devices

    NASA Astrophysics Data System (ADS)

    Holubek, T.; Casalbuoni, S.; Gerstl, S.; Grau, A.; Jauregui, D. Saez de; KlŠser, M.; Schneider, Th.; Motowidlo, L.

    Superconductive insertion devices (IDs) allow higher fields for a given gap and period length compared to the classical permanent magnet IDs. This technological concept enables an increase in the brilliance and/or the photon energy. The workhorse for superconducting magnets are multifilament NbTi wires, which are nowadays also used for superconducting insertion devices. Even higher magnetic fields can be reached by using a conductor with enhanced critical current density.Here, we propose a possible application for superconducting undulators, wound with NbTi wire with artificial pinning centers, developed by SupraMagnetics, Inc. We report the critical current characteristic, Jc(B), of a short wire measured in a liquid helium bath, and the load-line of a racetrack coil, designed to simulate the field configuration on the conductor as in a superconducting undulator. Based on the measured load line, we report on the simulations of the magnetic field on axis and of the spectrum in a third generation light source of a possible undulator wound with a wire having similar properties to the measured one.

  9. 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.

  10. Insertion of Totally Implantable Central Venous Access Devices by Surgeons

    PubMed Central

    An, Hyeonjun; Ryu, Chun-Geun; Jung, Eun-Joo; Kang, Hyun Jong; Paik, Jin Hee; Yang, Jung-Hyun

    2015-01-01

    Purpose The aim of this study is to evaluate the results for the insertion of totally implantable central venous access devices (TICVADs) by surgeons. Methods Total 397 patients, in whom TICVADs had been inserted for intravenous chemotherapy between September 2008 and June 2014, were pooled. This procedure was performed under local anesthesia in an operation room. The insertion site for the TICVAD was mainly in the right-side subclavian vein. In the case of breast cancer patients, the subclavian vein opposite the surgical site was used for insertion. Results The 397 patients included 73 males and 324 females. Primary malignant tumors were mainly colorectal and breast cancer. The mean operation time was 54 minutes (18-276 minutes). Operation-related complications occurred in 33 cases (8.3%). Early complications developed in 15 cases with catheter malposition and puncture failure. Late complications, which developed after 24 hours, included inflammation in 6 cases, skin necrosis in 6 cases, hematoma in 3 cases, port malfunction in 1 case, port migration in 1 case, and intractable pain at the port site in 1 case. Conclusion Insertion of a TICVAD under local anesthesia by a surgeon is a relatively safe procedure. Meticulous undermining of the skin and carefully managing the TICVAD could minimize complications. PMID:25960974

  11. Correction of hypermobile flatfoot in children by molded insert.

    PubMed

    Bordelon, R L

    1980-11-01

    One hundred feet in 50 children between the ages of 3 and 9 years with a diagnosis of idiopathic hypermobile flatfoot had a custom-molded insert ordered. A specific method of casting, correcting the various components of the deformity was utilized. An 1/8-inch polypropolene insert was fabricated from the positive cast. The insert was worn in leather shoes with a long counter, steel shank, and Thomas heel. The flatfoot was evaluated and classified by measurement of the talometatarsal angle on a standing lateral X-ray. The insert was fabricated so that the standing lateral talometatarsal angle was corrected to neutral with the insert on the foot and the foot in the shoe. The preliminary reports indicate that a correction can be obtained at the rate of 0.41 degrees per month or approximately 5 degrees per year. There was no significant loss of motion of the foot or the ankle. Perhaps this regimen may be utilized in those children with a hypermobile flatfoot for whom treatment is advised. PMID:7319430

  12. Mechanisms of integral membrane protein insertion and folding

    PubMed Central

    2014-01-01

    The biogenesis, folding, and structure of α-helical membrane proteins (MPs) are important to understand because they underlie virtually all physiological processes in cells including key metabolic pathways, such as the respiratory chain and the photosystems, and the transport of solutes and signals across membranes. Nearly all MPs require translocons—often referred to as protein-conducting channels—for proper insertion into their target membrane. Remarkable progress toward understanding the structure and functioning of translocons has been made during the past decade. Here we review and assess this progress critically. All available evidence indicates that MPs are equilibrium structures that achieve their final structural states by folding along thermodynamically controlled pathways. The main challenge for cells is the targeting and membrane insertion of highly hydrophobic amino acid sequences. Targeting and insertion are managed in cells principally by interactions between ribosomes and membrane-embedded translocons. Our review examines the biophysical and biological boundaries of membrane protein insertion and the folding of polytopic membrane proteins in vivo. A theme of the review is the under-appreciated role of basic thermodynamic principles in MP folding and assembly. Thermodynamics not only dictates the final folded structure, it is the driving force for the evolution of the ribosome-translocon system of assembly. We conclude the review with a perspective suggesting a new view of translocon-guided MP insertion. PMID:25277655

  13. Biomechanical comparisons of unidirectional and bidirectional Kirschner-wire insertion.

    PubMed

    Zohman, G L; Tsenter, M; Kabo, J M; Meals, R A

    1992-11-01

    Peak axial loads on entry and peak pull-out loads were measured for Kirschner-wires (K-wire) inserted into canine metacarpals. The wires were placed using a standard, unidirectional drill and using an oscillating, bidirectional drill that reversed spin direction every 120 degrees. Seven trocar-tip wires were compared with seven diamond-tip wires using each drilling method. The trocar-tip wires demonstrated equal peak axial loads on entry when drilled at comparable speeds unidirectionally and bidirectionally. Pull-out load of the trocar tip after bidirectional insertion was less than that for unidirectional insertion but was still in the clinically useful range. The diamond-tip wire exhibited higher peak axial load on entry and lower peak pull-out load when drilled bidirectionally. Bidirectional insertion of a trocar-tip K-wire offers fixation characteristics comparable with those with unidirectional insertion and will potentially reduce the risk of soft-tissue damage associated with winding-up and avulsion of vital structures on the spinning shaft of a unidirectional driver. PMID:1395308

  14. GAPDH regulates cellular heme insertion into inducible nitric oxide synthase

    PubMed Central

    Chakravarti, Ritu; Aulak, Kulwant S.; Fox, Paul L.; Stuehr, Dennis J.

    2010-01-01

    Heme proteins play essential roles in biology, but little is known about heme transport inside mammalian cells or how heme is inserted into soluble proteins. We recently found that nitric oxide (NO) blocks cells from inserting heme into several proteins, including cytochrome P450s, hemoglobin, NO synthases, and catalase. This finding led us to explore the basis for NO inhibition and to identify cytosolic proteins that may be involved, using inducible NO synthase (iNOS) as a model target. Surprisingly, we found that GAPDH plays a key role. GAPDH was associated with iNOS in cells. Pure GAPDH bound tightly to heme or to iNOS in an NO-sensitive manner. GAPDH knockdown inhibited heme insertion into iNOS and a GAPDH mutant with defective heme binding acted as a dominant negative inhibitor of iNOS heme insertion. Exposing cells to NO either from a chemical donor or by iNOS induction caused GAPDH to become S-nitrosylated at Cys152. Expressing a GAPDH C152S mutant in cells or providing a drug to selectively block GAPDH S-nitrosylation both made heme insertion into iNOS resistant to the NO inhibition. We propose that GAPDH delivers heme to iNOS through a process that is regulated by its S-nitrosylation. Our findings may uncover a fundamental step in intracellular heme trafficking, and reveal a mechanism whereby NO can govern the process. PMID:20921417

  15. Insertion site mapping for repeated elements in Mycobacterium tuberculosis.

    PubMed

    Moganeradj, Kartyk; Abubakar, Ibrahim; McHugh, Timothy D; Sonnenberg, Pam; Arnold, Catherine

    2013-02-15

    Insertion elements not only act as genetic markers for differentiation of bacteria but their movement in bacterial genomes likely plays an essential role in changing the physical and biochemical traits of the organisms when adapting to new environments. Genomic Insertion Site mapping of transposable elements could shed light on the putative altered function of adjacent genes. In the era of whole genome sequencing where repeat elements are difficult to sequence with short read technologies and in the absence of high throughput technologies especially in poorer resource settings, an alternative approach to their characterisation is needed. A rapid and simple method of insertion site mapping that uses Insertion Sequence 6110 (IS6110) fluorescent amplified fragment length polymorphism (FAFLP) PCR as a foundation and then uses additional selective bases to reduce the number of fragments generated was developed. This was applied to Mycobacterium tuberculosis H37Rv sequenced strain to compare the experimental data with the in silico results. This was successfully achieved for all but two of the sixteen fragments generated by FAFLP and demonstrated that, by using this technique, insertion sites can be mapped onto the genomes of M. tuberculosis. PMID:23262032

  16. Sorting genomes by reciprocal translocations, insertions, and deletions.

    PubMed

    Qi, Xingqin; Li, Guojun; Li, Shuguang; Xu, Ying

    2010-01-01

    The problem of sorting by reciprocal translocations (abbreviated as SBT) arises from the field of comparative genomics, which is to find a shortest sequence of reciprocal translocations that transforms one genome Pi into another genome Gamma, with the restriction that Pi and Gamma contain the same genes. SBT has been proved to be polynomial-time solvable, and several polynomial algorithms have been developed. In this paper, we show how to extend Bergeron's SBT algorithm to include insertions and deletions, allowing to compare genomes containing different genes. In particular, if the gene set of Pi is a subset (or superset, respectively) of the gene set of Gamma, we present an approximation algorithm for transforming Pi into Gamma by reciprocal translocations and deletions (insertions, respectively), providing a sorting sequence with length at most OPT + 2, where OPT is the minimum number of translocations and deletions (insertions, respectively) needed to transform Pi into Gamma; if Pi and Gamma have different genes but not containing each other, we give a heuristic to transform Pi into Gamma by a shortest sequence of reciprocal translocations, insertions, and deletions, with bounds for the length of the sorting sequence it outputs. At a conceptual level, there is some similarity between our algorithm and the algorithm developed by El Mabrouk which is used to sort two chromosomes with different gene contents by reversals, insertions, and deletions. PMID:20431155

  17. Lithium insertion mechanism in tin-based spinel sulfides

    SciTech Connect

    Branci, C.; Sarradin, J.; Olivier-Fourcade, J.; Jumas, J.C.

    1999-10-01

    Cu{sub 2}MSn{sub 3}S{sub 8} (M = Fe, Co) spinel compounds have been studied as cathode materials in Li/LiPF{sub 6}(1M)-PC-EC/spinel sulfide cells. To understand the lithium insertion mechanism, both pristine compounds and chemically inserted products were characterized by X-ray Powder Diffraction (XPD), {sup 119}Sn Moessbauer Spectroscopy (MS), and X-ray Absorption Spectroscopy (XAS) at the sulfur K-edge. To accommodate the inserted lithium ions, two steps of reduction have been observed. First, for a low lithium content (up to 2 Li per formula), a copper reduction is occurring. Moreover, for the iron-based compound the copper atoms extraction is associated with a migration of iron atoms from the 16c octahedral sites toward the 8a tetrahedral sites. Second, for a high lithium content (above 2 Li per formula) a complex and simultaneous reduction of tin(IV) toward tin(II) and tin(0) has been clearly observed as well as a distortion of sulfur coordination polyhedra during the insertion process. From an electrochemical point of view, these spinels are rather interesting as model compounds regarding the lithium insertion study.

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

    PubMed Central

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

    2012-01-01

    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. PMID:22534608

  19. 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. PMID:22534608

  20. A High-Throughput Arabidopsis Reverse Genetics System

    PubMed Central

    Sessions, Allen; Burke, Ellen; Presting, Gernot; Aux, George; McElver, John; Patton, David; Dietrich, Bob; Ho, Patrick; Bacwaden, Johana; Ko, Cynthia; Clarke, Joseph D.; Cotton, David; Bullis, David; Snell, Jennifer; Miguel, Trini; Hutchison, Don; Kimmerly, Bill; Mitzel, Theresa; Katagiri, Fumiaki; Glazebrook, Jane; Law, Marc; Goff, Stephen A.

    2002-01-01

    A collection of Arabidopsis lines with T-DNA insertions in known sites was generated to increase the efficiency of functional genomics. A high-throughput modified thermal asymetric interlaced (TAIL)-PCR protocol was developed and used to amplify DNA fragments flanking the T-DNA left borders from ?100,000 transformed lines. A total of 85,108 TAIL-PCR products from 52,964 T-DNA lines were sequenced and compared with the Arabidopsis genome to determine the positions of T-DNAs in each line. Predicted T-DNA insertion sites, when mapped, showed a bias against predicted coding sequences. Predicted insertion mutations in genes of interest can be identified using Arabidopsis Gene Index name searches or by BLAST (Basic Local Alignment Search Tool) search. Insertions can be confirmed by simple PCR assays on individual lines. Predicted insertions were confirmed in 257 of 340 lines tested (76%). This resource has been named SAIL (Syngenta Arabidopsis Insertion Library) and is available to the scientific community at www.tmri.org. PMID:12468722

  1. Dopant Modulated Li Insertion in Si for Battery Anodes

    SciTech Connect

    Long, Brandon R.; Chan, Maria K. Y.; Greeley, Jeffrey P.; Gewirth, Andrew A.

    2011-09-29

    We examine the effects of p-type and n-type dopants on the lithiation of crystalline Si as related to Li-ion batteries. In situ Raman spectroscopy and electrochemistry are used to investigate two crystallographic faces, (100) and (111), for boron (B) and phosphorus (P) dopants, to monitor the insertion of Li and the associated transition to amorphous Si. Density functional theory calculations are used to investigate the lithiation of doped and undoped crystalline Si bulk and surface models. The experimental and computational results suggest that lithiation voltages are different for P-doped and B-doped Si. The B-doped surfaces are found to insert Li at higher voltages than P- and undoped surfaces but result in less Li insertion. These results provide an understanding of the effects on dopants on the lithiation of silicon which may ultimately aid in the development of alternate anode materials for Li-ion batteries.

  2. 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.

  3. Improved technique for insertion of intra-aortic balloon pump.

    PubMed

    Cutler, B S; Vander Salm, T J

    1980-06-01

    Polytetrafluoroethylene (PTFE, Gortex) grafts have several advantages over woven Dacron grafts for the insertion of intra-aortic balloons (IABP). Because of the smooth inner surface of the PTFE graft, an 8-mm graft may be used instead of the 10-mm minimum size with Dacron. The pliability of the PTFE graft makes it easier to sew with fine suture material, and its comformability permits a "blood tight" seal to be obtained with a single Dacron umbilical tape tie. We have used PTFE grafts for IABP insertion in 24 patients, with no infectious or thrombotic complications. This graft material seems to offer substantial technical advantages over woven Dacron for the insertion of IABP and subsequent arterial repair. PMID:7387373

  4. Characterization of the performance of shoe insert materials.

    PubMed

    Lewis, G; Tan, T; Shiue, Y S

    1991-08-01

    It has been widely reported that shoe inserts are an effective interventional modality either for the relief of discomfort to the feet associated with a variety of orthopedic disorders or conditions or simply for comfort. Results from many types of experimental tests have been used to obtain the shock absorption capacity of shoe insert materials. The authors contend in this study that, while shock absorption is a highly desirable property, it is by no means the only that should be used to characterize these materials. Thus, a new index of performance of these materials is proposed. This index is computed from data, obtained in a simple experimental test, on both the shock absorption and energy return performances of the insert material. PMID:1920103

  5. 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.

  6. Hysteresis during lithium insertion in hydrogen-containing carbons

    SciTech Connect

    Zheng, T.; Dahn, J.R.; McKinnon, W.R.

    1996-07-01

    The authors studied lithium insertion in hydrogen-containing carbons heated at temperatures near 700 C. High capacities with large hysteresis (lithium insertion into these carbons at nearly 0 V and removal at nearly 1 V) were shown to be proportional to the hydrogen content of the samples. It is believed that the lithium atoms may bind on hydrogen-terminated edges of hexagonal carbon fragments, causing a change in the bond from sp{sup 2} to sp{sup 3}. The authors have carefully studied the electrochemical insertion of lithium in hydrogen-containing carbons using a variety of charge-discharge rates and cycling temperatures. These measurements allow the hysteresis to be quantified. A simple model, which treats the bonding change as an activated process, is used to model the hysteresis in the cells qualitatively.

  7. Electrochemical insertion of lithium into polyacrylonitrile-based disordered carbons

    SciTech Connect

    Jung, Y.; Suh, M.C.; Lee, S.I.; Shim, S.C.; Kwak, J.; Lee, H.; Kim, M.

    1997-12-01

    Electrochemical lithium insertion into polyacrylonitrile (PAN)-based disordered carbons was studied using the techniques of discharge/charge tests, cyclic voltammetry, and {sup 7}Li nuclear magnetic resonance (NMR) spectroscopy. The PAN-based carbons were prepared by vacuum pyrolysis of PAN at 500, 800, and 1,000 C. They showed charge capacities between 254 and 380 mAh/g in the first cycle. {sup 7}Li NMR spectra showed two kinds of lithium insertion sites in the PAN-based carbons: a reversible site where lithium is removed in the subsequent charge process and an irreversible site where lithium remains intact. The NMR results suggest that lithium in fully Li-inserted PAN-based carbons has an ionic character, and reversible site lithium resides between negatively charged carbon layers.

  8. Feasibility of a free-standing insertable heat source

    NASA Astrophysics Data System (ADS)

    Munger, A. C.; Mohler, J. H.; Kelly, M. D.

    The feasibility of a free-standing insertable heat source to replace an existing heat powder charge has been demonstrated. Successful development of this design eliminates the need for three laser weld joints, a high pressure interface and various manufacturing processes in the next assembly. Previous work yielded a component consisting of a stainless steel tube loaded with Fe/KC104 heat powder and a pyrotechnic ignitor. Although this design is adequate for the function, the advantages of an insertable heat source prompted the present development. The initial concept was to load a heat powder into a thin-walled container that could be inserted later. Varying air gaps prevented proper heat transmission. A freestanding thermite compact pressed to the inside contour of the stainless steel tube was then developed. This compact was attached to an ignitor which could then be threaded into the next assembly. Various thermite materials were evaluated.

  9. Bone scanning in the adductor insertion avulsion syndrome ("thigh splints").

    PubMed

    Charkes, N D; Siddhivarn, N; Schneck, C D

    1987-12-01

    Shin splints is a defined clinical entity resulting from extreme tension on muscles inserting on the tibia, resulting in periosteal elevation which is detectable by bone scanning. The clinical equivalent in the thigh has been described. We found scintigraphic changes in the femurs of seven short, female, basic trainees at the Fort Dix Army base, most of whom were referred for stress fractures elsewhere in the lower extremities. The scan findings were generally noted in the upper or mid femurs, always involved the anteromedial cortex, and were bilateral in five of the seven subjects. The abnormalities were linear and suggested periosteal elevation, and did not have the typical appearance of stress fracture. Since the findings correspond to the insertion of one or more adductor muscle groups, the descriptive term "adductor insertion avulsion syndrome" or "thigh splints" is proposed for this entity. PMID:3681443

  10. 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.

  11. Antibiotics Before Removal of Percutaneously Inserted Central Venous Catheters Reduces Clinical Sepsis in Premature Infants

    PubMed Central

    Reynolds, Gail E.; Tierney, Sarah B.

    2015-01-01

    OBJECTIVES: Evaluate the incidence of postcatheter removal clinical sepsis when antibiotics were infused prior to the removal of percutaneously inserted central venous catheters (PICCs). METHODS: A retrospective chart review of premature neonates (n = 196) weighing ?1250 g at birth with 218 PICC line removals in the presence or absence of antibiotics at a tertiary level neonatal intensive care unit (NICU) between January 1, 2010, and May 31, 2012. Charts were reviewed looking for the presence of clinical sepsis defined as a sepsis workup including white blood cell count, differential, C-reactive protein, blood and/or cerebral spinal fluid (CSF), and urine cultures along with at least 48 hours of antibiotic therapy given within 72 hours after removal of a PICC line. Antibiotics were considered present at line removal if given within 12 hours before catheter removal either electively or at completion of a planned course. RESULTS: When antibiotics were given within 12 hours before PICC line removal, only 2% of the line removal episodes (1/48) resulted in a neonate developing clinical sepsis versus 13% (21/165) when no antibiotics were given prior to removal (p = 0.03, Fisher's exact test). Despite the increased use of elective antibiotics with line removal, there was no increase in total antibiotic usage due to the overall decrease in episodes of clinical sepsis or changes in antibiogram susceptibility patterns. CONCLUSIONS: There was an 11% absolute decrease and a 6-fold relative decrease in postcatheter removal clinical sepsis events in premature neonates who received antibiotics prior to PICC line removal. PMID:26170772

  12. Spot welding of steel and aluminum using insert sheet

    SciTech Connect

    Oikawa, H.; Saito, T.; Yoshimura, T.

    1994-12-31

    Automobile industries have been increasingly interested in the use of aluminum and thus joining of steel and aluminum becomes of importance. The joining of the two types of metal raises a problem of brittle welds caused by the formation of intermetallic compounds. The authors solved the problem by using an insert sheet. This paper deals with the resistance spot welding of steel and aluminum sheets using insert sheets. The insert sheet used in the present development was a steel/aluminum clad sheet of the 0.8 mm thickness with 50% steel and 50% aluminum. The clad sheet was produced by warm rolling of steel and aluminum with a direct resistance heating process. Steel to be warm rolled was of EDDQ of the 0.4 mm thickness and aluminum was of JIS A1050 of 0.6 mm thickness. The mechanical properties of the insert clad sheets were in between those of the steel sheets and the aluminum sheets, while the clad sheets showed much better formability than the aluminum sheets. Resistance spot welding was conducted for 0.8 mm thick EDDQ steel sheets and 1.0 mm thick aluminum alloy (AL-5.5%Mg) sheets under the welding force of 1.96 kN, welding current ranging between 4.2 and 20.1 kA, and welding time from 0.5 to 10 cycles. The steel was spot welded to the steel side of the insert sheet while the aluminum was welded to the aluminum side. What the authors investigated were the applicable welding current range, nugget diameter, tensile shear strength, U-tension strength, and macro- and microstructures. In conclusion, steel sheets can be spot welded to aluminum sheets without difficulty by using clad sheets as insert materials while the strength level of the dissimilar metal spot welds is close to that of aluminum joints.

  13. Ultrasonic airborne insertion loss measurements at normal incidence (L).

    PubMed

    Farley, Jayrin; Anderson, Brian E

    2010-12-01

    Transmission loss and insertion loss measurements of building materials at audible frequencies are commonly made using plane wave tubes or as a panel between reverberant rooms. These measurements provide information for noise isolation control in architectural acoustics and in product development. Airborne ultrasonic sound transmission through common building materials has not been fully explored. Technologies and products that utilize ultrasonic frequencies are becoming increasingly more common, hence the need to conduct such measurements. This letter presents preliminary measurements of the ultrasonic insertion loss levels for common building materials over a frequency range of 28-90 kHz using continuous-wave excitation. PMID:21218864

  14. 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.

  15. Method of modifying a volume mesh using sheet insertion

    DOEpatents

    Borden, Michael J. (Albuquerque, NM); Shepherd, Jason F. (Albuquerque, NM)

    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.

  16. Metallocene Catalytic Insertion Polymerization of 1-Silene to Polycarbosilanes

    NASA Astrophysics Data System (ADS)

    Tian, Yuelong; Ge, Min; Zhang, Weigang; Lv, Xiaoxu; Yu, Shouquan

    2015-11-01

    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.

  17. 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. PMID:4072987

  18. Insertion and confinement of air bubbles inside a liquid marble.

    PubMed

    Sun, Guanqing; Sheng, Yifeng; Ngai, To

    2016-01-14

    Nanoparticles at the air/liquid interface can serve as solid separating barriers to form stable foams or liquid marbles depending on the wettability of the nanoparticles. This paper presents an effect that enables the insertion and confinement of air bubbles inside a liquid marble, based on encapsulating an air bubble surrounded by surfactant molecules or hydrophilic particles. We have demonstrated that more than one bubble can be inserted and trapped inside one liquid marble so that liquid marbles can be divided into several separate compartments. The findings presented here may stimulate fundamental studies of this novel bubble-marble phenomenon, as well as developments of various practical applications. PMID:26489449

  19. Sequence and insertion sites of murine melanoma-associated retrovirus.

    PubMed

    Li, M; Huang, X; Zhu, Z; Gorelik, E

    1999-11-01

    We previously showed that B16 melanoma cells produce ecotropic melanoma-associated retrovirus (MelARV) which encodes a melanoma-associated antigen recognized by MM2-9B6 monoclonal antibody. The biological significance of MelARV in melanoma formation remains unknown. We found that infection of normal melanocytes with MelARV resulted in malignant transformation. It is likely that MelARV emerged from the defective Emv-2 provirus, a single copy of ecotropic provirus existing in the genome of C57BL/6 mice. In the present study, we cloned and sequenced the full-length MelARV genome and its insertion sites and we completed sequencing of the Emv-2 provirus. Our data show that MelARV has a typical full-length retroviral genome with high homology (98.54%) to Emv-2, indicating a close relationship between both viruses. MelARV probably emerged as a result of recombination between Emv-2 and an endogenous nonecotropic provirus. Some observed differences in the gag and pol regions of MelARV might account for the restoration of productivity and infectivity of a novel retrovirus that somatically emerged during melanoma formation. MelARV does not contain any oncogene and therefore might induce transformation by insertional mutagenesis. We sequenced two insertion sites of MelARV. The first insertion site represents the 3' coding region of the c-maf proto-oncogene at 67.0 centimorgans (cM) on chromosome 8. The c-maf proto-oncogene encodes a basic leucine zipper protein homologous to c-fos and c-jun. Insertion of MelARV in BL6 melanoma cells resulted in the up-regulation of c-maf. It is noteworthy that the Emv-2 provirus is also inserted into a noncoding region at 61.0 cM on the same chromosome 8. The second insertion site is the 3' noncoding region of the DNA polymerase gamma (PolG) gene on chromosome 7. The expression of PolG was not affected by the MelARV insertion. Further investigation of the biological significance of MelARV in melanoma formation is being undertaken. PMID:10516025

  20. Tail-anchored membrane protein insertion into the endoplasmic reticulum

    PubMed Central

    Hegde, Ramanujan S.; Keenan, Robert J.

    2013-01-01

    Membrane proteins are inserted into the endoplasmic reticulum by two highly conserved parallel pathways. The well-studied co-translational pathway uses the signal recognition particle and its receptor for targeting, and the Sec61 translocon for membrane integration. A recently discovered post-translational pathway uses an entirely different set of factors involving transmembrane domain-selective cytosolic chaperones and an accompanying receptor at the endoplasmic reticulum. Elucidation of the structural and mechanistic basis of this post-translational membrane protein insertion pathway highlights general principles shared between the two pathways and key distinctions unique to each. PMID:22086371

  1. 78 FR 37214 - Transcontinental Gas Pipe Line Company, LLC; Notice of Availability of the Environmental...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-20

    ... Brunswick County; One new meter station, line heaters, and pig \\1\\ receiver at the terminus of the Brunswick Lateral; and \\1\\ A ``pig'' is a tool that is inserted into and moves through the pipeline, and is used...

  2. Structure of Inserted Bacteriophage Mu-1 DNA and Physical Mapping of Bacterial Genes by Mu-1 DNA Insertion

    PubMed Central

    Hsu, Ming-Ta; Davidson, Norman

    1972-01-01

    It is shown, by electron microscope observation of the structures of heteroduplexes, that Mu-1 DNA inserted into the bacterial episomes Flac and F8[1] is collinear with, rather than a circulation permutation of, the DNA of the mature Mu-1 bacteriophage. Observation of the position of the inserted Mu defines a point within the gene that has been inactivated (the lacI gene for Flac and a transfer gene in F8[1] in these particular instances). These examples illustrate a new, general method for physical gene mapping. The episome with Mu DNA inserted into F8[1] [i.e., F8[1](Mu)], although derived from a single colony, is heterogeneous in that a self-renatured sample shows a nonhomology loop of length 3.0 kb. This nonhomology loop, which has previously been observed in mature Mu-1 DNA, is due to an inversion. Images PMID:4562742

  3. Viral insertion in Evi12 causes expression of aberrant Grp94 mRNAs containing the viral gag myristylation motif

    SciTech Connect

    Akker, Eric van den; Aarts, Lambertus H.J.; Delwel, Ruud

    2007-09-30

    Ecotropic Virus Integration site 12 (Evi12) is a common virus insertion site (cVIS) in retrovirally induced murine models of leukemia and lymphoma, suggesting an important role for this locus in these hematopoietic disorders. Evi12 is located near the promoter of the ER chaperone protein and Hsp90 family member Grp94. Here we show that viral insertion in Evi12 results in the expression of aberrant Grp94 transcripts in Cas-Br-MuLV as well as in AKXD induced hematopoietic tumors, demonstrating that Grp94 is a common viral target gene. While most transcripts encode for truncated forms of Grp94, transcripts containing viral gag sequences were detected in the leukemia cell line NFS107. Interestingly, these fusion transcripts encode for myristylated viral-Grp94 fusion proteins that localize to the plasma membrane. Combined with recent evidence that myristylated forms of Hsp90 transform cells, our data suggest that myristylation of target genes may be an important mechanism in retrovirally mediated oncogenesis. Since retroviral insertion in Evi12 also affects the expression of a recently identified novel gene Grp94 neighboring nucleotidase (Gnn), located at the other side of Evi12, it appears that proviral insertion can lead to deregulation of two genes present in the same locus.

  4. 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

  5. Resorbable polymeric inserts as a means of enhancing fixation of fractures of porotic bones. I. Stability of mechanical properties and viscoelasticity of the inserts aged in vitro.

    PubMed

    Mainil-Varlet, P; Cordey, J; Gogolewski, S

    1997-02-01

    A titanium screw-poly(L-lactide) medullary insert system intended for use in the fixation of porotic bone fractures was subjected to in vitro ageing at 37 degrees C and pH = 7.4 for up to 6 months. The pull-out force of the screw from the insert and the viscoelastic properties of the insert were measured. The pull-out force did not change up to 6 months of ageing, indicating that the polymeric insert provides a good purchase for the screws. The viscoelastic properties of the insert provides of bone and did not change substantially with temperatures up to 52 degrees C. This indicates that the medullary insert would maintain its properties in vivo even if the body temperature was increased. The molecular weight of the insert at 6 months was reduced by 60%, but the molecular weight reduction did not affect the mechanical properties of the insert. PMID:9068889

  6. Magnesium oxide inserts for the LECO Carbon Analyzer

    SciTech Connect

    Bagaasen, L.M.; Jensen, C.M.

    1991-01-16

    LECO carbon analysis of plutonium metal and plutonium oxide at the Rocky Flats Plant generates several hundred kilograms of high silica residues each year. The plutonium in these residues is difficult and expensive to recover using production dissolution processes. A magnesium oxide (MgO) insert has been developed that significantly lowers the plutonium recovery costs without adversely affecting accuracy of the carbon analysis.

  7. PiggyBac Transgene Insertion Site GIZA Strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genetically modified New World screwworms were analyzed by inverse PCR and several sequences of genomic DNA flanking the transposon insertion sites were obtained. One transgenic strain, designated GIZA, exhibited the properties of a lethal genetic mutation. Classic genetic crosses of these insects r...

  8. 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. PMID:26723612

  9. Insert earphone modeling and measurement by IEC-60711 coupler.

    PubMed

    Huang, Chen-Hung; Pawar, S; Hong, Zih-Jyun; Huang, Jin

    2011-02-01

    In this study, an analytical model based on the equivalent circuit method is developed to simulate the frequency response of an insert earphone. This earphone incorporates a miniature loudspeaker commonly used in computer, communication, consumer, and car electronics. Through the laser triangulation method, electroacoustic parameters of a miniature loudspeaker are obtained. Several earphone design configurations are analyzed in accordance with the open and closed states of front leakage hole, vent, and back leakage hole. To validate the analysis, an insert earphone that is attached to IEC-60711 coupler and a specially designed fixture tube is experimentally measured for frequency response using electroacoustic equipment in the air. Simulation and experimental results show good agreement over the complete audible frequency range. Analysis indicates that states of front leakage hole, vent, and back leakage hole of an insert earphone have significant effects on frequency response. The front leakage hole affects the low frequency response, whereas the vent affects the fundamental resonance. Detailed analysis has been provided to further improve the design of insert earphones. PMID:21342831

  10. 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

  11. Stability analysis of chickpea large genomic DNA inserts in Agrobacterium.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Agrobacterium tumefaciens-mediated transformation of large DNA inserts directly into plants facilitates the transfer of gene clusters and flanking regulatory elements. It is recommended that the integrity of large genomic fragments in Agrobacterium be verified prior to plant transformation. In this ...

  12. 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.

  13. 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.

  14. Indentation and needle insertion properties of the human eye

    PubMed Central

    Matthews, A; Hutnik, C; Hill, K; Newson, T; Chan, T; Campbell, G

    2014-01-01

    Purpose Characterization of the biomechanical properties of the human eye has a number of potential utilities. One novel purpose is to provide the basis for development of suitable tissue-mimicking material. The purpose of this study was to determine the indentation and needle insertion characteristics on human eye globes and tissue strips. Methods An indenter assessed the elastic response of human eye globes and tissue strips under increasing compressive loads. Needle insertion determined the force (N) needed to penetrate various areas of the eye wall. Results The results demonstrated that globes underwent slightly greater indentation at the midline than at the central cornea, and corneal strips indented twofold more than scleral strips, although neither difference was significant (P=0.400 and P=0.100, respectively). Significant differences were observed among various areas of needle insertion (P<0.001). Needle insertion through the anterior sclera (adjacent to the limbus) and posterior sclera (adjacent to the optic nerve) required the greatest amount of force (0.954 and 1.005?N, respectively). The force required to penetrate the central cornea (0.518?N) was significantly lower than all other areas except the midline sclera (0.700?N) Conclusion These data form the basis for further research into the development of a tissue-mimicking human eye construct with potential utility as a model for use in ophthalmology research and surgical teaching. PMID:24810571

  15. Contrast-free endoscopic stent insertion in malignant biliary obstruction

    PubMed Central

    De Palma, Giovanni D; Lombardi, Giovanni; Rega, Maria; Simeoli, Immacolata; Masone, Stefania; Siciliano, Saverio; Maione, Francesco; Salvatori, Francesca; Balzano, Antonio; Persico, Giovanni

    2007-01-01

    AIM: To present a case series of MRCP-guided endoscopic biliary stent placement, performed entirely without contrast injection. METHODS: Contrast-free endoscopic biliary drainage was attempted in 20 patients with malignant obstruction, unsuitable for resection on the basis of tumor extent or medical illness. MRCP images were used to confirm the diagnosis of tumor, to exclude other biliary diseases and to demonstrate the stenoses as well as dilation of proximal liver segments. The procedure was carried out under conscious sedation. Patients were placed in the left lateral decubitus position. The endoscope was inserted, the papilla identified and cannulated by a papillotome. A guide wire was inserted and guided deeply into the biliary tree, above the stenosis, by fluoroscopy. A papillotomy approximately 1 cm. long was performed and the papillotome was exchanged with a guiding-catheter. A 10 Fr, Amsterdam-type plastic stent, 7 to 15 cm long, was finally inserted over the guide wire/guiding catheter by a pusher tube system. RESULTS: Successful stent insertion was achieved in all patients. There were no major complications. Successful drainage, with substantial reduction in bilirubin levels, was achieved in all patients. CONCLUSION: This new method of contrast-free endoscopic stenting in malignant biliary obstruction is a safe and effective method of palliation. However, a larger, randomized study comparing this new approach with the standard procedure is needed to confirm the findings of the present study. PMID:17663512

  16. 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.

  17. Effects of lithium insertion on thermal conductivity of silicon nanowires

    NASA Astrophysics Data System (ADS)

    Xu, Wen; Zhang, Gang; Li, Baowen

    2015-04-01

    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.

  18. Placental and cord insertion pathologies: screening, diagnosis, and management.

    PubMed

    Wiedaseck, Susan; Monchek, Ruth

    2014-01-01

    Placenta previa, low-lying placenta, and placenta accreta are aberrations in the normal development of the placenta. Diagnosis in the prenatal period is essential because each of these pathologies can have a profound impact on the management of labor, birth, and the third stage. In recent years, there has been an increase in the occurrence of these placental abnormalities, with the increase in the cesarean rate considered to be a main cause of this phenomenon. Comprehensive risk assessment, combined with recent advances in ultrasonography, can provide earlier detection of impaired placental implantation. Umbilical cord insertion pathologies are also of concern. Velamentous cord insertion is a defect in the insertion site of the umbilical cord resulting from the atrophy of portions of the developing placenta. In this condition, the blood vessels of the umbilical cord are not protected by Wharton's jelly, resulting in a potential for increased risk of breakage when the amniotic membranes rupture. Vasa previa is a velamentous insertion of the umbilical cord in which the blood vessels are present over the cervical os. If these blood vessels rupture during labor, it can have catastrophic effects on the fetus. Prenatal diagnosis of this condition can allow the certified nurse-midwife/certified midwife (CNM/CM) to plan for the safe birth of the newborn and avoid fetal hemorrhage. This article provides a review of risk factors, diagnosis pathophysiology, and management options for these conditions, thus enabling the CNM/CM to provide safe, effective care and management. PMID:24751147

  19. 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.

  20. 31. Photocopy of map. Insert from Master Plan Notes Prepared ...

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

    31. Photocopy of map. Insert from Master Plan Notes Prepared by Environmental Planning and Design, Pittsburgh, Pa. Original at Missouri Botanical Garden. 1972 MASTER PLAN 32. 'THE TREE,' SCULPTURE BY ALEXANDER CALDER, 1966 - Missouri Botanical Garden, 2345 Tower Grove Avenue, Saint Louis, Independent City, MO

  1. Coordination-insertion copolymerization of allyl monomers with ethylene.

    PubMed

    Ito, Shingo; Kanazawa, Masafumi; Munakata, Kagehiro; Kuroda, Jun-ichi; Okumura, Yoshikuni; Nozaki, Kyoko

    2011-02-01

    Coordination-insertion copolymerization of allyl monomers with ethylene was developed by using a palladium/phosphine-sulfonate catalyst. A variety of allyl monomers, including allyl acetate, allyl alcohol, protected allylamines, and allyl halides, were copolymerized with ethylene to form highly linear copolymers that possess in-chain -CH(2)CH(CH(2)FG)- units. PMID:21204540

  2. 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.

  3. 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.

  4. Transposon insertion site profiling chip (TIP-chip)

    PubMed Central

    Wheelan, Sarah J.; Scheifele, Lisa Z.; Martínez-Murillo, Francisco; Irizarry, Rafael A.; Boeke, Jef D.

    2006-01-01

    Mobile elements are important components of our genomes, with diverse and significant effects on phenotype. Not only can transposons inactivate genes by direct disruption and shuffle the genome through recombination, they can also alter gene expression subtly or powerfully. Currently active transposons are highly polymorphic in host populations, including, among hundreds of others, L1 and Alu elements in humans and Ty1 elements in yeast. For this reason, we wished to develop a simple genome-wide method for identifying all transposons in any given sample. We have designed a transposon insertion site profiling chip (TIP-chip), a microarray intended for use as a high-throughput technique for mapping transposon insertions. By selectively amplifying transposon flanking regions and hybridizing them to the array, we can locate all transposons present in a sample. We have tested the TIP-chip extensively to map Ty1 retrotransposon insertions in yeast and have achieved excellent results in two laboratory strains as well as in evolved Ty1 high-copy strains. We are able to identify all of the theoretically detectable transposons in the FY2 lab strain, with essentially no false positives. In addition, we mapped many new transposon copies in the high-copy Ty1 strain and determined its Ty1 insertion pattern. PMID:17101968

  5. 30. Photocopy of map. Insert, p. 138, Master Plan Report, ...

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

    30. Photocopy of map. Insert, p. 138, Master Plan Report, 1960, Vol. II. Layton, Layton and Rohrbach, General Planning and Resource Consultants. 1960 MASTER PLAN, SHOWING PROPOSED LOCATION OF LINNAEAN HOUSE - Missouri Botanical Garden, 2345 Tower Grove Avenue, Saint Louis, Independent City, MO

  6. Computerized Planning of Prostate Cryosurgery Using Variable Cryoprobe Insertion Depth

    PubMed Central

    Rossi, Michael R.; Tanaka, Daigo; Shimada, Kenji; Rabin, Yoed

    2010-01-01

    The current study presents a computerized planning scheme for prostate cryosurgery using a variable insertion-depth strategy. This study is a part of an ongoing effort to develop computerized tools for cryosurgery. Based on typical clinical practices, previous automated planning schemes have required that all cryoprobes be aligned at a single insertion depth. The current study investigates the benefit of removing this constraint, in comparison with results based on uniform insertion-depth planning as well as the so-called “pullback procedure”. Planning is based on the so-called “bubble-packing method”, and its quality is evaluated with bioheat transfer simulations. This study is based on five 3D prostate models, reconstructed from ultrasound imaging, and cryoprobe active length in the range of 15 mm to 35 mm. The variable insertion depth technique is found to consistently provide superior results when compared to the other placement methods. Furthermore, it is shown that both the optimal active length and the optimal number of cryoprobes vary among prostate models, based on the size and shape of the target region. Due to its low computational cost, the new scheme can be used to determine the optimal cryoprobe layout for a given prostate model in real time. PMID:19111714

  7. Insertion and deletion mutagenesis of the human cytomegalovirus genome

    SciTech Connect

    Spaete, R.R.; Mocarski, E.S.

    1987-10-01

    Studies on human cytomegalovirus (CMV) have been limited by a paucity of molecular genetic techniques available for manipulating the viral genome. The authors have developed methods for site-specific insertion and deletion mutagenesis of CMV utilizing a modified Escherichia coli lacZ gene as a genetic marker. The lacZ gene was placed under the control of the major ..beta.. gene regulatory signals and inserted into the viral genome by homologous recombination, disrupting one of two copies of this ..beta.. gene within the L-component repeats of CMV DNA. They observed high-level expression of ..beta..-galactosidase by the recombinant in a temporally authentic manner, with levels of this enzyme approaching 1% of total protein in infected cells. Thus, CMV is an efficient vector for high-level expression of foreign gene products in human cells. Using back selection of lacZ-deficient virus in the presence of the chromogenic substrate 5-bromo-4-chloro-3-indolyl ..beta..-D-galactoside, they generated random endpoint deletion mutants. Analysis of these mutant revealed that CMV DNA sequences flanking the insert had been removed, thereby establishing this approach as a means of determining whether sequences flanking a lacZ insertion are dispensable for viral growth. In an initial test of the methods, they have shown that 7800 base pairs of one copy of L-component repeat sequences can be deleted without affecting viral growth in human fibroblasts.

  8. 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.

  9. African origin of human-specific polymorphic Alu insertions.

    PubMed Central

    Batzer, M A; Stoneking, M; Alegria-Hartman, M; Bazan, H; Kass, D H; Shaikh, T H; Novick, G E; Ioannou, P A; Scheer, W D; Herrera, R J

    1994-01-01

    Alu elements are a family of interspersed repeats that have mobilized throughout primate genomes by retroposition from a few "master" genes. Among the 500,000 Alu elements in the human genome are members of the human-specific subfamily that are not fixed in the human species; that is, not all chromosomes carry an Alu element at a particular locus. Four such polymorphic human-specific Alu insertions were analyzed by a rapid, PCR-based assay that uses primers that flank the insertion point to determine genotypes based on the presence or absence of the Alu element. These four polymorphic Alu insertions were shown to be absent from the genomes of a number of nonhuman primates, consistent with their arising as human genetic polymorphisms sometime after the human/African ape divergence. Analysis of 664 unrelated individuals from 16 population groups from around the world revealed substantial levels of variation within population groups and significant genetic differentiation among groups. No significant associations were found among the four loci, consistent with their location on different chromosomes. A maximum-likelihood tree of population relationships showed four major groupings consisting of Africa, Europe, Asia/Americas, and Australia/New Guinea, which is concordant with similar trees based on other loci. A particularly useful feature of the polymorphic Alu insertions is that the ancestral state is known to be the absence of the Alu element, and the presence of the Alu element at a particular chromosomal site reflects a single, unique event in human evolution. A hypothetical ancestral group can then be included in the tree analysis, with the frequency of each insertion set to zero. The ancestral group connected to the maximum-likelihood tree within the African branch, which suggests an African origin of these polymorphic Alu insertions. These data are concordant with other diverse data sets, which lends further support to the recent African origin hypothesis for modern humans. Polymorphic Alu insertions represent a source of genetic variation for studying human population structure and evolution. Images PMID:7991620

  10. 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.

  11. A Transposable Element Insertion Confers Xenobiotic Resistance in Drosophila

    PubMed Central

    Mateo, Lidia; Ullastres, Anna; González, Josefa

    2014-01-01

    The increase in availability of whole genome sequences makes it possible to search for evidence of adaptation at an unprecedented scale. Despite recent progress, our understanding of the adaptive process is still very limited due to the difficulties in linking adaptive mutations to their phenotypic effects. In this study, we integrated different levels of biological information to pinpoint the ecologically relevant fitness effects and the underlying molecular and biochemical mechanisms of a putatively adaptive TE insertion in Drosophila melanogaster: the pogo transposon FBti0019627. We showed that other than being incorporated into Kmn1 transcript, FBti0019627 insertion also affects the polyadenylation signal choice of CG11699 gene. Consequently, only the short 3?UTR transcript of CG11699 gene is produced and the expression level of this gene is higher in flies with the insertion. Our results indicated that increased CG11699 expression leads to xenobiotic stress resistance through increased ALDH-III activity: flies with FBti0019627 insertion showed increased survival rate in response to benzaldehyde, a natural xenobiotic, and to carbofuran, a synthetic insecticide. Although differences in survival rate between flies with and without the insertion were not always significant, when they were, they were consistent with FBti0019627 mediating resistance to xenobiotics. Taken together, our results provide a plausible explanation for the increase in frequency of FBti0019627 in natural populations of D. melanogaster and add to the limited number of examples in which a natural genetic mutation has been linked to its ecologically relevant phenotype. Furthermore, the widespread distribution of TEs across the tree of life and conservation of stress response pathways across organisms make our results relevant not only for Drosophila, but for other organisms as well. PMID:25122208

  12. Functional importance of the NH2-terminal insertion sequence of lung surfactant protein B

    PubMed Central

    Frey, Shelli L.; Pocivavsek, Luka; Waring, Alan J.; Walther, Frans J.; Hernandez-Juviel, Jose M.; Ruchala, Piotr

    2010-01-01

    Lung surfactant protein B (SP-B) is required for proper surface activity of pulmonary surfactant. In model lung surfactant lipid systems composed of saturated and unsaturated lipids, the unsaturated lipids are removed from the film at high compression. It is thought that SP-B helps anchor these lipids closely to the monolayer in three-dimensional cylindrical structures termed “nanosilos” seen by atomic force microscopy imaging of deposited monolayers at high surface pressures. Here we explore the role of the SP-B NH2 terminus in the formation and stability of these cylindrical structures, specifically the distribution of lipid stack height, width, and density with four SP-B truncation peptides: SP-B 1–25, SP-B 9–25, SP-B 11–25, and SP-B 1–25Nflex (prolines 2 and 4 substituted with alanine). The first nine amino acids, termed the insertion sequence and the interface seeking tryptophan residue 9, are shown to stabilize the formation of nanosilos while an increase in the insertion sequence flexibility (SP-B 1–25Nflex) may improve peptide functionality. This provides a functional understanding of the insertion sequence beyond anchoring the protein to the two-dimensional membrane lining the lung, as it also stabilizes formation of nanosilos, creating reversible repositories for fluid lipids at high compression. In lavaged, surfactant-deficient rats, instillation of a mixture of SP-B 1–25 (as a monomer or dimer) and synthetic lung lavage lipids quickly improved oxygenation and dynamic compliance, whereas SP-B 11–25 surfactants showed oxygenation and dynamic compliance values similar to that of lipids alone, demonstrating a positive correlation between formation of stable, but reversible, nanosilos and in vivo efficacy. PMID:20023175

  13. Evaluation of the endodontic apical seal after post insertion by synchrotron radiation microtomography

    NASA Astrophysics Data System (ADS)

    Contardo, L.; De Luca, M.; Biasotto, M.; Longo, R.; Olivo, A.; Pani, S.; Di Lenarda, R.

    2005-08-01

    The commonly used methods for evaluating the endodontic apical seal, such as longitudinal and transversal section and diaphanization, show some operative difficulties and intrinsic limitation. This study suggests and describes a new method of analysis using a synchrotron radiation microtomography to analyse the root apex after post insertion, creating a three-dimensional image and analysing sections of the specimen every 5 ?m. The study was performed at SYRMEP beam line at the Electra Synchrotron in Trieste using monochromatic X-rays of 32 KeV. Eleven monoradicular teeth were prepared using NiTi GT Rotary files instruments to an apical size 20 with conicity .06 and divided in four groups: in G1 ( n=4) and G2 ( n=2), the specimens were endodontically filled with guttapercha and a zinc-oxide sealer, in G3 ( n=3) and G4 ( n=2) guttapercha and a silicon-based sealer were used. An endodontic post was inserted in specimens of groups 1 and 3 following the manufacturer's instructions. Specimens were analysed using monochromatic X-rays of 32 KeV. A CCD detector with pixel dimension pf 5×5 ?m 2 was used for the acquisition process. Seven hundred and twenty projections were performed over 180° range using a high-resolution rotator. The projections were reconstructed using standard algorithms for tomographic reconstruction. The apical infiltration was evaluated by verifying if black spots were detectable on the images. The specimens of groups 3 and 4 showed a better apical seal than the ones of groups 1 and 2. Post insertion, when a ZOE-based sealer is used, increases the apical gap even if it does not seem to be clinically relevant and sufficient to be a cause of endodontic failure. The new method for analysis appeared to be effective to detect small defects in endodontic obturation, analysing guttapercha-sealer and sealer-dentin interfaces.

  14. X-ray absorption spectroscopy of lithium insertion and de-insertion in copper birnessite nanoparticle electrodes.

    PubMed

    Pelliccione, Christopher J; Li, Yue Ru; Marschilok, Amy C; Takeuchi, Kenneth J; Takeuchi, Esther S

    2016-01-20

    The combination of ex situ X-ray diffraction (XRD) and X-ray absorption spectroscopy (XAS) measurements on 2D layered copper birnessite cathode materials for lithium ion battery applications provides detailed insight into both bulk-crystalline and localized atomic structural changes resulting from electrochemically driven lithium insertion and de-insertion. Copper birnessite electrodes that had been galvanostatically discharged and charged were measured with XRD to determine the accompanying long-range crystalline structure changes, while Mn and Cu K-edge XAS measurements provided a detailed view of the Mn and Cu oxidation state changes along with variations of the local neighboring atom environments around the Mn and Cu centers. While not detectable with XRD spectra, through XAS measurements it was determined that the copper ions (Cu(2+)) are reduced to form amorphous nano-sized Cu metal, and can be oxidized back to Cu(2+). The reversible nature of the interconversion provides a rationale to the enhanced discharge capacity of copper birnessite relative to the analogous copper-free birnessite materials. The manganese oxide octahedra comprising the 2D layers in the original copper birnessite crystal structure disperse during lithium insertion, and revert back close to their original orientation after lithium de-insertion. During electrochemical oxidation or reduction the layered birnessite structure does not collapse, even though significant local disordering around Mn and Cu centers is directly observed. PMID:26735498

  15. Analysis of Primary Stability of Dental Implants Inserted in Different Substrates Using the Pullout Test and Insertion Torque

    PubMed Central

    Oliscovicz, Nathalia Ferraz; Shimano, Antônio Carlos; Marcantonio Junior, Élcio; Lepri, César Penazzo; dos Reis, Andréa Candido

    2013-01-01

    The aim of the study was to evaluate mechanical behavior of implants inserted in three substrates, by measuring the pullout strength and the relative stiffness. 32 implants (Master Porous-Conexao, cylindrical, external hexagon, and surface treatment) were divided into 4 groups (n = 8): pig rib bone, polyurethane Synbone, polyurethane Nacional 40 PCF, and pinus wood. Implants were installed with the exact distance of 5?mm of another implant. The insertion torque (N·cm) was quantified using the digital Kratos torque meter and the pullout test (N) was performed by an axial traction force toward the long axis of the implant (2?min/mm) through mount implant devices attached to a piece adapted to a load cell of 200?Kg of a universal testing machine (Emic DL10000). Data of insertion torque and maximum pullout force were submitted to one-way ANOVA and Bonferroni tests (? = 0.05). Polyurethane Nacional 40 PCF and pinus wood showed the highest values of insertion torque and pullout force, with significant statistical difference (P < 0.05) with other groups. The analysis showed stiffness materials with the highest values for primary stability. PMID:23431301

  16. LINE retrotransposition and host DNA repair machinery

    PubMed Central

    Yamaguchi, Katsumi; Kajikawa, Masaki; Okada, Norihiro

    2015-01-01

    Long interspersed elements (LINEs), or non-long-terminal repeat (LTR) retrotransposons, are mobile genetic elements that exist in the genomic DNA of most eukaryotes, comprising a considerable portion of the host chromosomes. LINEs constitute endogenous mutagens that cause insertional mutations in host chromosomes and have a large impact on host genome evolution. Despite their importance, however, the molecular mechanism of LINE retrotransposition is not fully understood. Several studies suggest that host proteins that participate in the repair of DNA breaks modulate LINE retrotransposition. Recently, we provided evidence that there are 2 distinct pathways—annealing and direct—that join the 5?-end of LINEs to host chromosomal DNA. These pathways appear to be used distinctively by zebrafish LINEs and the human L1 in DT40 cells. In HeLa cells, only the annealing pathway appears to be used. This implies that different characteristics of the 2 LINEs and also host factors dictate which pathway is selected. Here, we discuss the 5?-end-joining pathways of LINE retrotransposition and propose that the pathways of LINE integration adopt certain host repair factors. PMID:26942045

  17. 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.

  18. Analysis of cell poration by femtosecond laser for particle insertion by optical manipulation

    NASA Astrophysics Data System (ADS)

    Muhammad, Waleed; Kim, Jung-Dae; Lee, Yong-Gu

    2012-10-01

    The introduction and subsequent expression of external DNA inside single living mammalian cell (transfection) can be achieved by photoporation with femtosecond laser. After photoporation, external DNA can be introduced by trapping and successive insertion of DNA coated nanoparticle in the cell using optical tweezers. To maximize the transfection efficiency, one of the major aspects is that the photoporated cell should not be damaged and cell membrane should heal itself immediately or after sometime while the cells are healed in the CO2 incubator. Furthermore, the size of hole created as a result of photoporation should be more than the size of DNA coated nanoparticle to be inserted inside the cell. In this paper, an analysis has been done on single cell of important breast cancer cell lines named MCF-7 and MDAMB231. Size of holes created in cell membrane after photoporation has been measured and the required optimum energy with sustained cell life were determined. Using this analysis, most favorable conditions for maximum transfection efficiency can be determined.

  19. New chicane magnet design for insertion device straights at the Advanced Light Source

    SciTech Connect

    Marks, Steve; Schlueter, Ross; Anderson, David; Gath, William; Jung, Jin-Young; Robin, David; Steier, Christoph; Stevens, Troy

    2001-12-10

    A chicane magnet incorporating counter-rotating permanent magnet pairs together with trim coils has been designed for use in the Advanced Light Source (ALS) straights in conjunction with two insertion devices. In particular, this design is being developed for use in the existing beam line (BL) 4 elliptically polarizing undulator (EPU) straight and in the BL11 EPU straight, currently under design and construction. The purpose of the chicane is to provide a fixed angular separation between two successive EPU photon fans, and to correct steering perturbations resulting from EPU polarization state changes. Polarization changes occur on the time scale of one second; associated steering corrections must be accomplished in less than a second. Hysteresis associated with conventional iron core electromagnets prevents fast steering correction to the required precision. This consideration motivated the iron-free design presented here.

  20. Influence of inserting a thin fullerene layer on pentacene organic thin-film transistor

    NASA Astrophysics Data System (ADS)

    Li, Yu-Chang; Lin, Yu-Ju; Wei, Chia-Yu; Chou, Dei-Wei; Tsao, Chun-Ho; Wang, Yeong-Her

    2012-03-01

    The performance of organic thin-film transistors (TFTs) with a pentacene/fullerene(C60)/pentacene (PCP) sandwich structure is presented. Using a 3.5 nm-thick C60 layer inserted between the pentacene films, the obtained hole mobility is improved by more than six times. By applying atomic force microscopy, x-ray diffraction, Raman spectrum, and transmission line method analysis, one can reasonably infer that the smoother surface of the pentacene film covered with thin C60 layer delays the phase transformation of the upper pentacene film, resulting in stronger intermolecular coupling and the reduction of channel resistance of the PCP TFTs from 3.03 to 1.72 MΩ, and, therefore, improving the device performance.

  1. 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

  2. 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. PMID:26706072

  3. Cytogenetic and Sequence Analyses of Mitochondrial DNA Insertions in Nuclear Chromosomes of Maize.

    PubMed

    Lough, Ashley N; Faries, Kaitlyn M; Koo, Dal-Hoe; Hussain, Abid; Roark, Leah M; Langewisch, Tiffany L; Backes, Teresa; Kremling, Karl A G; Jiang, Jiming; Birchler, James A; Newton, Kathleen J

    2015-11-01

    The transfer of mitochondrial DNA (mtDNA) into nuclear genomes is a regularly occurring process that has been observed in many species. Few studies, however, have focused on the variation of nuclear-mtDNA sequences (NUMTs) within a species. This study examined mtDNA insertions within chromosomes of a diverse set of Zea mays ssp. mays (maize) inbred lines by the use of fluorescence in situ hybridization. A relatively large NUMT on the long arm of chromosome 9 (9L) was identified at approximately the same position in four inbred lines (B73, M825, HP301, and Oh7B). Further examination of the similarly positioned 9L NUMT in two lines, B73 and M825, indicated that the large size of these sites is due to the presence of a majority of the mitochondrial genome; however, only portions of this NUMT (~252 kb total) were found in the publically available B73 nuclear sequence for chromosome 9. Fiber-fluorescence in situ hybridization analysis estimated the size of the B73 9L NUMT to be ~1.8 Mb and revealed that the NUMT is methylated. Two regions of mtDNA (2.4 kb and 3.3 kb) within the 9L NUMT are not present in the B73 mitochondrial NB genome; however, these 2.4-kb and 3.3-kb segments are present in other Zea mitochondrial genomes, including that of Zea mays ssp. parviglumis, a progenitor of domesticated maize. PMID:26333837

  4. 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

  5. Feasibility Study of Free-Hand Technique for Pedicle Screw Insertion at C7 without Fluoroscopy-Guidance

    PubMed Central

    Lee, Gun Woo; Kim, Ho-Joong; Yeom, Jin S.; Uh, Jae-Hyung; Park, Jong-Ho; Lee, Ji-Hoon; Kim, Dong-Wook

    2016-01-01

    Study Design Retrospective interventional study. Purpose To introduce a free-hand pedicle screw (PS) insertion technique without fluoroscopic guidance in the C7 vertebra and evaluate the procedure's feasibility and radiologic outcomes. Overview of Literature Although PS insertion at C7 has been recognized as a critical procedure in posterior cervical fusion surgery, conventional techniques for C7 PS have several limitations. Methods Thirty two patients (64 screws) who underwent PS insertion in C7 with the novel technique were included in this study. Postoperative clinical and radiological outcomes were evaluated. Special attention was paid to the presence of any problems in the screw position including cortical breaches of the PS and encroachment of the PS into the spinal canal or the vertebral foramen. This novel technique for PS insertion in C7 without fluoroscopy guidance had three key elements. First, the ideal PS entry point was chosen near the C6–7 facet joint using preoperative images. Second, the convergent angle distance was measured at axial computed tomography (CT) imaging, which defined the distance between the tip of C7 spinous process and the extended line passing through the pedicle axis from the ideal entry point. Third, the cranial-caudal angle distance was measured in sagittal CT images, which defined the distance between the tip of the C7 spinous process and the extended line passing through the pedicle axis. Results Cortical breach on postoperative CT images was observed in three screws. All violated only the lateral wall of the affected pedicle. The breached screws occurred in the initial five cases. Postoperative neurologic deterioration was not observed in any patient, regardless of cortical breaching. Conclusions The novel technique successfully allows for C7 PS to be placed and is associated with a low rate of cortical breach. PMID:26949456

  6. Site-specific transposition of insertion sequence IS630.

    PubMed Central

    Tenzen, T; Matsutani, S; Ohtsubo, E

    1990-01-01

    IS630 is a 1.15-kilobase sequence in Shigella sonnei that, unlike many mobile elements, seems not to mediate cointegration between different replicons. To assess its transposition, we constructed composite elements containing inverted copies of IS630 flanking a drug resistance gene. We found that these composite elements transposed to plasmid ColE1 in Escherichia coli. DNA sequencing showed that transposition was, in all cases, to the dinucleotide sequence 5'-TA-3'. There were two preferred insertion sites which corresponded to the TA sequences in the inverted repeats of a 13-base-pair stem region of the [rho]-dependent transcription terminator. IS630 is flanked by TA, and nucleotide substitution by in vitro mutagenesis at these ends did not affect transposition activity of a composite element or its ability to insert preferentially into TA within the 13-base-pair inverted repeat sequences or to duplicate the target sequence. PMID:2163390

  7. 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.

  8. Cylinder liner insert and method of making engine block therewith

    SciTech Connect

    Field, N.L.; Winter, B.P.

    1990-02-27

    This patent describes a cylinder liner insert to be cast-in-place in a light alloy cylinder block for an internal combustion engine. It comprises: at least two generally cylindrical cylinder liners each of which includes an upper end and a lower end, an interior surface for defining a combustion chamber of the engine and an exterior surface adapted to successfully interface with a light alloy during a casting process; and bridge means for joining the at least two cylinder liners along aligned portions of the exterior surfaces of the cylinder liners. Also described is a method of making a light alloy cylinder block for an internal combustion engine including a cylinder liner insert which is cast-in-place.

  9. Micro-optical insertion system for WDM transceiver

    NASA Astrophysics Data System (ADS)

    Bakala, Bretislav; Jerabek, Vitezslav; Mares, David; Prajzler, Vaclav

    2015-01-01

    The paper deals with the design of an micro-optical part of a WDM transceiver based on the circle volume holographic grating triplexer (VHGT) topology. The work is focused on detail description of the bidirectional micro-optical insertion system, especially on the coupling in the transmission direction of the WDM transceiver optical part. This micro-optical wavelength distribution system is imagine transmission system consisted of collimating cylindrical lenses set in the circle topology with the VHGT filter placed in the middle of the circle. The topology is described by ray matrices equations, calculated using analytic and numeric methods and optically measured. Considering simulation and measurement results of micro-optical insertion system the influence of these characteristics on ray path attenuation, diffraction efficiency and transceiver performance is estimated. The paper also discusses limits and conditions of the laser asymmetrical beam focus into the input/output optical fiber.

  10. Objective evaluation of insert material for diabetic and athletic footwear.

    PubMed

    Brodsky, J W; Kourosh, S; Stills, M; Mooney, V

    1988-12-01

    Five of the most commonly used materials for shoe inserts (soft Plastazote, medium Pelite, PPT, Spenco, and Sorbothane) were objectively evaluated in the laboratory to characterize their behavior in the following three specific functions that correspond to clinical use: (1) the effect on the materials of repeated compression. (2) the effect of a combination of repetitive shear and compression. (3) the force-distribution (force-attenuation) properties of these materials, both when new and after repeated compression. The last function represents a model for relief of pressure beneath plantar bony prominences, a topic of special concern for the insensitive foot. All materials were effective in reducing transmitted force over the simulated bony prominence with a rank order of effectiveness. Other factors considered were: amount and rate of permanent deformation offset by considerations of enhanced moldability when comparing the neoprene and urethane materials with the polyethylene foams. The ideal insert represents a combination of material to achieve both durability and moldability. PMID:3229697

  11. Insertional hypermutation in mineral oil-induced plasmacytomas

    PubMed Central

    Knittel, Gero; Metzner, Mirjam; Beck-Engeser, Gabriele; Kan, Ada; Ahrends, Tomasz; Eilat, Dan; Huppi, Konrad; Wabl, Matthias

    2014-01-01

    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 10 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. PMID:24975032

  12. 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.

  13. Biological insertion of computationally designed short transmembrane segments

    PubMed Central

    Baeza-Delgado, Carlos; von Heijne, Gunnar; Marti-Renom, Marc A.; Mingarro, Ismael

    2016-01-01

    The great majority of helical membrane proteins are inserted co-translationally into the ER membrane through a continuous ribosome-translocon channel. The efficiency of membrane insertion depends on transmembrane (TM) helix amino acid composition, the helix length and the position of the amino acids within the helix. In this work, we conducted a computational analysis of the composition and location of amino acids in transmembrane helices found in membrane proteins of known structure to obtain an extensive set of designed polypeptide segments with naturally occurring amino acid distributions. Then, using an in vitro translation system in the presence of biological membranes, we experimentally validated our predictions by analyzing its membrane integration capacity. Coupled with known strategies to control membrane protein topology, these findings may pave the way to de novo membrane protein design. PMID:26987712

  14. CUDA accelerated simulation of needle insertions in deformable tissue

    NASA Astrophysics Data System (ADS)

    Patriciu, Alexandru

    2012-10-01

    This paper presents a stiff needle-deformable tissue interaction model. The model uses a mesh-less discretization of continuum; avoiding thus the expensive remeshing required by the finite element models. The proposed model can accommodate both linear and nonlinear material characteristics. The needle-deformable tissue interaction is modeled through fundamental boundaries. The forces applied by the needle on the tissue are divided in tangent forces and constraint forces. The constraint forces are adaptively computed such that the material is properly constrained by the needle. The implementation is accelerated using NVidia CUDA. We present detailed analysis of the execution timing in both serial and parallel case. The proposed needle insertion model was integrated in a custom software that loads DICOM images, generate the deformable model, and can simulate different insertion strategies.

  15. 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

  16. Isolation and characterization of functional insertion sequences of rhizobia.

    PubMed

    Hernandez-Lucas, Ismael; Ramirez-Trujillo, Jose Augusto; Gaitan, Miguel Angel; Guo, Xianwu; Flores, Margarita; Martinez-Romero, Esperanza; Perez-Rueda, Ernesto; Mavingui, Patrick

    2006-08-01

    Rhizobia are a group of bacteria that form nodules on the roots of legume host plants. The sequenced genomes of the rhizobia are characterized by the presence of many putative insertion sequences (IS) elements. However, it is unknown whether these IS elements are functional and it is therefore relevant to assess their transposition activity. In this work, several functional insertion sequences belonging to the IS1256, IS3, IS5, IS166, and IS21 families were captured from Rhizobium tropici, Rhizobium sp. NGR234 and Sinorhizobium meliloti, using pGBG1 as a trapping system. In silico analysis shows that homologs of rhizobia mobile elements are present in distantly related genomes, suggesting that Rhizobium IS elements are prone to genetic transfer. PMID:16842354

  17. 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.

  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. Metallocene Catalytic Insertion Polymerization of 1-Silene to Polycarbosilanes.

    PubMed

    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

  20. 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

  1. Large-scale, saturating insertional mutagenesis of the mouse genome

    PubMed Central

    Gragerov, Alexander; Horie, Kyoji; Pavlova, Maria; Madisen, Linda; Zeng, Hongkui; Gragerova, Galina; Rhode, Alex; Dolka, Io; Roth, Patricia; Ebbert, Amanda; Moe, Stephanie; Navas, Christopher; Finn, Eric; Bergmann, John; Vassilatis, Demetrios K.; Pavlakis, George N.; Gaitanaris, George A.

    2007-01-01

    We describe the construction of a large-scale, orderly assembly of mutant ES cells, generated with retroviral insertions and having mutational coverage in >90% of mouse genes. We also describe a method for isolating ES cell clones with mutations in specific genes of interest from this library. This approach, which combines saturating random mutagenesis with targeted selection of mutations in the genes of interest, was successfully applied to the gene families of G protein-coupled receptors (GPCRs) and nuclear receptors. Mutant mouse strains in 60 different GPCRs were generated. Applicability of the technique for the GPCR genes, which on average represent fairly small targets for insertional mutagenesis, indicates the general utility of our approach for the rest of the genome. The method also allows for increased scale and automation for the large-scale production of mutant mice, which could substantially expedite the functional characterization of the mouse genome. PMID:17720809

  2. 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

  3. 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.

  4. Bone metabolism induced by denture insertion in positron emission tomography.

    PubMed

    Suenaga, H; Chen, J; Yamaguchi, K; Sugazaki, M; Li, W; Swain, M; Li, Q; Sasaki, K

    2016-03-01

    18F-fluoride positron emission tomogra-phy (PET) can identify subtle functional variation prior to the major structural change detectable by X-ray. This study aims to investigate the mechanobiological bone reaction around the abutment tooth and in the residual ridge, induced by insertion of removable partial denture (RPD) within two different groups of patients: patients without denture experience (Group 1) and patients with denture experience before (Group 2), using 18F-fluoride PET imaging technique. 18F-fluoride PET/computerised tomography (CT) scan was performed to examine the bone metabolic change in mandible before and after the RPD treatment. Region of interests (ROIs) were placed in alveolar bone around abutment tooth and in residual bone beneath the RPD. Standardised uptake value (SUV), reflecting the accumulation of 18F-fluoride, was measured for each ROI. In all subjects of Group 1, SUVs after insertion were higher than before in both alveolar bone and residual bone, while there was less significant change in SUV in subjects of Group 2. This study demonstrated using longitudinal 18F-fluoride PET scans to effectively examine the bone metabolic change in mandible induced by occlusal loading after RPD insertion. Using this technique, within the six subjects in this study, it was shown that bone metabolism around abutment tooth and residual ridge increased after RPD insertion in case of first-time denture user, while there was no big change in the patient with experience of denture before. This study revealed the effectiveness of applying PET to evaluate bone metabolic activity as mechanobiolo-gical reaction. PMID:26431672

  5. Management of subcalcaneal pain and Achilles tendonitis with heel inserts

    PubMed Central

    Maclellan, G. E.; Vyvyan, Barbara

    1981-01-01

    Soft tissue symptoms in the leg due to sporting activity are commonly associated with the force of heel strike. Conventional training shoes compromise between comfort and performance; few models are suitably designed for both considerations. Using a visco-elastic polymer insert the symptoms of heel pain and Achilles tendonitis have been largely or completely abolished in a preliminary study. Imagesp117-ap117-bp117-cp118-a PMID:7272653

  6. 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.

  7. 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.

  8. Particles as Wilson lines of the gravitational field

    NASA Astrophysics Data System (ADS)

    Freidel, L.; Kowalski-Glikman, J.; Starodubtsev, A.

    2006-10-01

    Since the work of Mac-Dowell-Mansouri it is well known that gravity can be written as a gauge theory for the de Sitter group. In this paper we consider the coupling of this theory to the simplest gauge invariant observables that is, Wilson lines. The dynamics of these Wilson lines is shown to reproduce exactly the dynamics of relativistic particles coupled to gravity, the gauge charges carried by Wilson lines being the mass and spin of the particles. Insertion of Wilson lines breaks in a controlled manner the diffeomorphism symmetry of the theory and the gauge degree of freedom are transmuted to particles degree of freedom.

  9. 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

  10. 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.

  11. 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.

  12. Schistosome dystrophin and dystrobrevin proteins contain large insertions.

    PubMed

    Tan, Sipin; Johnston, David A; Roberts, Roland G

    2007-10-26

    Dystrophin and dystrobrevin are distantly related proteins that form a heterodimeric membrane-bound complex in all metazoa so far studied. The signature feature of the dystrophin/dystrobrevin/dystrotelin superfamily is a highly conserved cluster of domains whose integrity and size are maintained in all known members- more than 60 sequences so far reported. We were therefore surprised to find that the analogous regions of the Schistosoma mansoni proteins bear multiple large insertions amounting to 100% (dystrophin) and 35% (dystrobrevin) of their expected size. We isolated orthologous sequences from Schistosoma haematobium and Schistosoma bovis, and found that the insertions have a 10-fold higher rate of change (Ka/Ks value) than their respective host sequences, suggesting a much lower degree of functional constraint. A survey of known S. mansoni protein sequences shows that only two other proteins (both acetylcholine receptors) share these properties. We discuss the implications of the positions and sizes of these insertions for the structure and function of the dystrophins and dystrobrevins. PMID:17764661

  13. Apparatus to test insertion and removal torque of bone screws.

    PubMed

    Koistinen, A; Santavirta, S; Lappalainen, R

    2003-01-01

    This paper describes affordable equipment for testing bone screw torque, corresponding to ASTM standard F543-00 for testing metallic medical bone screws. Correct testing of thin and long bone screws is essential due to screw failures during insertion and removal of the screws. Furthermore, insertion torque is an important factor in predicting fixation strength, screw pull-out force and effects of surface treatment of screws. The capability of the custom-built tester was determined using polytetrafluoroethylene and wood disc samples and bone screws. Bovine cortical bones allowed testing to the failure limit, i.e. the torque increased in long screws to the fracture limit. For 2.7 and 3.5 mm thick self-tapping cortical bone screws, the failure torques were 30-50 per cent higher than the minimum values required by the standard (1.0 and 2.3 N m respectively). The equipment provided reproducible results and fulfilled the ASTM standard very well. Preliminary testing with amorphous diamond coated bone screws showed good durability of the coating and on average 10-15 per cent lower torque values compared with uncoated screws. The equipment can be used to measure insertion and removal torques as described in the standard. Furthermore, it also allows testing of normal screws and bolts. PMID:14702987

  14. Localized control of the orbit in the RHIC insertions

    SciTech Connect

    Ohnuma, S.

    1992-08-01

    It is proposed here that, for RHIC92 insertions, we remove the corrector from Ql and the beam position monitor (BPM) from Q2 in order to alleviate difficulties associated with the physical layout of the quadrupole triplet (Ql-Q2-Q3). Furthermore, it is suggested that there should be both (horizontal and vertical) types of BPMs at each end of the free space between Q3 and Q4 and between Q7 and Q8 so that one can measure the direction of the closed orbit. With this model, a localized control of the beam position and angle at the interaction point (IP) with either four or six correctors has been investigated. Similarly, a control of the orbit within an insertion for minimizing the orbit displacements at seven (or eight) BPM locations with nine (or ten) correctors in each transverse direction has been studied. Examples are given for the beta at IP = 2m, 10m, 20m, and 200m. It is shown that the design value of the integrated field strength of 0.3 T-m for each corrector should be sufficient for the tasks considered here except for some cases with extreme parameter values. At the same time, it is emphasized that the overall correction of the closed orbit for the entire ring (arcs and insertions) should be re-examined for RHIC92 lattice with the proposed arrangement of correctors and BPMS.

  15. 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

  16. Fluorinated Aromatic Amino Acids Distinguish Cation-? Interactions from Membrane Insertion.

    PubMed

    He, Tao; Gershenson, Anne; Eyles, Stephen J; Lee, Yan-Jiun; Liu, Wenshe R; Wang, Jiangyun; Gao, Jianmin; Roberts, Mary F

    2015-07-31

    Cation-? interactions, where protein aromatic residues supply ? systems while a positive-charged portion of phospholipid head groups are the cations, have been suggested as important binding modes for peripheral membrane proteins. However, aromatic amino acids can also insert into membranes and hydrophobically interact with lipid tails. Heretofore there has been no facile way to differentiate these two types of interactions. We show that specific incorporation of fluorinated amino acids into proteins can experimentally distinguish cation-? interactions from membrane insertion of the aromatic side chains. Fluorinated aromatic amino acids destabilize the cation-? interactions by altering electrostatics of the aromatic ring, whereas their increased hydrophobicity enhances membrane insertion. Incorporation of pentafluorophenylalanine or difluorotyrosine into a Staphylococcus aureus phosphatidylinositol-specific phospholipase C variant engineered to contain a specific PC-binding site demonstrates the effectiveness of this methodology. Applying this methodology to the plethora of tyrosine residues in Bacillus thuringiensis phosphatidylinositol-specific phospholipase C definitively identifies those involved in cation-? interactions with phosphatidylcholine. This powerful method can easily be used to determine the roles of aromatic residues in other peripheral membrane proteins and in integral membrane proteins. PMID:26092728

  17. 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. PMID:18501945

  18. Insertional mutagenesis and clonal dominance: biological and statistical considerations.

    PubMed

    Fehse, B; Roeder, I

    2008-01-01

    Improvements of (retroviral) gene transfer vectors, stem cell isolation and culture techniques as well as transduction protocols eventually resulted not only in the successful genetic modification of cells capable of reconstituting the haematopoietic system in various animal models, but also human beings. This was a conditio sine qua non for the successful application of gene therapy for inherited diseases as meanwhile achieved for severe combined immune deficiencies (SCID-X1, ADA-SCID) and chronic granulomatous disease (CGD). Unexpectedly, in long-term animal experiments as well as in the follow up of patients from the CGD trial, haematopoietic clones bearing insertions in certain gene loci became dominant, which was most apparent in the myeloid blood compartment. Accumulating data strongly suggest that this clonal dominance was due to some growth and/or survival advantage conferred by gene-activating or -suppressing effects of the integrated retroviral vector (insertional mutagenesis). Importantly, such induced clonal dominance seems not to lead to malignant transformation of affected cell clones inadvertently. The latter finding has become the basis for the concept of 'induced haematopoietic stem cells', a potentially powerful tool to investigate genes involved in the regulation of mechanisms underlying competitive advantages of stem cells, but also in the multi-step nature of malignant transformation. Here we discuss promises and open issues of this concept as well as the important question of common insertion sites statistics and its pitfalls. PMID:17972922

  19. Enzymatic functionalization of a nanobody using protein insertion technology.

    PubMed

    Crasson, O; Rhazi, N; Jacquin, O; Freichels, A; Jérôme, C; Ruth, N; Galleni, M; Filée, P; Vandevenne, M

    2015-10-01

    Antibody-based products constitute one of the most attractive biological molecules for diagnostic, medical imagery and therapeutic purposes with very few side effects. Their development has become a major priority of biotech and pharmaceutical industries. Recently, a growing number of modified antibody-based products have emerged including fragments, multi-specific and conjugate antibodies. In this study, using protein engineering, we have functionalized the anti-hen egg-white lysozyme (HEWL) camelid VHH antibody fragment (cAb-Lys3), by insertion into a solvent-exposed loop of the Bacillus licheniformis β-lactamase BlaP. We showed that the generated hybrid protein conserved its enzymatic activity while the displayed nanobody retains its ability to inhibit HEWL with a nanomolar affinity range. Then, we successfully implemented the functionalized cAb-Lys3 in enzyme-linked immunosorbent assay, potentiometric biosensor and drug screening assays. The hybrid protein was also expressed on the surface of phage particles and, in this context, was able to interact specifically with HEWL while the β-lactamase activity was used to monitor phage interactions. Finally, using thrombin-cleavage sites surrounding the permissive insertion site in the β-lactamase, we reported an expression system in which the nanobody can be easily separated from its carrier protein. Altogether, our study shows that insertion into the BlaP β-lactamase constitutes a suitable technology to functionalize nanobodies and allows the creation of versatile tools that can be used in innovative biotechnological assays. PMID:25852149

  20. 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

  1. 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

  2. A medical simulator for subcutaneous contraceptive implant insertion.

    PubMed

    Jardin, A; Pham, M T; Mallet, A; Redarce, T; Dupuis, O

    2008-11-01

    New contraceptive methods like the subcutaneous implant offers a new kind of comfort for women with an efficiency similar to the contraceptive pill. Unfortunately the few numbers of unintended pregnancies that have been reported are generally due to a bad insertion of the implant. In order to give more security to patients, we have designed, in close collaboration with physicians, a new kind of medical simulator. This paper focuses on a device dedicated to a specific subcutaneous implant but it is worth noting that this simulator is relatively generic since it will be used for other subcutaneous techniques or other implant instruments. This simulator can be used for two purposes: one for training novice physicians in the correct manipulation and the other for physician certification which will help determine if they are capable of inserting the implant in vivo. This paper describes the approach which has led to the design of this simulator. It describes its functionalities, its several components but also methods used to analyze the manipulation of the implant insertion inside the patient. Finally first experimental results are reported and discussed. The system used in this paper makes possible to carry out training in a constraint-free context and provides the first mean of visualizing a maneuver that, until now, has been performed blindly. PMID:18417413

  3. 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

  4. Behavior of diphtheria toxin T domain containing substitutions that block normal membrane insertion at Pro345 and Leu307: control of deep membrane insertion and coupling between deep insertion of hydrophobic subdomains.

    PubMed

    Zhao, Gang; London, Erwin

    2005-03-22

    Diphtheria toxin T domain aids the translocation of toxin A chain across membranes. T domain has two hydrophobic layers/subdomains that can insert deeply into membranes: helices TH8 and 9, which form a transmembrane hairpin, and helices TH5-7, which form a nonclassical, nontransmembrane structure. Substitutions were made at Pro345, a residue located near the turn between TH8 and 9. P345 is critical for toxicity and pore formation by the T domain. Fluorescence methods showed that hairpin-disrupting Gly or Glu substitutions at 345 did not insert into lipid bilayers as deeply as the wild-type protein, and consistent with previous studies, these mutations reduced pore formation activity as assayed by a novel biotin-streptavidin-based influx assay. Introducing Pro at positions 347 or 353 not only failed to compensate for substitutions at P345, but also they further disrupted deep insertion and/or pore formation. Substitution of P345 with Asn, a residue that promotes helical hairpin formation almost as well as Pro, resulted in somewhat more normal insertion and pore formation than other substitutions. Importantly, a P345E substitution disrupted deep insertion of TH5-7. This suggests that TH8 and 9 and TH5-7 undergo some sort of coordinated insertion into the lipid bilayer and/or that the membrane-inserted T domain has a distinct tertiary structure in which TH5-7 interact with TH8 and 9 instead of consisting of noninteracting hydrophobic segments. Intriguingly, a L307R substitution in TH6, which disrupted deep insertion of TH7, had only a weak effect on pore formation and deep insertion of TH8 and 9. This suggests that the TH8 and 9 region can insert independently of TH5-7 to some degree and that TH8 and 9 insertion may occur early in T-domain insertion. PMID:15766279

  5. Tools for Targeted Genome Engineering of Established Drosophila Cell Lines.

    PubMed

    Cherbas, Lucy; Hackney, Jennifer; Gong, Lei; Salzer, Claire; Mauser, Eric; Zhang, Dayu; Cherbas, Peter

    2015-12-01

    We describe an adaptation of ?C31 integrase-mediated targeted cassette exchange for use in Drosophila cell lines. Single copies of an attP-bounded docking platform carrying a GFP-expression marker, with or without insulator elements flanking the attP sites, were inserted by P-element transformation into the Kc167 and Sg4 cell lines; each of the resulting docking-site lines carries a single mapped copy of one of the docking platforms. Vectors for targeted substitution contain a cloning cassette flanked by attB sites. Targeted substitution occurs by integrase-mediated substitution between the attP sites (integrated) and the attB sites (vector). We describe procedures for isolating cells carrying the substitutions and for eliminating the products of secondary off-target events. We demonstrate the technology by integrating a cassette containing a Cu(2+)-inducible mCherry marker, and we report the expression properties of those lines. When compared with clonal lines made by traditional transformation methods, which lead to the illegitimate insertion of tandem arrays, targeted insertion lines give more uniform expression, lower basal expression, and higher induction ratios. Targeted substitution, though intricate, affords results that should greatly improve comparative expression assays-a major emphasis of cell-based studies. PMID:26450921

  6. Tools for Targeted Genome Engineering of Established Drosophila Cell Lines

    PubMed Central

    Cherbas, Lucy; Hackney, Jennifer; Gong, Lei; Salzer, Claire; Mauser, Eric; Zhang, Dayu; Cherbas, Peter

    2015-01-01

    We describe an adaptation of φC31 integrase–mediated targeted cassette exchange for use in Drosophila cell lines. Single copies of an attP-bounded docking platform carrying a GFP-expression marker, with or without insulator elements flanking the attP sites, were inserted by P-element transformation into the Kc167 and Sg4 cell lines; each of the resulting docking-site lines carries a single mapped copy of one of the docking platforms. Vectors for targeted substitution contain a cloning cassette flanked by attB sites. Targeted substitution occurs by integrase-mediated substitution between the attP sites (integrated) and the attB sites (vector). We describe procedures for isolating cells carrying the substitutions and for eliminating the products of secondary off-target events. We demonstrate the technology by integrating a cassette containing a Cu2+-inducible mCherry marker, and we report the expression properties of those lines. When compared with clonal lines made by traditional transformation methods, which lead to the illegitimate insertion of tandem arrays, targeted insertion lines give more uniform expression, lower basal expression, and higher induction ratios. Targeted substitution, though intricate, affords results that should greatly improve comparative expression assays—a major emphasis of cell-based studies. PMID:26450921

  7. Investigation Of Plasma Discharges Within Maja-PF Device Operated With Tungsten Inserts In The Central Electrode

    SciTech Connect

    Malinowski, K.; Skladnik-Sadowska, E.; Jakubowski, L.; Sadowski, M. J.; Ladygina, M.

    2008-03-19

    The paper presents results of recent experimental studies with a MAJA-PF plasma-focus facility, as carried out for two different configurations of the end-plate of the central electrode. The main aim was to investigate an influence of the shape and material of the electrode end-plate on the formation and structure of the plasma column as well as to record and identify spectral lines of tungsten. The first version of the end-plate was made of Cu, while the second one had an insert made of a Cu-W alloy. Optical spectra were recorded by means of a Mechelle 900 spectrometer, operated with some delay in relation to a current peculiarity and with exposition times varied from 200 ns to 10 {mu}s. For shots with the Cu electrode spectral lines of deuterium (working gas) and carbon ions were recorded, but tungsten lines were not identified. It was estimated that the electron concentration changed from (8 divide 10)x10{sup 17} cm{sup -3} to about 2x10{sup 17} cm{sup -3} in 5 {mu}s after the current dip. In experiments with the second electrode version, among numerous spectral lines there were identified distinct WI and WII lines. Estimates of the tungsten ion concentration were impossible because of close positions of other lines. Diagrams of changes in the electron density are of importance for applications of DPM streams.

  8. Single-phase heat transfer improved by helical inserts in tubes

    NASA Technical Reports Server (NTRS)

    Gutsein, M. U.

    1970-01-01

    Helical-vane insert creates a single, well defined helical flow passage resulting in more reliable predictions and extrapolations of the thermal and hydraulic performance. Insert core houses instrumentation which does not disturb the flow.

  9. Insertion of tear proteins into a meibomian lipids film.

    PubMed

    Miano, F; Calcara, M; Millar, T J; Enea, V

    2005-07-25

    The eyelid meibomian gland secretions form the outer layer of the tear film. That layer functions as a lubricant during a blink, and as a barrier against intrusion of foreign bodies. The lipid film is also exposed to proteins present in the aqueous phase that may adsorb there, and thus form an integral part of the surface of the tear film, or possibly, cause disruption to the outermost layer. Therefore, the adsorption of tear proteins to the meibomian lipid layer was object of the present investigation. A model tear was set up coating a pendant drop of saline with a film of meibomian lipids and measuring variations of the interfacial pressure after the injection of tear proteins into the aqueous subphase at their physiological concentration. All tear proteins adsorbed at the interface causing the initial surface pressure to increase. For each protein, a limiting surface pressure at which a given protein was no longer able to insert into the lipid layer was found. Among the proteins tested, lipocalin was the most surface active one and inserted into the lipid layer in the whole range of surface pressure exerted by the meibomian lipid mixture. Lactoferrin, lysozyme and IgA also interacted with the lipids whereas albumin interacted more weakly. The timescale of the protein insertion into the lipid layer was of the order of 10(2) s. It was hypothesized that protein adsorption at the interface could be associated with structural changes. Indeed, the enzymatic activity of lysozyme was maintained in the presence of an outermost meibomian lipid layer that prevented its denaturation while exposure at the air/aqueous interface induced significant lysozime degradation. meibomian lipid composition is therefore functional to maintain tear proteins activity. PMID:16006106

  10. 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.

  11. The endogenous ecotropic murine retroviruses Emv-16 and Emv-17 are both capable of producing new proviral insertions in the mouse genome.

    PubMed

    Szabo, C; Kim, Y K; Mark, W H

    1993-09-01

    New germ line proviral insertions are acquired at a high frequency by the progeny of SWR/J-RF/J hybrid female mice that carry the endogenous ecotropic murine leukemia proviruses Emv-16 and Emv-17. The tight linkage of these RF/J strain proviral loci has prevented genetic segregation of the retroviral genomes. Hence, it is not known whether both of these proviruses are capable of giving rise to new proviral insertions. We have molecularly cloned Emv-16 and Emv-17 and have characterized them in vitro and in vivo. Restriction enzyme analysis of the recombinant clones revealed that the proviral genomes are very similar to each other and closely resemble the wild-type AKR virus. A comparison of the flanking cellular DNA suggests that the Emv-16 and Emv-17 loci did not arise by simple duplication of a viral insertion site within the RF/J genome but most likely are independent integration events. Both proviruses produce infectious virus when transfected into NIH 3T3 cells, indicating that they are nondefective retroviruses. Exogenous infection of SWR/J mice with either Emv-16 or Emv-17 leads to viremia in the host animals, and in both cases, progeny of viremic females acquire new proviral insertions. The ability of these retroviruses to generate novel retroviral integration sites in the mouse genome provides a simple method for inducing insertional mutations in mice. PMID:8394469

  12. 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. PMID:26590729

  13. A new gap separation mechanism for APS insertion devices

    NASA Astrophysics Data System (ADS)

    Trakhtenberg, E. M.; Tcheskidov, V. G.; Den Hartog, P. K.; Deriy, B. N.; Erdmann, M.; Makarov, O. A.; Moog, E. R.

    2000-06-01

    A new gap separation mechanism for use with the standard Advanced Photon Source (APS) 3.3-cm-period undulator magnetic structures has been designed and built, and the first system has been installed in the APS storage ring. The system allows a minimum magnetic gap of 10 mm for use with the APS 8-mm insertion device vacuum chambers. The mechanism is a bolted steel frame structure with a simple 4-motor mechanical drive train. The control system uses servomotors with incremental rotary encoders and virtual absolute linear encoders.

  14. The design and analysis of transposon insertion sequencing experiments.

    PubMed

    Chao, Michael C; Abel, Sören; Davis, Brigid M; Waldor, Matthew K

    2016-01-19

    Transposon insertion sequencing (TIS) is a powerful approach that can be extensively applied to the genome-wide definition of loci that are required for bacterial growth under diverse conditions. However, experimental design choices and stochastic biological processes can heavily influence the results of TIS experiments and affect downstream statistical analysis. In this Opinion article, 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 the computational analysis of TIS data. PMID:26775926

  15. Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants

    PubMed Central

    2011-01-01

    A method was developed to identify insertional mutants of Chlamydomonas reinhardtii disrupted for selected target genes. The approach relies on the generation of thousands of transformants followed by PCR-based screenings that allow for identification of strains harboring the introduced marker gene within specific genes of interest. Our results highlight the strengths and limitations of two independent screens that differed in the nature of the marker DNA used (PCR-amplified fragment containing the plasmid-free marker versus entire linearized plasmid with the marker) and in the strategies used to maintain and store transformants. PMID:21794168

  16. 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.

  17. 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.

  18. A new gap separation mechanism for APS insertion devices.

    SciTech Connect

    Trakhtenberg, E. M.; Tcheskidov, V.; Den Hartog, P. K.; Deriy, B.; Erdmann, M.; Makarov, O.; Moog, E. R.

    1999-10-25

    A new gap separation mechanism for use with the standard Advanced Photon Source (APS) 3.3-cm-period undulator magnetic structures has been designed and built and the first system has been installed in the APS storage ring. The system allows a minimum magnetic gap of 10 mm for use with the APS 8-mm insertion device vacuum chambers. The mechanism is a bolted steel frame structure with a simple 4-motor mechanical drive train. The control system uses servomotors with incremental rotary encoders and virtual absolute linear encoders.

  19. 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.

  20. 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. PMID:25800058

  1. Promise versus reality: optimism bias in package inserts for tuberculosis diagnostics.

    PubMed

    Denkinger, Claudia M; Grenier, Jasmine; Minion, Jessica; Pai, Madhukar

    2012-07-01

    Laboratorians and clinicians often rely on package inserts of diagnostic tests to assess their accuracy. We compared test accuracy for tuberculosis diagnostics reported in 19 package inserts against estimates in published meta-analyses and found that package inserts generally report overoptimistic accuracy estimates. However, package inserts of most tests approved by the U.S. Food and Drug Administration (FDA) or endorsed by the World Health Organization provide more realistic estimates that agree with meta-analyses. PMID:22573592

  2. 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. PMID:15859082

  3. 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.

  4. Improving Peripherally Inserted Central Catheter (PICC) care on a Trauma and Orthopaedics ward

    PubMed Central

    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.

  5. Cholecystoduodenal fistula: A complication of inserted self-expandable metallic bilitary stents

    SciTech Connect

    Nishida, Hirotoshi; Inoue, Hiroki; Ueno, Kazuto; Nagata, Yukitaka; Kato, Takeshi; Miyazono, Nobuaki; Nakajo, Masayuki

    1998-05-15

    We encountered a case of hepatic hilar cholangiocarcinoma resulting in cholecystoduodenal fistula after insertion of self-expandable metallic biliary stents (EMBSs). To our knowledge, there has been no report of cholecystoduodenal fistula after insertion of EMBSs. This case suggests that immediate gallbladder decompression may be necessary if acute cholecystitis occurs after insertion of EMBSs.

  6. Cholecystoduodenal Fistula: A Complication of Inserted Self-Expandable Metallic Biliary Stents

    SciTech Connect

    Nishida, Hirotoshi; Inoue, Hiroki; Ueno, Kazuto; Nagata, Yukitaka; Kato, Takeshi; Miyazono, Nobuaki; Nakajo, Masayuki

    1998-05-15

    We encountered a case of hepatic hilar cholangiocarcinoma resulting in cholecystoduodenal fistula after insertion of self-expandable metallic biliary stents (EMBSs). To our knowledge, there has been no report of cholecystoduodenal fistula after insertion of EMBSs. This case suggests that immediate gallbladder decompression may be necessary if acute cholecystitis occurs after insertion of EMBSs.

  7. 21 CFR 876.5830 - Hemodialyzer with disposable insert (Kiil type).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 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...

  8. 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...

  9. 21 CFR 872.3900 - Posterior artificial tooth with a metal insert.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Posterior artificial tooth with a metal insert... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is...

  10. 21 CFR 872.3900 - Posterior artificial tooth with a metal insert.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Posterior artificial tooth with a metal insert... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is...

  11. 21 CFR 872.3900 - Posterior artificial tooth with a metal insert.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Posterior artificial tooth with a metal insert... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is...

  12. 21 CFR 872.3900 - Posterior artificial tooth with a metal insert.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Posterior artificial tooth with a metal insert... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is...

  13. 21 CFR 872.3900 - Posterior artificial tooth with a metal insert.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Posterior artificial tooth with a metal insert... SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Prosthetic Devices § 872.3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is...

  14. 46 CFR 42.50-10 - Load line certificates for nonadherent foreign flag vessels.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... amended (46 U.S.C. 85-85g), and the Load Line Regulations in 46 CFR part 42: By (Insert full official... CFR part 42. This certificate remains in force until ____,2 subject to annual revalidation in... 46 Shipping 2 2010-10-01 2010-10-01 false Load line certificates for nonadherent foreign...

  15. Pyoderma gangrenosum after totally implanted central venous access device insertion

    PubMed Central

    Inan, Ihsan; Myers, Patrick O; Braun, Rolf; Hagen, Monica E; Morel, Philippe

    2008-01-01

    Background Pyoderma gangrenosum is an aseptic skin disease. The ulcerative form of pyoderma gangrenosum is characterized by a rapidly progressing painful irregular and undermined bordered necrotic ulcer. The aetiology of pyoderma gangrenosum remains unclear. In about 70% of cases, it is associated with a systemic disorder, most often inflammatory bowel disease, haematological disease or arthritis. In 25–50% of cases, a triggering factor such as recent surgery or trauma is identified. Treatment consists of local and systemic approaches. Systemic steroids are generally used first. If the lesions are refractory, steroids are combined with other immunosuppressive therapy or to antimicrobial agents. Case presentation A 90 years old patient with myelodysplastic syndrome, seeking regular transfusions required totally implanted central venous access device (Port-a-Cath®) insertion. Fever and inflammatory skin reaction at the site of insertion developed on the seventh post-operative day, requiring the device's explanation. A rapid progression of the skin lesions evolved into a circular skin necrosis. Intravenous steroid treatment stopped the necrosis' progression. Conclusion Early diagnosis remains the most important step to the successful treatment of pyoderma gangrenosum. PMID:18325095

  16. Emissive signature for HMMWV run flat insert modeling and simulation

    NASA Astrophysics Data System (ADS)

    Gardetto, W.; Gorsich, D.; Kurec, A.; Mabesa, J.; Murray, D.

    2007-04-01

    Analyzing thermodynamic patterns during product development can easily be characterized by various "Modeling and Simulation" software programs to observe Emissive Signatures. Baseline temperatures are referenced as an adjusted "Blackbody" value and used to compare differential temperature changes during dynamometer testing. An infrared spectrum distinguishes pattern profiles unique to the product for both thermodynamic performance and to accurately validate test materials. A collaborative CRADA1 effort has been established between the US Army RDECOMTARDEC and Drive Dynamics LLC of Dallas, TX on the development of an advanced Run Flat Insert System for military wheeled vehicles. Mapping of measured infrared thermometer values help in locating and determining whether or not material temperatures are within design limits. Prior testing by the US Army Physical Simulation Team has established a baseline Emissive Signature for HMMWV wheel assemblies at specific loads and speeds. As advanced Run Flat Insert Systems are developed for increased load capacities using structurally engineered profiles the Emissive Signature can be used to compare to baseline wheel assemblies and aid in establishing Run Flat performance and longevity.

  17. Single gene insertion drives bioalcohol production by a thermophilic archaeon

    SciTech Connect

    Basen, M; Schut, GJ; Nguyen, DM; Lipscomb, GL; Benn, RA; Prybol, CJ; Vaccaro, BJ; Poole, FL; Kelly, RM; Adams, MWW

    2014-12-09

    Bioethanol production is achieved by only two metabolic pathways and only at moderate temperatures. Herein a fundamentally different synthetic pathway for bioalcohol production at 70 degrees C was constructed by insertion of the gene for bacterial alcohol dehydrogenase (AdhA) into the archaeon Pyrococcus furiosus. The engineered strain converted glucose to ethanol via acetate and acetaldehyde, catalyzed by the host-encoded aldehyde ferredoxin oxidoreductase (AOR) and heterologously expressed AdhA, in an energy-conserving, redox-balanced pathway. Furthermore, the AOR/AdhA pathway also converted exogenously added aliphatic and aromatic carboxylic acids to the corresponding alcohol using glucose, pyruvate, and/or hydrogen as the source of reductant. By heterologous coexpression of a membrane-bound carbon monoxide dehydrogenase, CO was used as a reductant for converting carboxylic acids to alcohols. Redirecting the fermentative metabolism of P. furiosus through strategic insertion of foreign genes creates unprecedented opportunities for thermophilic bioalcohol production. Moreover, the AOR/AdhA pathway is a potentially game-changing strategy for syngas fermentation, especially in combination with carbon chain elongation pathways.

  18. Flavonoid insertion into cell walls improves wood properties.

    PubMed

    Ermeydan, Mahmut A; Cabane, Etienne; Masic, Admir; Koetz, Joachim; Burgert, Ingo

    2012-11-01

    Wood has an excellent mechanical performance, but wider utilization of this renewable resource as an engineering material is limited by unfavorable properties such as low dimensional stability upon moisture changes and a low durability. However, some wood species are known to produce a wood of higher quality by inserting mainly phenolic substances in the already formed cell walls--a process so-called heartwood formation. In the present study, we used the heartwood formation in black locust (Robinia pseudoacacia) as a source of bioinspiration and transferred principles of the modification in order to improve spruce wood properties (Picea abies) by a chemical treatment with commercially available flavonoids. We were able to effectively insert hydrophobic flavonoids in the cell wall after a tosylation treatment for activation. The chemical treatment reduced the water uptake of the wood cell walls and increased the dimensional stability of the bulk spruce wood. Further analysis of the chemical interaction of the flavonoid with the structural cell wall components revealed the basic principle of this bioinspired modification. Contrary to established modification treatments, which mainly address the hydroxyl groups of the carbohydrates with hydrophilic substances, the hydrophobic flavonoids are effective by a physical bulking in the cell wall most probably stabilized by ?-? interactions. A biomimetic transfer of the underlying principle may lead to alternative cell wall modification procedures and improve the performance of wood as an engineering material. PMID:23027798

  19. Application of plantar pressure assessment in footwear and insert design.

    PubMed

    Mueller, M J

    1999-12-01

    This clinical perspective describes the application of plantar pressure assessment in footwear and insert design. First, the rationale and evidence for using pressure assessment to assist in the design of footwear for patients with diabetes is described. I discuss 2 important measures obtained from pressure assessment: peak pressure, because it represents the magnitude of potential mechanical stresses that can contribute to skin breakdown, and contact area, because this identifies the treatment areas. Using measures obtained from pressure assessment, guidelines are presented to maximize contact area of the insert to the foot and reduce highest peak pressures on the skin, with the goal of preventing skin breakdown. Second, a rationale and guidelines are presented for the application of plantar pressure assessment in the evaluation and design of footwear for people without impairments (i.e., the general public). Finally, future applications of pressure assessment to improve the design and fit of shoes are discussed. Benefits and limitations of using pressure assessment to assist in footwear design are addressed throughout. PMID:10612072

  20. Transient expression and insertional mutagenesis of Puccinia triticina using biolistics.

    PubMed

    Webb, Craig A; Szabo, Les J; Bakkeren, Guus; Garry, Clarke; Staples, Richard C; Eversmeyer, Merle; Fellers, John P

    2006-07-01

    The fungal genus Puccinia contains more than 4,000 species. Puccinia triticina, causal agent of wheat leaf rust, is an economically significant, biotrophic basidiomycete. Little is known about the molecular biology of this group, and tools for understanding gene function have not yet been established. A set of parameters was established for the transient transformation of urediniospores. The expression of three heterologous promoters (actin, elongation factor 1-alpha, and Hss1, Heat Shock 70 protein), derived from Puccinia graminis, was evaluated along with the potential for insertional mutagenesis. The UidA (GUS) gene was used as a marker for transient expression. When transferred into P. triticina urediniospores, transient expression was observed across four helium pressures using one size of gold and three sizes of tungsten microprojectiles. Each of the three promoters displayed strong transient expression in germinated urediniospores; however, higher numbers of GUS-positive urediniospores were observed when either the actin or Hss1 promoters were used. Possible concomitant insertional mutagenesis of several avirulence genes was selected in wheat cultivars harboring the cognate resistance genes. Using a linearized cloning plasmid, stable integration into the genome was achieved as demonstrated by PCR and sequencing analysis. PMID:16284743

  1. Control system for insertion devices at the Advanced Photon Source

    SciTech Connect

    Makarov, O.A.; Den Hartog, P.; Moog, E.R.; Smith, M.L.

    1997-09-01

    Eighteen insertion devices (IDs) are installed at the Advanced Photon Source (APS), and three more are scheduled for installation by the end of this year. A distributed control system for insertion devices at the APS storage ring was created with the Experimental Physics and Industrial Control System (EPICS). The basic components of this system are operator interfaces (OPIs), input output controllers (IOCs), and a local area network that allows the OPI and IOC to communicate. The IOC operates under the VxWorks OS with an EPICS database and a sequencer. The sequencer runs an ID control program written in State Notation Language. The OPI is built with the EPICS tool MEDM and provides display screens with input and output fields and buttons for gap control of the IDs. Global commands like ``open all IDs`` are C-shell scripts invoked from the display menu. The algorithms for control and protection of the ID and ID vacuum chamber and the accuracy of gap control are discussed.

  2. Systematic discovery of complex insertions and deletions in human cancers.

    PubMed

    Ye, Kai; Wang, Jiayin; Jayasinghe, Reyka; Lameijer, Eric-Wubbo; McMichael, Joshua F; Ning, Jie; McLellan, Michael D; Xie, Mingchao; Cao, Song; Yellapantula, Venkata; Huang, Kuan-Lin; Scott, Adam; Foltz, Steven; Niu, Beifang; Johnson, Kimberly J; Moed, Matthijs; Slagboom, P Eline; Chen, Feng; Wendl, Michael C; Ding, Li

    2016-01-01

    Complex insertions and deletions (indels) are formed by simultaneously deleting and inserting DNA fragments of different sizes at a common genomic location. Here we present a systematic analysis of somatic complex indels in the coding sequences of samples from over 8,000 cancer cases using Pindel-C. We discovered 285 complex indels in cancer-associated genes (such as PIK3R1, TP53, ARID1A, GATA3 and KMT2D) in approximately 3.5% of cases analyzed; nearly all instances of complex indels were overlooked (81.1%) or misannotated (17.6%) in previous reports of 2,199 samples. In-frame complex indels are enriched in PIK3R1 and EGFR, whereas frameshifts are prevalent in VHL, GATA3, TP53, ARID1A, PTEN and ATRX. Furthermore, complex indels display strong tissue specificity (such as VHL in kidney cancer samples and GATA3 in breast cancer samples). Finally, structural analyses support findings of previously missed, but potentially druggable, mutations in the EGFR, MET and KIT oncogenes. This study indicates the critical importance of improving complex indel discovery and interpretation in medical research. PMID:26657142

  3. Mass insertions vs. mass eigenstates calculations in flavour physics

    NASA Astrophysics Data System (ADS)

    Dedes, A.; Paraskevas, M.; Rosiek, J.; Suxho, K.; Tamvakis, K.

    2015-06-01

    We present and prove a theorem of matrix analysis, the Flavour Expansion Theorem (or FET), according to which, an analytic function of a Hermitian matrix can be expanded polynomially in terms of its off-diagonal elements with coefficients being the divided differences of the analytic function and arguments the diagonal elements of the Hermitian matrix. The theorem is applicable in case of flavour changing amplitudes. At one-loop level this procedure is particularly natural due to the observation that every loop function in the Passarino-Veltman basis can be recursively expressed in terms of divided differences. FET helps to algebraically translate an amplitude written in mass eigenbasis into flavour mass insertions, without performing diagrammatic calculations in flavour basis. As a non-trivial application of FET up to a third order, we demonstrate its use in calculating strong bounds on the real parts of flavour changing mass insertions in the up- squark sector of the MSSM from neutron Electric Dipole Moment (nEDM) measurements, assuming that CP-violation arises only from the CKM matrix.

  4. Sodium Ion Insertion in Hollow Carbon Nanowires for Battery Applications

    SciTech Connect

    Cao, Yuliang; Xiao, Lifen; Sushko, Maria L.; Wang, Wei; Schwenzer, Birgit; Xiao, Jie; Nie, Zimin; Saraf, Laxmikant V.; Yang, Zhenguo; Liu, Jun

    2012-07-11

    Hollow Carbon Nanowires (HCNWs) were prepared through pyrolyzation of hollow polyaniline nanowires precursor. The HCNWs used as anode material for Na-ion batteries delivers a high reversible capacity of 251 mAh g{sup -1} and 82.2% capacity retention over 400 charge/discharge cycles between 1.2 and 0.01 V (vs. Na{sup +}/Na) at a constant current of 50 mA g{sup -1} (0.2 C). Excellent cycling stability is also observed at even higher charge-discharge rate. A high reversible capacity of 149 mAh g{sup -1} also can be obtained at a current rate of 500 mA g{sup -1} (2C). The good Na ion insertion property is attributed to the short diffusion distance in the HCNWs, and the large interlayer distance (0.37 nm) between the graphitic sheets, which agrees with the interlayered distance predicted by theoretical calculation to enable Na ion insertion in carbon materials.

  5. Stochastic models of sequence evolution including insertion-deletion events.

    PubMed

    Miklós, István; Novák, Adám; Satija, Rahul; Lyngsø, Rune; Hein, Jotun

    2009-10-01

    Comparison of sequences that have descended from a common ancestor based on an explicit stochastic model of substitutions, insertions and deletions has risen to prominence in the last decade. Making statements about the positions of insertions-deletions (abbr. indels) is central in sequence and genome analysis and is called alignment. This statistical approach is harder conceptually and computationally, than competing approaches based on choosing an alignment according to some optimality criteria. But it has major practical advantages in terms of testing evolutionary hypotheses and parameter estimation. Basic dynamic approaches can allow the analysis of up to 4-5 sequences. MCMC techniques can bring this to about 10-15 sequences. Beyond this, different or heuristic approaches must be used. Besides the computational challenges, increasing realism in the underlying models is presently being addressed. A recent development that has been especially fruitful is combining statistical alignment with the problem of sequence annotation, making statements about the function of each nucleotide/amino acid. So far gene finding, protein secondary structure prediction and regulatory signal detection has been tackled within this framework. Much progress can be reported, but clearly major challenges remain if this approach is to be central in the analyses of large incoming sequence data sets. PMID:19221170

  6. Approaches to querying bacterial genomes with transposon-insertion sequencing

    PubMed Central

    Barquist, Lars; Boinett, Christine J.; Cain, Amy K.

    2013-01-01

    In this review, we discuss transposon-insertion sequencing, variously known in the literature as TraDIS, Tn-seq, INSeq, and HITS. By monitoring a large library of single transposon-insertion mutants with high-throughput sequencing, these methods can rapidly identify genomic regions that contribute to organismal fitness under any condition assayable in the laboratory with exquisite resolution. We discuss the various protocols that have been developed and methods for analysis. We provide an overview of studies that have examined the reproducibility and accuracy of these methods, as well as studies showing the advantages offered by the high resolution and dynamic range of high-throughput sequencing over previous methods. We review a number of applications in the literature, from predicting genes essential for in vitro growth to directly assaying requirements for survival under infective conditions in vivo. We also highlight recent progress in assaying non-coding regions of the genome in addition to known coding sequences, including the combining of RNA-seq with high-throughput transposon mutagenesis. PMID:23635712

  7. Lithium insertion in carbons prepared from phosphorus-containing polymers

    NASA Astrophysics Data System (ADS)

    Xiang, Hong-Qi; Fang, Shi-Bi; Jiang, Ying-Yan

    Phosphorus-containing carbon has been prepared by the pyrolysis of phenolic resin containing chemically bonded phosphorus atoms, which is synthesized through the esterification of phenolic hydroxyl group by phosphoric acid. The electrochemical insertion of lithium in as-prepared carbon is also investigated. It is found that, at a pyrolysis temperature of 500°C, the addition of phosphoric acid in the precursor lowers the reversible capacity of the resultant carbon, but at 900°C, the addition of phosphoric acid in the precursor resin increases the reversible capacity of resultant carbon. Especially, the phosphorus-containing carbon prepared by the above method at 900°C gives a capacity beyond the theoretical value of graphite and reasonable discharge/charge curves. The phosphorus-containing carbon prepared by the pyrolysis at 500°C shows a similar electrochemical behavior to that of pure carbon and the effect of phosphorus atoms is little. Its reversible capacity is mainly determined by the content of hydrogen atoms rather than phosphorus atoms. However, at a relatively high pyrolysis temperature, 900°C, more phosphorus atoms are bonded with carbon atoms and are introduced to the microcrystallite structure of carbon. Hence, phosphorus element exerts a strike effect on the electrochemical behavior of carbon and, because of its different electronic effect from carbon element, more lithium species are inserted into carbon matrix.

  8. Launch vehicle and power level impacts on electric GEO insertion

    NASA Technical Reports Server (NTRS)

    Oleson, Steven R.; Myers, Roger M.

    1996-01-01

    Solar Electric Propulsion (SEP) has been shown to increase net geosynchronous spacecraft mass when used for station keeping and final orbit insertion. The impact of launch vehicle selection and power level on the benefits of this approach were examined for 20 and 25 kW systems launched using the Ariane 5, Atlas IIAR, Long March, Proton, and Sea Launch vehicles. Two advanced on-board propulsion technologies, 5 kW ion and Hall thruster systems, were used to establish the relative merits of the technologies and launch vehicles. GaAs solar arrays were assumed. The analysis identifies the optimal starting orbits for the SEP orbit raising/plane changing while considering the impacts of radiation degradation in the Van Allen belts, shading, power degradation, and oblateness. This use of SEP to provide part of the orbit insertion results in net mass increases of 15 - 38% and 18 - 46% for one to two month trip times, respectively, over just using SEP for 15 years of north/south station keeping. SEP technology was shown to have a greater impact on net masses of launch vehicles with higher launch latitudes when avoidance of solar array and payload degradation is desired. This greater impact of SEP could help reduce the plane changing disadvantage of high latitude launch sites. Comparison with results for 10 and 15 kW systems show clear benefits of incremental increases in SEP power level, suggesting that an evolutionary approach to high power SEP for geosynchronous spacecraft is possible.

  9. Early metamorphic insertion technology for insect flight behavior monitoring.

    PubMed

    Verderber, Alexander; McKnight, Michael; Bozkurt, Alper

    2014-01-01

    Early Metamorphosis Insertion Technology (EMIT) is a novel methodology for integrating microfabricated neuromuscular recording and actuation platforms on insects during their metamorphic development. Here, the implants are fused within the structure and function of the neuromuscular system as a result of metamorphic tissue remaking. The implants emerge with the insect where the development of tissue around the electronics during pupal development results in a bioelectrically and biomechanically enhanced tissue interface. This relatively more reliable and stable interface would be beneficial for many researchers exploring the neural basis of the insect locomotion with alleviated traumatic effects caused during adult stage insertions. In this article, we implant our electrodes into the indirect flight muscles of Manduca sexta. Located in the dorsal-thorax, these main flight powering dorsoventral and dorsolongitudinal muscles actuate the wings and supply the mechanical power for up and down strokes. Relative contraction of these two muscle groups has been under investigation to explore how the yaw maneuver is neurophysiologically coordinated. To characterize the flight dynamics, insects are often tethered with wires and their flight is recorded with digital cameras. We also developed a novel way to tether Manduca sexta on a magnetically levitating frame where the insect is connected to a commercially available wireless neural amplifier. This set up can be used to limit the degree of freedom to yawing "only" while transmitting the related electromyography signals from dorsoventral and dorsolongitudinal muscle groups. PMID:25079130

  10. Novel insertion sequence-like element IS982 in lactococci.

    PubMed

    Yu, W; Mierau, I; Mars, A; Johnson, E; Dunny, G; McKay, L L

    1995-05-01

    A novel insertion sequence-like (IS) element, designated IS982, was found on the lactose plasmid, pSK11L, from Lactococcus lactis subsp. cremoris SK11 and was located between the origin of replication and the oligopeptide transport gene cluster. The 1003-base pair (bp) IS982 was flanked by 18-bp perfect inverted repeats. IS982 contained an open reading frame encoding a putative transposase of 296 amino acids. An almost identical IS-like element (99% DNA sequence identity) was cloned and partially sequenced from the chromosome of Lactococcus lactis subsp. cremoris Wg2 with 17-bp perfect inverted repeats. Southern analysis indicated that in 12 lactococcal strains examined, IS982 was present with copy numbers ranging from 1 to at least 20. IS982 displayed sequence homology to the putative IS element RSBst-alpha from Bacillus stearothermophilus CU21, IS982 and RSBst-alpha were not related to other known insertion sequences and may represent a new family of IS elements. PMID:7568469

  11. Structure and variability of recently inserted Alu family members.

    PubMed Central

    Batzer, M A; Kilroy, G E; Richard, P E; Shaikh, T H; Desselle, T D; Hoppens, C L; Deininger, P L

    1990-01-01

    The HS subfamily of Alu sequences is comprised of a group of nearly identical members. Individual subfamily members share 97.7% nucleotide identity with each other and 98.9% nucleotide identity with the HS consensus sequence. Individual subfamily members are on the average 2.8 million years old, and were probably derived from a single source 'master' gene sometime after the human/great ape divergence. The recent Alu family member insertions provide a better image of the structure of Alu retroposons before they have had the opportunity to change significantly. All of the HS subfamily members are flanked by perfect direct repeats as a result of insertion at staggered nicks. The 'master' gene from which the HS subfamily members were derived had an oligo-dA rich tail at least 40 bases long. The 'master' gene is very rich in CpG dinucleotides, but nucleotide substitutions within subfamily members accumulated in a random manner typical for Alu sequence with CpG substitutions occurring 9.2 fold faster than non-CpG substitutions. PMID:2175877

  12. Detecting Alu insertions from high-throughput sequencing data.

    PubMed

    David, Matei; Mustafa, Harun; Brudno, Michael

    2013-09-01

    High-throughput sequencing technologies have allowed for the cataloguing of variation in personal human genomes. In this manuscript, we present alu-detect, a tool that combines read-pair and split-read information to detect novel Alus and their precise breakpoints directly from either whole-genome or whole-exome sequencing data while also identifying insertions directly in the vicinity of existing Alus. To set the parameters of our method, we use simulation of a faux reference, which allows us to compute the precision and recall of various parameter settings using real sequencing data. Applying our method to 100 bp paired Illumina data from seven individuals, including two trios, we detected on average 1519 novel Alus per sample. Based on the faux-reference simulation, we estimate that our method has 97% precision and 85% recall. We identify 808 novel Alus not previously described in other studies. We also demonstrate the use of alu-detect to study the local sequence and global location preferences for novel Alu insertions. PMID:23921633

  13. Fracture of crystalline silicon nanopillars during electrochemical lithium insertion

    PubMed Central

    Lee, Seok Woo; McDowell, Matthew T.; Berla, Lucas A.; Nix, William D.; Cui, Yi

    2012-01-01

    From surface hardening of steels to doping of semiconductors, atom insertion in solids plays an important role in modifying chemical, physical, and electronic properties of materials for a variety of applications. High densities of atomic insertion in a solid can result in dramatic structural transformations and associated changes in mechanical behavior: This is particularly evident during electrochemical cycling of novel battery electrodes, such as alloying anodes, conversion oxides, and sulfur and oxygen cathodes. Silicon, which undergoes 400% volume expansion when alloying with lithium, is an extreme case and represents an excellent model system for study. Here, we show that fracture locations are highly anisotropic for lithiation of crystalline Si nanopillars and that fracture is strongly correlated with previously discovered anisotropic expansion. Contrary to earlier theoretical models based on diffusion-induced stresses where fracture is predicted to occur in the core of the pillars during lithiation, the observed cracks are present only in the amorphous lithiated shell. We also show that the critical fracture size is between about 240 and 360 nm and that it depends on the electrochemical reaction rate. PMID:22371565

  14. Single gene insertion drives bioalcohol production by a thermophilic archaeon

    PubMed Central

    Basen, Mirko; Schut, Gerrit J.; Nguyen, Diep M.; Lipscomb, Gina L.; Benn, Robert A.; Prybol, Cameron J.; Vaccaro, Brian J.; Poole, Farris L.; Kelly, Robert M.; Adams, Michael W. W.

    2014-01-01

    Bioethanol production is achieved by only two metabolic pathways and only at moderate temperatures. Herein a fundamentally different synthetic pathway for bioalcohol production at 70 °C was constructed by insertion of the gene for bacterial alcohol dehydrogenase (AdhA) into the archaeon Pyrococcus furiosus. The engineered strain converted glucose to ethanol via acetate and acetaldehyde, catalyzed by the host-encoded aldehyde ferredoxin oxidoreductase (AOR) and heterologously expressed AdhA, in an energy-conserving, redox-balanced pathway. Furthermore, the AOR/AdhA pathway also converted exogenously added aliphatic and aromatic carboxylic acids to the corresponding alcohol using glucose, pyruvate, and/or hydrogen as the source of reductant. By heterologous coexpression of a membrane-bound carbon monoxide dehydrogenase, CO was used as a reductant for converting carboxylic acids to alcohols. Redirecting the fermentative metabolism of P. furiosus through strategic insertion of foreign genes creates unprecedented opportunities for thermophilic bioalcohol production. Moreover, the AOR/AdhA pathway is a potentially game-changing strategy for syngas fermentation, especially in combination with carbon chain elongation pathways. PMID:25368184

  15. Control system for insertion devices at the advanced photon source

    SciTech Connect

    Makarov, Oleg A.; Den Hartog, Patric; Moog, Elizabeth R.; Smith, Martin L.

    1997-07-01

    Eighteen insertion devices (IDs) are installed at the Advanced Photon Source (APS), and three more are scheduled for installation by the end of this year. A distributed control system for insertion devices at the APS storage ring was created with the Experimental Physics and Industrial Control System (EPICS). The basic components of this system are operator interfaces (OPIs), input output controllers (IOCs), and a local area network that allows the OPI and IOC to communicate. The IOC operates under the VxWorks OS with an EPICS database and a sequencer. The sequencer runs an ID control program written in State Notation Language. The OPI is built with the EPICS tool MEDM and provides display screens with input and output fields and buttons for gap control of the IDs. Global commands like 'open all IDs' are C-shell scripts invoked from the display menu. The algorithms for control and protection of the ID and ID vacuum chamber and the accuracy of gap control are discussed.

  16. Control system for insertion devices at the advanced photon source

    SciTech Connect

    Makarov, O.A.; Den Hartog, P.; Moog, E.R.; Smith, M.L.

    1997-07-01

    Eighteen insertion devices (IDs) are installed at the Advanced Photon Source (APS), and three more are scheduled for installation by the end of this year. A distributed control system for insertion devices at the APS storage ring was created with the Experimental Physics and Industrial Control System (EPICS). The basic components of this system are operator interfaces (OPIs), input output controllers (IOCs), and a local area network that allows the OPI and IOC to communicate. The IOC operates under the VxWorks OS with an EPICS database and a sequencer. The sequencer runs an ID control program written in State Notation Language. The OPI is built with the EPICS tool MEDM and provides display screens with input and output fields and buttons for gap control of the IDs. Global commands like {open_quotes}open all IDs{close_quotes} are C-shell scripts invoked from the display menu. The algorithms for control and protection of the ID and ID vacuum chamber and the accuracy of gap control are discussed. {copyright} {ital 1997 American Institute of Physics.}

  17. Experimental and Numerical Analysis of Inserts in Sandwich Structures

    NASA Astrophysics Data System (ADS)

    Bunyawanichakul, P.; Castanie, B.; Barrau, J.-J.

    2005-05-01

    In aeronautics, sandwich structures are widely used for secondary structures like flaps or landing gear doors. In the case of landing gear doors, the junction is made by a local reinforcement called an insert. This insert is made by a resin molded in the Nomex™ sandwich core. Such structures are still designed mainly using test results and the lack of an efficient numerical model remains a problem. The purpose of this study is on the one hand to perform experiments in order to be able to identify the failure modes and on the other hand to propose an efficient numerical model. Pull-out tests with cycling were conducted and 3D displacement measured by optical methods. The potential failure modes are numerous (delamination, local fiber breaking, skin/core debonding, core crushing, core shear buckling, potting failure, etc.). Experiments demonstrated that, for the lower loads, the non-linearity and the hysteresis are mainly due to core shear buckling. From this observation, the nonlinear behavior of the core is identified by a 3 point-bending test. The shear-modulus damage law is then implemented on a non-linear finite element model and an acceptable correlation of the tests is achieved. As a consequence, some improvements of the technology will be proposed, manufactured and tested.

  18. Seamless insertion of real pulmonary nodules in chest CT exams

    NASA Astrophysics Data System (ADS)

    Pezeshk, Aria; Sahiner, Berkman; Zeng, Rongping; Wunderlich, Adam; Chen, Weijie; Petrick, Nicholas

    2014-03-01

    The availability of large medical image datasets is critical in many applications such as training and testing of computer aided diagnosis (CAD) systems, evaluation of segmentation algorithms, and conducting perceptual studies. However, collection of large repositories of clinical images is hindered by the high cost and difficulties associated with both the accumulation of data and establishment of the ground truth. To address this problem, we are developing an image blending tool that allows users to modify or supplement existing datasets by seamlessly inserting a real lesion extracted from a source image into a different location on a target image. In this study we focus on the application of this tool to pulmonary nodules in chest CT exams. We minimize the impact of user skill on the perceived quality of the blended image by limiting user involvement to two simple steps: the user first draws a casual boundary around the nodule of interest in the source, and then selects the center of desired insertion area in the target. We demonstrate examples of the performance of the proposed system on samples taken from the Lung Image Database Consortium (LIDC) dataset, and compare the noise power spectrum (NPS) of blended nodules versus that of native nodules in simulated phantoms.

  19. Molecular Details of Bax Activation, Oligomerization, and Membrane Insertion*

    PubMed Central

    Bleicken, Stephanie; Classen, Mirjam; Padmavathi, Pulagam V. L.; Ishikawa, Takashi; Zeth, Kornelius; Steinhoff, Heinz-Jürgen; Bordignon, Enrica

    2010-01-01

    Bax and Bid are pro-apoptotic members of the Bcl-2 protein family. Upon cleavage by caspase-8, Bid activates Bax. Activated Bax inserts into the mitochondrial outer membrane forming oligomers which lead to membrane poration, release of cytochrome c, and apoptosis. The detailed mechanism of Bax activation and the topology and composition of the oligomers are still under debate. Here molecular details of Bax activation and oligomerization were obtained by application of several biophysical techniques, including atomic force microscopy, cryoelectron microscopy, and particularly electron paramagnetic resonance (EPR) spectroscopy performed on spin-labeled Bax. Incubation with detergents, reconstitution, and Bid-triggered insertion into liposomes were found to be effective in inducing Bax oligomerization. Bid was shown to activate Bax independently of the stoichiometric ratio, suggesting that Bid has a catalytic function and that the interaction with Bax is transient. The formation of a stable dimerization interface involving two Bcl-2 homology 3 (BH3) domains was found to be the nucleation event for Bax homo-oligomerization. Based on intermolecular distance determined by EPR, a model of six adjacent Bax molecules in the oligomer is presented where the hydrophobic hairpins (helices α5 and α6) are equally spaced in the membrane and the two BH3 domains are in close vicinity in the dimer interface, separated by >5 nm from the next BH3 pairs. PMID:20008353

  20. Automated synthesis, insertion and detection of polyps for CT colonography

    NASA Astrophysics Data System (ADS)

    Sezille, Nicolas; Sadleir, Robert J. T.; Whelan, Paul F.

    2003-03-01

    CT Colonography (CTC) is a new non-invasive colon imaging technique which has the potential to replace conventional colonoscopy for colorectal cancer screening. A novel system which facilitates automated detection of colorectal polyps at CTC is introduced. As exhaustive testing of such a system using real patient data is not feasible, more complete testing is achieved through synthesis of artificial polyps and insertion into real datasets. The polyp insertion is semi-automatic: candidate points are manually selected using a custom GUI, suitable points are determined automatically from an analysis of the local neighborhood surrounding each of the candidate points. Local density and orientation information are used to generate polyps based on an elliptical model. Anomalies are identified from the modified dataset by analyzing the axial images. Detected anomalies are classified as potential polyps or natural features using 3D morphological techniques. The final results are flagged for review. The system was evaluated using 15 scenarios. The sensitivity of the system was found to be 65% with 34% false positive detections. Automated diagnosis at CTC is possible and thorough testing is facilitated by augmenting real patient data with computer generated polyps. Ultimately, automated diagnosis will enhance standard CTC and increase performance.