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1

Arabidopsis T-DNA insertional lines for CDC25 are hypersensitive to hydroxyurea but not to zeocin or salt stress  

PubMed Central

Background and Aims In yeasts and animals, cyclin-dependent kinases are key regulators of cell cycle progression and are negatively and positively regulated by WEE1 kinase and CDC25 phosphatase, respectively. In higher plants a full-length orthologue of CDC25 has not been isolated but a shorter gene with homology only to the C-terminal catalytic domain is present. The Arabidopis thaliana;CDC25 can act as a phosphatase in vitro. Since in arabidopsis, WEE1 plays an important role in the DNA damage/DNA replication checkpoints, the role of Arath;CDC25 in conditions that induce these checkpoints or induce abiotic stress was tested. Methods arath;cdc25 T-DNA insertion lines, Arath;CDC25 over-expressing lines and wild type were challenged with hydroxyurea (HU) and zeocin, substances that stall DNA replication and damage DNA, respectively, together with an abiotic stressor, NaCl. A molecular and phenotypic assessment was made of all genotypes Key Results There was a null phenotypic response to perturbation of Arath;CDC25 expression under control conditions. However, compared with wild type, the arath;cdc25 T-DNA insertion lines were hypersensitive to HU, whereas the Arath;CDC25 over-expressing lines were relatively insensitive. In particular, the over-expressing lines consistently outgrew the T-DNA insertion lines and wild type when challenged with HU. All genotypes were equally sensitive to zeocin and NaCl. Conclusions Arath;CDC25 plays a role in overcoming stress imposed by HU, an agent know to induce the DNA replication checkpoint in arabidopsis. However, it could not enhance tolerance to either a zeocin treatment, known to induce DNA damage, or salinity stress.

Spadafora, Natasha D.; Doonan, John H.; Herbert, Robert J.; Bitonti, M. Beatrice; Wallace, Emily; Rogers, Hilary J.; Francis, Dennis

2011-01-01

2

Identification and characterization of T-DNA inserts by T-DNA fingerprinting  

Microsoft Academic Search

A T-DNA fingerprinting method is presented based on amplified fragmentlength polymorphism with an anchored polymerase chain reaction step. Thismethod allows discrimination between different T-DNA inserts in stablytransformed plants. The technique was evaluated by analyzing 51 transgenicArabidopsis lines that had been characterized in detail by genomicblotting. Comparison of the obtained fingerprints with the availableintegration information demonstrated that fingerprints were correlated tothe

I. Theuns; P. Windels; S. De Buck; A. Depicker; E. Van Bockstaele; M. De Loose

2002-01-01

3

Generation and Characterization of Arabidopsis T-DNA Insertion Mutants.  

PubMed

Transfer DNA (T-DNA) insertion mutants are often used in forward and reverse genetics to reveal the molecular mechanisms of a particular biological process in plants. To generate T-DNA insertion mutants, T-DNA must be inserted randomly in the genome through transformation mediated by Agrobacterium tumefaciens. During generation of a T-DNA insertion mutant, Agrobacterium competent cells are first prepared and plasmids containing the T-DNA introduced into Agrobacterium cells. Agrobacterium containing T-DNA vectors are then used to transform T-DNA into Arabidopsis. After screening and identifying T-DNA insertion mutants with interesting phenotypes, genomic DNA is extracted from the mutants and used to isolate the T-DNA flanking sequences. To finally determine the mutated genes causing the specific phenotype in the T-DNA insertion mutants, cosegregation analysis and complementation or recapitulation analysis are needed. In this chapter, we describe detailed protocols for generation and characterization of T-DNA insertion mutants. PMID:24057370

Qu, Li-Jia; Qin, Genji

2014-01-01

4

SNP discovery and genetic mapping of T-DNA insertional mutants in Fragaria vesca L  

Microsoft Academic Search

As part of a program to develop forward and reverse genetics platforms in the diploid strawberry [Fragaria vesca L.; (2n = 2x = 14)] we have generated insertional mutant lines by T-DNA mutagenesis using pCAMBIA vectors. To characterize the T-DNA insertion\\u000a sites of a population of 108 unique single copy mutants, we utilized thermal asymmetric interlaced PCR (hiTAIL-PCR) to amplify\\u000a the flanking region surrounding

J. J. Ruiz-Rojas; D. J. Sargent; V. Shulaev; A. W. Dickerman; J. Pattison; S. H. Holt; A. Ciordia; Richard E. Veilleux

2010-01-01

5

Generation and Characterization of the Western Regional Research Center Brachypodium T-DNA Insertional Mutant Collection  

PubMed Central

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 optimized our Agrobacterium tumefaciens-mediated high-efficiency transformation method and generated 8,491 Brachypodium T-DNA lines. We used inverse PCR to sequence the DNA flanking the insertion sites in the mutants. Using these flanking sequence tags (FSTs) we were able to assign 7,389 FSTs from 4,402 T-DNA mutants to 5,285 specific insertion sites (ISs) in the Brachypodium genome. More than 29% of the assigned ISs are supported by multiple FSTs. T-DNA insertions span the entire genome with an average of 19.3 insertions/Mb. The distribution of T-DNA insertions is non-uniform with a larger number of insertions at the distal ends compared to the centromeric regions of the chromosomes. Insertions are correlated with genic regions, but are biased toward UTRs and non-coding regions within 1 kb of genes over exons and intron regions. More than 1,300 unique genes have been tagged in this population. Information about the Western Regional Research Center Brachypodium insertional mutant population is available on a searchable website (http://brachypodium.pw.usda.gov) designed to provide researchers with a means to order T-DNA lines with mutations in genes of interest.

Gordon, Sean P.; Guttman, Mara E.; Thilmony, Roger; Lazo, Gerard R.; Gu, Yong Q.; Vogel, John P.

2012-01-01

6

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

PubMed

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 (RB) end of the T-DNA is largely preserved whereas the left border (LB) 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

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

2012-12-02

7

Insertional mutagenesis in Arabidopsis thaliana : isolation of a T-DNA-linked mutation that alters leaf morphology  

Microsoft Academic Search

We investigated the potential of the Agrobacterium tumefaciens T-DNA as an insertional mutagen in Arabidopsis thaliana. Arabidopsis lines transformed with different T-DNA vectors were generated using a leaf disc infection procedure adapted for efficient selection on either kanamycin or hygromycin medium. A standardized screening procedure was developed for the detection of recessive mutations in T2 populations of regenerated and\\/or transformed

M. Van Lijsebettens; R. Vanderhaeghen; M. Van Montagu

1991-01-01

8

Characterization of T-DNA insertion patterns in the genome of rice blast fungus Magnaporthe oryzae  

Microsoft Academic Search

Agrobacterium tumefaciens-mediated transformation (ATMT) has been proven to be a powerful strategy for gene disruption in plants and fungi. Patterns\\u000a associated with transferred DNA (T-DNA) integration in plants and yeast have been studied comprehensively, whereas no detailed\\u000a analysis of T-DNA integration has been reported yet in filamentous fungi. Here, we reported the T-DNA insertion patterns in\\u000a the genome of filamentous

Guihua Li; Zhuangzhi Zhou; Guifu Liu; Fucong Zheng; Chaozu He

2007-01-01

9

T-DNA insertion mutagenesis in Arabidopsis: going back and forth  

Microsoft Academic Search

T-DNA insertion mutagenesis in Arabidopsis has been instrumental in advancing our knowledge of the physiology, biochemistry and development of plants. In just a few years plant biology, the late starter of molecular genetics, has placed itself at the forefront of science in large measure as a result of the more than 20 000 T-DNA transformants that have been generated in

Ricardo Azpiroz-Leehan; Kenneth A. Feldmann

1997-01-01

10

Probing the roles of LRR RLK genes in Arabidopsis thaliana roots using a custom T-DNA insertion set.  

PubMed

Leucine-rich repeat receptor-like protein kinases (LRR RLKs) represent the largest group of Arabidopsis RLKs with approximately 235 members. A minority of these LRR RLKs have been assigned to diverse roles in development, pathogen resistance and hormone perception. Using a reverse genetics approach, a collection of homozygous T-DNA insertion lines for 69 root expressed LRR RLK genes was screened for root developmental defects and altered response after exposure to environmental, hormonal/chemical and abiotic stress. The obtained data demonstrate that LRR RLKs play a role in a wide variety of signal transduction pathways related to hormone and abiotic stress responses. The described collection of T-DNA insertion mutants provides a valuable tool for future research into the function of LRR RLK genes. PMID:21431781

ten Hove, Colette A; Bochdanovits, Zoltán; Jansweijer, Vera M A; Koning, Fenne G; Berke, Lidija; Sanchez-Perez, Gabino F; Scheres, Ben; Heidstra, Renze

2011-03-24

11

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

Microsoft Academic Search

A large collection of T-DNA insertion transformants of Arabidopsis thaliana has been generated at the Institute of Agronomic Research, Versailles, France. The molecular characterisation of the insertion sites is currently performed by sequencing genomic regions flanking the inserted T-DNA (FST). The almost complete sequence of the nuclear genome of A.thaliana provides the framework for organising FSTs in a genome oriented

Franck Samson; Véronique Brunaud; Sandrine Balzergue; B. Dubreucq; L. Lepiniec; G. Pelletier; Michel Caboche; Alain Lecharny

2002-01-01

12

Cytological characterization of four meiotic mutants of Arabidopsis isolated from T-DNA-transformed lines  

Microsoft Academic Search

A secondary screen of the Feldmann collection of T-DNA transformed Arabidopsis lines identified several meiotic mutants. We used a spreading technique combined with DAPI staining in a detailed cytogenetic analysis of meiotic chromosome behaviour in four of these mutants, all of which are putatively T-DNA tagged and therefore candidates for molecular and functional analysis of the mutated genes. Two of

K. J. Ross; P. Fransz; S. J. Armstrong; I. Vizir; B. Mulligan; F. C. H. Franklin; G. H. Jones

1997-01-01

13

Analyses of single-copy Arabidopsis T-DNA-transformed lines show that the presence of vector backbone sequences, short inverted repeats and DNA methylation is not sufficient or necessary for the induction of transgene silencing  

Microsoft Academic Search

In genetically transformed plants, transgene silen- cing has been correlated with multiple and complex insertions of foreign DNA, e.g. T-DNA and vector backbone sequences. Occasionally, single-copy transgenes also suffer transgene silencing. We have compared integration patterns and T-DNA\\/plant DNA junctions in a collection of 37 single-copy T-DNA-transformed Arabidopsis lines, of which 13 displayed silencing. Vector sequences were found integrated in

Trine J. Meza; Biljana Stangeland; Inderjit S. Mercy; Magne Skarn; Dag A. Nymoen; Anita Berg; Melinka A. Butenko; Anne-Mari Hakelien; Camilla Haslekas; Leonardo A. Meza-Zepeda; Reidunn B. Aalen

2002-01-01

14

Analyses of single-copy Arabidopsis T-DNA-transformed lines show that the presence of vector backbone sequences, short inverted repeats and DNA methylation is not sufficient or necessary for the induction of transgene silencing  

PubMed Central

In genetically transformed plants, transgene silencing has been correlated with multiple and complex insertions of foreign DNA, e.g. T-DNA and vector backbone sequences. Occasionally, single-copy transgenes also suffer transgene silencing. We have compared integration patterns and T-DNA/plant DNA junctions in a collection of 37 single-copy T-DNA-transformed Arabidopsis lines, of which 13 displayed silencing. Vector sequences were found integrated in five lines, but only one of these displayed silencing. Truncated T-DNA copies, positioned in inverse orientation to an intact T-DNA copy, were discovered in three lines. The whole nptII gene with pnos promoter was present in the truncated copy of one such line in which heavy silencing has been observed. In the two other lines no silencing has been observed over five generations. Thus, vector sequences and short additional T-DNA sequences are not sufficient or necessary to induce transgene silencing. DNA methylation of selected restriction endonuclease sites could not be correlated with silencing. Our collection of T-DNA/plant DNA junctions has also been used to evaluate current models of T-DNA integration. Data for some of our lines are compatible with T-DNA integration in double-strand breaks, while for others initial invasion of plant DNA by the left or by the right T-DNA end seem important.

Meza, Trine J.; Stangeland, Biljana; Mercy, Inderjit S.; Skarn, Magne; Nymoen, Dag A.; Berg, Anita; Butenko, Melinka A.; Hakelien, Anne-Mari; Haslekas, Camilla; Meza-Zepeda, Leonardo A.; Aalen, Reidunn B.

2002-01-01

15

Characterization of a T-DNA promoter trap line of Arabidopsis thaliana uncovers a cryptic bi-directional promoter.  

PubMed

Investigation of the transgenic Arabidopsis promoter trap line GFP-868 that showed GFP expression only in anthers revealed the T-DNA insertion at 461bp upstream to the hypothetical gene At4g10596 with the GFP reporter gene in head-to-head orientation to the At4g10596 gene. The expression of the At4g10596 gene in wild type and in GFP-868 plant homozygous for T-DNA insertion was comparable and found in all tissues tested, while the GFP expression was restricted to anthers of the GFP-868 plants suggesting that the 461bp fragment separating the two genes in the GFP-868 line is functioning as bi-directional promoter. This 461bp fragment was cloned upstream to the GUS gene in two orientations to test for bi-directional promoter activity. Transgenic Arabidopsis plants carrying either of these constructs showed GUS activity in anthers indicating that this fragment behaves as bi-directional promoter specific to anthers. These results were also supported by the presence of cis-acting motifs such as TATA box and POLLEN1LELAT52 (AGAAA) within the 461bp sequence in both orientations. However, transcripts corresponding to the upstream sequences beyond -461 nucleotides were not detected in the wild type suggesting that this 461bp fragment is a cryptic promoter. The significance of the promoter trap approach and the usefulness of this type of promoter are discussed. PMID:23612249

Pratibha, Pritu; Singh, Sunil Kumar; Sharma, Isha; Kumar, Ravi; Srinivasan, Ramamurthy; Bhat, Shripad Ramachandra; Ahuja, Paramvir Singh; Sreenivasulu, Yelam

2013-04-21

16

Development of a highly effective T-DNA inserted mutant screening method in a Chinese cabbage ( Brassica rapa L. spp. pekinensis ) reverse genetics system  

Microsoft Academic Search

We present a highly effective T-DNA inserted gene screening method as part of a reverse genetics model system using the Chinese\\u000a cabbage (Brassica rapa L. spp. pekinensis). Three-step two-dimensional (2D) matrix strategies are potentially accurate and useful for the identification of specific\\u000a T-DNA inserted mutants from a large population. To construct our Chinese cabbage model, we utilized a forward genetics

Gi-Ho Lee; Yoon-Jee Kang; Seul-Ki Yi; Suk-Bin Lim; Young-Doo Park

2010-01-01

17

Multi-functional T-DNA/Ds tomato lines designed for gene cloning and molecular and physical dissection of the tomato genome.  

PubMed

In order to make the tomato genome more accessible for molecular analysis and gene cloning, we have produced 405 individual tomato (Lycopersicon esculentum) lines containing a characterized copy of pJasm13, a multifunctional T-DNA/modified Ds transposon element construct. Both the T-DNA and the Ds element in pJasm13 harbor a set of selectable marker genes to monitor excision and reintegration of Ds and additionally, target sequences for rare cutting restriction enzymes (I-PpoI, SfiI, NotI) and for site-specific recombinases (Cre, FLP, R). Blast analysis of flanking genomic sequences of 174 T-DNA inserts revealed homology to transcribed genes in 69 (40%), of which about half are known or putatively identified as genes and ESTs. The map position of 140 individual inserts was determined on the molecular genetic map of tomato. These inserts are distributed over the 12 chromosomes of tomato, allowing targeted and non-targeted transposon tagging, marking of closely linked genes of interest and induction of chromosomal rearrangements including translocations or creation of saturation-deletions or inversions within defined regions linked to the T-DNA insertion site. The different features of pJasm13 were successfully tested in tomato and Arabidopsis thaliana, thus providing a new tool for molecular/genetic dissection studies, including molecular and physical mapping, mutation analysis and cloning strategies in tomato and potentially, in other plants as well. PMID:12602893

Gidoni, D; Fuss, E; Burbidge, A; Speckmann, G J; James, S; Nijkamp, D; Mett, A; Feiler, J; Smoker, M; de Vroomen, M J; Leader, D; Liharska, T; Groenendijk, J; Coppoolse, E; Smit, J J M; Levin, I; de Both, M; Schuch, W; Jones, J D G; Taylor, I B; Theres, K; van Haaren, M J J

2003-01-01

18

Major Chromosomal Rearrangements Induced by T-DNA Transformation in Arabidopsis  

Microsoft Academic Search

We show that major chromosomal rearrangements can occur upon T-DNA transformation of Arabidopsis thaliana. In the ACL4 line, two T-DNA insertion loci were found; one is a tandem T-DNA insert in a head- to-head orientation, and the other is a truncated insert with only the left part of the T-region. The four flanking DNA regions were isolated and located on

Philippe Nacry; Christine Camilleri; Beatrice Courtial; Michel Caboche; David Bouchez

19

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

PubMed Central

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.

An, Gynheung

2013-01-01

20

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

PubMed

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

Kim, Song Lim; Choi, Minkyung; Jung, Ki-Hong; An, Gynheung

2013-08-21

21

Oryza Tag Line, a phenotypic mutant database for the G?noplante rice insertion line library  

PubMed Central

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.

Larmande, Pierre; Gay, Celine; Lorieux, Mathias; Perin, Christophe; Bouniol, Matthieu; Droc, Gaetan; Sallaud, Christophe; Perez, Pascual; Barnola, Isabelle; Biderre-Petit, Corinne; Martin, Jerome; Morel, Jean Benoit; Johnson, Alexander A. T.; Bourgis, Fabienne; Ghesquiere, Alain; Ruiz, Manuel; Courtois, Brigitte; Guiderdoni, Emmanuel

2008-01-01

22

Localization of T-DNA Insertions in Petunia by Fluorescence in Situ Hybridization: Physical Evidence for Suppression of Recombination.  

PubMed Central

Using fluorescence in situ hybridization (FISH) with metaphase preparations, we localized a 4-kb single-copy T-DNA sequence in a group of petunia transformants. The selected T-DNAs previously had been shown to be linked to the phenotypic marker FI on chromosome II. Linkage analysis had revealed that recombination around the FI locus is suppressed in a wide cross relative to an inbred recombination assay. The localization of six FI-linked T-DNAs and the FI locus itself, using FISH, revealed a number of aspects of recombination in petunia: (1) the central region of chromosome II showed at least a 10-fold suppression of recombination in wide crosses relative to the distal region; (2) recombination in wide hybrids over two-thirds of the chromosome was extremely low; and (3) recombination between completely homologous chromosomes in an inbred cross also was suppressed in the central region. In addition, the T-DNAs were not evenly distributed along the chromosome, suggesting a possible preference for a distal position for T-DNA integration. Implications for such a preference are discussed.

Ten Hoopen, R; Robbins, TP; Fransz, PF; Montijn, BM; Oud, O; Gerats, A; Nanninga, N

1996-01-01

23

Single-copy T-DNA insertions in Arabidopsis are the predominant form of integration in root-derived transgenics, whereas multiple insertions are found in leaf discs  

Microsoft Academic Search

Different patterns of T-DNA integration in Arabidopsis were obtained that depended on whether a root or a leaf-disc transformation method was used. An examination of 82 individual transgenic Arabidopsis plants, derived from 15 independent Agrobacterium-mediated transformations in which different cointegrate and binary constructs were used, indicated that the transformation method had a significant influence on the type and copy number

Christoph Grevelding; Verena Fantes; Elke Kemper; Jeff Schell; Robert Masterson

1993-01-01

24

ATIDB: Arabidopsis thaliana insertion database  

Microsoft Academic Search

Insertional mutagenesis techniques, including transposon- and T-DNA-mediated mutagenesis, are key resources for systematic identification of gene function in the model plant species Arabidopsis thaliana. We have developed a database (http:\\/\\/ atidb.cshl.org\\/) for archiving, searching and analyz- ing insertional mutagenesis lines. Flanking sequences from approximately 10 500 insertion lines (including transposon and T-DNA insertions) from several tagging programs in Arabidopsis were

Xiaokang Pan; Hong Liu; Jonathan Clarke; Jonathan Jones; Mike Bevan; Lincoln Stein

2003-01-01

25

A T-DNA insertion knockout of the bifunctional lysine-ketoglutarate reductase/saccharopine dehydrogenase gene elevates lysine levels in Arabidopsis seeds.  

PubMed

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

Zhu, X; Tang, G; Granier, F; Bouchez, D; Galili, G

2001-08-01

26

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

PubMed Central

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

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

2001-01-01

27

Major chromosomal rearrangements induced by T-DNA transformation in Arabidopsis.  

PubMed Central

We show that major chromosomal rearrangements can occur upon T-DNA transformation of Arabidopsis thaliana. In the ACL4 line, two T-DNA insertion loci were found; one is a tandem T-DNA insert in a head-to-head orientation, and the other is a truncated insert with only the left part of the T-region. The four flanking DNA regions were isolated and located on the Arabidopsis chromosomes; for both inserts, one side of the T-DNA maps to chromosome 2, whereas the other side maps to chromosome 3. Both chromosome 3 flanking regions map to the same location, despite a 1.4-kb deletion at this point, whereas chromosome 2 flanking regions are located 40 cM apart on the bottom arm of chromosome 2. These results strongly suggest a reciprocal translocation between chromosomes 2 and 3, with the breakpoints located at the T-DNA insertion sites. The interchanged fragments roughly correspond to the 20-cM distal ends of both chromosomes. Moreover, a large inversion, spanning 40 cM on the genetic map, occurs on the bottom arm of chromosome 2. This was confirmed by genetic analyses that demonstrated a strong reduction of recombination in the inverted region. Models for T-DNA integration and the consequences for T-DNA tagging are discussed in light of these results.

Nacry, P; Camilleri, C; Courtial, B; Caboche, M; Bouchez, D

1998-01-01

28

An Arabidopsis thaliana T-DNA mutagenized population (GABI-Kat) for flanking sequence tag-based reverse genetics  

Microsoft Academic Search

The GABI-Kat population of T-DNA mutagenized Arabidopsis thaliana lines with sequence-characterized insertion sites is used extensively for efficient progress in plant functional genomics. Here we provide details about the establishment of the material, demonstrate the population's functionality and discuss results from quality control studies. T-DNA insertion mutants of the accession Columbia (Col-0) were created by Agrobacterium tumefaciens-mediated transformation. To allow

Mario G. Rosso; Yong Li; Nicolai Strizhov; Bernd Reiss; Koen Dekker; Bernd Weisshaar

2003-01-01

29

Transoesophageal echocardiography and central line insertion.  

PubMed

We investigated the potential utility of transoesophageal echocardiography (TOE) in facilitating central venous catheter (CVC) insertion in patients undergoing cardiac surgery. Thirty five patients undergoing elective cardiac surgery and CVC insertion were prospectively included in the observational, single-centre clinical investigation. Following induction of general anaesthesia and tracheal intubation, the TOE probe was inserted and the bicaval view obtained prior to CVC insertion (site at discretion of the anaesthesiologist). Prospectively collected data included site and sequence of CVC insertion attempts, information regarding ease of guidewire insertion, whether or not guidewire was visualized via TOE, and other pertinent information. In 1 patient, the TOE bicaval view could not be readily obtained because of right atrial (RA) distortion. In 31 patients, the TOE bicaval view was obtained and CVC access was successful at the site of first choice (guidewire visualized in all). Three patients had noteworthy CVC insertions. In one, CVC insertion was difficult despite visualization ofguidewire in the RA. In another, multiple guidewire insertions met with substantial resistance and without visualization of guidewire in the RA. One patient was found to have an unanticipated large mobile superior vena cava thrombus that extended into the RA, which changed clinical management by prompting initial CVC insertion into the femoral vein (potentially avoiding morbidity associated with thrombus dislodgement). Our prospective observational clinical study indicates that routine use of TOE during CVC insertion may help avoid potential complications associated with this intervention. If both CVC insertion and TOE are going to be used in the same patient, the benefits of TOE should be maximized by routine visualization of the bicaval view during guidewire insertion. PMID:17644885

Chaney, Mark A; Minhaj, Mohammed M; Patel, Komal; Muzic, David

2007-07-01

30

An intron sense suppression construct targeting soybean FAD2-1 requires a double-stranded RNA-producing inverted repeat T-DNA insert.  

PubMed

We demonstrate that the transformation of soybean (Glycine max) with sense suppression constructs using intron sequences from the fatty acid oleyl Delta12 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

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

2010-04-27

31

Systematic Reverse Genetic Screening of T-DNA Tagged Genes in Rice for Functional Genomic Analyses: MADS-box Genes as a Test Case  

Microsoft Academic Search

; We have generated 47 DNA pools and 235 subpools from 21,049 T-DNA insertion lines of rice. DNA pools of 500-1,000 lines were adequate for screening a T-DNA inser- tion within a 2-kb region. To examine the efficacy of the DNA pools, we selected MADS-box genes, which play an important role in controlling various aspects of plant devel- opment. A

Shinyoung Lee; Joonyul Kim; Jun-Seock Son; Jongmin Nam; Dong-Hoon Jeong; Keunsub Lee; Seonghoe Jang; Jihye Yoo; Jinwon Lee; Dong-Yeon Lee; Hong-Gyu Kang

2003-01-01

32

Forward Genetics Screening of Medicago truncatula Tnt1 Insertion Lines.  

PubMed

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

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

2013-01-01

33

Vacuolar cation/H+ exchange, ion homeostasis, and leaf development are altered in a T-DNA insertional mutant of AtNHX1, the Arabidopsis vacuolar Na+/H+ antiporter.  

PubMed

The function of vacuolar Na+/H+ antiporter(s) in plants has been studied primarily in the context of salinity tolerance. By facilitating the accumulation of Na+ away from the cytosol, plant cells can avert ion toxicity and also utilize vacuolar Na+ as osmoticum to maintain turgor. As many genes encoding these antiporters have been cloned from salt-sensitive plants, it is likely that they function in some capacity other than salinity tolerance. The wide expression pattern of Arabidopsis thaliana sodium proton exchanger 1 (AtNHX1) in this study supports this hypothesis. Here, we report the isolation of a T-DNA insertional mutant of AtNHX1, a vacuolar Na+/H+ antiporter in Arabidopsis. Vacuoles isolated from leaves of the nhx1 plants had a much lower Na+/H+ and K+/H+ exchange activity. nhx1 plants also showed an altered leaf development, with reduction in the frequency of large epidermal cells and a reduction in overall leaf area compared to wild-type plants. The overexpression of AtNHX1 in the nhx1 background complemented these phenotypes. In the presence of NaCl, nhx1 seedling establishment was impaired. These results place AtNHX1 as the dominant K+ and Na+/H+ antiporter in leaf vacuoles in Arabidopsis and also suggest that its contribution to ion homeostasis is important for not only salinity tolerance but development as well. PMID:14535887

Apse, Maris P; Sottosanto, Jordan B; Blumwald, Eduardo

2003-10-01

34

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

PubMed Central

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.

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

2012-01-01

35

Technical note: development of a tool to insert abomasal infusion lines into dairy cows.  

PubMed

A tool was developed to aid in ruminal insertion of abomasal infusion lines into dairy cows. The tool consisted of 2 pieces cut from polyvinyl chloride pipe. The first piece of pipe, the insertion tool, contained a groove that held the flexible plastic flange that is on the end of the infusion line. The insertion tool containing the flange was inserted into the ruminal cannula, through the sulcus omasi, and into the abomasum. The second piece of pipe, the delivery tool, was threaded through the insertion tool, and it was used to dislodge the flange from the insertion tool and into the abomasum. PMID:16960071

Gressley, T F; Reynal, S M; Colmenero, J J Olmos; Broderick, G A; Armentano, L E

2006-10-01

36

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

PubMed Central

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.

2013-01-01

37

Identification of transferred DNA insertions within Arabidopsis genes involved in signal transduction and ion transport.  

PubMed Central

The transferred DNA (T-DNA) of Agrobacterium tumefaciens serves as an insertional mutagen once integrated into a host plant's genome. As a means of facilitating reverse genetic analysis in Arabidopsis thaliana, we have developed a method that allows one to search for plants carrying F-DNA insertions within any sequenced Arabidopsis gene. Using PCR, we screened a collection of 9100 independent T-DNA-transformed Arabidopsis lines and found 17 T-DNA insertions within the 63 genes analyzed. The genes surveyed include members of various gene families involved in signal transduction and ion transport. As an example, data are shown for a T-DNA insertion that was found within CPK-9, a member of the gene family encoding calmodulin-domain protein kinases. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5

Krysan, P J; Young, J C; Tax, F; Sussman, M R

1996-01-01

38

Genomic rearrangements by LINE1 insertion-mediated deletion in the human and chimpanzee lineages  

Microsoft Academic Search

Long INterspersed Elements (LINE-1s or L1s) are abundant non-LTR retrotransposons in mammalian genomes that are capable of insertional mutagen- esis. They have been associated with target site deletions upon insertion in cell culture studies of retrotransposition. Here, we report 50 deletion events in the human and chimpanzee genomes dir- ectly linked to the insertion of L1 elements, resulting in the

Kyudong Han; Shurjo K. Sen; Jianxin Wang; Pauline A. Callinan; Jungnam Lee; Richard Cordaux; Ping Liang; Mark A. Batzer

2005-01-01

39

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 ...

40

Silent T-DNA genes in plant lines transformed by Agrobacterium tumefaciens are activated by grafting and by 5-azacytidine treatment  

Microsoft Academic Search

A shooty tumor induced by a shooter mutant of an octopine strain of Agrobacterium tumefaciens was cloned. One clone obtained (TS038) behaved aberrantly in that it grew as a shooty tumor tissue on phytohormone free medium, but did not contain octopine synthase activity. In line TS038 the genes for octopine synthase and for the enzymes involved in agropine and mannopine

G. M. S. Slogteren; P. J. J. Hooykaas; R. A. Schilperoort

1984-01-01

41

T-DNA border sequences required for crown gall tumorigenesis.  

PubMed Central

Similar 23-base-pair (bp) direct repeats occur at the ends of two adjacent but noncontiguous T-DNAs, TL and TR (left and right T-DNA), in the tumor-inducing plasmid pTiA6NC. Thus, three border repeats lie right and one lies left of TL, which carries the genes needed for tumor maintenance. To determine whether T-DNA transfer and integration (subsequently called T-DNA transmission) require sequences in addition to the 23-bp border repeat, we constructed a deletion removing the three potential TL right borders (the TL right border and both TR borders). Since this deletion severely attenuated virulence, we reintroduced restriction fragments containing the TL right border repeat at a new location to the right of TL and tested their ability to restore virulence. Fragments that carried the border repeat flanked by at least 67 bp of wild-type Ti plasmid sequences on the left and 1035 bp on the right restored virulence completely. Smaller fragments restored virulence significantly but not fully, even though the border repeat remained intact. Therefore, T-region sequences flanking the border repeat in the fully active fragments stimulated T-DNA integration. Fragments that restored virulence fully when inserted in the wild-type orientation stimulated virulence only slightly in the opposite orientation. Thus, the right border sequence promotes T-DNA transfer and integration best in one direction. Images

Peralta, E G; Ream, L W

1985-01-01

42

Trans-repression of gene activity upstream of T-DNA tagged RLK902 links Arabidopsis root growth inhibition and downy mildew resistance.  

PubMed

Receptor-like kinases (RLKs) constitute a large family of signal perception molecules in Arabidopsis. The largest group of RLKs is the leucine-rich repeat (LRR) class that has been described to function in development and defense. Of these, CLAVATA1 (CLV1) and ERECTA (ER) receptors function in maintaining shoot meristem homeostasis and organ growth, but LRR RLKs with similar function in the root remain unknown. For the interaction of Arabidopsis with the oomycete pathogen Hyaloperonospora arabidopsidis the involvement of LRR RLKs has not been demonstrated. A set of homozygous T-DNA insertion lines mutated in LRR RLKs was investigated to assess the potential role of these receptors in root meristem maintenance and compatibility. One mutant line, rlk902, was discovered that showed both reduced root growth and resistance to downy mildew in a recessive manner. The phenotypes of this mutated line could not be rescued by complementation, but are nevertheless linked to the T-DNA insertion. Microarray studies showed that gene expression spanning a region of approximately 84 kb upstream of the mutated gene was downregulated. The results suggest T-DNA mediated trans-repression of multiple genes upstream of the RLK902 locus links both phenotypes. PMID:21532992

ten Hove, Colette A; de Jong, Mark; Lapin, Dmitry; Andel, Annemiek; Sanchez-Perez, Gabino F; Tarutani, Yoshiaki; Suzuki, Yoshihito; Heidstra, Renze; van den Ackerveken, Guido

2011-04-21

43

Trans-Repression of Gene Activity Upstream of T-DNA Tagged RLK902 Links Arabidopsis Root Growth Inhibition and Downy Mildew Resistance  

PubMed Central

Receptor-like kinases (RLKs) constitute a large family of signal perception molecules in Arabidopsis. The largest group of RLKs is the leucine-rich repeat (LRR) class that has been described to function in development and defense. Of these, CLAVATA1 (CLV1) and ERECTA (ER) receptors function in maintaining shoot meristem homeostasis and organ growth, but LRR RLKs with similar function in the root remain unknown. For the interaction of Arabidopsis with the oomycete pathogen Hyaloperonospora arabidopsidis the involvement of LRR RLKs has not been demonstrated. A set of homozygous T-DNA insertion lines mutated in LRR RLKs was investigated to assess the potential role of these receptors in root meristem maintenance and compatibility. One mutant line, rlk902, was discovered that showed both reduced root growth and resistance to downy mildew in a recessive manner. The phenotypes of this mutated line could not be rescued by complementation, but are nevertheless linked to the T-DNA insertion. Microarray studies showed that gene expression spanning a region of approximately 84 kb upstream of the mutated gene was downregulated. The results suggest T-DNA mediated trans-repression of multiple genes upstream of the RLK902 locus links both phenotypes.

Lapin, Dmitry; Andel, Annemiek; Sanchez-Perez, Gabino F.; Tarutani, Yoshiaki; Suzuki, Yoshihito; Heidstra, Renze; van den Ackerveken, Guido

2011-01-01

44

Paramutation-Like Interaction of T-DNA Loci in Arabidopsis  

PubMed Central

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.

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

2012-01-01

45

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

PubMed Central

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

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

1982-01-01

46

T-DNA locus structure in a large population of soybean plants transformed using the Agrobacterium-mediated cotyledonary-node method.  

PubMed

Designing transformation experiments for either functional genomics or crop improvement requires knowledge of the transgene locus structure, number, transmission and expression resulting from a specific transformation method. We recently reported an improvement to the soybean [Glycine max (L.) Merrill] cotyledonary-node transformation method that resulted in the efficient production of transgenic plants. To characterize the transgene loci resulting from this method, we analysed 270 independent T0 plants and 95 randomly selected T1 progenies for T-DNA locus complexity using Southern analysis. The lines were transformed with Agrobacterium tumefaciens strains LBA4404 or EHA105 carrying the binary plasmids pGPTV, pTOK233, pCAMBIA1303 or pCAMBIA1309, and regenerated in medium supplemented with or without silver nitrate (AgNO3). Analysis in the T0 generation showed that the number of hpt-hybridizing fragments per plant ranged from 1-15, with 31.5% of the lines having a single hpt-hybridizing fragment. Each primary soybean transformant had, on average, 2.0 unlinked transgene loci and that half of the segregating loci in the T1 progenies were single, simple T-DNA insertions. Of the loci containing multiple T-DNA fragments, a low frequency had tandem and inverted repeat T-DNA structures. Integration of binary plasmid backbone sequences occurred in 37% of primary transformants. A. tumefaciens strain, binary plasmid and thiol treatment had no significant effect on transgene locus structure, numbers or expression. Interestingly, exposure of soybean explants to AgNO3 throughout shoot induction and elongation increased T-DNA locus complexity in the primary transformants and decreased silencing of gusA expression in the T1 generation. PMID:17134390

Olhoft, Paula M; Flagel, Lex E; Somers, David A

2004-07-01

47

High-frequency T-DNA-mediated gene tagging in plants.  

PubMed Central

An insertion element [transferred DNA (T-DNA)], transferred by soil agrobacteria into the nuclear genome of plants, was used for induction of gene fusions in Arabidopsis thaliana, Nicotiana tabacum, and Nicotiana plumbaginifolia. A promoterless aph(3')II (aminoglycoside phosphotransferase II) reporter gene was linked to the right end of the T-DNA and transformed into plants along with a plasmid replicon and a selectable hygromycin-resistance gene. Transcriptional and translational reporter gene fusions were identified by screening for APH(3')II enzyme activity in diverse tissues of transgenic plants. The frequency of gene fusions, estimated by determination of the copy number of T-DNA insertions, showed that on average 30% of T-DNA inserts induced gene fusions in Arabidopsis and Nicotiana. Gene fusions were rescued from plants by transformation of the T-DNA-linked plasmid and flanking plant DNA into Escherichia coli. By dissection of gene fusions and construction of chimeric genes, callus- and root-specific promoters were identified that showed an altered tissue specificity in the presence of a 3'-downstream-located 35S promoter. Transcript mapping of a gene fusion and expression of a non-frame transcriptional fusion of bacterial luciferase luxA and luxB genes demonstrated that dicistronic transcripts are translated in tobacco. Images

Koncz, C; Martini, N; Mayerhofer, R; Koncz-Kalman, Z; Korber, H; Redei, G P; Schell, J

1989-01-01

48

Identification and characterization of novel polymorphic LINE1 insertions through comparison of two human genome sequence assemblies  

Microsoft Academic Search

Mobile elements represent a relatively new class of markers for the study of human evolution. Long interspersed elements (LINEs) belong to a group of retrotransposons comprising approximately 21% of the human genome. Young LINE-1 (L1) elements that have integrated recently into the human genome can be polymorphic for insertion presence\\/absence in different human populations at particular chromosomal locations. To identify

Miriam K. Konkel; Jianxin Wang; Ping Liang; Mark A. Batzer

2007-01-01

49

Towards a novel efficient T-DNA-based mutagenesis and screening system using green fluorescent protein as a vital reporter in the industrially important fungus Trichoderma reesei  

Microsoft Academic Search

Agrobacterium-mediated T-DNA transfer has been proven to be an efficient strategy for insertional mutagenesis and elucidation of gene function\\u000a in filamentous fungi. The implementation of large-scale T-DNA insertional mutagenesis requires the development of high-efficient\\u000a transformation and high-throughput screening procedures. Here, using green fluorescent protein (GFP) as a vital marker, a\\u000a highly efficient T-DNA-based mutagenesis and screening system was developed in

Yaohua Zhong; Haina Yu; Xiaoli Wang; Yi Lu; Tianhong Wang

2011-01-01

50

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

PubMed

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

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

2005-03-03

51

Aberrant pre-mRNA maturation is caused by LINE insertions into introns of the white gene of Drosophila melanogaster.  

PubMed Central

Insertional mutagenesis screens have provided thousands of mutant alleles for analysing genes of varied functions in Drosophila melanogaster. We here document mechanisms of insertional mutagenesis by a LINE element, the I factor, by determining the molecular structure of RNAs produced from two alleles of the white gene of D.melanogaster, wIR1 and wIR6. These alleles result from insertion of the I factor into introns of the gene. We show that sequences present within the element direct aberrant splicing and termination events. When the I factor is inserted within the white first intron it may lead to the use of a cryptic 3' splice site which does not contain the dinucleotide AG. This splicing gives rise to a chimeric messenger RNA whose synthesis is controlled differently in tissues where the mutated gene is expressed. When the I factor is inserted within the white last intron it induces synthesis of truncated mRNAs. These results provide, for the first time, mechanisms for I factor insertional mutagenesis. They are discussed in the more general context of RNA processing in Drosophila and the evolution of eukaryotic gene introns. Images

Lajoinie, O; Drake, M E; Dastugue, B; Vaury, C

1995-01-01

52

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

PubMed Central

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.

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

2004-01-01

53

Symbiotic mutants deficient in nodule establishment identified after T-DNA transformation of Lotus japonicus  

Microsoft Academic Search

Nitrogen-fixing root nodules develop on legumes as a result of an interaction between host plants and soil bacteria collectively\\u000a referred to as rhizobia. The organogenic process resulting in nodule development is triggered by the bacterial microsymbiont,\\u000a but genetically controlled by the host plant genome. Using T-DNA insertion as a tool to identify novel plant genes that regulate\\u000a nodule ontogeny, we

L. Schauser; K. Handberg; N. Sandal; J. Stiller; T. Thykjær; E. Pajuelo; A. Nielsen; J. Stougaard

1998-01-01

54

High-Throughput Selection of Retrovirus Producer Cell Lines Leads to Markedly Improved Efficiency of Germ Line-Transmissible Insertions in Zebra Fish  

Microsoft Academic Search

Vesicular stomatitis virus glycoprotein G-pseudotyped mouse retroviral vectors have been used as mutagens for a large-scale insertional mutagenesis screen in the zebra fish. To reproducibly generate high-titer virus stocks, we devised a method for rapidly selecting cell lines that can yield high-titer viruses and isolated a producer cell line that yields virus at a high titer on zebra fish embryos.

Wenbiao Chen; Shawn Burgess; Greg Golling; Adam Amsterdam; Nancy Hopkins

2002-01-01

55

Generation of Single-Copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-Mediated Resolution System1  

PubMed Central

We investigated whether complex T-DNA loci, often resulting in low transgene expression, can be resolved efficiently into single copies by CRE/loxP-mediated recombination. An SB-loxP T-DNA, containing two invertedly oriented loxP sequences located inside and immediately adjacent to the T-DNA border ends, was constructed. Regardless of the orientation and number of SB-loxP-derived T-DNAs integrated at one locus, recombination between the outermost loxP sequences in direct orientation should resolve multiple copies into a single T-DNA copy. Seven transformants with a complex SB-loxP locus were crossed with a CRE-expressing plant. In three hybrids, the complex T-DNA locus was reduced efficiently to a single-copy locus. Upon segregation of the CRE recombinase gene, only the simplified T-DNA locus was found in the progeny, demonstrating DNA had been excised efficiently in the progenitor cells of the gametes. In the two transformants with an inverted T-DNA repeat, the T-DNA resolution was accompanied by at least a 10-fold enhanced transgene expression. Therefore, the resolution of complex loci to a single-copy T-DNA insert by the CRE/loxP recombination system can become a valuable method for the production of elite transgenic Arabidopsis thaliana plants that are less prone to gene silencing.

De Buck, Sylvie; Peck, Ingrid; De Wilde, Chris; Marjanac, Gordana; Nolf, Jonah; De Paepe, Annelies; Depicker, Ann

2007-01-01

56

Bacterial transposons are co-transferred with T-DNA to rice chromosomes during Agrobacterium-mediated transformation.  

PubMed

Agrobacterium tumefaciens is widely utilized for delivering a foreign gene into a plant's genome. We found the bacterial transposon Tn5393 in transgenic rice plants. Analysis of the flanking sequences of the transferred-DNA (T-DNA) identified that a portion of the Tn5393 sequence was present immediately next to the end of the T-DNA. Because this transposon was present in A. tumefaciens strain LBA4404, but not in EHA105 and GV3101, our findings indicated that Tn5393 was transferred from LBA4404 into the rice genome during the transformation process. We also noted that another bacterial transposon, Tn5563, is present in transgenic plants. Analyses of 331 transgenic lines revealed that 26.0% carried Tn5393 and 2.1% contained Tn5563. In most of the lines, an intact transposon was integrated into the T-DNA and transferred to the rice chromosome. More than one copy of T-DNA was introduced into the plants, often at a single locus. This resulted in T-DNA repeats of normal and transposon-carrying TDNA that generated deletions of a portion of the T-DNA, joining the T-DNA end to the bacterial transposon. Based on these data, we suggest that one should carefully select the appropriate Agrobacterium strain to avoid undesirable transformation of such sequences. PMID:22570148

Kim, Sung-Ryul; An, Gynheung

2012-05-07

57

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

PubMed Central

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.

Santos, Efren; Remy, Serge; Thiry, Els; Windelinckx, Saskia; Swennen, Rony; Sagi, Laszlo

2009-01-01

58

Transcription of Agrobacterium rhizogenes A4 T-DNA  

Microsoft Academic Search

Two distinct segments of transferred DNA (T-DNA) were detected in tumor tissue incited by Agrobacterium rhizogenes strain A4 on Nicotiana glauca. RNA from these tumors was examined for the presence of T-DNA encoded transcripts. Six genetic loci involved in virulence and tumor morphology have been identified by transposon mutagenesis: rol A-D on the leftward T-DNA (TL-DNA) and tms1 and 2

Brian H. Taylor; Frank F. White; Eugene W. Nester; Milton P. Gordon

1985-01-01

59

The experiences of patients and nurses with a nurse-led peripherally inserted central venous catheter line service  

Microsoft Academic Search

Nurse-led Peripherally Inserted Central Venous Catheter (PICC) services are becoming commonplace in cancer centres across the UK. Research has shown that these services are cost-effective and are associated with fewer catheter-related complications, including infection and thrombosis, than skin-tunnelled catheters. This exploratory study aimed to explore patients' and nurses' experiences of a nurse-led PICC line service. Recorded interviews were conducted with

Catherine Oakley; Elizabeth Wright; Emma Ream

2000-01-01

60

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

PubMed Central

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.

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

2012-01-01

61

Suppression of transfer of non-T-DNA ‘vector backbone’ sequences by multiple left border repeats in vectors for transformation of higher plants mediated by Agrobacterium tumefaciens  

Microsoft Academic Search

Vectors for transformation of higher plants mediated by Agrobacterium tumefaciens were modified so that one, two or three additional copies of the left border (LB) sequences were inserted close to the original LB of the T-DNA. A gene for ß-glucuronidase (gusA) was placed outside the T-DNA to monitor the transfer to plants of 'vector backbone' sequences. The expression of GUS

Yoshiki Kuraya; Shozo Ohta; Miyuki Fukuda; Yukoh Hiei; Nobuhiko Murai; Kazuyuki Hamada; Jun Ueki; Hidemasa Imaseki; Toshihiko Komari

2004-01-01

62

Segregation distortion of T-DNA markers linked to the self-incompatibility (S) locus in Petunia hybrida.  

PubMed Central

In plants with a gametophytic self-incompatibility system the specificity of the pollen is determined by the haploid genotype at the self-incompatibility (S) locus. In certain crosses this can lead to the exclusion of half the gametes from the male parent carrying a particular S-allele. This leads to pronounced segregation distortion for any genetic markers that are linked to the S-locus. We have used this approach to identify T-DNA insertions carrying a maize transposable element that are linked to the S-locus of Petunia hybrida. A total of 83 T-DNA insertions were tested for segregation distortion of the selectable marker used during transformation with Agrobacterium. Segregation distortion was observed for 12 T-DNA insertions and at least 8 of these were shown to be in the same linkage group by intercrossing. This indicates that differential transmission of a single locus (S) is probably responsible for all of these examples of T-DNA segregation distortion. The identification of selectable markers in coupling with a functional S-allele will allow the preselection of recombination events around the S-locus in petunia. Our approach provides a general method for identifying transgenes that are linked to gametophytic self-incompatibility loci and provides an opportunity for transposon tagging of the petunia S-locus.

Harbord, R M; Napoli, C A; Robbins, T P

2000-01-01

63

Retroviral Elements and Their Hosts: Insertional Mutagenesis in the Mouse Germ Line  

PubMed Central

The inbred mouse is an invaluable model for human biology and disease. Nevertheless, when considering genetic mechanisms of variation and disease, it is important to appreciate the significant differences in the spectra of spontaneous mutations that distinguish these species. While insertions of transposable elements are responsible for only ~0.1% of de novo mutations in humans, the figure is 100-fold higher in the laboratory mouse. This striking difference is largely due to the ongoing activity of mouse endogenous retroviral elements. Here we briefly review mouse endogenous retroviruses (ERVs) and their influence on gene expression, analyze mechanisms of interaction between ERVs and the host cell, and summarize the variety of mutations caused by ERV insertions. The prevalence of mouse ERV activity indicates that the genome of the laboratory mouse is presently behind in the “arms race” against invasion.

Maksakova, Irina A; Romanish, Mark T; Gagnier, Liane; Dunn, Catherine A; van de Lagemaat, Louie N.; Mager, Dixie L

2006-01-01

64

Reversal of cystic fibrosis phenotype in a cultured ?508 cystic fibrosis transmembrane conductance regulator cell line by oligonucleotide insertion  

PubMed Central

Cystic fibrosis (CF) is a lethal genetic disorder that is due to mutations in the gene encoding the cAMP-activated anion CF transmembrane conductance regulator (CFTR) channel. A three-nucleotide base deletion (TTT), encoding phenylalanine in position 508 of the translatable CFTR sequence (accompanied by a C to T replacement immediately 5? to the deletion), accounts for ?75% of cases of the disease. In the present study, an oligonucleotide complex (CF4–CF6, 2?-0-methyl RNA–unmodified RNA oligonucleotide duplex, respectively) was used to restore CFTR function by insertion of missing bases in ?508 CFTR mRNA from a cultured (?508) cell line. cAMP-activated whole-cell currents and Cl– transport were detected in CF4–CF6-treated, but not control ?508, cells by patch-clamp and 6-methoxy-N-(3-sulfopropyl)quinolinium fluorescence (SPQ) quenching analyses, respectively. Further, the nucleotide addition in the deleted region of ?508 CFTR was determined after amplification by RT-PCR. Insertion of UGU and replacement of U by C immediately 5? to the deletion site in ?508 mRNA appear to have taken place, with phenotypic but not genotypic reversion in tissue culture of treated cells. The mechanism of insertion of nucleotides has yet to be determined.

Zamecnik, Paul C.; Raychowdhury, Malay K.; Tabatadze, David R.; Cantiello, Horacio F.

2004-01-01

65

Epigenetic control of Agrobacterium T-DNA integration  

PubMed Central

To genetically transform plants, Agrobacterium transfers its T-DNA into the host cell and integrates it into the plant genome, resulting in neoplastic growths. Over the past two decades, a great deal has been learned about the molecular mechanism by which Agrobacterium produces T-DNA and transports it into the host nucleus. However, T-DNA integration, which is the limiting, hence, the most critical step of the transformation process, largely remains an enigma. Increasing evidence suggests that Agrobacterium utilizes the host DNA repair machinery to facilitate T-DNA integration. Meanwhile, it is well known that chromatin modifications, including the phosphorylation of histone H2AX, play an important role in DNA repair. Thus, by implication, such epigenetic codes in chromatin may also have a considerable impact on T-DNA integration, although the direct evidence to demonstrate this hypothesis is still lacking. In this review, we summarize the recent advances in our understanding of Agrobacterium T-DNA integration and discuss the potential link between this process and the epigenetic information in the host chromatin.

Magori, Shimpei; Citovsky, Vitaly

2011-01-01

66

Characterization at nucleotide resolution of the homogeneously staining region sites of insertion in two cancer cell lines  

PubMed Central

The mechanisms of formation of intrachromosomal amplifications in tumours are still poorly understood. By using quantitative polymerase chain reaction, DNA sequencing, chromosome walking, in situ hybridization on metaphase chromosomes and whole-genome analysis, we studied two cancer cell lines containing an MYC oncogene amplification with acquired copies ectopically inserted in rearranged chromosomes 17. These intrachromosomal amplifications result from the integration of extrachromosomal DNA molecules. Replication stress could explain the formation of the double-strand breaks involved in their insertion and in the rearrangements of the targeted chromosomes. The sequences of the junctions indicate that homologous recombination was not involved in their formation and support a non-homologous end-joining process. The replication stress-inducible common fragile sites present in the amplicons may have driven the intrachromosomal amplifications. Mechanisms associating break-fusion-bridge cycles and/or chromosome fragmentation may have led to the formation of the uncovered complex structures. To our knowledge, this is the first characterization of an intrachromosomal amplification site at nucleotide resolution.

Gibaud, Anne; Vogt, Nicolas; Brison, Olivier; Debatisse, Michelle; Malfoy, Bernard

2013-01-01

67

Characterization at nucleotide resolution of the homogeneously staining region sites of insertion in two cancer cell lines.  

PubMed

The mechanisms of formation of intrachromosomal amplifications in tumours are still poorly understood. By using quantitative polymerase chain reaction, DNA sequencing, chromosome walking, in situ hybridization on metaphase chromosomes and whole-genome analysis, we studied two cancer cell lines containing an MYC oncogene amplification with acquired copies ectopically inserted in rearranged chromosomes 17. These intrachromosomal amplifications result from the integration of extrachromosomal DNA molecules. Replication stress could explain the formation of the double-strand breaks involved in their insertion and in the rearrangements of the targeted chromosomes. The sequences of the junctions indicate that homologous recombination was not involved in their formation and support a non-homologous end-joining process. The replication stress-inducible common fragile sites present in the amplicons may have driven the intrachromosomal amplifications. Mechanisms associating break-fusion-bridge cycles and/or chromosome fragmentation may have led to the formation of the uncovered complex structures. To our knowledge, this is the first characterization of an intrachromosomal amplification site at nucleotide resolution. PMID:23821669

Gibaud, Anne; Vogt, Nicolas; Brison, Olivier; Debatisse, Michelle; Malfoy, Bernard

2013-07-02

68

Retroviral insertional activation of the c-myb proto-oncogene in a Marek's disease T-lymphoma cell line.  

PubMed Central

Marek's disease virus (MDV) is an avian herpesvirus that causes, in chickens, a lymphoproliferative disease characterized by malignant transformation of T lymphocytes. The rapid onset of polyclonal tumors indicates the existence of MDV-encoded oncogenic products. However, the molecular basis of MDV-induced lymphoproliferative disease and latency remains largely unclear. Several lines of evidence suggest that MDV and Rous-associated virus (RAV) might cooperate in the development of B-cell lymphomas induced by RAV. Our present results indicate for the first time that MDV and RAV might also act synergistically in the development of T-cell lymphomas. We report an example of an MDV-transformed T-lymphoblastoid cell line (T9) expressing high levels of a truncated C-MYB protein as a result of RAV integration within one c-myb allele. The chimeric RAV-c-myb mRNA species initiated in the 5' long terminal repeat of RAV are deprived of sequences corresponding to c-myb exons 1 to 3. The attenuation of MDV oncogenicity has been strongly related to structural changes in the MDV BamHI-D and BamHI-H DNA fragments. We have established that both DNA restriction fragments are rearranged in the T9 MDV-transformed cells. Our results suggest that retroviral insertional activation of the c-myb proto-oncogene is a critical factor involved in the maintenance of the transformed phenotype and the tumorigenic potential of this T-lymphoma cell line.

Le Rouzic, E; Perbal, B

1996-01-01

69

Controlled insertional mutagenesis using a LINE-1 (ORFeus) gene-trap mouse model.  

PubMed

A codon-optimized mouse LINE-1 element, ORFeus, exhibits dramatically higher retrotransposition frequencies compared with its native long interspersed element 1 counterpart. To establish a retrotransposon-mediated mouse model with regulatable and potent mutagenic capabilities, we generated a tetracycline (tet)-regulated ORFeus element harboring a gene-trap cassette. Here, we show that mice expressing tet-ORFeus broadly exhibit robust retrotransposition in somatic tissues when treated with doxycycline. Consistent with a significant mutagenic burden, we observed a reduced number of double transgenic animals when treated with high-level doxycycline during embryogenesis. Transgene induction in skin resulted in a white spotting phenotype due to somatic ORFeus-mediated mutations that likely disrupt melanocyte development. The data suggest a high level of transposition in melanocyte precursors and consequent mutation of genes important for melanoblast proliferation, differentiation, or migration. These findings reveal the utility of a retrotransposon-based mutagenesis system as an alternative to existing DNA transposon systems. Moreover, breeding these mice to different tet-transactivator/reversible tet-transactivator lines supports broad functionality of tet-ORFeus because of the potential for dose-dependent, tissue-specific, and temporal-specific mutagenesis. PMID:23818630

O'Donnell, Kathryn A; An, Wenfeng; Schrum, Christina T; Wheelan, Sarah J; Boeke, Jef D

2013-07-01

70

Transcription of T-DNA in octopine and nopaline crown gall tumours is inhibited by low concentrations of alpha-amanitin.  

PubMed Central

Highly purified and physiologically active nuclei were isolated from four different octopine and nopaline crown gall lines. These nuclei exhibited a high endogenous RNA synthesizing activity involving all three RNA-polymerases I, II and III. Isolated nuclei were shown by Southern blotting to synthesize T-DNA specific RNA. This synthesis was shown to be sensitive to actinomycin D and therefore to be DNA-dependent. The transcription of the T-DNA was also inhibited for more than 90% by low concentrations of alpha-amanitin (0.7 micrograms/ml) indicating that the T-DNA, although from bacterial origin, is transcribed by the host RNA polymerase II. Images

Willmitzer, L; Schmalenbach, W; Schell, J

1981-01-01

71

Assessment of the efficiency of cotransformation of the T-DNA of disarmed binary vectors derived from Agrobacterium tumefaciens and the T-DNA of A. rhizogenes  

Microsoft Academic Search

Co-transfer of Agrobacterium rhizogenes T-DNA and T-DNA from the A. tumefaciens binary vector pBin19 (Bevan, 1984) was studied in detail using Nicotiana rustica. High frequencies of co-transfer of T-DNA's were observed, even when no selection pressure was exerted. Increased levels of pBin19 T-DNA were found in hairy root cultures with selection at higher levels of kanamycin sulphate (50–200 µg ml-1).

John D. Hamill; Andrea Prescott; Cathie Martin

1987-01-01

72

Analysis of T-DNA integration and generative segregation in transgenic winter triticale (x Triticosecale Wittmack)  

PubMed Central

Background While the genetic transformation of the major cereal crops has become relatively routine, to date only a few reports were published on transgenic triticale, and robust data on T-DNA integration and segregation have not been available in this species. Results Here, we present a comprehensive analysis of stable transgenic winter triticale cv. Bogo carrying the selectable marker gene HYGROMYCIN PHOSPHOTRANSFERASE (HPT) and a synthetic green fluorescent protein gene (gfp). Progeny of four independent transgenic plants were comprehensively investigated with regard to the number of integrated T-DNA copies, the number of plant genomic integration loci, the integrity and functionality of individual T-DNA copies, as well as the segregation of transgenes in T1 and T2 generations, which also enabled us to identify homozygous transgenic lines. The truncation of some integrated T-DNAs at their left end along with the occurrence of independent segregation of multiple T-DNAs unintendedly resulted in a single-copy segregant that is selectable marker-free and homozygous for the gfp gene. The heritable expression of gfp driven by the maize UBI-1 promoter was demonstrated by confocal laser scanning microscopy. Conclusions The used transformation method is a valuable tool for the genetic engineering of triticale. Here we show that comprehensive molecular analyses are required for the correct interpretation of phenotypic data collected from the transgenic plants.

2012-01-01

73

Analysis of two P-element enhancer-trap insertion lines that show expression in the giant fibre neuron of Drosophila melanogaster.  

PubMed

The giant fibre system (GFS) of Drosophila is a simple neural circuit that mediates escape responses in adult flies. Here we report the initial characterization of two genes that are preferentially expressed in the GFS. Two P-element insertion lines, carrying the GAL4 transcriptional activator, were identified that exhibited pronounced expression in elements of the GFS and relatively low levels elsewhere within the adult central nervous system. Genomic DNA flanking the P-element insertion site was recovered from each of these lines, sequenced, and nearby transcripts identified and confirmed to exhibit GFS expression by in situ hybridization. This analysis revealed that these P-elements were in previously characterized genes. Line P[GAL4]-A307 has an insert in the gene short stop for which we have identified a novel transcript, while line P[GAL4]-141 has an insert in the transcription factor ken and barbie. Here we show that ken and barbie mutants have defects in escape behaviour, behavioural responses to visual stimuli and synaptic functions in the GFS. We have therefore revealed a neural role for a transcription factor that previously had no implicated neural function. PMID:16879616

Allen, M J; Drummond, J A; Sweetman, D J; Moffat, K G

2006-07-27

74

Increased frequency of N-region insertion in a murine pre-B-cell line infected with a terminal deoxynucleotidyl transferase retroviral expression vector.  

PubMed Central

The role of terminal deoxynucleotidyl transferase (TdT) in the insertion of N regions into the junctional sites of immunoglobulin genes was investigated. Pre-B-cell lines capable of continuous rearrangement of immunoglobulin light-chain genes and differing only in the presence or apparent absence of TdT were derived by infecting cells with a TdT retroviral expression vector or a control vector. The cell lines were then superinfected with a retrovirus-based artificial immunoglobulin gene rearrangement substrate. The substrate was allowed to rearrange in the cell lines and the rearranged proviruses were rescued from the cell lines. Nucleotide sequence analysis of the V-J junctions of the proviral rearranged genes showed a fivefold greater frequency of N-region insertion in proviruses rescued from the TdT+ cell lines than in those rescued from the TdT- cell lines, so that at least 50% of the rearrangements that occurred in the presence of TdT had N regions. It is thus evident that TdT can stimulate N-region insertion, and the enzyme is presumably directly responsible for adding nucleotides at V-J and other immunoglobulin and T-cell receptor gene junctions. Images

Landau, N R; Schatz, D G; Rosa, M; Baltimore, D

1987-01-01

75

Silencing in Arabidopsis T-DNA Transformants: The Predominant Role of a Gene-Specific RNA Sensing Mechanism versus Position Effects  

Microsoft Academic Search

Pronounced variability of transgene expression and transgene silencing are commonly observed among independent plant lines transformed with the same construct. Single-copy T-DNA lines harboring reporter genes of various kind and number under the control of a strong promoter were established in Arabidopsis thaliana for a comprehensive analysis of transgene expression. Characterization of 132 independent transgenic lines revealed no case of

Daniel Schubert; Berthold Lechtenberg; Alexandra Forsbach; Mario Gils; Sultan Bahadur; Renate Schmidt

2004-01-01

76

Agrobacterium rhizogenes inserts T-DNA into the genomes of the host plant root cells  

Microsoft Academic Search

Agrobacterium rhizogenes, which induces hairy root disease of dicotyledonous plants1, is closely related to Agrobacterium tumefaciens, the causative agent of crown gall disease1-3. Virulence in both species is conferred by large plasmids4-7. Infected plant tissue synthesizes novel metabolites, opines8-11, that are not found in normal plant tissues. The pattern of opines synthesized is determined by the type of virulence plasmid

Mary-Dell Chilton; David A. Tepfer; Annik Petit; Chantal David; Francine Casse-Delbart; Jacques Tempé

1982-01-01

77

The Right Border Region of pTiT37 T-DNA is Intrinsically More Active than the Left Border Region in Promoting T-DNA Transformation  

Microsoft Academic Search

Deletions of border regions of T-DNA on Agrobacterium Ti plasmid or mini-T plasmid have shown the right border region of pTiT37 T-DNA to be more active than the left border region in promoting T-DNA transformation. In this study we examine the possibility that the apparent difference in activity of left and right border regions may be due to position or

George C. Jen; Mary-Dell Chilton

1986-01-01

78

Cardiac tamponade and successful pericardiocentesis in an extremely low birth weight neonate with percutaneously inserted central venous line: a case report  

Microsoft Academic Search

BACKGROUND: Pericardial effusion and cardiac tamponade are rare but life-threatening complications of percutaneosuly inserted central line (PICL) use in extremely low birth weight (ELBW) neonates, with an incidence reported between 0.07% and 2% of PICLs placement. Timely diagnosis and pericardiocentesis has been proven to be life-saving. CASE PRESENTATION: The patient was a 620 g birth weight neonate who presented with

Alfredo Pizzuti; Emilia Parodi; Paola Abbondi; Mario Frigerio

2010-01-01

79

Association of the Agrobacterium T-DNA-protein complex with plant nucleosomes  

Microsoft Academic Search

Agrobacterium represents the only natural example of transkingdom transfer of genetic information, from bacteria to plants. Before the bacterial transferred DNA (T- DNA) can integrate into the plant genome, it should be targeted to and bind the host chromatin. However, the T-DNA association with the host chromatin has not been demonstrated. Here, we study T-DNA binding to plant nucleosomes in

Benoît Lacroix; Abraham Loyter; Vitaly Citovsky

2008-01-01

80

Analysis of crosstalk of coupled transmission lines by inserting additional traces grounded with vias on printed circuit boards  

Microsoft Academic Search

In analog and digital electronic systems, crosstalk between traces on the printed circuit boards (PCB) can degrade the performance of equipment operations. To reduce crosstalk between two parallel traces of which one is called aggressor trace and the other one is called victim trace, two additional traces grounded by vias are inserted and the aggressor trace is in between the

Shulan Li; Yuan'an Liu; Zhanhai Song; Hefei Hu

2003-01-01

81

An insertional mutagenesis programme with an enhancer trap for the identification and tagging of genes involved in abiotic stress tolerance in the tomato wild-related species Solanum pennellii.  

PubMed

Salinity and drought have a huge impact on agriculture since there are few areas free of these abiotic stresses and the problem continues to increase. In tomato, the most important horticultural crop worldwide, there are accessions of wild-related species with a high degree of tolerance to salinity and drought. Thus, the finding of insertional mutants with other tolerance levels could lead to the identification and tagging of key genes responsible for abiotic stress tolerance. To this end, we are performing an insertional mutagenesis programme with an enhancer trap in the tomato wild-related species Solanum pennellii. First, we developed an efficient transformation method which has allowed us to generate more than 2,000 T-DNA lines. Next, the collection of S. pennelli T(0) lines has been screened in saline or drought conditions and several presumptive mutants have been selected for their salt and drought sensitivity. Moreover, T-DNA lines with expression of the reporter uidA gene in specific organs, such as vascular bundles, trichomes and stomata, which may play key roles in processes related to abiotic stress tolerance, have been identified. Finally, the growth of T-DNA lines in control conditions allowed us the identification of different development mutants. Taking into account that progenies from the lines are being obtained and that the collection of T-DNA lines is going to enlarge progressively due to the high transformation efficiency achieved, there are great possibilities for identifying key genes involved in different tolerance mechanisms to salinity and drought. PMID:21647638

Atarés, Alejandro; Moyano, Elena; Morales, Belén; Schleicher, Peter; García-Abellán, José Osvaldo; Antón, Teresa; García-Sogo, Begoña; Perez-Martin, Fernando; Lozano, Rafael; Flores, Francisco Borja; Moreno, Vicente; Bolarin, María del Carmen; Pineda, Benito

2011-06-07

82

Comparison of operation efficiency for the insert task when using stereoscopic images with additional lines, stereoscopic images, and a manipulator with force feedback  

NASA Astrophysics Data System (ADS)

It has been reported that operation efficiency for the teleoperations using stereoscopic video images is lower than when using the naked eye in real environments. Here, the authors tried to improve the human-machine interface of this particular system to achieve higher operation efficiency for stereoscopic video imags by adding other information. An experiment was carried out under the four following conditions: when the insert task was performed by subjects using conventional stereoscopic video imags, when the centering lines of the cylindrical objects and holes were added to the conventional stereoscopic video images, when the force feedback was provided to the system manipulator as one object touched another object, and when both of the additional centering lines and force feedback were provided. The subject's task was to inset a cylindrical object into a round hole. The completion time was measured from the time of the starting signal to the time when the object was inserted into the hole. Completion time, when additional lines were given, was shorter than when the force feedback was provided, and when no additional information was provided. It was concluded that additional visual information contributed more to the recognition of the space rather than providing additional information about surface phenomena.

Matsunaga, Katsuya; Shidoji, Kazunori; Matsubara, Kenjiro

1999-05-01

83

Molecular Structure and Regulatory Potential of a T-DNA Integration Site in Petunia  

Microsoft Academic Search

The genomic structure surrounding a T-DNA integration site in a transgenic petunia plant, which shows deregulation of a root-specific promoter, was investigated. We have already demonstrated that T-DNA integration in this transformant (P13) had occurred close to a scaffold\\/matrix attachment region (S\\/MAR). A major question regarding the observed promoter leakiness was whether the T-DNA had integrated into the centre or

Antje Dietz-Pfeilstetter; Nicola Arndt; Volker Kay; Jürgen Bode

2003-01-01

84

Interpreting the biological relevance of bioinformatic analyses with T-DNA sequence for protein allergenicity.  

PubMed

Global regulatory agencies require bioinformatic sequence analysis as part of their safety evaluation for transgenic crops. Analysis typically focuses on encoded proteins and adjacent endogenous flanking sequences. Recently, regulatory expectations have expanded to include all reading frames of the inserted DNA. The intent is to provide biologically relevant results that can be used in the overall assessment of safety. This paper evaluates the relevance of assessing the allergenic potential of all DNA reading frames found in common food genes using methods considered for the analysis of T-DNA sequences used in transgenic crops. FASTA and BLASTX algorithms were used to compare genes from maize, rice, soybean, cucumber, melon, watermelon, and tomato using international regulatory guidance. Results show that BLASTX for maize yielded 7254 alignments that exceeded allergen similarity thresholds and 210,772 alignments that matched eight or more consecutive amino acids with an allergen; other crops produced similar results. This analysis suggests that each nontransgenic crop has a much greater potential for allergenic risk than what has been observed clinically. We demonstrate that a meaningful safety assessment is unlikely to be provided by using methods with inherently high frequencies of false positive alignments when broadly applied to all reading frames of DNA sequence. PMID:22668749

Harper, B; McClain, S; Ganko, E W

2012-06-02

85

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

Microsoft Academic Search

The non-long-terminal-repeat (non-LTR) retrotransposons (also called long interspersed repetitive ele- ments (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

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

2004-01-01

86

Association of the Agrobacterium T-DNA-protein complex with plant nucleosomes.  

PubMed

Agrobacterium represents the only natural example of transkingdom transfer of genetic information, from bacteria to plants. Before the bacterial transferred DNA (T-DNA) can integrate into the plant genome, it should be targeted to and bind the host chromatin. However, the T-DNA association with the host chromatin has not been demonstrated. Here, we study T-DNA binding to plant nucleosomes in vitro and show that it is mediated by bacterial and host proteins associated with the T-DNA. The main factor that determines nucleosomal binding of the T-DNA is the cellular VirE2-interacting protein 1 (VIP1), which functions as a molecular link between the T-DNA-associated bacterial virulence protein VirE2 and core histones. The presence of both VIP1 and VirE2 is required for association of the T-DNA with mononucleosomes in which the DNA molecule exists as a tripartite complex DNA-VirE2-VIP1. Furthermore, this nucleosome-associated ternary complex can bind another bacterial virulence factor, VirF, which is an F-box protein known to target both VirE2 and VIP1 for proteasomal degradation and uncoat the T-DNA. PMID:18832163

Lacroix, Benoît; Loyter, Abraham; Citovsky, Vitaly

2008-10-01

87

?-Thalassemia Due to Intronic LINE-1 Insertion in the ?-Globin Gene (HBB): Molecular Mechanisms Underlying Reduced Transcript Levels of the ?-GlobinL1 Allele.  

PubMed

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 ?-globinL1 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 ?-globinL1 allele were hypermethylated; however, treatment with a demethylating agent did not restore the impaired transcription. A histone deacetylase inhibitor partially reactivated the ?-globinL1 transcription despite permanent ?-globinL1 promoter CpG methylation. This result indicates that the decreased rate of transcription from the ?-globinL1 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

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

2013-08-13

88

Insertion of a nuclear factor kappa B DNA nuclear-targeting sequence potentiates suicide gene therapy efficacy in lung cancer cell lines.  

PubMed

Lung cancer currently causes the majority of cancer-related deaths worldwide and new treatments are in high demand. Gene therapy could be a promising treatment but currently lacks sufficient efficiency for clinical use, primarily due to limited cellular and nuclear DNA delivery. In the present study, we investigated whether it was possible to exploit the endogenous nuclear-shuttling activity by the nuclear factor kappa B (NF?B) system, which is highly prominent in many cancers as well as lung cancer. We observed that insertion of a DNA nuclear-targeting sequence (DTS) recognized by NF?B could improve plasmid nuclear delivery and enhance the therapeutic effect of a validated transcriptionally cancer-targeted suicide gene therapy system. A clear correlation between the number of inserted NF?B-binding sites and the therapeutic effect of the suicide system was observed in both small cell lung cancer (SCLC) and non-SCLC cell lines. The effect was observed to be due to elevated nuclear translocation of the suicide gene-encoding plasmids. The results show that a significant improvement of gene therapeutic efficiency can be obtained by increasing the intracellular trafficking of therapeutic DNA. This is to our knowledge the first time a DTS strategy has been implemented for suicide gene therapy. PMID:22898898

Cramer, F; Christensen, C L; Poulsen, T T; Badding, M A; Dean, D A; Poulsen, H S

2012-08-17

89

The angiotensin converting enzyme insertion/deletion polymorphism alters the response of muscle energy supply lines to exercise.  

PubMed

The presence of a silencing sequence (the I-allele) in the gene for the upstream regulator of blood flow, angiotensin I-converting enzyme (ACE), is associated with superior endurance performance and its trainability. We tested in a retrospective study with 36 Caucasian men of Swiss descent whether carriers of the ACE I-allele demonstrate a modified adaptive response of energy supply lines in knee extensor muscle, and aerobic fitness, to endurance training based on 6 weeks of supervised bicycle exercise or 6 months of self-regulated running (p value

Vaughan, David; Huber-Abel, Felicitas A; Graber, Franziska; Hoppeler, Hans; Flück, Martin

2013-02-09

90

Structure and expression of DNA transferred to tobacco via transformation of protoplasts with Ti-plasmid DNA: co-transfer of T-DNA and non T-DNA sequences  

Microsoft Academic Search

The T-DNA structure and organization in tissues obtained via transformation of tobacco protoplasts with Ti-plasmid DNA was found to be completely different from the T-DNA introduced via Agrobacterium tumefaciens. It is often fragmented. Overlapping copies of T-DNA, having various sizes, as well as separated fragments of T-DNA were detected. The border sequences of 23 basepairs (bp), flanking the T-region in

Frans A. Krens; Ruud M. W. Mans; Truus M. S. Slogteren; J. Harry C. Hoge; George J. Wullems; Robbert A. Schilperoort

1985-01-01

91

Genome-wide insertional mutagenesis of Arabidopsis thaliana.  

PubMed

Over 225,000 independent Agrobacterium transferred DNA (T-DNA) insertion events in the genome of the reference plant Arabidopsis thaliana have been created that represent near saturation of the gene space. The precise locations were determined for more than 88,000 T-DNA insertions, which resulted in the identification of mutations in more than 21,700 of the approximately 29,454 predicted Arabidopsis genes. Genome-wide analysis of the distribution of integration events revealed the existence of a large integration site bias at both the chromosome and gene levels. Insertion mutations were identified in genes that are regulated in response to the plant hormone ethylene. PMID:12893945

Alonso, José M; Stepanova, Anna N; Leisse, Thomas J; Kim, Christopher J; Chen, Huaming; Shinn, Paul; Stevenson, Denise K; Zimmerman, Justin; Barajas, Pascual; Cheuk, Rosa; Gadrinab, Carmelita; Heller, Collen; Jeske, Albert; Koesema, Eric; Meyers, Cristina C; Parker, Holly; Prednis, Lance; Ansari, Yasser; Choy, Nathan; Deen, Hashim; Geralt, Michael; Hazari, Nisha; Hom, Emily; Karnes, Meagan; Mulholland, Celene; Ndubaku, Ral; Schmidt, Ian; Guzman, Plinio; Aguilar-Henonin, Laura; Schmid, Markus; Weigel, Detlef; Carter, David E; Marchand, Trudy; Risseeuw, Eddy; Brogden, Debra; Zeko, Albana; Crosby, William L; Berry, Charles C; Ecker, Joseph R

2003-08-01

92

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

PubMed Central

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

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

2003-01-01

93

Improved efficiency for T-DNA-mediated transformation and plasmid rescue in Arabidopsis thaliana  

Microsoft Academic Search

A vector was constructed for the isolation of gene fusions to thelacZ reporter gene following T-DNA integration into the genome ofArabidopsis thaliana. To facilitate the generation of taggedA. thaliana plants, we established a modified method for high-frequency transformation ofA. thaliana byAgrobacterium tumefaciens. The main modification required was to inhibit the methylation of T-DNA in the transformed calli. Apparently, cytosine residues

A. Mandal; V. Lång; W. Orczyk; E. T. Palva

1993-01-01

94

A new PCR system for Agrobacterium tumefaciens detection based on amplification of T-DNA fragment.  

PubMed

The design of the PCR system presented in this work is based on the knowledge of the molecular character of the crown gall disease. The virulence of Agrobacterium tumefaciens requires the presence of a big (up to 235,000 bp) plasmid Ti (pTi-tumour inducing plasmid). This plasmid carries the so-called T-DNA fragment (T-DNA-transferred DNA), which integrates into cell chromosomes of the infected plants and subsequently changes the plant morphology nad metabolism. In cannot be excluded that after T-DNA integration the presence of Agrobacterium is not necessary for the development of pathological changes. Thus, T-DNA is the only sign that must be present both in virulent bacteria and in infected plants in any stadium of the disease and even before the infection. This is why T-DNA was chosen as the target region for PCR amplification. Primers flanking a 220 bp fragment of one of the conservative regions responsible for Agrobacterium pathogenicity, namely tms2 gene coding for indolacetamide amidohydrolase (the second step of auxin biosynthesis) were designed as the optimal for PCR amplification. The PCR amplification reactions were performed for matrixes isolated from cultures of reference strains giving one predicted product for each sample. First attempts of T-DNA detection in infected soils and plants were performed. We hope that the presented new PCR system for Agrobacterium tumefaciens detection will help to fight the crown gall disease in the nearest future. PMID:9429288

Sachadyn, P; Kur, J

1997-01-01

95

Arabidopsis VIRE2 INTERACTING PROTEIN2 is required for Agrobacterium T-DNA integration in plants.  

PubMed

Agrobacterium tumefaciens-mediated genetic transformation is an efficient tool for genetic engineering of plants. VirE2 is a single-stranded DNA binding Agrobacterium protein that is transported into the plant cell and presumably protects the T-DNA from degradation. Using a yeast two-hybrid system, we identified Arabidopsis thaliana VIRE2-INTERACTING PROTEIN2 (VIP2) with a NOT domain that is conserved in both plants and animals. Furthermore, we provide evidence supporting VIP2 interaction with VIP1, a basic domain/leucine zipper motif-containing protein required for nuclear import and integration of T-DNA. Virus-induced gene silencing of VIP2 in Nicotiana benthamiana and characterization of the Arabidopsis vip2 mutant (At vip2) demonstrate that VIP2 is required for Agrobacterium-mediated stable transformation but not for transient transformation. Assays based upon a promoter-trap vector and quantification of T-DNA integration further confirmed VIP2 involvement in T-DNA integration. Interestingly, VIP2 transcripts were induced to a greater extent over prolonged periods after infection with a T-DNA transfer-competent Agrobacterium strain compared with the transfer-deficient Agrobacterium strain. Transcriptome analyses of At vip2 suggest that VIP2 is likely a transcriptional regulator, and the recalcitrancy to transformation in At vip2 is probably due to the combination of muted gene expression response upon Agrobacterium infection and repression of histone genes resulting in decreased T-DNA integration events. PMID:17496122

Anand, Ajith; Krichevsky, Alexander; Schornack, Sebastian; Lahaye, Thomas; Tzfira, Tzvi; Tang, Yuhong; Citovsky, Vitaly; Mysore, Kirankumar S

2007-05-11

96

Gallery 31: mys inserted in chromosomes  

NSDL National Science Digital Library

Repetitive DNA can have preferred insertion sites. In this example, yellow represents the distribution of mys (a type of LINE) over a mouse genome where chromosomes are orange. There are more mys inserted in the sex (X) chromosomes.

2009-12-26

97

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

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.

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

2010-01-01

98

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

PubMed Central

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.

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

99

A germ-line insertion in the Birt-Hogg-Dub? (BHD) gene gives rise to the Nihon rat model of inherited renal cancer  

PubMed Central

A rat model of hereditary renal carcinoma (RC) was found in a rat colony of the Sprague–Dawley strain in Japan and named the “Nihon” rat. In heterozygotes, RCs, predominantly the clear cell type, develop from early preneoplastic lesions, which began to appear as early as 3 weeks of age, to adenocarcinomas by the age of 6 months. The Nihon rat is an example of a Mendelian dominantly inherited predisposition for development of RCs like the Eker (Tsc2 gene mutant) rat. We have previously shown that the Nihon mutation was tightly linked to genes that are located on the distal part of rat chromosome 10. The order of the genes is the Eker (Tsc2 gene (human 16p13.3)–Il3 gene–Nihon gene–Llgl1 locus– Myhse gene. We now describe a germ-line mutation in the Birt–Hogg–Dubé gene (Bhd) (human 17p11.2) caused by the insertion of a single nucleotide in the Nihon rat, resulting in a frameshift and producing a stop codon 26 aa downstream. We found that the homozygous mutant condition was lethal at an early stage of fetal life in the rat. We detected a high frequency of loss of heterozygosity (LOH) in primary RCs (10/11) at the Bhd locus and found a point mutation (nonsense) in one LOH-negative case, fitting Knudson's “two-hit” model. The Nihon rat may therefore provide insights into a tumor-suppressor gene that is related to renal carcinogenesis and an animal model of human BHD syndrome.

Okimoto, Kazuo; Sakurai, Junko; Kobayashi, Toshiyuki; Mitani, Hiroaki; Hirayama, Youko; Nickerson, Michael L.; Warren, Michelle B.; Zbar, Berton; Schmidt, Laura S.; Hino, Okio

2004-01-01

100

The Genomic Pattern of tDNA Operon Expression in E. coli  

PubMed Central

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.

Ardell, David H; Kirsebom, Leif A

2005-01-01

101

Altered Growth and Wood Characteristics in Transgenic Hybrid Aspen Expressing Agrobacterium tumefaciens T-DNA Indoleacetic Acid-Biosynthetic Genes.  

PubMed Central

A key regulator of cambial growth is the plant hormone indoleacetic acid (IAA). Here we report on altered wood characteristics and growth patterns in transgenic hybrid aspen (Populus tremula L. x Populus tremuloides Michx.) expressing Agrobacterium tumefaciens T-DNA IAA-biosynthetic iaaM and iaaH genes. Eighteen lines simultaneously expressing both genes were regenerated. Of these, four lines, verified to be transgenic by northern blot analysis, were selected and raised under controlled growth conditions. All four lines were affected in their growth patterns, including alterations in height and stem diameter growth, internode elongation, leaf enlargement, and degree of apical dominance. Two transgenic lines, showing the most distinct phenotypic deviation from the wild type, were characterized in more detail for free and conjugated IAA levels and for wood characteristics. Both lines showed an altered IAA balance, particularly in mature leaves and roots where IAA levels were elevated. They also exhibited changes in wood anatomy, most notably a reduction in vessel size, an increase in vessel density, and changes in ray development. Thus, the recent development of techniques for gene transfer to forest trees enabled us to investigate the influence of an altered IAA balance on xylem development in an intact experimental system. In addition, the results demonstrate the possibility of manipulating wood properties in a forest tree through controlled changes of IAA concentration and distribution.

Tuominen, H.; Sitbon, F.; Jacobsson, C.; Sandberg, G.; Olsson, O.; Sundberg, B.

1995-01-01

102

Activation of a Plant Gene by T-DNA Tagging: Auxin-Independent Growth in Vitro  

Microsoft Academic Search

A transferred DNA (T-DNA) tagging vector with the potential to produce dominant mutations was used with cocultured Agrobacterium tumefaciens and protoplasts to tag genes involved in the action of the plant growth substance auxin. Transgenic calli were selected for their ability to grow in the absence of auxin in the culture media. From one experiment, 12 calli that displayed this

Hiroaki Hayashi; Inge Czaja; Helge Lubenow; Jeff Schell; Richard Walden

1992-01-01

103

Enhanced Plant Cell Transformation by Addition of Host Genes Involved in T-DNA Integration.  

National Technical Information Service (NTIS)

Adding at least one gene involved in plant host cell T-DNA integration enhances transformation by Agrobacterium. The histone H2A gene encoded by the Arabidopsis RAT5 gene increases transformation frequencies of plants, most likely by causing overexpressio...

K. S. Mysore S. B. Gelvin

2004-01-01

104

Multiple Independent Defective Suppressor-mutator Transposon Insertions in Arabidopsis: A Tool for Functional Genomics  

Microsoft Academic Search

A new system for insertional mutagenesis based on the maize Enhancer\\/Suppressor-mutator ( En\\/Spm ) element was introduced into Arabidopsis. A single T-DNA construct carried a nonautonomous defective Spm (d Spm ) element with a phosphinothricin herbicide resistance ( BAR ) gene, a transposase expression cassette, and a counterselectable gene. This construct was used to select for stable d Spm transpositions.

Alain F. Tissier; Sylvestre Marillonnet; Victor Klimyuk; Kanu Patel; Miguel Angel Torres; George Murphy; Jonathan D. G. Jones

1999-01-01

105

Insertion element IS 1296 in Mycoplasma mycoides subsp. mycoides small colony identifies a European clonal line distinct from African and Australian strains  

Microsoft Academic Search

Strains of Mycoplasma mycoides subsp. mycoides small colony (SC) type, the agent of contagious bovine pleuropneumonia (CBPP), were analysed with respect to the polymorphism of distribution of a newly discovered insertion element, lS1296, on the chromosome. Analysis of 64 strains isolated from Europe, Africa and Australia, including four vaccine strains and the type strain PG1, revealed ten different IS patterns,

Xiaoxing Cheng; Jacques Nicolet; F. Poumarat; Jose Regalla; Franqois Thiaucourt; J. Frey

1995-01-01

106

Silencing in Arabidopsis T-DNA Transformants: The Predominant Role of a Gene-Specific RNA Sensing Mechanism versus Position Effects  

PubMed Central

Pronounced variability of transgene expression and transgene silencing are commonly observed among independent plant lines transformed with the same construct. Single-copy T-DNA lines harboring reporter genes of various kind and number under the control of a strong promoter were established in Arabidopsis thaliana for a comprehensive analysis of transgene expression. Characterization of 132 independent transgenic lines revealed no case of silencing as a result of site of T-DNA integration. Below a certain number of identical transgenes in the genome, gene copy number and expression were positively correlated. Expression was high, stable over all generations analyzed, and of a comparable level among independent lines harboring the same copy number of a particular transgene. Conversely, RNA silencing was triggered if the transcript level of a transgene surpassed a gene-specific threshold. Transcript level–mediated silencing effectively accounts for the pronounced transgene expression variability seen among transformants. It is proposed that the RNA sensing mechanism described is a genome surveillance system that eliminates RNA corresponding to excessively transcribed genes, including transgenes, and so plays an important role in genome defense.

Schubert, Daniel; Lechtenberg, Berthold; Forsbach, Alexandra; Gils, Mario; Bahadur, Sultan; Schmidt, Renate

2004-01-01

107

DNA sequences homologous to the T DNA region of Agrobacterium tumefaciens are present in diverse Rhizobium species  

Microsoft Academic Search

DNA sequences homologous to the T DNA region of the octopine-type Ti plasmid from Agrobacterium tumefaciens are present in different Rhizobium species. Plasmid DNA from each of two R. leguminosarum, two R. meliloti, and four slow-growing Rhizobium strains examined contain restriction endonuclease fragments that hybridize with the T DNA region, or with DNA sequences at or near the adjacent Ti

R. G. Hadley; A. A. Szalay

1982-01-01

108

T-DNA is organized predominantly in inverted repeat structures in plants transformed with Agrobacterium tumefaciens C58 derivatives  

Microsoft Academic Search

The detailed structural organization of DNA sequences transferred to the plant genome via Agrobacterium tumefaciens has been determined in 11 transgenic tomato plants that carry the transferred DNA (T-DNA) at a single genetic locus. The majority (seven) of these plants were found to carry multiple copies of T-DNA arranged in inverted repeat structures. Such a high frequency of inverted repeats

Richard Jorgensen; Christine Snyder; Jonathan D. G. Jones

1987-01-01

109

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

PubMed Central

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

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

1995-01-01

110

Dihalocarbene Insertion Experiment  

ERIC Educational Resources Information Center

|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)|

Goh, S. H.

1975-01-01

111

Dihalocarbene Insertion Experiment  

ERIC Educational Resources Information Center

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)

Goh, S. H.

1975-01-01

112

Chest tube insertion  

MedlinePLUS

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

113

Arabidopsis VIRE2 INTERACTING PROTEIN2 Is Required for Agrobacterium T-DNA Integration in Plants  

Microsoft Academic Search

Agrobacterium tumefaciens-mediated genetic transformation is an efficient tool for genetic engineering of plants. VirE2 is a single-stranded DNA binding Agrobacterium protein that is transported into the plant cell and presumably protects the T-DNA from degradation. Using a yeast two-hybrid system, we identified Arabidopsis thaliana VIRE2-INTERACTING PROTEIN2 (VIP2) with a NOT domain that is conserved in both plants and animals. Furthermore,

Ajith Anand; Alexander Krichevsky; Sebastian Schornack; Thomas Lahaye; Tzvi Tzfira; Yuhong Tang; Vitaly Citovsky; K. S. Mysore

2007-01-01

114

Agrobacterium may delay plant nonhomologous end-joining DNA repair via XRCC4 to favor T-DNA integration.  

PubMed

Agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (T-DNA) into the plant genome. Several plant factors that play a role in Agrobacterium attachment to plant cells and transport of T-DNA to the nucleus have been identified, but the T-DNA integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (NHEJ)-mediated double-strand DNA break (DSB) repair. Here, we report a negative role of X-ray cross complementation group4 (XRCC4), one of the key proteins required for NHEJ, in Agrobacterium T-DNA integration. Downregulation of XRCC4 in Arabidopsis and Nicotiana benthamiana increased stable transformation due to increased T-DNA integration. Overexpression of XRCC4 in Arabidopsis decreased stable transformation due to decreased T-DNA integration. Interestingly, XRCC4 directly interacted with Agrobacterium protein VirE2 in a yeast two-hybrid system and in planta. VirE2-expressing Arabidopsis plants were more susceptible to the DNA damaging chemical bleomycin and showed increased stable transformation. We hypothesize that VirE2 titrates or excludes active XRCC4 protein available for DSB repair, thus delaying the closure of DSBs in the chromosome, providing greater opportunity for T-DNA to integrate. PMID:23064322

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

2012-10-12

115

Two large-insert soybean genomic libraries constructed in a binary vector: applications in chromosome walking and genome wide physical mapping  

Microsoft Academic Search

Large DNA insert libraries in binary T-DNA vectors can assist in the isolation of the gene(s) underlying a quantitative trait\\u000a locus (QTL). Binary vectors facilitate the transfer of large-insert DNA fragments containing a QTL from E. coli to Agrobacterium sp. and then to plants. We constructed two soybean large-insert libraries from cv. Forrest in the pCLD04541 (V41) binary\\u000a vector after

K. Meksem; K. Zobrist; E. Ruben; D. Hyten; T. Quanzhou; H. B. Zhang; D. A. Lightfoot

2000-01-01

116

[Cotransformation of aspen and birch with three T-DNA regions from two different replicons in one Agrobacterium tumefaciens strain].  

PubMed

The cointegration rate into the aspen and birch genomes of foreign genes from a binary vector and a disarmed Ti plasmid pCBE21 carried by the same Agrobacterium tumefaciens strain was studied. The cotransformation rate for the genes within the Ti plasmid varied from 30 to 100%; while the transformation rate for the gene from T(L) region was twofold higher as compared with the T(R) region. On the average, the gene transfer from all three T-DNAs was recorded in 10.9% of the transgenic lines. For the vector pBI121, the cotransformation rates for the genes from both regions of pCBE21 T-DNA were higher as compared with the vector pGS. In addition, a concurrent transfer of the genes from the Ti plasmid T(L) and T(R) regions was recorded only after the transformation with the vector pBI121. These results can be used for constructing woody plants containing several genes. PMID:21261057

Lebedev, V G; Shestibratov, K A; Shadrina, T E; Bulatova, I V; Abramochkin, D G; Miroshnikov, A I

2010-11-01

117

High Efficiency Transgene Segregation in Co-Transformed Maize Plants using an Agrobacterium Tumefaciens 2 T-DNA Binary System  

Microsoft Academic Search

For regulatory issues and research purposes it would be desirable to have the ability to segregate transgenes in co-transformed maize. We have developed a highly efficient system to segregate transgenes in maize that was co-transformed using an Agrobacterium tumefaciens 2 T-DNA binary system. Three vector treatments were compared in this study; (1) a 2 T-DNA vector, where the selectable marker

Michael Miller; Laura Tagliani; Ning Wang; Benjamin Berka; Dennis Bidney; Zuo-Yu Zhao

2002-01-01

118

The non-conserved region of cucumopine-type Agrobacterium rhizogenes T-DNA is responsible for hairy root induction  

Microsoft Academic Search

The T-DNA regions of three strains of Ri plasmids 1855, 8196, 2659 (agropine, mannopine and cucumopine type respectively) share two highly conserved regions flanking a non-homologous central part [1,2]. We have cloned segments of the cucumopine Ri plasmid 2659 T-DNA in the binary vector system Bin 19 and infected carrot discs with recombinant Agrobacterium strains. We show here that the

M. C. Failla; F. Maimone; A. De Paolis; P. Costantino; M. Cardarelli

1990-01-01

119

Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors.  

PubMed

Signal transducer and activator of transcription 3 (STAT3) is an oncogenic protein that is constitutively activated in numerous cancer cell lines and human cancers. Another STAT family member, STAT1, possesses cancer-inhibitory properties and can promote apoptosis in tumor cells upon activation. To better characterize these important cancer related genes, we tagged STAT3 and STAT1 loci with fluorescent protein (FP) sequences (RFP and GFP respectively) by targeted integration via zinc finger nuclease (ZFN) - mediated homologous recombination in A549 cells that express aberrantly activated STAT3. We inserted the FP transgenes at the N-terminus of the STAT3 locus and at the C-terminus of the STAT1 locus. The integration resulted in endogenous expression of fluorescent STAT3 and STAT1 chimeric fusion proteins. When stimulated with IL-6 or IFN-?, the cells showed robust nuclear translocation of RFP-STAT3 or STAT1-GFP, respectively. Pre-incubation of cells with a known specific STAT3 inhibitor showed that IFN-?-induced translocation of STAT1-GFP was not impaired. STAT3 activates multiple downstream targets such as genes involved in cell cycle progression - e.g. cyclin D1. To detect changes in expression of endogenous cyclin D1, we used ZFN technology to insert a secreted luciferase reporter behind the cyclin D1 promoter and separated the luciferase and cyclin D1 coding regions by a 2A sequence to induce a translational skip. The luciferase insertion was made in the RFP-STAT3/STAT1-GFP cell line to have all three reporters in a single cell line. Addition of a STAT3 inhibitor led to suppression of cyclin D1 promoter activity and cell growth arrest. The triple-modified cell line provides a simple and convenient method for high-content screening and pre-clinical testing of potential STAT3 inhibitors in live cells while ensuring that the STAT1 pathway is not affected. This approach of reporting endogenous gene activities using ZFN technology could be applied to other cancer targets. PMID:23950841

Samsonov, Andrey; Zenser, Nathan; Zhang, Fan; Zhang, Hongyi; Fetter, John; Malkov, Dmitry

2013-07-09

120

Tagging of Genomic STAT3 and STAT1 with Fluorescent Proteins and Insertion of a Luciferase Reporter in the Cyclin D1 Gene Provides a Modified A549 Cell Line to Screen for Selective STAT3 Inhibitors  

PubMed Central

Signal transducer and activator of transcription 3 (STAT3) is an oncogenic protein that is constitutively activated in numerous cancer cell lines and human cancers. Another STAT family member, STAT1, possesses cancer-inhibitory properties and can promote apoptosis in tumor cells upon activation. To better characterize these important cancer related genes, we tagged STAT3 and STAT1 loci with fluorescent protein (FP) sequences (RFP and GFP respectively) by targeted integration via zinc finger nuclease (ZFN) - mediated homologous recombination in A549 cells that express aberrantly activated STAT3. We inserted the FP transgenes at the N-terminus of the STAT3 locus and at the C-terminus of the STAT1 locus. The integration resulted in endogenous expression of fluorescent STAT3 and STAT1 chimeric fusion proteins. When stimulated with IL-6 or IFN-?, the cells showed robust nuclear translocation of RFP-STAT3 or STAT1-GFP, respectively. Pre-incubation of cells with a known specific STAT3 inhibitor showed that IFN-?-induced translocation of STAT1-GFP was not impaired. STAT3 activates multiple downstream targets such as genes involved in cell cycle progression - e.g. cyclin D1. To detect changes in expression of endogenous cyclin D1, we used ZFN technology to insert a secreted luciferase reporter behind the cyclin D1 promoter and separated the luciferase and cyclin D1 coding regions by a 2A sequence to induce a translational skip. The luciferase insertion was made in the RFP-STAT3/STAT1-GFP cell line to have all three reporters in a single cell line. Addition of a STAT3 inhibitor led to suppression of cyclin D1 promoter activity and cell growth arrest. The triple-modified cell line provides a simple and convenient method for high-content screening and pre-clinical testing of potential STAT3 inhibitors in live cells while ensuring that the STAT1 pathway is not affected. This approach of reporting endogenous gene activities using ZFN technology could be applied to other cancer targets.

Samsonov, Andrey; Zenser, Nathan; Zhang, Fan; Zhang, Hongyi; Fetter, John; Malkov, Dmitry

2013-01-01

121

Application of PCR ribotyping and tDNA-PCR for Klebsiella pneumoniae identification.  

PubMed

PCR analysis of 16S-23S internal transcribed spacer (PCR ribotyping) and tRNA intergenic spacer (tDNA-PCR) were evaluated for their effectiveness in identification of clinical strains of Klebsiella pneumoniae and differentiation with related species. For this purpose both methods were applied to forty-three clinical isolates biochemically identified as K. pneumoniae subsp. pneumoniae isolated from patients clinical specimens attended at five hospitals in three Brazilian cities. References strains of K. pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae, K. oxytoca, K. planticola and Enterobacter aerogenes were also analyzed. Both PCR methods showed specific patterns for each species. A conserved PCR ribotype pattern was observed for all clinical K. pneumoniae isolates, while differing from other related analyzed species. tDNA-PCR revealed five distinct patterns among the K. pneumoniae clinical isolates studied, demonstrating a predominant group with 90.6% of isolates presenting the same pattern of K. pneumoniae type strain. Both PCR-based methods were not able to differentiate K. pneumoniae subspecies. On the basis of the results obtained, both methods were efficient to differentiate the Klebsiella species analyzed, as well as E. aerogenes. Meanwhile tDNA-PCR revealed different tRNA arrangements in K. pneumoniae, suggesting intra-species heterogeneity of their genome organization, the polymorphism of the intergenic spacers between 16S and 23S rRNA genes appears to be highly conserved whithin K. pneumoniae clinical isolates, showing that PCR ribotyping can be an useful tool for identification of K. pneumoniae isolates. PMID:17992365

Lopes, Ana Catarina S; Rodrigues, Juliana Falcão; Clementino, Maysa B M; Miranda, Catia A C; Nascimento, Ana Paula A; de Morais Júnior, Marcos Antônio

2007-12-11

122

Genetic transformation in two potato cultivars with T-DNA from disarmed Agrobacterium  

Microsoft Academic Search

Derivatives of potato (Solanum tuberosum cv.'s ‘Maris Bard’ and ‘Desiree’) transformed with disarmed T-DNA from genetically engineered Agrobacterium tumefaciens strains were isolated. The transformed plants were recovered from shoot-forming tumours induced by infection of wounds with mixedcultures of shoot-inducing A. tumefaciens strains T37 and either Agrobacterium strain LBA1834(pRAL1834), (Hille et al. 1983) or LBA4404(pBIN6; pRAL4404), (Bevan 1984). Two small-scale feasibility

G. Ooms; M. M. Burrell; A. Karp; M. Bevan; J. Hille

1987-01-01

123

Marine riser insert sleeves  

SciTech Connect

Drilling operations are run in a riser pipe, eg., 20 inches in diameter, which extends from a ship to an anchored wellhead assembly on the ocean floor. Various strings of casing for lining the borehole wall are run down through the interior of the riser pipe. The first string of casing run is of relatively large diameter, eg., 9- 5/8 inches in diameter. Each successive string of casing which is run in this particular well is progressively smaller in diameter. Ideally, one would need a new and smaller riser in order to maintain about the same drilling fluid upward velocity. A high upward fluid velocity can be maintained by placing sleeve inserts in the annulus space between the drill string and the riser pipe interior wall. Additional strings of sleeves may be run inside the original sleeve to obtain a multiple effective sizing of the riser pipe.

Waller, M.R.

1980-11-04

124

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

PubMed

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

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

2013-06-24

125

Agrobacterium tumefaciens -mediated transformation for random insertional mutagenesis in Colletotrichum lagenarium  

Microsoft Academic Search

Random insertional mutagenesis using a marker DNA fragment is an effective method for identifying fungal genes relevant to morphogenesis, metabolism, and so on. Agrobacterium tumefaciens-mediated transformation (AtMT) has long been used as a tool for the genetic modification of a wide range of plant species. Recent study has indicated that A. tumefaciens could transfer T-DNA not only to plant cells

Gento Tsuji; Satoshi Fujii; Naoki Fujihara; Chika Hirose; Seiji Tsuge; Tomonori Shiraishi; Yasuyuki Kubo

2003-01-01

126

Fractal Dimension Analysis in Self-Assembled Poly(dA)·poly(dT) DNA Network on Mica Surface  

NASA Astrophysics Data System (ADS)

Characteristics of the self-assembled poly(dA)·poly(dT) DNA network adhered on the mica substrate are experimentally investigated based on the AFM observations and the fractal dimension analysis. Artificial B-type double stranded DNA, which consists of 50 base pairs of adenine and thymine, is specially prepared for the experiment. The manufacturing process of DNA network is done in the aqueous solution of poly(dA)·poly(dT) DNA, and the systematical experimental runs are made for various concentration of the solution. It is found that the 2D fractal dimension strongly depends on the fabrication process of the DNA network.

Kawano, Satoyuki

127

Rapid Assay of Foreign Gene Expression in Leaf Discs Transformed by Agrobacterium tumefaciens: Role of T-DNA Borders in the Transfer Process  

Microsoft Academic Search

We have developed a sensitive leaf disc transformation procedure for studying early and\\/or transient T-DNA expression during Agrobacterium tumefaciens-mediated transformation of plant cells. Using this system, we have examined the function of T-DNA border sequences on the early expression of T-DNA genes and on the stable integration of those genes in infected cells. Deletion of the right border from the

R. B. Horsch; H. J. Klee

1986-01-01

128

Localization of the T-DNA on marker chromosomes in transformed tobacco cells by in situ hybridization  

Microsoft Academic Search

Chromosome and molecular analyses were conducted on tobacco cells which had been transformed by the T-DNA of the Ti-plasmid. These analyses showed that there were specific chromosome rearrangements in the transformed cells (marker chromosomes). There was a positive correlation between the number of marker chromosomes per cell and the oncogenic potential of the transformed cells. However, we show, using the

A. Mouras; I. Negrutiu

1989-01-01

129

Disassembly of synthetic Agrobacterium T-DNA-protein complexes via the host SCF(VBF) ubiquitin-ligase complex pathway.  

PubMed

One the most intriguing, yet least studied, aspects of the bacterium-host plant interaction is the role of the host ubiquitin/proteasome system (UPS) in the infection process. Increasing evidence indicates that pathogenic bacteria subvert the host UPS to facilitate infection. Although both mammalian and plant bacterial pathogens are known to use the host UPS, the first prokaryotic F-box protein, an essential component of UPS, was identified in Agrobacterium. During its infection, which culminates in genetic modification of the host cell, Agrobacterium transfers its T-DNA--as a complex (T-complex) with the bacterial VirE2 and host VIP1 proteins--into the host cell nucleus. There the T-DNA is uncoated from its protein components before undergoing integration into the host genome. It has been suggested that the host UPS mediates this uncoating process, but there is no evidence indicating that this activity can unmask the T-DNA molecule. Here we provide support for the idea that the plant UPS uncoats synthetic T-complexes via the Skp1/Cullin/F-box protein VBF pathway and exposes the T-DNA molecule to external enzymatic activity. PMID:23248273

Zaltsman, Adi; Lacroix, Benoît; Gafni, Yedidya; Citovsky, Vitaly

2012-12-17

130

Disassembly of synthetic Agrobacterium T-DNA-protein complexes via the host SCFVBF ubiquitin-ligase complex pathway  

PubMed Central

One the most intriguing, yet least studied, aspects of the bacterium–host plant interaction is the role of the host ubiquitin/proteasome system (UPS) in the infection process. Increasing evidence indicates that pathogenic bacteria subvert the host UPS to facilitate infection. Although both mammalian and plant bacterial pathogens are known to use the host UPS, the first prokaryotic F-box protein, an essential component of UPS, was identified in Agrobacterium. During its infection, which culminates in genetic modification of the host cell, Agrobacterium transfers its T-DNA—as a complex (T-complex) with the bacterial VirE2 and host VIP1 proteins—into the host cell nucleus. There the T-DNA is uncoated from its protein components before undergoing integration into the host genome. It has been suggested that the host UPS mediates this uncoating process, but there is no evidence indicating that this activity can unmask the T-DNA molecule. Here we provide support for the idea that the plant UPS uncoats synthetic T-complexes via the Skp1/Cullin/F-box protein VBF pathway and exposes the T-DNA molecule to external enzymatic activity.

Zaltsman, Adi; Lacroix, Benoit; Gafni, Yedidya; Citovsky, Vitaly

2013-01-01

131

Inserting the Même prosthesis.  

PubMed

The Même implant may result in fewer capsular contractures. However, difficulty with insertion of this prosthesis may discourage the surgeon and result in the abandonment of a useful prosthesis. A simple technique has been evolved which allows for easy insertion and placement of this implant. PMID:6701223

Dolsky, R L

1984-03-01

132

The LHC insertion magnets  

Microsoft Academic Search

The Large Hadron Collider comprises eight insertions, four of which are dedicated to the LHC experiments while the others are used for the major collider systems. The various functions of the insertions are fulfilled by a variety of magnet systems, most of them based on the technology of NbTi superconductors cooled by superfluid helium at 1.9 K. A number of

R. Ostojic

2002-01-01

133

Increased frequency of homologous recombination and T-DNA integration in Arabidopsis CAF-1 mutants.  

PubMed

Chromatin assembly factor 1 (CAF-1) is involved in nucleo some assembly following DNA replication and nucleotide excision repair. In Arabidopsis thaliana, the three CAF-1 subunits are encoded by FAS1, FAS2 and, most likely, MSI1, respectively. In this study, we asked whether genomic stability is altered in fas1 and fas2 mutants that are lacking CAF-1 activity. Depletion of either subunit increased the frequency of somatic homologous recombination (HR) in planta approximately 40-fold. The frequency of transferred DNA (T-DNA) integration was also elevated. A delay in loading histones onto newly replicated or repaired DNA might make these DNA stretches more accessible, both to repair enzymes and to foreign DNA. Furthermore, fas mutants exhibited increased levels of DNA double-strand breaks, a G2-phase retardation that accelerates endoreduplication, and elevated levels of mRNAs coding for proteins involved in HR-all factors that could also contribute to upregulation of HR frequency in fas mutants. PMID:17110925

Endo, Masaki; Ishikawa, Yuichi; Osakabe, Keishi; Nakayama, Shigeki; Kaya, Hidetaka; Araki, Takashi; Shibahara, Kei-ichi; Abe, Kiyomi; Ichikawa, Hiroaki; Valentine, Lisa; Hohn, Barbara; Toki, Seiichi

2006-11-16

134

Increased frequency of homologous recombination and T-DNA integration in Arabidopsis CAF-1 mutants  

PubMed Central

Chromatin assembly factor 1 (CAF-1) is involved in nucleo some assembly following DNA replication and nucleotide excision repair. In Arabidopsis thaliana, the three CAF-1 subunits are encoded by FAS1, FAS2 and, most likely, MSI1, respectively. In this study, we asked whether genomic stability is altered in fas1 and fas2 mutants that are lacking CAF-1 activity. Depletion of either subunit increased the frequency of somatic homologous recombination (HR) in planta ?40-fold. The frequency of transferred DNA (T-DNA) integration was also elevated. A delay in loading histones onto newly replicated or repaired DNA might make these DNA stretches more accessible, both to repair enzymes and to foreign DNA. Furthermore, fas mutants exhibited increased levels of DNA double-strand breaks, a G2-phase retardation that accelerates endoreduplication, and elevated levels of mRNAs coding for proteins involved in HR—all factors that could also contribute to upregulation of HR frequency in fas mutants.

Endo, Masaki; Ishikawa, Yuichi; Osakabe, Keishi; Nakayama, Shigeki; Kaya, Hidetaka; Araki, Takashi; Shibahara, Kei-ichi; Abe, Kiyomi; Ichikawa, Hiroaki; Valentine, Lisa; Hohn, Barbara; Toki, Seiichi

2006-01-01

135

Agrobacterium-Mediated Transformation of Fusarium oxysporum: An Efficient Tool for Insertional Mutagenesis and Gene Transfer.  

PubMed

ABSTRACT Agrobacterium tumefaciens-mediated transformation (ATMT) has long been used to transfer genes to a wide variety of plants and has also served as an efficient tool for insertional mutagenesis. In this paper, we report the construction of four novel binary vectors for fungal transformation and the optimization of an ATMT protocol for insertional mutagenesis, which permits an efficient genetic manipulation of Fusarium oxysporum and other phytopathogenic fungi to be achieved. Employing the binary vectors, carrying the bacterial hygromycin B phosphotrans-ferase gene (hph) under the control of the Aspergillus nidulans trpC promoter as a selectable marker, led to the production of 300 to 500 hygromycin B resistant transformants per 1 x 10(6) conidia of F. oxysporum, which is at least an order of magnitude higher than that previously accomplished. Transformation efficiency correlated strongly with the duration of cocultivation of fungal spores with Agrobacterium tumefaciens cells and significantly with the number of Agrobacteruium tumefaciens cells present during the cocultivation period (r = 0.996; n = 3; P < 0.01). All transformants tested remained mitotically stable, maintaining their hygromycin B resistance. Growing Agrobacterium tumefaciens cells in the presence of acetosyringone (AS) prior to cocultivation shortened the time required for the formation of transformants but decreased to 53% the percentage of transformants containing a single T-DNA insert per genome. This increased to over 80% when Agrobacterium tumefaciens cells grown in the absence of AS were used. There was no correlation between the average copy number of T-DNA per genome and the colony diameter of the transformants, the period of cocultivation or the quantity of Agrobacterium tumefaciens cells present during cocultivation. To isolate the host sequences flanking the inserted T-DNA, we employed a modified thermal asymmetric interlaced PCR (TAIL-PCR) technique. Utilizing just one arbitrary primer resulted in the successful amplification of desired products in 90% of those transformants analyzed. The insertion event appeared to be a random process with truncation of the inserted T-DNA, ranging from 1 to 14 bp in size, occurring on both the right and left border sequences. Considering the size and design of the vectors described here, coupled with the efficiency and flexibility of this ATMT protocol, it is suggested that ATMT should be regarded as a highly efficient alternative to other DNA transfer procedures in characterizing those genes important for the pathogenicity of F. oxysporum and potentially those of other fungal pathogens. PMID:18944391

Mullins, E D; Chen, X; Romaine, P; Raina, R; Geiser, D M; Kang, S

2001-02-01

136

Transient GFP expression in Nicotiana plumbaginifolia suspension cells: the role of gene silencing, cell death and T-DNA loss  

Microsoft Academic Search

The transient nature of T-DNA expression was studied with a gfp reporter gene transferred to Nicotiana plumbaginifolia suspension cells fromAgrobacterium tumefaciens. Individual GFP-expressing protoplasts were isolated after 4 days' co-cultivation. The protoplasts were cultured without selection and 4 weeks later the surviving proto-calluses were again screened for GFP expression. Of the proto-calluses initially expressing GFP, 50% had lost detectable GFP

Richard Weld; Jack Heinemann; Colin Eady

2001-01-01

137

A procedure for the establishment of Glomus mosseae in dual culture with Ri T-DNA-transformed carrot roots  

Microsoft Academic Search

A stepwise procedure was investigated to determine the optimal conditions for the establishment of Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe in dual in vitro culture with Ri T-DNA-transformed roots of Daucus carota L. Glomus mosseae spores germinated best in 10?mm Tris or MES-buffered medium at pH?values just above neutral. Growth of hyphae from germinated spores was much greater

David D. Douds Jr.

1997-01-01

138

Agrobacterium vitis nopaline Ti plasmid pTiAB4: relationship to other Ti plasmids and T-DNA structure  

Microsoft Academic Search

The Ti plasmid of the Agrobacterium vitis nopaline-type strain AB4 was subcloned and mapped. Several regions of the 157 kb Ti plasmid are similar or identical to parts of the A. vitis octopine\\/cucumopine (o\\/c)-type Ti plasmids, and other regions are homologous to the nopaline-type Ti plasmid pTiC58. The T-DNA of pTiAB4 is a chimaeric structure of recent origin: the left

Léon Otten; Patrice Ruffray

1994-01-01

139

Fractal Dimension Analysis in Self-Assembled Poly(dA)·poly(dT) DNA Network on Mica Surface  

Microsoft Academic Search

Characteristics of the self-assembled poly(dA)·poly(dT) DNA network adhered on the mica substrate are experimentally investigated based on the AFM observations and the fractal dimension analysis. Artificial B-type double stranded DNA, which consists of 50 base pairs of adenine and thymine, is specially prepared for the experiment. The manufacturing process of DNA network is done in the aqueous solution of poly(dA)·poly(dT)

Satoyuki Kawano

2005-01-01

140

Infectivity of monoaxenic subcultures of the arbuscular mycorrhizal fungus Glomus versiforme associated with Ri-T-DNA-transformed carrot root  

Microsoft Academic Search

Glomus versiforme was associated in vitro with Ri-T-DNA-transformed carrot root and after 4 months of cultivation, numerous axenic arbuscular\\u000a mycorrhizal (AM) propagules were obtained. Three successive generations of spores and mycorrhizal root pieces were obtained\\u000a by reassociating a 4-month-old root piece with a new carrot root. A biological test was conducted to assess the infectivity\\u000a of the three generations of

C. Plenchette; S. Declerck; T. A. Diop; D. G. Strullu

1996-01-01

141

T-DNA transfer and integration in the ectomycorrhizal fungus Suillus bovinus using hygromycin B as a selectable marker  

Microsoft Academic Search

The T-DNA of Agrobacterium tumefaciens can be transferred to plants, yeasts, fungi and human cells. Using this system, dikaryotic mycelium of the ectomycorrhizal fungus Suillus bovinus was transformed with recombinant hygromycin B phosphotransferase (hph) and enhanced green fluorescent protein (EGFP) genes fused with a heterologous fungal promoter and CaMV 35S terminator. Transformation resulted in hygromycin B-resistant clones, which were mitotically

Mubashir Hanif; Alejandro Pardo; Markus Gorfer; Marjatta Raudaskoski

2002-01-01

142

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

PubMed Central

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

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

1985-01-01

143

Agrobacterium aiming for the host chromatin: Host and bacterial proteins involved in interactions between T-DNA and plant nucleosomes.  

PubMed

Agrobacterium genetically transforms its hosts by transferring a segment of DNA (T-DNA) into the host cell and integrating it into the host genome. Integration requires a close interaction between T-DNA, which is packaged into a nucleoprotein complex (T-complex) by bacterial virulence (Vir) proteins, and the host chromatin. This interaction is facilitated by the host protein VIP 1, which binds both to the major protein component of the T-complex, VirE2, and to the core histones. Recently, VIP1 has been demonstrated to mediate the interaction between plant nucleosomes and VirE2-DNA complexes (i.e., synthetic T-complex-like structures) in vitro. Here, we discuss major implications of these observations-such as the possible role of core histone modifications, proteasomal uncoating of the T-complex mediated by the bacterial F-box protein VirF, and the need for changes in chromatin structure to render it accessible to the T-DNA integration-for the process of chromatin targeting of foreign DNA and its integration into the eukaryotic genome. PMID:19513263

Lacroix, Benoît; Citovsky, Vitaly

2009-01-01

144

T-DNA and opine synthetic loci in tumors incited by Agrobacterium tumefaciens A281 on soybean and alfalfa plants  

SciTech Connect

A report is given of the molecular characterization of transferred DNA (T-DNA) in leguminous tumors incited by Agrobacterium tumefaciens A281 harboring the tumor-inducing plasmid pTiBo542. The T-DNA is composed of two regions named T/sub L/ (left portion)-DNA and T/sub R/ (right portion)-DNA, in accordance with the nomenclature for the octopine strains. T/sub L/-DNA is defined by several internal HindIII restriction fragments totaling 10.8 kilobase pairs (kbp) in uncloned soybean and alfalfa tumors. Alfalfa tumor DNA may contain one more HindIII fragment at the left end of T/sub L/-DNA that does soybean tumor DNA. T/sub R/-DNA has a 5.8-kbp BamHI-EcoRi internal fragment. All borders other than the left border of T/sub L/-DNA appear to be the same within the detection limits of Southern blot hybridization experiments. The two T-DNA regions are separated by 16 to 19 kbp of DNA not stably maintained in tumors. The distance from the left border of T/sub L/-DNA to the right border of T/sub R/-DNA is approximately 40 kbp. Loci for the mannityl opines are situated in T/sub R/-DNA, based on genetic and biochemical criteria.

Hood, E.E.; Chilton, W.S.; Chilton, M.D.; Fraley, R.T.

1986-12-01

145

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

PubMed Central

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

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

2012-01-01

146

Cloning quantitative trait loci by insertional mutagenesis  

Microsoft Academic Search

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

M. Soller; J. S. Beckmann

1987-01-01

147

Nanopattern insert molding  

NASA Astrophysics Data System (ADS)

A new method was investigated to produce nanopatterns on polymeric surfaces with high resolution, good productivity, and low cost. It has certain advantages when compared with such conventional techniques as nanoimprint lithography (NIL), hot embossing, and injection molding. Polyvinyl alcohol (PVA) was utilized for preparation of the stamp with nanopatterns on its surface. The nanoimprinted PVA film was inserted into the cavity and the polymer melt was injected into the mold. Nanopatterns with pillars smaller than 100 nm were produced on the polymeric surface. The water soluble PVA film was used as the inserted template to overcome the difficulties of releasing the nanopatterned film from the substrate.

Kim, S. H.; Jeong, J. H.; Youn, J. R.

2010-05-01

148

Nanopattern insert molding  

Microsoft Academic Search

A new method was investigated to produce nanopatterns on polymeric surfaces with high resolution, good productivity, and low cost. It has certain advantages when compared with such conventional techniques as nanoimprint lithography (NIL), hot embossing, and injection molding. Polyvinyl alcohol (PVA) was utilized for preparation of the stamp with nanopatterns on its surface. The nanoimprinted PVA film was inserted into

S H Kim; J H Jeong; J R Youn

2010-01-01

149

Insertion in Persian  

ERIC Educational Resources Information Center

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…

Kambuziya, Aliyeh Kord-e Zafaranlu; Dehghan, Masoud

2011-01-01

150

Generation of selectable marker-free sheath blight resistant transgenic rice plants by efficient co-transformation of a cointegrate vector T-DNA and a binary vector T-DNA in one Agrobacterium tumefaciens strain  

Microsoft Academic Search

Co-transformation of Oryza sativa L. var. Pusa Basmati1 was done using an Agrobacterium\\u000a tumefaciens strain harbouring a single-copy cointegrate vector and a multi-copy binary vector in the same cell. The T-DNA of the cointegrate\\u000a vector pGV2260::pSSJ1 carried the hygromycin phosphotransferase (hph) and ?-glucuronidase (gus) genes. The binary vector pCam-chi11, without a plant selectable marker gene, harboured the rice chitinase (chi11)

Rajasekaran Sripriya; Vengoji Raghupathy; Karuppannan Veluthambi

2008-01-01

151

Agrobacterium tumefaciens-Mediated Transformation of Aspergillus fumigatus: an Efficient Tool for Insertional Mutagenesis and Targeted Gene Disruption  

PubMed Central

Agrobacterium tumefaciens was used to transform Aspergillus fumigatus by either random or site-directed integration of transforming DNA (T-DNA). Random mutagenesis via Agrobacterium tumefaciens-mediated transformation (ATMT) was accomplished with T-DNA containing a hygromycin resistance cassette. Cocultivation of A. fumigatus conidia and Agrobacterium (1:10 ratio) for 48 h at 24°C resulted in high frequencies of transformation (>100 transformants/107 conidia). The majority of transformants harbored a randomly integrated single copy of T-DNA and were mitotically stable. We chose alb1, a polyketide synthase gene, as the target gene for homologous integration because of the clear phenotype difference between the white colonies of ?alb1 mutant strains and the bluish-green colonies of wild-type strains. ATMT with a T-DNA-containing alb1 disruption construct resulted in 66% albino transformants. Southern analysis revealed that 19 of the 20 randomly chosen albino transformants (95%) were disrupted by homologous recombination. These results suggest that ATMT is an efficient tool for transformation, random insertional mutagenesis, and gene disruption in A. fumigatus.

Sugui, Janyce A.; Chang, Yun C.; Kwon-Chung, K. J.

2005-01-01

152

Virulence genes, borders, and overdrive generate single-stranded T-DNA molecules from the A6 Ti plasmid of Agrobacterium tumefaciens.  

PubMed Central

Agrobacterium tumefaciens transfers the T-DNA portion of its Ti plasmid to the nuclear genome of plant cells. Upon cocultivation of A. tumefaciens A348 with regenerating tobacco leaf protoplasts, six distinct single-stranded T-DNA molecules (T strands) were generated in addition to double-stranded T-DNA border cleavages which we have previously reported (K. Veluthambi, R.K. Jayaswal, and S.B. Gelvin, Proc. Natl. Acad. Sci. USA 84:1881-1885, 1987). The T region of an octopine-type Ti plasmid has four border repeats delimiting three T-DNA regions defined as T left (TL), T center (TC), and T right (TR). The six T strands generated upon induction corresponded to the TL, TC, TR, TL + TC, TC + TR, and TL + TC + TR regions, suggesting that the initiation and termination of T-strand synthesis can occur at each of the four borders. Most TL + TC + TR T-strand molecules corresponded to the top T-DNA strand, whereas the other five T strands corresponded to the bottom T-DNA strand. Generation of T strands required the virA, virG, and virD operons. Extra copies of vir genes, harbored on cosmids within derivatives of A. tumefaciens A348, enhanced production of T strands. The presence of right and left border repeats in their native orientation is important for the generation of full-length T strands. When a right border repeat was placed in the opposite orientation, single-stranded T-DNA molecules that corresponded to the top strand were generated. Deletion of overdrive, a sequence that flanks right border repeats and functions as a T-DNA transmission enhancer, reduced the level of T-strand generation. Induction of A. tumefaciens cells by regenerating tobacco protoplasts increased the copy number of the Ti plasmid relative to the bacterial chromosome. Images

Veluthambi, K; Ream, W; Gelvin, S B

1988-01-01

153

T-DNA direct repeat and 35S promoter methylation affect transgene expression but do not cause silencing in transgenic sweet orange  

Microsoft Academic Search

In transgenic plants, multiple T-DNA copies and cytosine methylation usually correlate with a loss of transgene expression\\u000a while methylation of promoter is associated with transgene silencing. Here, six independent GFP transgenic ‘Bingtang’ sweet orange (Citrus sinensis (L.) Osb.) plants were analyzed with copy number, T-DNA repeat, cytosine methylation and transgene expression. The fluorescence\\u000a of transgenic plants was normal and bright

Jing Fan; Xin Liu; Shi-Xiao Xu; Qiang Xu; Wen-Wu Guo

154

Genetic analysis of the individual T-DNA genes of Agrobacterium tumefaciens ; further evidence that two genes are involved in indole-3-acetic acid synthesis  

Microsoft Academic Search

The T-DNA genes of Ti plasmids of Agrobacterium tumefaciens can induce tumorous growth on a wide range of dicotyledonous plants. We subcloned the individual onc genes of the pTiC58 T-DNA and reintroduced them in the T-region of the Ti plasmid gene vector pGV3850 (from which the onc genes had been removed (Zambryski et al. 1983)). These experiments were designed to

D. Inzé; A. Follin; M. Van Lijsebettens; C. Simoens; C. Genetello; M. Van Montagu; J. Schell

1984-01-01

155

Two T-DNA's co-transformed into Brassica napus by a double Agrobacterium tumefaciens infection are mainly integrated at the same locus  

Microsoft Academic Search

Summary Hypocotyl explants of threeBrassica napus varieties were infected with two nopaline typeAgrobacterium strains each carrying a distinct disarmed T-DNA containing different selectable markers. Selection was done for only one of the markers, after which the regenerated plants were screened for the presence of the second marker. High co-transformation frequencies of both T-DNA's were obtained (39%–85% of the transformants). Where

Marc De Block; Dirk Debrouwer

1991-01-01

156

[Optimization of T-dNA insertional mutagenesis and analysis of mutants of Magnaporthe grisea].  

PubMed

The rice blast fungus Magnaporthe grisea causes one of the most destructive diseases of rice around the world. Significant progresses have been made recently in genomics studies of the fungus, opening new era of the functional genomics which requires to generate a large scale of gene knockout mutants. It has been demonstrated that T-DNA insertional mutagenesis is a powerful tool of functional genomics not only for plants but also for fungi. In this paper, we optimized the conditions for T-DNA insertional mutagenesis of M. grisea using Agrobacterium tumefaciens-mediated transformation (ATMT) approach. We employed the binary vector pBHtl constructed by Dr. S. Kang's laboratory at the Pennsylvania State University, which carries the bacterial hygromycin B phosphotransferase gene (hph) under the control of the Aspergillus nidulans trpC promoter as a selectable marker to transform the conidia of M. grisea. We optimized the conditions for T-DNA insertional mutagenesis including the medium, dosage of hygromycin B, cefotaxime and carbenicillin to select the transformants and inhibit the growth of A. tumefaciens after co-culturing. The dosage to inhibit non-transformants could vary from 200-600microg/mL among different M. grisea isolates so that the optimal dosage of the antibiotics should be decided according to isolates. Rice polished agar medium was found the best selection medium which would facilitate the mutant sporulation and minimize the contamination chance. In average, about 500 transformants could be obtained when transforming 1 x 10(6) spores at the optimum condition, among which 85% had T-DNA insertion detected by polymerase chain reaction (PCR) and thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR). Fifteen out of 1520 transformants showed mutation in colony morphology. Within 58 randomly selected mutants, it was found that there were 4 sporulation-decreased mutants, 8 less germination mutants and 9 appressorium defective mutants. Several virulent mutants to C101LAC(Pi-1)and 75-1-127(Pi-9)were also obtained which would facilitate cloning the corresponding avirulence genes. PMID:15969057

Li, Hong-Yu; Pan, Chu-Yi; Chen, Han; Zhao, Chang-Jiang; Lu, Guo-Dong; Wang, Zong-Hua

2003-07-01

157

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 ...

158

Induction and growth properties of carrot roots with different complements of Agrobacterium rhizogenes T-DNA.  

PubMed

Single and multiple infections of carrot discs were carried out with Agrobacterium strains harbouring different segments of pRi1855 TL-DNA cloned in the binary vector Bin 19 and with a strain carrying the TR-DNA from the same Ri plasmid. Roots induced by the various co-inoculations were cultured and their growth patterns were followed. Abundant roots could be induced by TL-DNA rol genes A, B and C as a single insert (rolA + B + C) and by rolB alone provided an extended segment beyond its 5' non-coding region was included in the construction. A depression of rooting capability was caused by the inclusion of rolC together with rolB (rolB + C). In all cases co-inoculation with the Agrobacterium carrying TR-DNA-borne auxin genes was necessary for root induction since none of the rol constructions was in itself capable of eliciting any response; an exceeding majority of these roots were however shown to contain rol genes but no TR-DNA. Rooting was also elicited if rol constructions were co-inoculated with a strain carrying TL-DNA genes 13 and 14 (ORF13 + 14) instead of the TR-DNA strain. These roots were shown to contain both rol genes and ORF13 + 14. Striking differences in growth properties were shown by roots containing different complements of TL-DNA genes. Typical hairy root traits, high growth rate, branching and, most noticeably, absence of geotropism, were shown by roots containing rolB alone, while roots with rolA + B + C were geotropic as normal carrot roots. Hairy root traits were conferred to rolA + B + C roots by the concomitant presence of ORF13 + 14 and by the addition of auxin to the culture medium. A model is presented which attempts to rationalize the growth patterns by assigning interplaying roles to the various TL-DNA genes involved. PMID:2562759

Capone, I; Spanò, L; Cardarelli, M; Bellincampi, D; Petit, A; Costantino, P

1989-07-01

159

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

PubMed

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

Kunitake, Emi; Tani, Shuji; Sumitani, Jun-Ichi; Kawaguchi, Takashi

2011-12-14

160

Impedance calculation for ferrite inserts  

SciTech Connect

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.

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

2005-01-01

161

Marine riser insert sleeves  

Microsoft Academic Search

Drilling operations are run in a riser pipe, eg., 20 inches in diameter, which extends from a ship to an anchored wellhead assembly on the ocean floor. Various strings of casing for lining the borehole wall are run down through the interior of the riser pipe. The first string of casing run is of relatively large diameter, eg., 9- 5\\/8

Waller

1980-01-01

162

A selectively terminable transgenic rice line expressing human lactoferrin.  

PubMed

Human lactoferrin (hLF) is a multifunctional milk protein which could be utilized for promoting human health. Transgenic rice has been used as a bioreactor for mass production of recombinant hLF. However, one major concern over such transgenic rice is the risk of its unintended spreading into environment and into our food supplies. Here we report the development of selectively terminable transgenic rice expressing human lactoferrin in seeds. These transgenic rice plants could be selectively terminated by bentazon, a common herbicide used for rice weed control. The hLF expression cassette was constructed into a T-DNA containing the RNA interference cassette suppressing the expression of the rice gene CYP81A6 which detoxifies herbicide bentazon, and the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) cassette which confers to glyphosate tolerance. A transgenic line, named as G281, was identified for its high sensitivity to bentazon, high tolerance to glyphosate, and high expression of hLF. Southern analysis suggested G281 is a single copy insertion event. Field tests demonstrated that G281 plants can be completely killed by a single spray of bentazon at 1000 mg/L, which is safe to regular rice and represents only half of the dose recommended by manufacturer for rice field weed control. Therefore, any G281 contaminations in regular rice could be selectively terminated to make sure it will not enter food or feed supplies. PMID:20433928

Lin, Chaoyang; Nie, Peng; Lu, Wei; Zhang, Qing; Li, Jing; Shen, Zhicheng

2010-04-28

163

Insertion device vacuum system designs  

SciTech Connect

Synchrotron light source insertion device vacuum systems now in operation and systems proposed for the future are reviewed. An overview of insertion devices is given and four generic vacuum chamber designs, transition section design and pumping considerations are discussed. Examples of vacuum chamber systems are presented.

Hoyer, E.

1988-05-01

164

Completion of CS insert fabrication  

Microsoft Academic Search

The central solenoid (CS) model coil program is in progress with an international collaboration under the frame of the ITER-EDA. The purpose of the CS insert coil is to test the performance of the ITER-CS conductor. The CS insert coil is installed in the bore of the CS model coil and tested at a magnetic flux density of 13 T.

M. Sugimoto; T. Isono; Y. Nunoya; N. Koizumi; H. Nakajima; T. Kato; M. Matsukawa; K. Hamada; K. Matsui; G. Nishijima; Y. Tsuchiya; K. Sawada; Y. Takahashi; T. Ando; H. Tsuji; T. Ichihara; T. Minato; T. Sasaki

2000-01-01

165

Sink Inserts for Flood Prevention  

NASA Astrophysics Data System (ADS)

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.

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

2004-09-01

166

Nucleotide exchange and excision technology (NExT) DNA shuffling: a robust method for DNA fragmentation and directed evolution.  

PubMed

DNA shuffling is widely used for optimizing complex properties contained within DNA and proteins. Demonstrated here is the amplification of a gene library by PCR using uridine triphosphate (dUTP) as a fragmentation defining exchange nucleotide with thymidine, together with the three other nucleotides. The incorporated uracil bases were excised using uracil-DNA-glycosylase and the DNA backbone subsequently cleaved with piperidine. These end-point reactions required no adjustments. Polyacrylamide urea gels demonstrated adjustable fragmentation size over a wide range. The oligonucleotide pool was reassembled by internal primer extension to full length with a proofreading polymerase to improve yield over Taq. We present a computer program that accurately predicts the fragmentation pattern and yields all possible fragment sequences with their respective likelihood of occurrence, taking the guesswork out of the fragmentation. The technique has been demonstrated by shuffling chloramphenicol acetyltransferase gene libraries. A 33% dUTP PCR resulted in shuffled clones with an average parental fragment size of 86 bases even without employment of a fragment size separation, and revealed a low mutation rate (0.1%). NExT DNA fragmentation is rational, easily executed and reproducible, making it superior to other techniques. Additionally, NExT could feasibly be applied to several other nucleotide analogs. PMID:16061932

Müller, Kristian M; Stebel, Sabine C; Knall, Susanne; Zipf, Gregor; Bernauer, Hubert S; Arndt, Katja M

2005-08-01

167

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 ...

168

Comments on DAPTACEL Package Insert  

Center for Biologics Evaluation and Research (CBER)

Text Version... The sponsor is seeking to revise the GBS statement in the warnings and precautions and post- marketing experience sections of the package insert ... More results from www.fda.gov/downloads/biologicsbloodvaccines/vaccines

169

The orf13 T-DNA Gene of Agrobacterium rhizogenes Confers Meristematic Competence to Differentiated Cells1  

PubMed Central

Plant infections by the soil bacterium Agrobacterium rhizogenes result in neoplastic disease with the formation of hairy roots at the site of infection. Expression of a set of oncogenes residing on the stably integrated T-DNA is responsible for the disease symptoms. Besides the rol (root locus) genes, which are essential for the formation of hairy roots, the open reading frame orf13 mediates cytokinin-like effects, suggesting an interaction with hormone signaling pathways. Here we show that ORF13 induced ectopic expression of KNOX (KNOTTED1-like homeobox) class transcription factors, as well as of several genes involved in cell cycle control in tomato (Lycopersicon esculentum). ORF13 has a retinoblastoma (RB)-binding motif and interacted with maize (Zea mays) RB in vitro, whereas ORF13, bearing a point mutation in the RB-binding motif (ORF13*), did not. Increased cell divisions in the vegetative shoot apical meristem and accelerated formation of leaf primordia were observed in plants expressing orf13, whereas the expression of orf13* had no influence on cell division rates in the shoot apical meristem, suggesting a role of RB in the regulation of the cell cycle in meristematic tissues. On the other hand, ectopic expression of LeT6 was not dependent on a functional RB-binding motif. Hormone homeostasis was only altered in explants of leaves, whereas in the root no effects were observed. We suggest that ORF13 confers meristematic competence to cells infected by A. rhizogenes by inducing the expression of KNOX genes and promotes the transition of infected cells from the G1 to the S phase by binding to RB.

Stieger, Pia A.; Meyer, Alain D.; Kathmann, Petra; Frundt, Corinne; Niederhauser, Isabel; Barone, Mario; Kuhlemeier, Cris

2004-01-01

170

An essential virulence protein of Agrobacterium tumefaciens , VirB4, requires an intact mononucleotide binding domain to function in transfer of T-DNA  

Microsoft Academic Search

The 11 gene products of the Agrobacterium tumefaciens virB operon, together with the VirD4 protein, are proposed to form a membrane complex which mediates the transfer of T-DNA to plant cells. This study examined one putative component of that complex, VirB4. A deletion of the virB4 gene on the Ti plasmid pTiA6NC was constructed by replacing the virB4 gene with

Karla Jean Fullner; Kathryn M. Stephens; Eugene W. Nester

1994-01-01

171

The T-DNA ORF8 of the cucumopine-type Agrobacterium rhizogenes Ri plasmid is involved in auxin response in transgenic tobacco  

Microsoft Academic Search

In A. rhizogenes, various Ri plasmids are found that show some differences in terms of genetic organization as compared to the reference plasmid pRiA4. On this plasmid as well on the pRi2659 and pRi8196, the T-DNA genes rolA, rolB, and rolC (ORFs10, 11, and 12) appear to sufice to induce root formation in most assayed plant species. However, the most

Akila Ouartsi; Danielle Clérot; Alain D Meyer; Yves Dessaux; Jean Brevet; Mercè Bonfill

2004-01-01

172

Mycelium development and architecture, and spore production of Scutellospora reticulata in monoxenic culture with Ri T-DNA transformed carrot roots  

Microsoft Academic Search

Mycelium development and architecture and spore production were studied in Scutellospora reticulata from single-spore isolates grown with Ri T-DNA transformed carrot root-organ culture in monoxenic system. Culture establishment, anastomosis occurrence and auxiliary cell development also were examined. Seventy percent of the pregerminated disinfected spores colonized the transformed carrot roots. After 8 mo, the average spore production was 56 (24–130) per

F. A. De Souza; S. Declerck

2003-01-01

173

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

Microsoft Academic Search

Hairy roots are generated by integration of T-DNA in host plant genome from root inducing (Ri) plasmid of Agrobacterium rhizogenes and have been utilized for production of secondary metabolites in different plant systems. In Catharanthus roseus, hairy roots are known to show different morphologies, growth patterns, and alkaloid contents. It is also known that during\\u000a transformation, there is a differential

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

2010-01-01

174

Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri T-DNA gene integration  

Microsoft Academic Search

Hairy root cultures of Catharanthus roseus var. Prabal were established by infecting the leaves with Agrobacterium rhizogenes agropine-type A4 strain. Two hundred and fifty independent root clones were evaluated for growth, morphology, number of integration of Ri T-DNA genes and alkaloid contents. On the basis of growth pattern, type of branching and number of lateral roots we were able to separate

Jyoti Batra; Ajaswrata Dutta; Digvijay Singh; Sushil Kumar; Jayanti Sen

2004-01-01

175

Role of the host cell cycle in the Agrobacterium -mediated genetic transformation of Petunia : Evidence of an S-phase control mechanism for T-DNA transfer  

Microsoft Academic Search

Chimeric ß-glucuronidase (GUS) gene expression in an efficientAgrobacterium-mediated transformation system utilising mesophyll cells ofPetunia hybrida synchronized with cell cycle phase-specific inhibitors (mimosine and colchicine) was used to show the absolute requirement of S-phase for transfer and\\/or integration of the transferred DNA (T-DNA). Flow-cytometric analysis of nuclear DNA content and immunohistological detection of bromodeoxyuridine (BrdUrd) incorporation showed that, prior to phytohormone

Estelle Villemont; Frédéric Dubois; Rajbir S. Sangwan; Gérard Vasseur; Yvan Bourgeois; Brigitte S. Sangwan-Norreel

1997-01-01

176

Detection of transformed cells in crown gall tumors using the GUS reporter gene and correlation of GUS stained cells with T-DNA gene activity  

SciTech Connect

Crown gall tumors are a mixture of transformed hormone producing cells and normal cells. Until now it has not been possible to directly visualize these cell types in situ. We have constructed strains of Agrobacterium tumefaciens that carry the 35S-{beta}-glucuronidase (GUS) reporter gene in either wild type or mutant Ti plasmids. Using histochemical staining for GUS activity, blue (GUS positive) sectors are observed in tumor sections. In order to demonstrate that the blue sectors actually represent cells expressing other T-DNA genes, we have looked for T-DNA gene encoded enzyme activity in the stained and unstained sectors. The blue sectors accumulate octopine (a product of the octopine synthase gene on the T-DNA) while the white (GUS negative) sectors do not. We conclude that the use of the GUS reporter gene provides a sensitive and reliable method for visualizing transformation events in plant tissues. A comparison of the proportion of transformed and nontransformed cells in wild type tumors vs. tumors deficient in auxin or cytokinin encoding genes will be discussed.

Black, R.C. (Pennsylvania State Univ., Media (USA)); Labriola, J.; Binns, A.N. (Univ. of Pennsylvania, Philadelphia (USA))

1990-05-01

177

Cystic fibrosis in the mouse by targeted insertional mutagenesis  

Microsoft Academic Search

Cystic flbrosis is a fatal genetic disorder which afflicts 50,000 people worldwide. A viable animal model would be invaluable for investigating and combating this disease. The mouse cystic flbrosis transmembrane conductance regulator gene was disrupted in embryonal stem cells using an insertional gene targeting vector. Germ-line chimaeras were derived and the offspring of heterozygous crosses studied. These homozygous mutant mice

Julia R. Dorin; Paul Dickinson; Eric W. F. W. Alton; Stephen N. Smith; Duncan M. Geddes; Barbara J. Stevenson; Wendy L. Kimber; Stewart Fleming; Alan R. Clarke; Martin L. Hooper; Louise Anderson; Rosa S. P. Beddington; David J. Porteous

1992-01-01

178

The Basques according to polymorphic Alu insertions  

Microsoft Academic Search

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

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

2001-01-01

179

Quantum Dynamics of Insertion Reactions  

Microsoft Academic Search

We describe quantum-dynamical calculations of insertion reactions involving metastable C, N and O atoms with hydrogen molecules. We use a time independent hyperspherical body-frame formalism. Reaction probabilities, rovibrational distributions, integral and differential cross sections and product translational energy distributions have been computed and compared with recent experimental data.

Pascal Honvault; Jean-Michel Launay

180

Insertional mutagenesis in transgenic mice  

SciTech Connect

The molecular analysis of mutations in the mouse is an ideal way of establishing structure-function relationships between individual genes and complex developmental phenotypes. Insertional mutations, caused by the integration of a transgene, are advantageous because the mutant locus is tagged' with the exogenously added DNA and can be accessed with existing cloning and molecular analysis techniques. The authors have implemented a large-scale insertional mutagenesis program at the Oak Ridge National Laboratory in an effort to identify numerous mutations with developmental phenotypes. Several of these mutations are currently being subjected to an in-depth molecular analysis involving the characterization of the structure and expression of the mutant locus. Additionally, the authors are mapping transgene integration sites with in situ hybridization procedures, which will result in additional well-characterized molecular access points for the mouse genome.

Woychik, R.P.; Bultman, S.J.; Schrick, J.J.; Moyer, J.H.; Beatty, B.R.; Godfrey, V.L.; Selby, P.B. (Oak Ridge National Lab., TN (United States))

1991-03-11

181

Insert type milling cutter evaluation  

SciTech Connect

The purpose of this test was to obtain data on the major manufacturers of Insert Type Milling Cutters. Each manufacturers' representative was contacted and informed of the pending test. They were told of the type of material to be machined and the parameters of the operation. The manufacturers were asked to supply us with a cutter and inserts; they recommended to meet our needs for machining 304 stainless steel as specified in the Machining Data Handbook. Only three manufacturers responded; Carboloy, Kennametal, and Valenite., and cutters from each were tested. Each cutter was exposed to the exact same conditions, varying only where the cutter parameters dictated, to assure equality. The test results show that the Valenite cutter and inserts would require the least amount of maintenance, the greatest amount of wear life, and the best finishes. This cutter also required the least amount of horsepower in each cut. This is also advantageous from a power consumption outlook. The Valenite used a maximum of 8.01 Hp, which is 5968.48 W of power, where the Kennametal cutter used a maximum of 17.443 Hp, which is 13,012.48 W of power consumed. This is almost two times the energy consumption.

Gonzales, J.F.

1980-01-01

182

BayGenomics: a resource of insertional mutations in mouse embryonic stem cells  

Microsoft Academic Search

The BayGenomics gene-trap resource (http:\\/\\/ baygenomics.ucsf.edu) provides researchers with access to thousands of mouse embryonic stem (ES) cell lines harboring characterized insertional mutations in both known and novel genes. Each cell line contains an insertional mutation in a specific gene. The identity of the gene that has been interrupted can be determined from a DNA sequence tag. Approximately 75% of

Doug Stryke; Michiko Kawamoto; Conrad C. Huang; Susan J. Johns; Leslie A. King; Courtney A. Harper; Elaine C. Meng; Roy E. Lee; Alice Yee; Larry L'italien; Pao-tien Chuang; Stephen G. Young; William C. Skarnes; Patricia C. Babbitt; Thomas E. Ferrin

2003-01-01

183

Retargeting sleeping beauty transposon insertions by engineered zinc finger DNA-binding domains.  

PubMed

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

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

2012-07-10

184

Insertional Mutagenesis Using Tnt1 Retrotransposon in Potato1[OPEN  

PubMed Central

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.

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

2013-01-01

185

P systems with minimal insertion and deletion  

Microsoft Academic Search

In this paper, we consider insertion–deletion P systems with priority of deletion over insertion. We show that such systems with one-symbol context-free insertion and deletion rules are able to generate Parikh sets of all recursively enumerable languages (PsRE). If a one-symbol one-sided context is added to the insertion or deletion rules, then all recursively enumerable languages can be generated. The

Artiom Alhazov; Alexander Krassovitskiy; Yurii Rogozhin; Sergey Verlan

2011-01-01

186

Insertion devices control development at SRRC  

Microsoft Academic Search

Insertion devices are the most important gadget of the third generation Taiwan Light Source at SRRC. These insertion devices include the turn-key systems and in-house developed systems in SRRC. The corresponding control configurations are different due to historical reason. However precision, fastness, simplicity, and uniformity in insertion devices control are the most desirable from practical viewpoint. The proposed uniform control

K. T. Pan; Jenny Chen; C. J. Wang; C. S. Chen; K. H. Hu; Chaoen Wang; C. H. Chang; C. S. Hwang; T. C. Fan; K. T. Hsu

1997-01-01

187

Automatic insertion of BIST hardware using VHDL  

Microsoft Academic Search

We present a system which automatically inserts BIST hardware to a circuit described in VHDL. An appropriate VHDL modeling style for automatic insertion of BIST hardware is investigated. Use of BILBO is primarily pursued in the system. Algorithmic and rule-based approaches are used in the insertion of BILBO. Test scheduling and control signal distribution are also performed by the system.

Kwanghyun Kim; Joseph G. Tront; Dong Sam Ha

1988-01-01

188

Effect of Agrobacterium rhizogenes on phenolic content of Mentha pulegium elite clonal line for phytoremediation applications  

Microsoft Academic Search

Mentha pulegium elite clonal line MPH-4 is known to contain high levels of endogenous phenolic compounds. These compounds are secondary metabolites of the phenylpropanoid pathway that aid in several metabolic processes, including stress response and hormone production. Agrobacterium rhizogenes is a soil bacterium utilized for its ability to transfer its Rol genes via the Ri plasmid on t-DNA. A. rhizogenes

Sarah Strycharz; Kalidas Shetty

2002-01-01

189

Detection of two Alu insertions in the CFTR gene  

Microsoft Academic Search

BackgroundLINE-1 (long interspersed element-1) or L1-mediated retrotransposition is a potent force in human genome evolution and an occasional cause of human genetic disease. Since the first report of two de novo L1 insertions in the F8 gene causing hemophilia A, more than 50 L1-mediated retrotranspositional events have been identified as causing human genetic disease. However, a significant bias has generally

Jian-Min Chen; Emmanuelle Masson; Milan Macek; Odile Raguénès; Tereza Piskackova; Brigitte Fercot; Libor Fila; David N. Cooper; Marie-Pierre Audrézet; Claude Férec

2008-01-01

190

Flublok (Influenza Vaccine) Package Insert  

Center for Biologics Evaluation and Research (CBER)

Text Version... Each of the three HAs is expressed in this cell line using a baculovirus vector (Autographa californica nuclear polyhedrosis virus), extracted from ... More results from www.fda.gov/downloads/biologicsbloodvaccines/vaccines

191

O-ring insertion tool  

NASA Astrophysics Data System (ADS)

A tool for installing an O-ring in an inwardly facing O-ring groove in an electrical connection includes a guide shaft and a contact member slidably and rotatably received in the guide shaft. The guide shaft includes an elongated cylindrical shaft portion and a flared end portion on the shaft portion. The contact member includes an attachment portion for slidably and rotatably securing the contact member on the shaft portion and a contact finger on the attachment portion which extends in a direction toward the flared end portion of the guide shaft. The tool is operable by assembling an O-ring on the shaft portion between the flared end portion and the contact finger, inserting the flared end portion into a connection and then manipulating the contact member to advance the O-ring into an O-ring groove in the connector with the contact finger.

Zannini, Frank

1991-09-01

192

Frictional insertion kinetics of bone biopsy needles.  

PubMed

Patients undergoing a percutaneous bone biopsy often complain of pain during needle insertion, despite local anesthesia. Bone biopsy needles are typically inserted with combined axial and twisting motions. These motions could cause pain through frictional heating or direct mechanical irritation. The hypothesis of this study is that the insertion energy of bone biopsy needles can be reduced by modifying the insertion kinetics or by adding a friction-lowering coating to the needles. Jamshidi bone biopsy needles were driven into a bone analog model by an MTS materials testing machine operating under axial and rotational displacement control. The load/torque recordings showed that, to significantly decrease insertion energy and peak resistance to needle insertion, axial velocity and angular frequency had to be decreased to one quarter of the baseline, typical-usage parameters. However the increased insertion time may not be acceptable clinically. The majority of the insertion energy was associated with the needle axial thrust rather than with needle twisting. Overcoming friction against the side of the needle consumed much more of the insertion energy than did the process of cutting per se. None of five needle coatings tested succeeded in appreciably lowering the insertion energy, and none achieved a substantial decrease in peak resisting force. PMID:11783735

Heiner, A D; Brown, T D; Rossin, V; Buckwalter, J A

2001-12-01

193

A High-Throughput Arabidopsis Reverse Genetics System  

Microsoft Academic Search

A collection of Arabidopsis lines with T-DNA insertions in known sites was generated to increase the efficiency of func- tional 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

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

2002-01-01

194

Insertion devices at the advanced photon source  

SciTech Connect

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

Moog, E.R.

1996-07-01

195

Legitimate Skew Clock Routing with Buffer Insertion  

Microsoft Academic Search

In this paper, we propose a new quick and effective legitimate skew clock routing with buffer insertion algorithm. We analyze\\u000a the optimal buffer position in the clock path, and conclude the sufficient condition and heuristic condition for buffer insertion\\u000a in clock net. During the routing process, this algorithm integrates buffer insertion and node merging together, and performs\\u000a them in parallel.

Xinjie Wei; Yici Cai; Meng Zhao; Xianlong Hong

2006-01-01

196

Identification of insertion hot spots for non-LTR retrotransposons: computational and biochemical application to Entamoeba histolytica  

Microsoft Academic Search

The genome of the human pathogen Entamoeba histolytica contains non-long terminal repeat (LTR) retrotransposons, the EhLINEs and EhSINEs, which lack targeted insertion. We investigated the importance of local DNA structure, and sequence preference of the element-encoded endonuclease (EN) in selecting target sites for retrotransposon insertion. Pre-insertion loci were tested compu- tationally to detect unique features based on DNA structure, thermodynamic

Prabhat K. Mandal; Kamal Rawal; Ram Ramaswamy; Alok Bhattacharya; Sudha Bhattacharya

2006-01-01

197

Nozzle insert for mixed mode fuel injector  

DOEpatents

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.

Lawrence, Keith E. (Peoria, IL)

2006-11-21

198

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

NASA Astrophysics Data System (ADS)

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

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

199

Mitochondrial DNA transfer to the nucleus generates extensive insertion site variation in maize.  

PubMed

Mitochondrial DNA (mtDNA) insertions into nuclear chromosomes have been documented in a number of eukaryotes. We used fluorescence in situ hybridization (FISH) to examine the variation of mtDNA insertions in maize. Twenty overlapping cosmids, representing the 570-kb maize mitochondrial genome, were individually labeled and hybridized to root tip metaphase chromosomes from the B73 inbred line. A minimum of 15 mtDNA insertion sites on nine chromosomes were detectable using this method. One site near the centromere on chromosome arm 9L was identified by a majority of the cosmids. To examine variation in nuclear mitochondrial DNA sequences (NUMTs), a mixture of labeled cosmids was applied to chromosome spreads of ten diverse inbred lines: A188, A632, B37, B73, BMS, KYS, Mo17, Oh43, W22, and W23. The number of detectable NUMTs varied dramatically among the lines. None of the tested inbred lines other than B73 showed the strong hybridization signal on 9L, suggesting that there is a recent mtDNA insertion at this site in B73. Different sources of B73 and W23 were examined for NUMT variation within inbred lines. Differences were detectable, suggesting either that mtDNA is being incorporated or lost from the maize nuclear genome continuously. The results indicate that mtDNA insertions represent a major source of nuclear chromosomal variation. PMID:18202357

Lough, Ashley N; Roark, Leah M; Kato, Akio; Ream, Thomas S; Lamb, Jonathan C; Birchler, James A; Newton, Kathleen J

2008-01-01

200

Mitochondrial DNA Transfer to the Nucleus Generates Extensive Insertion Site Variation in Maize  

PubMed Central

Mitochondrial DNA (mtDNA) insertions into nuclear chromosomes have been documented in a number of eukaryotes. We used fluorescence in situ hybridization (FISH) to examine the variation of mtDNA insertions in maize. Twenty overlapping cosmids, representing the 570-kb maize mitochondrial genome, were individually labeled and hybridized to root tip metaphase chromosomes from the B73 inbred line. A minimum of 15 mtDNA insertion sites on nine chromosomes were detectable using this method. One site near the centromere on chromosome arm 9L was identified by a majority of the cosmids. To examine variation in nuclear mitochondrial DNA sequences (NUMTs), a mixture of labeled cosmids was applied to chromosome spreads of ten diverse inbred lines: A188, A632, B37, B73, BMS, KYS, Mo17, Oh43, W22, and W23. The number of detectable NUMTs varied dramatically among the lines. None of the tested inbred lines other than B73 showed the strong hybridization signal on 9L, suggesting that there is a recent mtDNA insertion at this site in B73. Different sources of B73 and W23 were examined for NUMT variation within inbred lines. Differences were detectable, suggesting either that mtDNA is being incorporated or lost from the maize nuclear genome continuously. The results indicate that mtDNA insertions represent a major source of nuclear chromosomal variation.

Lough, Ashley N.; Roark, Leah M.; Kato, Akio; Ream, Thomas S.; Lamb, Jonathan C.; Birchler, James A.; Newton, Kathleen J.

2008-01-01

201

Measurement of Orthopedic Cortical Bone Screw Insertion Performance in Cadaver Bone and Model Materials  

Microsoft Academic Search

In this study, a method was developed for distinguishing insertion and driving performance between different self-tapping\\u000a bone screw designs. To measure screw starting load, torque and displacements, a test apparatus was developed utilizing a modified\\u000a drill press with the capability to measure in-line torque, axial compression load, and axial displacement. Specimens were\\u000a inserted into cadaver bone to measure a baseline

B. Kincaid; L. Schroder; J. Mason

2007-01-01

202

Mitochondrial DNA Transfer to the Nucleus Generates Extensive Insertion Site Variation in Maize  

Microsoft Academic Search

Mitochondrial DNA (mtDNA) insertions into nuclear chromosomes have been documented in a num- ber of eukaryotes. We used fluorescence in situ hybridization (FISH) to examine the variation of mtDNA insertions in maize. Twenty overlapping cosmids, representing the 570-kb maize mitochondrial genome, were individually labeled and hybridized to root tip metaphase chromosomes from the B73 inbred line. A minimum of 15

Ashley N. Lough; Leah M. Roark; Akio Kato; Thomas S. Ream; Jonathan C. Lamb; James A. Birchler; Kathleen J. Newton

2008-01-01

203

Effects of Single P-Element Insertions on Bristle Number and Viability in Drosophila Melanogaster  

PubMed Central

Single P-element mutagenesis was used to construct 1094 lines with P[lArB] inserts on all three major chromosomes in an isogenic background previously free of P elements. The effects of insertions on bristle number and on viability were assessed by comparison to 392 control lines. The variance and effects of P-element inserts on bristle number and viability were larger than those inferred from spontaneous mutations. The distributions of effects on bristle number were symmetrical and highly leptokurtic, such that a few inserts with large effects caused most of the increase in variance. The distribution of effects on viability were negatively skewed and platykurtic. On average, the effects of P-element insertions on bristle number were partly recessive and on viability were completely recessive. P-element inserts with large effects on bristle number tended to have reduced viability, but the correlation between the absolute value of the effects on bristle number and on viability was not strong. Fifty P-element inserts tagging quantitative trait loci (QTLs) with large effects on bristle number were mapped cytogenetically. Two P-element-induced scabrous alleles and five extramacrochaetae alleles were generated. Single P-element mutagenesis is a powerful method for identifying QTLs at the level of genetic locus.

Lyman, R. F.; Lawrence, F.; Nuzhdin, S. V.; Mackay, TFC.

1996-01-01

204

Penile prosthesis insertion for acute priapism.  

PubMed

Shunt surgery is not universally successful toward detumescence, may lead to erectile dysfunction, and can make eventual penile prosthesis insertion difficult. Penile prosthesis insertion during a priapistic episode alleviates ischemic pain, allows the patient to resume sexual function sooner, and prevents corporal scarring and shortening that makes subsequent prosthesis implantation difficult. PMID:23905940

Tausch, Timothy J; Mauck, Ryan; Zhao, Lee C; Morey, Allen F

2013-06-27

205

Ad insertion in automatically composed documents  

Microsoft Academic Search

We consider the problem of automatically inserting advertisements (ads) into machine composed documents. We explicitly analyze the fundamental tradeoff between expected revenue due to ad insertion and the quality of the corresponding composed documents. We show that the optimal tradeoff a publisher can expect may be expressed as an efficient-frontier in the revenue-quality space. We develop algorithms to compose documents

Niranjan Damera-Venkata; José Bento

2012-01-01

206

Buffer insertion for noise and delay optimization  

Microsoft Academic Search

Buffer insertion has successfully been applied to reduce delay in global interconnect paths; however, existing techniques only optimize delay and timing slack. With the increasing ratio of coupling to total capacitance and the use of aggressive dynamic logic circuit families, noise is becoming a major design bottleneck. We present comprehensive buffer insertion techniques for noise and delay optimization. Our experiments

Charles J. Alpert; Anirudh Devgan; Stephen T. Quay

1998-01-01

207

Insertion Sequence Diversity in Archaea  

PubMed Central

Insertion sequences (ISs) can constitute an important component of prokaryotic (bacterial and archaeal) genomes. Over 1,500 individual ISs are included at present in the ISfinder database (www-is.biotoul.fr), and these represent only a small portion of those in the available prokaryotic genome sequences and those that are being discovered in ongoing sequencing projects. In spite of this diversity, the transposition mechanisms of only a few of these ubiquitous mobile genetic elements are known, and these are all restricted to those present in bacteria. This review presents an overview of ISs within the archaeal kingdom. We first provide a general historical summary of the known properties and behaviors of archaeal ISs. We then consider how transposition might be regulated in some cases by small antisense RNAs and by termination codon readthrough. This is followed by an extensive analysis of the IS content in the sequenced archaeal genomes present in the public databases as of June 2006, which provides an overview of their distribution among the major archaeal classes and species. We show that the diversity of archaeal ISs is very great and comparable to that of bacteria. We compare archaeal ISs to known bacterial ISs and find that most are clearly members of families first described for bacteria. Several cases of lateral gene transfer between bacteria and archaea are clearly documented, notably for methanogenic archaea. However, several archaeal ISs do not have bacterial equivalents but can be grouped into Archaea-specific groups or families. In addition to ISs, we identify and list nonautonomous IS-derived elements, such as miniature inverted-repeat transposable elements. Finally, we present a possible scenario for the evolutionary history of ISs in the Archaea.

Filee, J.; Siguier, P.; Chandler, M.

2007-01-01

208

Membrane protein insertion in E. coli.  

PubMed

Integral membrane proteins typically span the lipid bilayer with hydrophobic alpha helices. These proteins can span the membrane once or multiple times with hydrophilic domains facing both sides of the membrane. In Escherichia coli, the insertion of proteins into the membrane is catalyzed by the Sec translocase and the YidC membrane insertase. YidC can function on its own to insert proteins or together with the Sec translocase to facilitate membrane protein insertion. In this chapter, we will describe the construction of a YidC depletion strain that can be used to examine whether YidC is required for membrane protein insertion. We will also present assays for determining whether a region of a membrane protein is inserted across the membrane. PMID:20419404

Yuan, Jijun; Dalbey, Ross E; Kuhn, Andreas

2010-01-01

209

Effects of Single P-Element Insertions on Olfactory Behavior in Drosophila Melanogaster  

PubMed Central

Single P-element (P[lArB]) insertional mutagenesis of an isogenic strain was used to identify autosomal loci affecting odor-guided behavior of Drosophila melanogaster. The avoidance response to benzaldehyde of 379 homozygous P[lArB] element-containing insert lines was evaluated quantitatively. Fourteen smell impaired (smi) lines were identified in which P[lArB] element insertion caused different degrees of hyposmia in one or both sexes. The smi loci map to different cytological locations and probably are novel olfactory genes. Enhancer trap analysis of the smi lines indicates that expression of at least 10 smi genes is controlled by olfactory tissue-specific promoter/enhancer elements.

Anholt, RRH.; Lyman, R. F.; Mackay, TFC.

1996-01-01

210

Central Solenoid Insert Technical Specification  

SciTech Connect

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.

Martovetsky, Nicolai N [ORNL; Smirnov, Alexandre [ORNL

2011-09-01

211

The Maize Genome Contains a Helitron Insertion  

PubMed Central

The maize mutation sh2-7527 was isolated in a conventional maize breeding program in the 1970s. Although the mutant contains foreign sequences within the gene, the mutation is not attributable to an interchromosomal exchange or to a chromosomal inversion. Hence, the mutation was caused by an insertion. Sequences at the two Sh2 borders have not been scrambled or mutated, suggesting that the insertion is not caused by a catastrophic reshuffling of the maize genome. The insertion is large, at least 12 kb, and is highly repetitive in maize. As judged by hybridization, sorghum contains only one or a few copies of the element, whereas no hybridization was seen to the Arabidopsis genome. The insertion acts from a distance to alter the splicing of the sh2 pre-mRNA. Three distinct intron-bearing maize genes were found in the insertion. Of most significance, the insertion bears striking similarity to the recently described DNA helicase–bearing transposable elements termed Helitrons. Like Helitrons, the inserted sequence of sh2-7527 is large, lacks terminal repeats, does not duplicate host sequences, and was inserted between a host dinucleotide AT. Like Helitrons, the maize element contains 5? TC and 3? CTRR termini as well as two short palindromic sequences near the 3? terminus that potentially can form a 20-bp hairpin. Although the maize element lacks sequence information for a DNA helicase, it does contain four exons with similarity to a plant DEAD box RNA helicase. A second Helitron insertion was found in the maize genomic database. These data strongly suggest an active Helitron in the present-day maize genome.

Lal, Shailesh K.; Giroux, Michael J.; Brendel, Volker; Vallejos, C. Eduardo; Hannah, L. Curtis

2003-01-01

212

Elliptically polarizing adjustable phase insertion device  

DOEpatents

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.

Carr, Roger (Redwood City, CA)

1995-01-01

213

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

PubMed

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

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

2005-05-20

214

Package insert Influenza Vaccine STN BL 125254  

Center for Biologics Evaluation and Research (CBER)

Text Version... 1 Grdcp2\\Cabinets\\RA Labeling\\Afluria\\USA\\Manuscripts\\Package Insert, Blank [fabe21]_ASU for 2013-2014 Influenza Season Version 36.0 ... More results from www.fda.gov/downloads/biologicsbloodvaccines/vaccines

215

Insertable fluid flow passage bridgepiece and method  

DOEpatents

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.

Jones, Daniel O. (Glenville, NV)

2000-01-01

216

A Investigation of Some Lithium Insertion Compounds  

Microsoft Academic Search

Lithium insertion compounds are of technical interest as storage-battery electrodes; they are also of scientific interest as they offer low-temperature synthetic routes to compounds having unusual atomic ordering and to formal valence states that may not be accessible at higher temperatures normally employed in solid-state reactions. In this study lithium insertion compounds have been investigated to understand structure-property relationships of

Geeta Ahuja

1991-01-01

217

Low-insertion loss Raman fiber modulator  

Microsoft Academic Search

Characteristics of the Raman-effect based modulator for high power Yb-doped fiber lasers have been analyzed by numerical simulation. Insertion loss, backreflection, and contrast ratio are considered as functions of the laser power and Ge concentration in fiber. The optimum GeO2 doping has been shown to improve the efficiency of the modulator at powers of 5-20 W, providing the insertion loss

I. Torres-Gomez; A. N. Starodumov; V. N. Filippov; Yu. O. Barmenkov; A. Martínez-Rios

2001-01-01

218

Low-insertion loss Raman fiber modulator  

Microsoft Academic Search

Characteristics of the Raman-effect based modulator for high power Yb-doped fiber lasers have been analyzed by numerical simulation. Insertion loss, backreflection, and contrast ratio are considered as functions of the laser power and Ge concentration in fiber. The optimum GeO2 doping has been shown to improve the efficiency of the modulator at powers of 5–20 W, providing the insertion loss

I. Torres-Gomez; A. N. Starodumov; V. N. Filippov; Yu. O. Barmenkov; A. Mart??nez-Rios

2001-01-01

219

Compliant Control of Threaded Fastener Insertion  

Microsoft Academic Search

Controlled compliance and accommodation matrixtechniques are considered for robust insertion ofthreaded fasteners. Errors in translational positioningare shown to be easily corrected. Errors in the angle oftilt between the threaded parts are shown to be muchmore difficult to correct and constrain the region ofconvergence for simple linear techniques.1 IntroductionAccording to a study by Nevins and Whitney[Nevins 80] the insertion and tightening

Edward J. Nicolson; Ronald S. Fearing

1993-01-01

220

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

PubMed

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

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

2013-08-14

221

Human Gene Targeting Favors Insertions Over Deletions  

PubMed Central

Abstract Gene targeting is a powerful technique for manipulating the human genome, but few studies have directly compared the targeting frequencies of various types of vector constructs. Here we show that similar targeting constructs are able to insert nucleotides at the homologous chromosomal target locus more efficiently than they can delete nucleotides, and combination insertion/deletion vectors appear to target at intermediate frequencies. This holds true for deletions ranging from 1 to 334 bp and insertions ranging from 1 to 1332 bp. In addition, vectors designed to inactivate the human hypoxanthine phosphoribosyltransferase gene (HPRT) by deleting nucleotides often produced rearrangements at the target locus that in many cases were due to insertions of multimerized vector constructs, effectively converting a deletion vector into an insertion vector. These findings were obtained when adeno-associated virus vectors were used to efficiently deliver single-stranded DNA targeting constructs, but the same phenomenon was also observed when transfecting linearized double-stranded plasmids. Thus human cells distinguish between deletion and insertion vectors and process their recombination intermediates differently, presumably at the heteroduplex stage, with implications for the design of gene-targeting vectors and the evolution of human genomes.

Hirata, Roli K.

2008-01-01

222

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

PubMed

An Agrobacterium tumefaciens -mediated transformation (ATMT) system was successfully developed for Colletotrichum truncatum, the causal agent of chili anthracnose. A. tumefaciens carrying a hygromycin phosphotransferase gene (hph) and a green fluorescent protein (gfp) gene was used to transform the conidiospores of two C. truncatum pathotypes F8-3B and BRIP26974. Optimum transformation efficiency was obtained when equal volumes of A. tumefaciens strain AGL1 carrying either pJF1 or pPK2 binary vector was used to transform C. truncatum conidiospores at 10(6) /ml and co-cultivated at 24 °C for three days. Southern blot analysis indicated that 87.5% of the transformants contained randomly inserted, single copies of the T-DNA. Infection and colonisation of chili fruit at the mature red stage with F8-3B-GFP and BRIP26974-GFP confirmed the maintenance of virulence within these transformed pathotypes. In situ studies of infection and colonisation of the susceptible genotype fruit using fluorescent microscopy and transformed isolates of C. truncatum expressing GFP revealed that the pathogen was able to colonise healthy fruit tissue intercellularly in an endophytic manner without producing secondary biotrophic infection structures. The developed transformation system will be used to study the function of pathogenicity genes in C. truncatum using both forward and reverse genetics approaches. PMID:22052577

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

2011-11-03

223

Generation of enhancer trap lines in Arabidopsis and characterization of expression patterns in the inflorescence.  

PubMed

Eleven thousand, three hundred and seventy enhancer/promoter trap lines in Arabidopsis were generated via T-DNA transformation utilizing the binary vector pD991 that contains a minimal promoter fused to the uidA reporter gene. Overall 31% of the lines generated exhibit a staining pattern in the inflorescence. Flanking DNA has been cloned from 15 lines exhibiting inflorescence staining patterns by either thermal asymmetric interlaced PCR (TAIL-PCR), inverse PCR (IPCR), or partial library construction. Seeds from these lines are available from the ABRC and NASC Arabidopsis stock centers and DNA pools are available from the ABRC. PMID:10230066

Campisi, L; Yang, Y; Yi, Y; Heilig, E; Herman, B; Cassista, A J; Allen, D W; Xiang, H; Jack, T

1999-03-01

224

Mechanisms for sodium insertion in carbon materials  

NASA Astrophysics Data System (ADS)

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.

Stevens, David Andrew

225

Peptide partitioning properties from direct insertion studies  

SciTech Connect

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.

Ulmschneider, Martin [University of Oxford; Smith, Jeremy C [ORNL; Ulmschneider, Jakob [University of Heidelberg

2010-06-01

226

Optimal anaesthetic depth for LMA insertion  

PubMed Central

Purpose: A fixed dose of propofol administered rapidly can be insufficient or in excess resulting in airway complications and haemodynamic disturbances. This study is designed to assess whether loss of motor response to jaw thrust can be a reliable clinical indicator of anaesthetic depth for laryngeal mask airway (LMA) insertion. Methods: One hundred and twenty ASA I and II patients scheduled for general anaesthesia on day care basis were randomly allocated into two groups. Following pre-oxygenation, anaesthesia was induced to accomplish LMA insertion either with a 3 mg/kg propofol (Group CD, n=60) or in dose to abolish jaw thrust response (Group JT, n=60). Mean arterial pressure (MAP) and heart rate were continuously monitored while LMA insertion conditions were recorded using 6 variable, 3 point score. Results: 85% patients developed apnea in group CD when compared to 2% in group JT, P<0.0001. Despite similar insertion score, propofol consumption was significantly more in group CD when compared to group JT. More than 20% fall of MAP from baseline was noted in group CD after induction but there was no significant hypotension at any time in group JT. Conclusion: Loss of motor response to jaw thrust provides satisfactory LMA insertion conditions.

Krishnappa, Sudeep; Kundra, Pankaj

2011-01-01

227

Cancer gene discovery: exploiting insertional mutagenesis.  

PubMed

Insertional mutagenesis has been used 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 are integrating viruses with the capacity to induce the deregulation of genes in the neighborhood of the insertion site. Retroviruses have been used 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 retrovirus-based approaches. Most recently, lentiviral vectors have appeared on the scene for use in cancer gene screens. Lentiviral vectors are replication-defective integrating vectors that have the advantage of being able to infect nondividing 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. Mol Cancer Res; 11(10); 1141-58. ©2013 AACR. PMID:23928056

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

2013-08-08

228

Nozzle cavity impingement/area reduction insert  

DOEpatents

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.

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

2002-01-01

229

Site-specific mutagenesis of the Ti plasmid by transformation of Agrobacterium tumefaciens with mutagenized T-DNA fragments cloned in E. coli plasmids  

Microsoft Academic Search

A DNA fragment covering the complete T-region of the Ti plasmid from Agrobacterium tumefaciens strain C58 was cloned in the Escherichia coli cosmid pHC79. This fragment was mutagenized by insertion of transposon Tn5. The isolated DNA from hybrid plasmids was used to transform cells of A. tumefaciens strain C58 applying the freeze-thaw method. Although the E. coli plasmids with the

Peter Zahm; Christine Hohmeyer; Klaus Geider

1984-01-01

230

Responses of an Arbuscular Mycorrhizal Fungus, Gigaspora margarita, to Exudates and Volatiles from the Ri T-DNA-Transformed Roots of Nonmycorrhizal and Mycorrhizal Mutants of Pisum sativum L Sparkle  

Microsoft Academic Search

Balaji, B., Poulin, M. J., Vierheilig, H., and Piché Y. 1995. Responses of an arbuscular mycorrhizal fungus, Gigaspora margarita, to exudates and volatiles from the Ri T-DNA-transformed roots of nonmycorrhizal and mycorrhizal mutants of Pisum sativum L Sparkle. Experimental Mycology 19, 275-283. Transformed root cultures were established from the nonmycorrhizal (Myc-) and mycorrhizal (Myc+) Pisum sativum L Sparkle mutants to

Boovaraghan Balaji; Marie Josée Poulin; Horst Vierheilig; Yves Piché

1995-01-01

231

Compact, low insertion-loss, sharp-rejection, and wide-band microstrip bandpass filters  

Microsoft Academic Search

This paper presents a new compact, low insertion-loss, sharp-rejection, and wide-band microstrip bandpass filter. A bandstop filter is introduced that uses a ring resonator with direct-connected orthogonal feed lines. A new bandpass filter based on the bandstop filter uses two tuning stubs to construct a wide-band passband with two sharp stopbands. Without coupling gaps between feed lines and rings, there

Lung-Hwa Hsieh; Kai Chang

2003-01-01

232

Insertion Loads and Forearm Muscle Activity During Flexible Hose Insertion Tasks  

Microsoft Academic Search

Objective: To quantify the physical demands of hose insertion tasks in automotive assembly operations and how they are affected by method and the mechanical interference between the hose and the flange. Background: Insertion tasks were identified by workers as physically demanding and can often lead to fatigue or losses in production attributable to pain or injury. Methods: Six male and

D. Christian Grieshaber; Thomas J. Armstrong

2007-01-01

233

Lithium insertion in tungsten oxide thin films  

NASA Astrophysics Data System (ADS)

Intercalation of lithium ions in amorphous tungsten oxide films and in monoclinic polycrystalline tungsten oxide films has been studied using electrochemical analyses and Raman spectroscopy. We present direct evidence for W5+ 5d- electron localization in amorphous tungsten oxide films upon lithium ion and electron intercalation, by comparing the Raman spectra of amorphous and crystalline tungsten oxide films. We observe that when lithium ions and electrons are inserted into the crystalline tungsten oxide films, the monoclinic structure of the material proceeds to progressively increased symmetry. On the other hand, when lithium ions and electrons are inserted into amorphous tungsten oxide films, the inserted electrons are localized in W5+ sites and polarize their surrounding lattice to form small polarons. The extra Raman peaks due to the W5+-O single bonds and W5+equalsO double bonds appear at 330 and 450 cm-1, respectively.

Lee, Se-Hee; Seong, Maeng J.; Ozcan, Esra; Tracy, C. Ed; Tepehan, Fatma Z.; Deb, Satyen K.

2001-11-01

234

Insertion loss of an acoustic enclosure  

NASA Astrophysics Data System (ADS)

Acoustical enclosures are the common arrangements in reducing airborne noise from shipboard machinery such as engines and generators. In this paper the theoretical models, established based on statistical energy analysis, are presented for predicting the insertion loss of acoustical enclosures in different frequency ranges. In addition to the consideration of resonant modal coupling between internal sound field and enclosure structural vibration, the nonresonant transmission though and the interaction between enclosure walls in the models are also included. It is shown that the insertion loss of enclosures is mainly controlled by the nonresonant modes in the intermediate frequency range. At high frequencies, the insertion loss of enclosures can be improved by increasing the sound absorption at the internal boundaries of enclosures. Experiments were carried out on two enclosures made of different materials. The measured results are compared with the predicted values and the good agreement between them is the initial demonstration of the validity and feasibility of the theoretical models. .

Ming, Ruisen; Pan, Jie

2004-12-01

235

Membrane Insertion of a Voltage Sensor Helix  

PubMed Central

Most membrane proteins contain a transmembrane (TM) domain made up of a bundle of lipid-bilayer-spanning ?-helices. TM ?-helices are generally composed of a core of largely hydrophobic amino acids, with basic and aromatic amino acids at each end of the helix forming interactions with the lipid headgroups and water. In contrast, the S4 helix of ion channel voltage sensor (VS) domains contains four or five basic (largely arginine) side chains along its length and yet adopts a TM orientation as part of an independently stable VS domain. Multiscale molecular dynamics simulations are used to explore how a charged TM S4 ?-helix may be stabilized in a lipid bilayer, which is of relevance in the context of mechanisms of translocon-mediated insertion of S4. Free-energy profiles for insertion of the S4 helix into a phospholipid bilayer suggest that it is thermodynamically favorable for S4 to insert from water to the center of the membrane, where the helix adopts a TM orientation. This is consistent with crystal structures of Kv channels, biophysical studies of isolated VS domains in lipid bilayers, and studies of translocon-mediated S4 helix insertion. Decomposition of the free-energy profiles reveals the underlying physical basis for TM stability, whereby the preference of the hydrophobic residues of S4 to enter the bilayer dominates over the free-energy penalty for inserting charged residues, accompanied by local distortion of the bilayer and penetration of waters. We show that the unique combination of charged and hydrophobic residues in S4 allows it to insert stably into the membrane.

Wee, Chze Ling; Chetwynd, Alan; Sansom, Mark S.P.

2011-01-01

236

RERTR-12 Insertion 2 Irradiation Summary Report  

SciTech Connect

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.

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

2012-09-01

237

Matching Sequences under Deletion/Insertion Constraints  

PubMed Central

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.

Sankoff, David

1972-01-01

238

The Insertional History of an Active Family of L1 Retrotransposons in Humans  

PubMed Central

As humans contain a currently active L1 (LINE-1) non-LTR retrotransposon family (Ta-1), the human genome database likely provides only a partial picture of Ta-1-generated diversity. We used a non-biased method to clone Ta-1 retrotransposon-containing loci from representatives of four ethnic populations. We obtained 277 distinct Ta-1 loci and identified an additional 67 loci in the human genome database. This collection represents ?90% of the Ta-1 population in the individuals examined and is thus more representative of the insertional history of Ta-1 than the human genome database, which lacked ?40% of our cloned Ta-1 elements. As both polymorphic and fixed Ta-1 elements are as abundant in the GC-poor genomic regions as in ancestral L1 elements, the enrichment of L1 elements in GC-poor areas is likely due to insertional bias rather than selection. Although the chromosomal distribution of Ta-1 inserts is generally a function of chromosomal length and gene density, chromosome 4 significantly deviates from this pattern and has been much more hospitable to Ta-1 insertions than any other chromosome. Also, the intra-chromosomal distribution of Ta-1 elements is not uniform. Ta-1 elements tend to cluster, and the maximal gaps between Ta-1 inserts are larger than would be expected from a model of uniform random insertion.

Boissinot, Stephane; Entezam, Ali; Young, Lynn; Munson, Peter J.; Furano, Anthony V.

2004-01-01

239

A Miniaturized Multilayered Marchand Balun Using Coupled Artificial Transmission Lines  

Microsoft Academic Search

A miniaturized Marchand balun is investigated in this letter. The Marchand balun is realized on a six-layered printed circuit board by integration of two coupled artificial transmission line sections. An additional artificial line is inserted in-between the coupled lines for compensating the asymmetry. With the help of the even odd mode analysis, the electrical characteristics of the coupled lines are

Tzyh-Ghuang Ma; Yu-Ting Cheng

2009-01-01

240

Anatomy and dimensions of rotator cuff insertions  

Microsoft Academic Search

The purpose of this study was to devise and implement an accurate and reproducible method of measuring the area and dimensions of the rotator cuff tendon insertions and their distance from the articular surface. Twenty fresh-frozen cadaveric upper-extremity specimens were divided into 2 groups of 10. In group 1 the specimens were dissected, leaving only the most distal rotator cuff

Jeffrey R Dugas; Deirdre A Campbell; Russell F Warren; Bruce H Robie; Peter J Millett

2002-01-01

241

Lymphatic Leak Complicating Central Venous Catheter Insertion  

Microsoft Academic Search

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.

Alex M. Barnacle; Tricia M. Kleidon

2005-01-01

242

Novel use of an "insertable" loop recorder  

PubMed Central

A patient with palpitations and suspected arrhythmia underwent Holter and external loop recorder monitoring. No arrhythmias were detected by these traditional monitoring methods. An insertable loop recorder (ILR) was placed on the patient’s chest and used as an extended loop recorder. An arrhythmia was ultimately recorded by the externally placed ILR leading to appropriate treatment.

Gimbel, J R

2003-01-01

243

Novel use of an "insertable" loop recorder.  

PubMed

A patient with palpitations and suspected arrhythmia underwent Holter and external loop recorder monitoring. No arrhythmias were detected by these traditional monitoring methods. An insertable loop recorder (ILR) was placed on the patient's chest and used as an extended loop recorder. An arrhythmia was ultimately recorded by the externally placed ILR leading to appropriate treatment. PMID:12748241

Gimbel, J R

2003-06-01

244

Speculum 'Self Insertion': A Pilot Study  

Microsoft Academic Search

Aim - This paper reports phase II of a pilot study that aimed to determine whether self-insertion of a speculum by women undergoing a pap smear would be more comfortable and lead to an improvement in satisfaction and a decrease in anxiety associated with this procedure. Background - Research demonstrates that pelvic examinations are considered by most women to be

Deborah Wright; Jennifer Fenwick; Pam Stephenson; Leanne Monterosso

2005-01-01

245

Methidium intercalator inserted into synthetic oligonucleotides.  

SciTech Connect

A new methidium intercalator phosphoramidite has been synthesized. Methidium incorporation into an oligonucleotide during the synthesis was confirmed by UV and MALDI TOF MS data. UV melting experiments showed enhanced stability of a duplex, containing internal methidium. Methidium phosphoramidite has been synthesized and used for insertion of intercalator into the deoxyoligonucleotides.

Timofeev, E. N.; Smirnov, I. P.; Haff, L. A.; Tishchenko, E. I.; Mirzabekov, A. D.; Florentiev, V. L.; Center for Mechanistic Biology and Biotechnology; Engelhardt Inst. of Molecular Biology; PerSeptive BioSystems Inc.

1996-01-01

246

Predicting cancellous bone failure during screw insertion.  

PubMed

Internal fixation of fractures often requires the tightening of bone screws to stabilise fragments. Inadequate application of torque can leave the fracture unstable, while over-tightening results in the stripping of the thread and loss of fixation. The optimal amount of screw torque is specific to each application and in practice is difficult to attain due to the wide variability in bone properties including bone density. The aim of the research presented in this paper is to investigate the relationships between motor torque and screw compression during powered screw insertion, and to evaluate whether the torque during insertion can be used to predict the ultimate failure torque of the bone. A custom test rig was designed and built for bone screw experiments. By inserting cancellous bone screws into synthetic, ovine and human bone specimens, it was established that variations related to bone density could be automatically detected through the effects of the bone on the rotational characteristics of the screw. The torque measured during screw insertion was found to be directly related to bone density and can be used, on its own, as a good predictor of ultimate failure torque of the bone. PMID:23466167

Reynolds, Karen J; Cleek, Tammy M; Mohtar, Aaron A; Hearn, Trevor C

2013-03-01

247

Status of the DIAMOND insertion devices  

Microsoft Academic Search

Seven insertion devices must be provided for the first DIAMOND beamlines on Day 1 of operations. A variety of devices have been selected, including four short period in-vacuum undulators, one 5 m conventional planar undulator, one 5 m helical undulator and a 3.5 T superconducting wiggler. Construction of several of these devices will soon commence. This paper will detail the

J. A. Clarke; N. Bliss; D. G. Clarke; F. E. Hannon; C. Hill; A. A. Muir; D. J. Scott; V. Francis

2003-01-01

248

Direct random insertion mutagenesis of Helicobacter pylori  

Microsoft Academic Search

Random insertion mutagenesis is a widely used technique for the identification of bacterial virulence genes. Most strategies for random mutagenesis involve cloning in Escherichia coli for passage of plasmids or for phenotypic selection. This can result in biased selection due to restriction or instability of the cloned DNA, or toxicity of the encoded products. We therefore created two mutant libraries

Ramon de Jonge; Dennis Bakker; Arnoud H. M van Vliet; Ernst J Kuipers; Christina M. J. E Vandenbroucke-Grauls; Johannes G Kusters

2003-01-01

249

Wire segmenting for improved buffer insertion  

Microsoft Academic Search

Buffer insertion seeks to place buffers on the wires of a signal netto minimize delay. Van Ginneken [Buffer Placement in Distributed RC-tree Networks for Minimal Elmore Delay] proposed an optimal dynamicprogramming solution (with extensions proposed by [7] [8][9] [12]) such that at most one buffer can be placed on a singlewire. This constraint can hurt solution quality, but it may

Charles J. Alpert; Anirudh Devgan

1997-01-01

250

Rack Insertion End Effector (RIEE) Automation.  

National Technical Information Service (NTIS)

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 leng...

N. Malladi

1993-01-01

251

Rack Insertion End Effector (RIEE) Guidance.  

National Technical Information Service (NTIS)

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 Ins...

N. S. Malladi

1994-01-01

252

Metal buffer layer inserted switchable mirrors  

Microsoft Academic Search

Thin buffer layers of hydrogen diffusive metals such as Ti, Nb, and V were inserted between a Mg4Ni thin film and a Pd top layer, which were prepared by DC magnetron sputtering. Their optical, electrochemical properties, and switching durability were investigated using both gasochromic and electrochromic switching methods. It has been proved that Ti, Nb, and V buffer layers can

S. Bao; K. Tajima; Y. Yamada; M. Okada; K. Yoshimura

2008-01-01

253

Lymphatic Leak Complicating Central Venous Catheter Insertion  

SciTech Connect

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.

Barnacle, Alex M., E-mail: alexbarnacle@yahoo.co.uk; Kleidon, Tricia M. [Great Ormond Street Hospital for Children, Department of Radiology (United Kingdom)

2005-12-15

254

Rotational vs. standard smooth laryngeal mask airway insertion in adults  

Microsoft Academic Search

OBJECTIVE: To compare the ease of insertion between rotational laryngeal mask airway (LMA) insertion and Brain's LMA insertion technique in terms of number of LMA insertion attempts, time duration of LMA insertion and complications: trauma, laryngospasm, and hypoxaemia.STUDY DESIGN: Randomized control study.PLACE AND DURATION OF STUDY: The Aga Khan University Hospital, Karachi, from September 2006 to May 2007.METHODOLOGY: One hundred

Dileep Kumar; Mueenullah Khan; Muhammad Ishaq

2012-01-01

255

MTID: a database of Sleeping Beauty transposon insertions in mice  

Microsoft Academic Search

The Sleeping Beauty (SB) transposon system pro- vides the first random insertional mutagen available for germline genetic screens in mice.In preparation for a large scale project to create, map and manage up to 5000 SB insertions, we have developed the Mouse Transposon Insertion Database (MTID; http:\\/\\/ mouse.ccgb.umn.edu\\/transposon\\/). Each insertion's genomic position, as well as the distance between the insertion and

Kevin Roberg-perez; Corey M. Carlson; David A. Largaespada

2003-01-01

256

Consolidation and disposal of PWR fuel inserts  

SciTech Connect

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

Wakeman, B.H. (Virginia Electric and Power Co., Glen Allen, VA (United States))

1992-08-01

257

MINI-. beta. insertion and luminosity for the RHIC lattice  

SciTech Connect

Requirement and implication of the mini-..beta.. insertion in RHIC lattice is studied. For the heavy ion operation, we find that: When the gradient is maintained at 67T/m, the free space for the experimental setup becomes +-4m at ..beta..* = 1.5m with maximum crossing angle of 3mrad at 100 GeV/amu. To maintain a minimum of +-5m free space for experimental setup, a higher gradient is needed. At 84.5T/m gradient, ..beta..* = 1m can be achieved. In this case, the beam size limitation is located at BC2, where 10cm coil i.d. is designed. When the machine is operating at the proton-heavy ion interaction mode, these common quadrupoles should be removed physically in order to let both beams crossing the center line at 3.5mrad. These mini-..beta.. insertion can increase the luminosity by a factor of 2.3. For the proton operational mode, we find that ..beta..* = 1m, which corresponds to ..beta.. = 1200m is also operable without the mini-..beta.. quadrupoles. When the mini-..beta.. quads are used, ..beta..* = 0.25 can be achieved. 1 ref., 3 tabs.

Lee, S.Y.

1988-01-01

258

Multiple modes on embedded inverted microstrip lines  

Microsoft Academic Search

Embedded transmission lines are commonly used in silicon radio frequency integrated circuits to reduce insertion loss and decrease circuit size. In this paper, experimental measurements and finite difference time domain (FDTD) analysis are used to show that embedded inverted microstrip lines are not suitable for use above a few GHz

George E. Ponchak; Manos M. Tentzeris

2006-01-01

259

Minimizing risks associated with peripherally inserted central catheters in the NICU.  

PubMed

Peripherally inserted central catheters (PICC lines) provide prolonged venous access, a means of administration of needed medications that would otherwise be irritating to peripheral vessels, and a means of assuring better parenteral nutrition to infants who are unable to tolerate enteral feedings. Not only do these central lines provide life-saving therapy, they are easily inserted, cost effective, and convenient. Although there are many benefits to the use of these catheters, physicians and nurses within the NICU must remain acutely aware of the risks involved with placement of PICC lines so that complications can be minimized. Prevention of sepsis in neonates is always a priority, and several measures can be implemented to reduce this risk, including scrupulous aseptic technique, knowledgeable selection of the insertion site, and consistent daily care. Other complications such as thrombosis, extravascular collection of fluid due to catheter migration or blockage, vessel perforation, and line leakage are all concerns when using this valuable tool in the care of infants. Careful catheter tip placement and conscientious ongoing monitoring can assist in reducing morbidity as well as mortality related to PICC lines. PMID:11198450

Camara, D

260

Investigation of flexible hose insertion forces and selected factors.  

PubMed

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

Drinkaus, P; Armstrong, T; Foulke, J

2008-03-12

261

Clique-inserted-graphs and spectral dynamics of clique-inserting  

Microsoft Academic Search

Motivated by studying the spectra of truncated polyhedra, we consider the clique-inserted-graphs. For a regular graph G of degree r>0, the graph obtained by replacing every vertex of G with a complete graph of order r is called the clique-inserted-graph of G, denoted as C(G). We obtain a formula for the characteristic polynomial of C(G) in terms of the characteristic

Fuji Zhang; Yi-Chiuan Chen; Zhibo Chen

2009-01-01

262

Effects of insertion conditions on tissue strain and vascular damage during neuroprosthetic device insertion  

Microsoft Academic Search

Long-term integration of neuroprosthetic devices is challenged by reactive responses that compromise the brain-device interface. The contribution of physical insertion parameters to immediate damage is not well described. We have developed an ex vivo preparation to capture real-time images of tissue deformation during device insertion using thick tissue slices from rat brains prepared with fluorescently labeled vasculature. Qualitative and quantitative

C. S. Bjornsson; S. J. Oh; Y. A. Al-Kofahi; Y. J. Lim; K. L. Smith; J. N. Turner; S. De; B. Roysam; W. Shain; S. J. Kim

2006-01-01

263

A small, insertable oven for boronization  

SciTech Connect

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}

Brouchous, D.A.; Diebold, D.A.; Doczy, M.L. [Department of Nuclear Engineering and Engineering Physics, University of Wisconsin, 1500 Johnson Drive, 337 ERB, Madison, Wisconsin 53706-1687 (United States)

1996-04-01

264

INSERTION DEVICE ACTIVITIES FOR NSLS-II.  

SciTech Connect

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.

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

2007-06-25

265

Targeted gene insertion for molecular medicine  

Microsoft Academic Search

Genomic insertion of a functional gene together with suitable transcriptional regulatory elements is often required for long-term\\u000a therapeutical benefit in gene therapy for several genetic diseases. A variety of integrating vectors for gene delivery exist.\\u000a Some of them exhibit random genomic integration, whereas others have integration preferences based on attributes of the targeted\\u000a site, such as primary DNA sequence and

Katrin Voigt; Zsuzsanna Izsvák; Zoltán Ivics

2008-01-01

266

Insertional Polymorphisms of Endogenous Feline Leukemia Viruses  

Microsoft Academic Search

The number, chromosomal distribution, and insertional polymorphisms of endogenous feline leukemia vi- ruses (enFeLVs) were determined in four domestic cats (Burmese, Egyptian Mau, Persian, and nonbreed) using fluorescent in situ hybridization and radiation hybrid mapping. Twenty-nine distinct enFeLV loci were detected across 12 of the 18 autosomes. Each cat carried enFeLV at only 9 to 16 of the loci, and

Alfred L. Roca; William G. Nash; Joan C. Menninger; William J. Murphy; Stephen J. O'Brien

2005-01-01

267

Insertion reactions in advanced electrochemical energy storage  

Microsoft Academic Search

A rechargeable electrochemical energy storage system employing an organic solvent-based electrolyte will be introduced: the lithium-ion cell. The electrode reactions base on Li+-insertion\\/-removal which ensure high rechargeability. The system possesses an operating voltage of -3.5 V and a high energy density. From a thermodynamic viewpoint such a cell is impossible because the used organic electrolyte is in contact with two

Jiirgen Otto Besenhard; Martin Winter

1998-01-01

268

Needle Insertion Simulator with Haptic Feedback  

Microsoft Academic Search

\\u000a We introduce a novel injection simulator with haptic feedback which provides realistic physical experience to the medical\\u000a user. Needle insertion requires very dexterous hands-on skills and fast and appropriate response to avoid dangerous situations\\u000a for patients. In order to train the injection operation, the proposed injection simulator has been designed to generate delicate\\u000a force feedback to simulate the needle penetration

Seungjae Shin; Hyunchul Cho; Laehyun Kim

269

Direct random insertion mutagenesis of Helicobacter pylori.  

PubMed

Random insertion mutagenesis is a widely used technique for the identification of bacterial virulence genes. Most strategies for random mutagenesis involve cloning in Escherichia coli for passage of plasmids or for phenotypic selection. This can result in biased selection due to restriction or instability of the cloned DNA, or toxicity of the encoded products. We therefore created two mutant libraries in the human pathogen Helicobacter pylori using a simple, direct mutagenesis technique, which does not require E. coli as intermediate. H. pylori total DNA was digested, circularized and digested again with a frequently cutting restriction enzyme, and the resulting fragments were ligated to a kanamycin antibiotic resistance cassette. Subsequently, the ligation mixture was transformed into the parental H. pylori strain 1061. Insertion of the kanamycin cassette by double homologous recombination into the genome of H. pylori 1061 resulted in approximately 2500 kanamycin resistant colonies. Heterogeneity of kanamycin cassette insertion was confirmed by Southern blotting. The isolation of two independent H. pylori mutants defective in production of urease from this library underlines the potential of this mutagenesis strategy. PMID:12401231

de Jonge, Ramon; Bakker, Dennis; van Vliet, Arnoud H M; Kuipers, Ernst J; Vandenbroucke-Grauls, Christina M J E; Kusters, Johannes G

2003-01-01

270

Rack Insertion End Effector (RIEE) guidance  

NASA Astrophysics Data System (ADS)

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.

Malladi, Narasimha S.

1994-10-01

271

DEVICE FOR CONTROLLING INSERTION OF ROD  

DOEpatents

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.

Beaty, B.J.

1958-10-14

272

Human LINE retrotransposons generate processed pseudogenes  

Microsoft Academic Search

Long interspersed elements (LINEs) are endogenous mobile genetic elements1-4 that have dispersed and accumulated in the genomes of higher eukaryotes via germline transposition, with up to 100,000 copies in mammalian genomes. In humans, LINEs are the major source of insertional mutagenesis, being involved in both germinal and somatic mutant phenotypes4. Here we show that the human LINE retrotransposons, which transpose

Cécile Esnault; Joël Maestre; Thierry Heidmann

2000-01-01

273

Hand-held Dopplers in central catheter insertion.  

PubMed

Peripherally inserted central catheters benefit many patients requiring repeated intravenous treatment. This study investigated the effect of using a hand-held Doppler to aid insertion where a patient's veins are not visible or palpable. PMID:9873346

MacRae, K

1998-11-01

274

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

PubMed Central

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.

Xu, X; Colombini, M

1997-01-01

275

Wireless cardiac action potential transmission with ultrasonically inserted silicon microprobes  

Microsoft Academic Search

This paper reports on the integration of ultrasonically inserted horn-shaped cardiac probes with wireless transmission of 3D cardiac action potential measurement for applications in ex vivo preparations such as monitoring the onset of ventricular fibrillation. Ultrasonically inserted silicon horn probes permit reduced penetration force during insertion, allowing silicon, a brittle material, to penetrate cardiac tissue. The probes also allow recording

C. J. Shen; A. Ramkumar; A. Lal; R. F. Gilmour Jr.

2011-01-01

276

An evaluation of consumable inserts for gas tungsten arc welding  

Microsoft Academic Search

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

A. E. Chavez; S. J. Marburger

1987-01-01

277

Reading between the LINEs: Human Genomic Variation Induced by LINE1 Retrotransposition  

Microsoft Academic Search

The insertion of mobile elements into the genome represents a new class of genetic markers for the study of human evolution. Long interspersed elements (LINEs) have amplified to a copy number of about 100,000 over the last 100 million years of mammalian evolution and comprise ?15% of the human genome. The majority of LINE-1 (L1) elements within the human genome

Fang-miin Sheen; Stephen T. Sherry; Gregory M. Risch; Myles Robichaux; Ivane Nasidze; Mark Stoneking; Mark A. Batzer; Gary D. Swergold

2000-01-01

278

CRUCIBLE LINING METHOD  

DOEpatents

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

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

1958-11-01

279

Chronic insertional Achilles tendinopathy: surgical outcomes  

PubMed Central

Summary Background and objective: insertional Achilles tendinopathy is a common condition among athletes and joggers. One fifth of the injuries involves the insertion of the tendon. The etiology is either due to mechanical overuse related to sports activity, or a systemic inflammatory disease. The clinical appearance includes pain and movement restriction. The primary treatment is conservative. The surgery referred to in this study (Calcaneal Osteotomy) is performed by decompression of the posterior margin of the calcaneus. If the tendon is degenerative, debridement is needed. There is controversy on the surgical outcome and the surgical approach. A retrospective analytic observational study. Methods: 20 patients who were diagnosed with IAT (21 feet) and were operated on between the years 2000–2007 by calcaneal osteotomy. Main outcome measures: the primary measure of success was diminished pain. It was demonstrated in the AOFAS score and in the VAS scale of pain. Results: the average grade in the AOFAS questionnaire improved by 20 points, and the average grading of pain in the VAS scale was decreased by 4.21. The median satisfaction rate was 8, the average was 5.81. 62% of the patients would repeat the surgery/recommend it. We found a significant relationship (p=0.022) between patients who avoided sports activity while suffering from insertional Achilles tendinopathy and the satisfaction rate from the surgery. Conclusions: using the calcaneal osteotomy technique as a surgical treatment for IAT is successful, and improves measures of pain and function. Low compliance with avoiding sports activity while suffering from an IAT might lead to a need for surgery.

Oshri, Yael; Palmanovich, Ezequiel; Brin, Yaron Shagra; Karpf, Ronen; Massarwe, Sabri; Kish, Benny; Nyska, Meir

2012-01-01

280

Cassini magnetometer observations during Saturn orbit insertion.  

PubMed

Cassini's successful orbit insertion has provided the first examination of Saturn's magnetosphere in 23 years, revealing a dynamic plasma and magnetic environment on short and long time scales. There has been no noticeable change in the internal magnetic field, either in its strength or its near-alignment with the rotation axis. However, the external magnetic field is different compared with past spacecraft observations. The current sheet within the magnetosphere is thinner and more extended, and we observed small diamagnetic cavities and ion cyclotron waves of types that were not reported before. PMID:15731444

Dougherty, M K; Achilleos, N; Andre, N; Arridge, C S; Balogh, A; Bertucci, C; Burton, M E; Cowley, S W H; Erdos, G; Giampieri, G; Glassmeier, K-H; Khurana, K K; Leisner, J; Neubauer, F M; Russell, C T; Smith, E J; Southwood, D J; Tsurutani, B T

2005-02-25

281

An extrachromosomal plasmid is the etiological precursor of kalDNA insertion sequences in the mitochondrial chromosome of senescent neurospora.  

PubMed

In the kalilo strains of N. intermedia, senescence is initiated by insertion of a 9.0 kb foreign nucleotide sequence, kalDNA, into mitochondrial DNA. A 9.0 kb linear DNA plasmid that is structurally homologous to the mitochondrial kalDNA insertion sequences exists in high copy numbers in close association with the nuclei of presenescent and senescent kalilo cells, but is not present in cells of long-lived normal strains. The free kalilo plasmid has not been detected in mitochondria, suggesting that the element does not contain a mitochondrial origin of replication. Unexpectedly, the nuclear plasmid, like the mitochondrial insertion element, follows a strict pattern of maternal inheritance. We surmise that the extramitochondrial plasmid is the etiological precursor of the kalDNA insertion sequences that appear in the mtDNAs of senescent cell lines and conclude that the kalilo element induces senescence because it is a mutator of mitochondrial genes. PMID:2946419

Bertrand, H; Griffiths, A J; Court, D A; Cheng, C K

1986-12-01

282

Effect of Insertion Speed on Tissue Response and Insertion Mechanics of a Chronically Implanted Silicon-Based Neural Probe  

Microsoft Academic Search

In this study, the effect of insertion speed on long- term tissue response and insertion mechanics was investigated. A dummy silicon parylene-coated probe was used in this context and implanted in the rat brain at 10 ?m\\/s (n = 6) or 100 ?m\\/s (n = 6) to a depth of 9 mm. The insertion mechanics were assessed by the dimpling

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

2011-01-01

283

Compact distributed RLC interconnect models - part IV: unified models for time delay, crosstalk, and repeater insertion  

Microsoft Academic Search

For pt. III see ibid., vol. 50, p. 1081-93 (2003). Using a new physical model for the transient response of a distributed resistance-inductance-capacitance (RLC) interconnect with a capacitive load, novel compact expressions have been derived for the 1) time delay, 2) peak crosstalk for coupled lines, 3) optimum number and size of repeaters, and 4) time delay for repeater-inserted distributed

Raguraman Venkatesan; Jeffrey A. Davis; James D. Meindl

2003-01-01

284

Lymphoid V(D)J Recombination: Nucleotide Insertion at Signal Joints as Well as Coding Joints  

Microsoft Academic Search

The coding regions of antigen receptor genes assembled by variable-diversity-joining region [V(D)J] recombination are known in many cases to have undergone deletions of several nucleotides and also to contain insertions of noncoded nucleotides at the recombined junction (the coding joint). By using extrachromosomal recombination substrates to transfect lymphoid cell lines, we show that the signal joint (the fusion of the

Michael R. Lieber; Joanne E. Hesse; Kiyoshi Mizuuchi; Martin Gellert

1988-01-01

285

Mapping Ds insertions in barley using a sequence-based approach  

Microsoft Academic Search

A transposon tagging system, based upon maize Ac\\/Ds elements, was developed in barley (Hordeum vulgare subsp. vulgare). The long-term objective of this project is to identify a set of lines with Ds insertions dispersed throughout the genome as a comprehensive tool for gene discovery and reverse genetics. AcTPase and Ds-bar elements were introduced into immature embryos of Golden Promise by

L. D. Cooper; L. Marquez-Cedillo; J. Singh; A. K. Sturbaum; S. Zhang; V. Edwards; K. Johnson; A. Kleinhofs; S. Rangel; V. Carollo; P. Bregitzer; P. G. Lemaux; P. M. Hayes

2004-01-01

286

Recessed impingement insert metering plate for gas turbine nozzles  

DOEpatents

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.

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

2002-01-01

287

Pediatric peripherally inserted central catheter placement: application of ultrasound technology.  

PubMed

Ultrasound-guided placement of peripherally inserted central catheters has been well documented for adults who require infusion therapy. This same technology is surfacing in the pediatric population to improve outcomes when confronted with the challenges of the smaller vascular system and chubbier body shapes. The scope of practice is addressed in coupling peripherally inserted central catheters with ultrasound imaging, and recommendations are identified for the advancement of nursing practice within the field of imaging technology and application. Obstacles related to successful insertion of peripherally inserted central catheters are defined, and the benefits of ultrasound-guided placement of peripherally inserted central catheters are reviewed. PMID:18025983

Nichols, Inez; Doellman, Darcy

288

Lithium Insertion Chemistry of Some Iron Vanadates  

SciTech Connect

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.

Patoux, Sebastien; Richardson, Thomas J.

2007-02-02

289

Proposed mini beta insertion for spear  

SciTech Connect

A significant gain in luminosity has been achieved by use of mini beta insertions implemented in several electron-positron storage rings (CESR, DORIS II, PEP, PETRA) during the last year. This concept consists of strong vertical focusing quadrupoles mounted as close as possible to the interaction point. Such an arrangement avoids the acceptance and chromaticity problems caused by an extreme focusing with the normal magnet structure, but they interfere with the particle detectors and if the detector has a magnetic field one has to protect the quadrupole from this field. In this paper a mini beta insertion for SPEAR is suggested which is rather similar to that developed for DORIS II. The essential element is a conventional iron magnet with moderate design values up to an energy of E = 4 GeV. The resulting optics allow vertical amplitude functions down to ..beta../sub y/* = 3 cm at the interaction points. Compared to the present SPEAR optics, the mini beta optics is only different in the interaction regions but unchanged in the whole arcs. This is of great importance to the injection system which does not need any modification. The resulting gain in luminosity is calculated to be approximately a factor of four. 8 references, 13 figures, 5 tables.

Wille, K.

1984-03-01

290

Helix insertion into bilayers and the evolution of membrane proteins  

PubMed Central

Polytopic ?-helical membrane proteins cannot spontaneously insert into lipid bilayers without assistance from polytopic ?-helical membrane proteins that already reside in the membrane. This raises the question of how these proteins evolved. Our current knowledge of the insertion of ?-helices into natural and model membranes is reviewed with the goal of gaining insight into the evolution of membrane proteins. Topics include: translocon-dependent membrane protein insertion, antibiotic peptides and proteins, in vitro insertion of membrane proteins, chaperone-mediated insertion of transmembrane helices, and C-terminal tail-anchored (TA) proteins. Analysis of the E. coli genome reveals several predicted C-terminal TA proteins that may be descendents of proteins involved in pre-cellular membrane protein insertion. Mechanisms of pre-translocon polytopic ?-helical membrane protein insertion are discussed.

2010-01-01

291

INCA: an expert system for process planning in PCB assembly line  

Microsoft Academic Search

A description is given of INCA, an expert system that tackles the problem of optimization of the automatic insertion of components on printed circuit board (PCB) in the production assembly line. The authors describe the present situation in the PCB assembly line for component insertion, why an expert system has been chosen and the benefits expected from its introduction, how

Patrizia Cavalloro; Emanuela Cividati

1988-01-01

292

Collagenase-aided Insertion of Intracortical Microelectrode Arrays: Evaluation of Insertion Force and Chronic Recording Performance  

Microsoft Academic Search

Typically intracortical electrodes are required to puncture the intact pia mater during insertion which in the process can lead to brain dimpling and trauma. Furthermore, there is interest in the development of more flexible substrates to reduce relative micromotion after implantation, but such device have difficulty penetrating the pia without buckling. In this paper a strategy for reducing the mechanical

Kunal J. Paralikar; Jonathan K. Lawrence; Ryan S. Clement

2006-01-01

293

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

Microsoft Academic Search

A simple strategy to identify and isolate new promoters suitable for driving the expression of selectable marker genes is described. By employing a Brassica napus hypocotyl transformation protocol and a promoterless gus::nptII tagging construct, a series of 20 kanamycin-resistant tagged lines was produced. Most of the regenerated plants showed hardly any GUS activity in leaf, stem and root tissues. However,

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

2003-01-01

294

Intermetallic insertion anodes for lithium batteries.  

SciTech Connect

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.

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

1999-11-12

295

A large-scale collection of phenotypic data describing an insertional mutant population to facilitate functional analysis of rice genes  

PubMed Central

In order to facilitate the functional analysis of rice genes, we produced about 50,000 insertion lines with the endogenous retrotransposon Tos17. Phenotypes of these lines in the M2 generation were observed in the field and characterized based on 53 phenotype descriptors. Nearly half of the lines showed more than one mutant phenotype. The most frequently observed phenotype was low fertility, followed by dwarfism. Phenotype data with photographs of each line are stored in the Tos17 mutant panel web-based database with a dataset of sequences flanking Tos17 insertion points in the rice genome (http://tos.nias.affrc.go.jp/). This combination of phenotypic and flanking sequence data will stimulate the functional analysis of rice genes.

Iwasaki, Yukimoto; Kitano, Hidemi; Itoh, Jun-Ichi; Maekawa, Masahiko; Murata, Kazumasa; Yatou, Osamu; Nagato, Yasuo; Hirochika, Hirohiko

2006-01-01

296

Functional characterization of Arabidopsis thaliana metallothionein-deficient mutants  

Microsoft Academic Search

In an effort to understand the functions of MT genes in plants, a reverse genetic approach was used in this study. Arabidopsis thaliana lines containing T-DNA insertions in each of the MT genes were identified. Because mutants that were deficient in a single MT showed few dramatic or interesting phenotypes, the insertion lines were crossed and used to develop mutant

Matheus Romanos Benatti

2010-01-01

297

Human endogenous retrovirus-H insertion screening.  

PubMed

Endogenous retroviruses (ERVs) and ERV-like sequences comprise 8% of the human genome. We aimed to analyze genome integration polymorphisms of human endogenous retrovirus (HERV)-H by the inter-retrotransposon amplified polymorphism (IRAP) technique using the sequences of LTR7A (450 bp), LTR7B (445 bp) and LTR7C (471 bp). Blood samples from 20 individuals (10 females and 10 males) of diverse ethnic origins were used for the determination of integration variations at the genomic level. Isolated genomic DNA was screened using 3 pairs of primers corresponding to LTR regions of the HERV-H gene. We observed insertion polymorphism patterns between 0-87% in all subjects. The findings obtained contribute to our understanding of the effects of HERV-H on variations within the human genome. PMID:23358623

Guliyev, Mehrab; Yilmaz, Sibel; Sahin, Kaniye; Marakli, Sevgi; Gozukirmizi, Nermin

2013-01-28

298

Stabilization of insertion electrodes for lithium batteries.  

SciTech Connect

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.

Thackeray, M. M.

1998-09-03

299

How Do I Overcome Difficulties in Insertion?  

PubMed Central

Demand for colonoscopy is increasing because it is an important tool not only for screening of colorectal neoplasm but also for resection of such lesions in early stage. Cecal intubation requires expertise on shortening of the examination time and improvement of the cecal intubation rate without causing pain to the patients. About 5% to 10% of patients still experience difficulties or failure of the cecal intubation. There are number of factors that affect the difficulty of the colonoscopy such as technical skill of the endoscopist, angulated sigmoid, redundant colon, advanced age, female gender, diverticular disease, and inadequate bowel preparation. In an effort to overcome these situations and to and aiding colonoscope insertion with reducing pain, various methods have introduced. Like this review discusses ways to approach patients with technically difficult colons for achieving the successful cecal intubation.

Jung, Yunho

2012-01-01

300

INSERTION DEVICE UPGRADE PLANS AT THE NSLS.  

SciTech Connect

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

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

2005-05-16

301

DNA polymerase beta ribonucleotide discrimination: insertion, misinsertion, extension, and coding.  

PubMed

DNA polymerases must select nucleotides that preserve Watson-Crick base pairing rules and choose substrates with the correct (deoxyribose) sugar. Sugar discrimination represents a great challenge because ribonucleotide triphosphates are present at much higher cellular concentrations than their deoxy-counterparts. Although DNA polymerases discriminate against ribonucleotides, many therapeutic nucleotide analogs that target polymerases have sugar modifications, and their efficacy depends on their ability to be incorporated into DNA. Here, we investigate the ability of DNA polymerase beta to utilize nucleotides with modified sugars. DNA polymerase beta readily inserts dideoxynucleoside triphosphates but inserts ribonucleotides nearly 4 orders of magnitude less efficiently than natural deoxynucleotides. The efficiency of ribonucleotide insertion is similar to that reported for other DNA polymerases. The poor polymerase-dependent insertion represents a key step in discriminating against ribonucleotides because, once inserted, a ribonucleotide is easily extended. Likewise, a templating ribonucleotide has little effect on insertion efficiency or fidelity. In contrast to insertion and extension of a ribonucleotide, the chemotherapeutic drug arabinofuranosylcytosine triphosphate is efficiently inserted but poorly extended. These results suggest that the sugar pucker at the primer terminus plays a crucial role in DNA synthesis; a 3'-endo sugar pucker facilitates nucleotide insertion, whereas a 2'-endo conformation inhibits insertion. PMID:20519499

Cavanaugh, Nisha A; Beard, William A; Wilson, Samuel H

2010-06-02

302

A Collection of Ds Insertional Mutants Associated With Defects in Male Gametophyte Development and Function in Arabidopsis thaliana  

Technology Transfer Automated Retrieval System (TEKTRAN)

Functional analyses of the Arabidopsis genome require analysis of the gametophytic generation, since 10% of the genes are expressed in the male gametophyte and 9% in the female gametophyte. Here we describe the genetic and molecular characterization of 67 Ds insertion lines that show reduced transmi...

303

Multipacting analysis of a quarter wave choke joint used for insertion of a demountable cathode into a SRF photoinjector  

Microsoft Academic Search

The multipacting phenomena in accelerating structures and coaxial lines are well documented and methods of mitigating or suppressing it are understood. The multipacting that occurs in a quarter wave choke joint designed to mount a cathode insertion stalk into a superconducting RF photoinjector has been analyzed via calculations and experimental measurements and the effect of introducing multipacting suppression grooves into

A. Burrill; I. Ben-Zvi; M. Cole; J. Rathke; P. Kneisel; R. Manus; R. Rimmer

2007-01-01

304

Rapid R2 retrotransposition leads to the loss of previously inserted copies via large deletions of the rDNA locus.  

PubMed

R2 non-long terminal repeat retrotransposable elements insert specifically into the 28S rRNA genes of a wide range of animals. These elements maintain long-term stable relationships with the host genome. By scoring the variation present at the 5' ends of individual R2 copies, lines of Drosophila simulans have been identified with high rates of R2 retrotransposition. Comparing the R2 elements present in the parents with that of their progeny after 1 or 30 generations in this report revealed that retrotransposition rates were higher through the female germ line compared with the male germ line. In addition, most events in females occur late in germ line development. Surprisingly, the gain of new R2 insertions by retrotranspositions was counterbalanced by deletions of preexisting R2 insertions. These deletions occurred by the loss of large segments of the rDNA units that contained on average an estimated 15 R2 elements. When monitored over single generations, the rate of loss of preexisting elements was higher than the rate of new insertions. However, the chromosomes with the largest deletions appear to be eliminated from the population because the rates of R2 insertions and deletions after 30 generations were approximately equal. These findings suggest that high rates of R2 retrotransposition do not necessarily lead to dramatic increases in the level of R2 insertions in the rDNA locus but can lead to a more rapid turnover of rDNA units. PMID:18003600

Zhang, Xian; Zhou, Jun; Eickbush, Thomas H

2007-11-13

305

An evaluation of consumable inserts for gas tungsten arc welding  

SciTech Connect

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.

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

1987-03-01

306

Design and evaluation of moxifloxacin hydrochloride ocular inserts.  

PubMed

The objective of the present investigation was to prepare and evaluate ocular inserts of moxifloxacin. An ocular insert was made from an aqueous dispersion of moxifloxacin, sodium alginate, polyvinyl alcohol, and dibutyl phthalate by the film casting method. The ocular insert (5.5 mm diameter) was cross-linked by CaCl2 and was coated with Eudragit S-100, RL-100, RS-100, E-100 or L-100. The in vitro drug drainage/permeation studies were carried out using an all-glass modified Franz diffusion cell. The drug concentration and mucoadhesion time of the ocular insert were found satisfactory. Cross-linking and coating with polymers extended the drainage from inserts. The cross-linked ocular insert coated with Eudragit RL-100 showed maximum drug permeation compared to other formulations. PMID:22472452

Pawar, Pravin K; Katara, Rajesh; Majumdar, Dipak K

2012-03-01

307

Fabrication of LIGA mold inserts using a modified procedure  

NASA Astrophysics Data System (ADS)

The LIGA process, invented in Germany and being further developed at Louisiana State University can be used to economically mass produce high aspect ratio microstructures (HARM) by molding. The template, or mold insert, is required to mold microstructure and is fabricated by a two step sequence of x-ray lithography and electroplating. A modified version of the LIGA process has been used to produce a mold insert. A sheet of PMMA is first patterned using x-ray lithography. After the lithography process is completed, the PMMA sheet is clamped to a nickel substrate and a subsequent electroplating step produces a mold insert. This processing sequence eliminates the need to establish a chemical bond between the resist and the substrate. The electroplated HARM were used as a mold insert in subsequent molding experiments. One mold insert which was fabricated is presented, along with the results of molding experiments using a similar insert.

Galhotra, Vikas; Marques, Christophe; Desta, Yohannes M.; Kelly, Kevin W.; Despa, Mircea; Pendse, Ajit; Collier, John

1996-09-01

308

Ocular inserts -- Advancement in therapy of eye diseases  

PubMed Central

The ocular insert represents a significant advancement in the therapy of eye disease. Ocular inserts are defined as sterile, thin, multilayered, drug-impregnated, solid or semisolid consistency devices placed into the cul-de-sac or conjuctival sac, whose size and shape are especially designed for ophthalmic application. They are composed of a polymeric support that may or may not contain a drug. The drug can later be incorporated as dispersion or a solution in the polymeric support. They offer several advantages as increased ocular residence and sustained release of medication into the eye. The insert includes a body portion sized to position within a lachrymal canaliculus of the eyelid. The inserts are classified according to their solubility as insoluble, soluble, or bioerodible inserts. The release of drug from the insert depends upon the diffusion, osmosis, and bioerosion of the drug, and this article is an attempt to present a brief about this newer drug delivery system.

Kumari, Anita; Sharma, Pramod K.; Garg, Vipin K.; Garg, Garima

2010-01-01

309

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

PubMed

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

Wojnar, Danuta G; Beaman, Margaret L

310

Cognitive-Behavioral Interventions for IV Insertion Pain  

Microsoft Academic Search

INSERTION OF AN IV CATHETER is a commonly performed and painful procedure. The use of cognitive-behavioral interventions (CBIs) may decrease pain by diverting the patient's attention to stimuli other than pain.•THIS RANDOMIZED, CONTROLLED TRIAL examined the effect of three CBIs—music, kaleidoscope, and guided imagery—on IV insertion pain in 324 patients.•NO STATISTICALLY SIGNIFICANT differences in IV insertion pain were found among

Ann F. Jacobson

2006-01-01

311

Relaxation phenomena in lithium-ion-insertion cells  

Microsoft Academic Search

Relaxation phenomena in lithium-ion-insertion cells are modeled. Simulation results are presented for a dual lithium-ion-insertion cell and for a cell using a lithium-foil negative electrode. A period of relaxation after a charge or discharge can cause appreciable changes in the distribution of material in the insertion electrodes. Local concentration cells in the solution phase and an open-circuit potential that depends

Thomas F. Fuller; M. Doyle; J. S. Newman

1994-01-01

312

A Investigation of Some Lithium Insertion Compounds  

NASA Astrophysics Data System (ADS)

Lithium insertion compounds are of technical interest as storage-battery electrodes; they are also of scientific interest as they offer low-temperature synthetic routes to compounds having unusual atomic ordering and to formal valence states that may not be accessible at higher temperatures normally employed in solid-state reactions. In this study lithium insertion compounds have been investigated to understand structure-property relationships of some V and Ni compounds and also to explore new cathode and anode materials for lithium secondary batteries. An investigation of the systems Li_{rm 1-x}VO _2 and LiV_{rm 1-y}M_{rm y} O_2, where M = Cr or Ti, has been carried out to determine a critical V-V separation R_{rm c} for the transition from strongly correlated (R > R_{rm c}, W < U) to weakly correlated (R < R _{rm c}, W > U) electrons in a V-3d band associated with octahedral -site V^{3+} ions sharing common octahedral-site edges with six like nearest neighbors in an oxide. A first-order transition on passing from the itinerant-electron regime R < R _{rm c} to the localized -electron regime R > R_{ rm c} has been established. The behavior of the phase transition in Li_{rm 1-x}VO_2 in the compositional range 0 <= x <= 0.12 has also been studied, and it has been shown that the transition temperature T_{rm t} is controlled more by the lattice energy change through the distortion than by the enthalpy of the transition. Li_{rm 1-x} NiO_2 and the spinel Li (Ni _2) O_4 have been prepared chemically for the first time; the 90 ^circ Ni-O-Ni interactions in these compounds are shown, from electronic and magnetic measurements, to yield properties that are quite different from the 180 ^circ Ni-O-Ni interactions in the perovskite LaNiO_3. Li _{rm 1-x}NiO_2 and the spinel Li (Ni_2) O_4 are found to be semiconductors, and both exhibit ferromagnetic interatomic-exchange interactions. Lithium insertion into NASICON frameworks Li_{ rm x}M_2(PO _4)_3 (M = Ti, Nb, Sb, Zr) was investigated with a view to prepare novel electrodes for lithium secondary batteries. The systems Li_{rm 1+x}Ti_2(PO _4)_3, Li_{rm x}NbTi(PO_4)_3 and Li_{rm x}SbTi(PO _4)_3 accept lithium to x = 2, 3, and 4 respectively; the latter two systems are reduced to an unusual valence state at one of the cations.

Ahuja, Geeta

313

In vivo oxidation in retrieved highly crosslinked tibial inserts.  

PubMed

The current study determined (I) the environment where oxidation in a series of retrieved, HXL UHMWPE tibial inserts occurred (in vivo or postexplant); and (II) the effect of fabrication variables (irradiation source, irradiation dose) and thermal processing after irradiation (annealing or remelting) on oxidation resistance. Hypotheses examined are (1) HXL UHMWPE tibial inserts have potential to oxidize in vivo, and (2) annealed HXL UHMWPE oxidizes at a higher rate in vivo than remelted HXL UHMWPE. Highly crosslinked UHMWPE tibial inserts (87), received by an IRB-approved retrieval laboratory from 20 surgeons at 10 institutions across the U.S., were analyzed from 2005 to 2011. Fourier transform infrared spectroscopy was used to determine oxidation and trans-vinylene index for each retrieved insert. Measured oxidation that was maximum subsurface was found in 56% of all HXL tibial inserts. This maximum oxidation correlated significantly with in vivo time, trans-vinylene index, and thermal processing after irradiation. Articular oxidation rate correlated with crosslinking irradiation dose and thermal processing after irradiation. Retrieved below-melt annealed tibial inserts had significantly higher articular oxidation rates than remelted tibial inserts (p < 0.001). Articular oxidation rates correlated positively with cross-linking irradiation dose and postirradiation thermal processing. Edge oxidation correlated with postirradiation thermal processing. Oxidation of HXL UHMWPE may have clinical implications for tibial inserts, since loss of UHMWPE toughness resulting from oxidation has led to fatigue damage in gamma-sterilized tibial inserts. PMID:22997097

Currier, B H; Van Citters, D W; Currier, J H; Carlson, E M; Tibbo, M E; Collier, J P

2012-09-21

314

Accurate consent for insertion and later removal of grommets.  

PubMed

Each year in the UK over 30,000 patients undergo insertion of grommets. The grommet insertion may cause many problems like persistent otorrhoea, scarred drum, retraction pockets and retention. The grommets may be extruded from the middle ears by the normal epithelial migration mechanism once they have served their purpose. These may become infected and require removal. We have analysed the Department of Health Hospital Episode Statistics relating to the insertion and removal of grommets (ventilation tubes). We have shown that 7.6 per cent of patients who have grommets inserted will have grommets removed. PMID:17403263

Sood, S; Waddell, A

2007-04-01

315

Does Feeding Tube Insertion and its Timing Improve Survival?  

PubMed Central

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.

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

2012-01-01

316

Markov Process Analysis of PCM Line Monitors  

Microsoft Academic Search

In PCM transmission systems, ensuring repeater timing and regenerative capabilities requires monitored enforcement of the ones density of the line waveform. One possible corrective monitor action is to pre-empt information bits and insert ones when necessary, and thus introduce errors into the pulse stream. The analysis of the resultant error rates for classes of digital and analog implementations of a

G. Foschini; A. Salazar; J. Smith

1974-01-01

317

Tunable Nonlinear Transmission Lines with switched varactors  

Microsoft Academic Search

Nonlinear Transmission Lines (NLTLs) have been used in many applications for waveform shaping and impedance engineering. To minimize insertion loss and jitter, adaptive impedance is desirable. Here we propose to use switched varactors on NLTL to achieve impedance-tuning. We present an analytical model to examine the impact of switch parasitics on the performance of the NLTL. We find that for

Fan Yu; K. G. Lyon; E. C. Kan

2009-01-01

318

Antibody binding loop insertions as diversity elements  

PubMed Central

In the use of non-antibody proteins as affinity reagents, diversity has generally been derived from oligonucleotide-encoded random amino acids. Although specific binders of high-affinity have been selected from such libraries, random oligonucleotides often encode stop codons and amino acid combinations that affect protein folding. Recently it has been shown that specific antibody binding loops grafted into heterologous proteins can confer the specific antibody binding activity to the created chimeric protein. In this paper, we examine the use of such antibody binding loops as diversity elements. We first show that we are able to graft a lysozyme-binding antibody loop into green fluorescent protein (GFP), creating a fluorescent protein with lysozyme-binding activity. Subsequently we have developed a PCR method to harvest random binding loops from antibodies and insert them at predefined sites in any protein, using GFP as an example. The majority of such GFP chimeras remain fluorescent, indicating that binding loops do not disrupt folding. This method can be adapted to the creation of other nucleic acid libraries where diversity is flanked by regions of relative sequence conservation, and its availability sets the stage for the use of antibody loop libraries as diversity elements for selection experiments.

Kiss, Csaba; Fisher, Hugh; Pesavento, Emanuele; Dai, Minghua; Valero, Rosa; Ovecka, Milan; Nolan, Rhiannon; Phipps, M. Lisa; Velappan, Nileena; Chasteen, Leslie; Martinez, Jennifer S.; Waldo, Geoffrey S.; Pavlik, Peter; Bradbury, Andrew R.M.

2006-01-01

319

Replicative resolution of integron cassette insertion  

PubMed Central

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

Loot, Celine; Ducos-Galand, Magaly; Escudero, Jose Antonio; Bouvier, Marie; Mazel, Didier

2012-01-01

320

APS Insertion Devices: Recent Developments and Results.  

PubMed

The Advanced Photon Source (APS) now has a total of 23 insertion devices (IDs). Over two-thirds of them are installed on the storage ring. The installed devices include 18, 27 and 55 mm-period undulators; an 85 mm-period wiggler; a 16 cm-period elliptical multipole wiggler; and many 33 mm-period undulators. Most of the IDs occupy storage-ring straight sections equipped with 8 mm vertical-aperture vacuum chambers. All of the IDs were measured magnetically at the APS and, in most cases, underwent a final magnetic tuning in order to minimize variation in the various integrals of the field through the ID over the full gap range. Special shimming techniques to correct magnetic field parameters in appropriate gap-dependent ways were developed and applied. Measurements of the closed-orbit distortion as a function of the ID gap variation have been completed, and results are in a good agreement with magnetic measurements. Spectral diagnostics of the ID radiation, including measurements of the absolute spectral flux, brilliance and polarization, show excellent agreement between calculated and measured results. Studies of the sensitivity of IDs to radiation exposure and measurements of the dose rate received by the IDs are in progress. PMID:15263477

Gluskin, E

1998-05-01

321

Roles of carboxyl groups in the transmembrane insertion of peptides  

PubMed Central

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

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

2011-01-01

322

Roles of carboxyl groups in the transmembrane insertion of peptides.  

PubMed

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

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

2011-08-23

323

Insertional mutagenesis in mice: new perspectives and tools  

Microsoft Academic Search

Insertional mutagenesis has been at the core of functional genomics in many species. In the mouse, improved vectors and methodologies allow easier genome-wide and phenotype-driven insertional mutagenesis screens. The ability to generate homozygous diploid mutations in mouse embryonic stem cells allows prescreening for specific null phenotypes prior to in vivo analysis. In addition, the discovery of active transposable elements in

Corey M. Carlson; David A. Largaespada

2005-01-01

324

Microfabrication using silicon mold inserts and hot embossing  

Microsoft Academic Search

We have successfully demonstrated the feasibility of fabricating three-dimensional microstructures by using a combined silicon mold insert and micro hot embossing process (SMIHE). Anisotropic silicon wet etching process has been used to define microstructures on top of a four inch silicon wafer. The whole wafer is then used as the mold insert in a micro hot pressing machine to duplicate

Liwei Lin; Chun-Jung Chiu; W. Bacher; Mathias Heckele

1996-01-01

325

Load beam unit replaceable inserts for dry coal extrusion pumps  

SciTech Connect

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.

Saunders, Timothy; Brady, John D.

2012-11-13

326

Mechanisms of insertional mutagenesis in human genes causing genetic disease  

Microsoft Academic Search

Examples of the insertion of < 10 bp of DNA sequence into human gene-coding regions causing genetic disease were collated in order to study the underlying causative mechanisms. The nature of these insertions was found to be consistent with several mechanisms of mutagenesis including: (1) slipped mispairing mediated by direct repeats or runs of identical bases and (2) the templated

David N. Cooper; Michael Krawczak

1991-01-01

327

Metal Pipe Joining with Aluminum Alloy by Ultrasonic Insert Casting  

Microsoft Academic Search

Insert casting of aluminum is widely used in industry. In order to realize better metallurgical bonding between metallic pipe and aluminum alloy castings, a new insert casting method with aided by high-power ultrasound has been developed. The bonding strength between the metal pipe and the cast aluminum alloy was evaluated by shear testing, EPMA analysis at the bonding interface was

J. Pan; M. Yoshida; G. Sasaki; H. Fukunaga

2000-01-01

328

Simultaneous routing and buffer insertion with restrictions on buffer locations  

Microsoft Academic Search

During the routing of global interconnects, macro blocks formuseful routing regions which allow wires to go through but forbidbuffers to be inserted. They give restrictions on bufferlocations. In this paper, we take these buffer location restrictionsinto consideration and solve the simultaneous mazerouting and buffer insertion problem. Given a block placementdefining buffer location restrictions and a pair of pins(a source and

Hai Zhou; D. F. Wong; I-Min Liu; Adnan Aziz

1999-01-01

329

Management of insertional Achilles tendinopathy through a Cincinnati incision  

Microsoft Academic Search

BACKGROUND: About 10% of patients not responding to 3–6 months of conservative management for insertional Achilles tendinopathy undergo surgery. Traditionally, surgery of the Achilles tendon is performed through longitudinal extensile incisions. Such surgery is prone to the complications of wound healing, wound breakdown and iatrogenic nerve injury. METHODS: We describe our current method of exposure of the Achilles tendon insertion

Michael R Carmont; Nicola Maffulli

2007-01-01

330

Piezo-Driven Vibrating Insertion Device for Microelectrode Array  

NASA Astrophysics Data System (ADS)

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

Noda, Takahiro; Kanzaki, Ryohei; Takahashi, Hirokazu

331

In vivo regulation of acetylcholinesterase insertion at the neuromuscular junction.  

PubMed

The efficiency of synaptic transmission between nerve and muscle depends on the number and density of acetylcholinesterase molecules (AChE) at the neuromuscular junction. However, little is known about the way this density is maintained and regulated in vivo. By using time lapse and quantitative fluorescence imaging assays in living mice, we demonstrated that insertion of new AChEs occurs within hours of saturating pre-existing AChEs with fasciculin2, a snake toxin that selectively labels AChE. In the absence of muscle postsynaptic activity or evoked nerve presynaptic neurotransmitter release, AChE insertion was decreased significantly, whereas direct stimulation of the muscle completely restored AChE insertion to control levels. This activity-dependent AChE insertion is mediated by intracellular calcium. In muscle stimulated in the presence of a Ca2+ channel blocker or calcium-permeable Ca2+ chelator, AChE insertion into synapses was significantly decreased, whereas ryanodine or ionophore A12387 treatment of blocked and unstimulated synapses significantly increased AChE insertion. These results demonstrated that synaptic activity is critical for AChE insertion and indicated that a rise in intracellular calcium either through voltage-gated calcium channels or from intracellular stores is critical for proper AChE insertion into the adult synapse. PMID:15998641

Martinez-Pena y Valenzuela, Isabel; Hume, Richard I; Krejci, Eric; Akaaboune, Mohammed

2005-07-05

332

Force Modeling for Needle Insertion into Soft Tissue  

Microsoft Academic Search

The modeling of forces during needle insertion into soft tissue is important for accurate surgical simulation, preoperative planning, and intelligent robotic assistance for percutaneous therapies. We present a force model for needle insertion and experimental procedures for acquiring data from ex vivo tissue to populate that model. Data were collected from bovine livers using a one-degree-of-freedom robot equipped with a

Allison M. Okamura; Christina Simone; M. D. O'Leary

2004-01-01

333

Error compensation in insertion-region magnets  

SciTech Connect

In heavy ion storage rings, intra-beam scattering (IBS) between high charge state ions results in a large beam emittance during storage. The ultimate machine performance depends on achieving the highest possible magnetic field quality and alignment accuracy in the insertion-region (IR) triplet magnets during low-{beta} operation when the beam size is the largest in the triplets. Therefore, effective compensation of magnet construction errors and misalignments is crucial. Heavy-ion beams (Au{sup 79+} will be accelerated and stored for 10 hours in the Relativistic Heavy Ion Collider (RHIC) at the energy of 100 GeV/u in two separated rings consisting of superconducting magnets. Due to strong IBS, the transverse beam emittance grows from 10{pi} mm-mr at injection to more than 40{pi} mm-mr at storage. Dipoles and riplets of quadrupoles of large bore are placed on both sides of the six interaction points (IP). In order to maximize the luminosity at two IPs with proposed experiments, the nearby triplets are designed to enable the collision {beta}-function to be reduced to {beta} = 1 m. Consequently, the rms transverse beam size becomes large at the triplets ({beta}{sub max} = 1400 m), increasing from {sigma} = 2.3 mm to 4.7 mm during the period of storage. At the end of storage, the 5{sigma} beam size becomes about 71% of the coil radius (65 mm). The goal if IR triplet error compensation is to ensure satisfactory magnetic field quality and beam long-term stability up to this 5{sigma} radius.

Wei, Jie

1995-12-31

334

Metallic glass mold insert for hot embossing of polymers  

NASA Astrophysics Data System (ADS)

Molding of micro components from thermoplastic polymers (TPs) has become a routinely used industrial production process. To find hard, ductile and durable material for mold insert and to fabricate the mold insert are two big challenges for the thermoplastic polymers fabrication techniques. We report that a Pd-based metallic glass (MG) mold insert was readily fabricated in its supercooled liquid region, and the atomic force microscope measurement and time-temperature-transformation analysis show that the metallic glass mold insert has very fine surface quality and long service life. We show that the metallic glasses, which have remarkable mechanical properties and excellent thermoplastic forming ability, are new ideal materials for hot embossing mold insert of thermoplastic polymers.

Ma, J.; Zhang, X.; Wang, W. H.

2012-07-01

335

Method for improving the durability of ion insertion materials  

DOEpatents

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.

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

2002-01-01

336

Movie Lines  

NSDL National Science Digital Library

This algebra lesson from Illuminations involves using linear equations and graphs in a real world context. Students will graph a line based on data points, find the equation of the line, identify y-intercept and slope, and extrapolate data. The material is appropriate for grades 9-12 and should require 1 class period to complete.

2011-01-11

337

A new magnetostatic wave delay line using YIG film  

Microsoft Academic Search

We have proposed a delay line of microstrip type consisting of a microstrip line fabricated on a composite substrate of two yttrium iron garnet films separated by a gadolinium gallium garnet layer. The dispersion relations are derived assuming magnetic walls at the edges of the strip. Experiments are carried out at S band with nonuniform magnetic field. Insertion loss less

Kensuke Okubo; Vishnu Priye; Makoto Tsutsumi

1997-01-01

338

HTS-based switched filter banks and delay lines  

SciTech Connect

For a number of communications and other applications, switched filter banks (some channelizers) and switched delay lines (phase shifters) are extremely useful since YBaCuO and TlCaBaCuO filters and delay lines have shown significant performance enhancements over their conventional counterparts, a purely superconducting version of the switched assemblies could result in additional improvements. A thermal switch has been developed that provides good isolation and insertion loss with adequate switching times to allow a monolithic approach to the switched lines and filter banks. Filter banks in the 8--11 GHz range have been demonstrated with insertion losses < 1 dB and out-of-band rejection greater than a package-limited 50 dB. Switched delay lines have been fabricated with insertion losses less than 0.3 dB/bit and peak phase deviations from linearity of less than 5 degrees over 30 GHz bandwidths.

Martens, J.S.; Hietala, V.M.; Ginley, D.S.; Tigges, C.P.; Plut, T.A. [Sandia National Labs., Albuquerque, NM (United States); Truman, J.K. [CVC Products, Inc., Rochester, NY (United States); Track, E.K. [Hypres, Inc., Elmsford, NY (United States); Young, K.H.; Young, R.T. [Energy Conversion Devices, Inc., Troy, MI (United States)

1992-09-01

339

HTS-based switched filter banks and delay lines  

SciTech Connect

For a number of communications and other applications, switched filter banks (some channelizers) and switched delay lines (phase shifters) are extremely useful since YBaCuO and TlCaBaCuO filters and delay lines have shown significant performance enhancements over their conventional counterparts, a purely superconducting version of the switched assemblies could result in additional improvements. A thermal switch has been developed that provides good isolation and insertion loss with adequate switching times to allow a monolithic approach to the switched lines and filter banks. Filter banks in the 8--11 GHz range have been demonstrated with insertion losses < 1 dB and out-of-band rejection greater than a package-limited 50 dB. Switched delay lines have been fabricated with insertion losses less than 0.3 dB/bit and peak phase deviations from linearity of less than 5 degrees over 30 GHz bandwidths.

Martens, J.S.; Hietala, V.M.; Ginley, D.S.; Tigges, C.P.; Plut, T.A. (Sandia National Labs., Albuquerque, NM (United States)); Truman, J.K. (CVC Products, Inc., Rochester, NY (United States)); Track, E.K. (Hypres, Inc., Elmsford, NY (United States)); Young, K.H.; Young, R.T. (Energy Conversion Devices, Inc., Troy, MI (United States))

1992-01-01

340

HTS-based switched filter banks and delay lines  

NASA Astrophysics Data System (ADS)

For communications and a number of other applications, switched filter banks (some channelizers) and switched delay lines (phase shifters) are extremely useful since YBaCuO and TlCaBaCuO filters and delay lines have shown significant performance enhancements over their conventional counterparts. A purely superconducting version of the switched assemblies could result in additional improvements. A thermal switch was developed that provides good isolation and insertion loss with adequate switching times to allow a monolithic approach to the switched lines and filter banks. Filter banks in the 8-11 GHz range were demonstrated with insertion losses less than 1 dB and out-of-band rejection greater than a package-limited 50 dB. Switched delay lines were fabricated with insertion losses less than 0.3 dB/bit and peak phase deviations from linearity of less than 5 degrees over 30 GHz bandwidths.

Martens, J. S.; Hietala, V. M.; Ginley, D. S.; Tigges, C. P.; Plut, T. A.; Truman, J. K.; Track, E. K.

341

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

DOEpatents

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

Burdgick, Steven Sebastian (Schenectady, NY)

2002-01-01

342

Positioning long lines: contrast versus plain radiography  

PubMed Central

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

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

2001-01-01

343

Role of suture diameter and vessel insertion position in the establishment of the middle cerebral artery occlusion rat model  

PubMed Central

The aim of the present study was to explore the role of suture diameter and vessel insertion position in the preparation of the middle cerebral artery occlusion (MCAO) rat model. A total of 84 Sprague-Dawley rats (weighing 250–300 g) were randomly divided to three groups: group A (type 1.0, suture diameter 0.16–0.17 mm and tip 0.21–0.22 mm); group B (type 2.0; suture diameter, 0.22–0.23 mm; tip, 0.27–0.28 mm); and group C (type 3.0; suture diameter, 0.28–0.29 mm; and tip, 0.33–0.34 mm). The animals in each group were then subdivided into two subgroups, one of which received a nylon line inserted through the external carotid artery (ECA insertion), while the other received the nylon line through the common carotid artery (CCA insertion) subsequent to a middle or lateral neck incision. The neurological deficit score was evaluated at 4, 8, 24, 48 and 72 h post-surgery. The ischemic brain tissue was stained by 2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of the infarct volume. The cerebral edema rate, cerebral infarction volume rate, relative standard deviation (RSD) of the cerebral infarction rate and the success rate were also assessed. The rectal temperature, PaO2, PaCO2, pH, blood pressure and blood glucose levels were controlled and did not vary between the group types. The results suggested that suture diameter and insertion route affected the infarct volume and success rate in the establishment of the suture MCAO rat model. Furthermore, the MCAO model with a 0.22–0.23 mm diameter suture and CCA insertion route provided the highest success rate in the SD rats.

TANG, QIQIANG; HAN, RUODONG; XIAO, HAN; SHI, LILI; SHEN, JILONG; LUN, QINGLI; LI, JUN

2013-01-01

344

Dissection of USP catalytic domains reveals five common insertion points.  

PubMed

Ubiquitin specific proteases (USPs) are the largest family of deubiquitinating enzymes with approximately 56 members in humans. USPs regulate a wide variety of cellular processes by their ability to remove (poly)ubiquitin from target proteins. Their enzymatic activity is encoded in a common catalytic core of approximately 350 amino acids, however many USPs show significantly larger catalytic domains. Here we have analysed human and yeast USP domains, combining bioinformatics with structural information. We reveal that all USP domains can be divided into six conserved boxes, and we map the conserved boxes onto the USP domain core structure. The boxes are interspersed by insertions, some of which as large as the catalytic core. The two most common insertion points place inserts near the distal ubiquitin binding site, and in many cases ubiquitin binding domains or ubiquitin-like folds are found in these insertions, potentially directly affecting catalytic function. Other inserted sequences are unstructured, and removal of these might aid future structural and functional analysis. Yeast USP domains have a different pattern of inserted sequences, suggesting that the insertions are hotspots for evolutionary diversity to expand USP functionality. PMID:19734957

Ye, Yu; Scheel, Hartmut; Hofmann, Kay; Komander, David

2009-07-17

345

Mechanism of U-insertion RNA Editing in Trypanosome Mitochondria: The Bimodal TUTase Activity of the Core Complex  

PubMed Central

Expression of the trypanosomal mitochondrial genome requires the insertion and deletion of uridylyl residues at specific sites in pre-mRNAs. RET2 terminal uridylyl transferase (TUTase) is an integral component of the RNA editing core complex (RECC) and is responsible for the guide RNA-dependent U-insertion reaction. By analyzing RNAi-based knock-in Trypanosoma brucei cell lines, purified editing complex and individual protein, we have investigated RET2’s association with the RECC. In addition, the U-insertion activity exhibited by RET2 as RECC subunit was compared with characteristics of the monomeric protein. We show that RET2 interaction with RECC is accomplished via a protein-protein contact between its middle domain and a structural subunit MP81. The recombinant RET2 catalyzes a faithful editing on gapped (pre-cleaved) double-stranded RNA substrates and this reaction requires an internal monophosphate group at the 5?-end of the mRNA 3?-cleavage fragment. However, RET2 processivity is limited to insertion of three Us. Incorporation into the RECC voids the internal phosphate requirement and allows filling of longer gaps similar to those observed in vivo. Remarkably, monomeric and RECC-embedded enzymes display a similar bimodal activity: the distributive insertion of a single uracil is followed with a processive extension limited by the number of guiding nucleotides. Based on the RNA substrate specificity of RET2 and purine-rich nature of U-insertion sites, we propose that the distributive +1 insertion creates a substrate for the processive gap-filling reaction. Upon base-pairing of the +1-extended 5?-cleavage fragment with a guiding nucleotide, this substrate is recognized by RET2 in a different mode as compared to the product of the initial nucleolytic cleavage. Therefore, RET2 distinguishes base-pairs in gapped RNA substrates which may constitute an additional checkpoint contributing to overall fidelity of the editing process.

Ringpis, Gene-Errol; Aphasizheva, Inna; Wang, Xiaorong; Huang, Lan; Lathrop, Richard H.; Hatfield, G. Wesley; Aphasizhev, Ruslan

2010-01-01

346

Poverty Lines  

NSDL National Science Digital Library

The World Bank has released the November 1997 issue of Poverty Lines, a newsletter that summarizes current research studies on poverty. The latest issue looks at how public works programs can help people who are poor.

1996-01-01

347

Partial focusing by a bulk metamaterial formed by a periodically loaded wire medium with impedance insertions  

NASA Astrophysics Data System (ADS)

In this paper, a uniaxial wire medium periodically loaded with metallic patches and lumped impedance insertions is proposed for partial focusing of electromagnetic radiation due to a magnetic line source. The analysis is based on the nonlocal homogenization model for a bi-layer mushroom structure with generalized additional boundary conditions for loaded vias, and it is extended to a multilayered configuration with the transfer matrix approach. The proposed structure exhibits a high transmission and is nearly insensitive to the losses. The analytical results are validated against full-wave numerical simulations.

Kaipa, Chandra S. R.; Yakovlev, Alexander B.

2012-12-01

348

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

PubMed Central

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.

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

1997-01-01

349

Inserting lenses into corneal epithelial pockets to improve vision  

US Patent & Trademark Office Database

Methods of correcting vision are described. The methods include inserting a lens into a corneal epithelial pocket or a pocket created between a corneal epithelium of an eye of a patient and Bowman's membrane of the eye. A lens is inserted into the pocket to correct vision. Certain methods include one or more steps of cooling the corneal epithelium, and applying an aqueous liquid to the eye. The lenses that are inserted into the pocket can include collagen, including recombinant collagen, synthetic polymeric materials, and combinations thereof.

2010-11-09

350

Motor coordination deficits in Alpk1 mutant mice with the inserted piggyBac transposon  

PubMed Central

Background ALPK1 (?-kinase 1) is a member of an unconventional alpha-kinase family, and its biological function remains largely unknown. Here we report the phenotypic characterization of one mutant line, in which the piggyBac (PB) transposon is inserted into the Alpk1 gene. Results The piggyBac(PB) insertion site in mutants was mapped to the first intron of the Alpk1 gene, resulting in the effective disruption of the intact Alpk1 transcript expression. The transposon-inserted Alpk1 homozygous mutants (Alpk1PB/PB) displayed severe defects in motor coordination in a series of behavioral analysis, including dowel test, hanging wire test, rotarod analysis and footprint analysis. However, the cerebellar architecture, Purkinje cell morphology and electrophysiology of the Purkinje cells appeared normal in mutants. The motor coordination deficits in the Alpk1PB/PB mice were rescued by transgenic mice expressing the full-length Alpk1-coding sequence under the control of the ubiquitous expression promoter. Conclusions Our results indicate that ALPK1 plays an important role in the regulation of motor coordination. Alpk1PB/PB mice would be a useful model to provide a clue to the better understanding of the cellular and molecular mechanisms of ALPK1 in the control of fine motor activities.

2011-01-01

351

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

PubMed Central

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

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

2001-01-01

352

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

PubMed

In this study, the effect of insertion speed on long-term tissue response and insertion mechanics was investigated. A dummy silicon parylene-coated probe was used in this context and implanted in the rat brain at 10??m/s (n = 6) or 100??m/s (n = 6) to a depth of 9?mm. The insertion mechanics were assessed by the dimpling distance, and the force at the point of penetration, at the end of the insertion phase, and after a 3-min rest period in the brain. After 6 weeks, the tissue response was evaluated by estimating the amount of gliosis, inflammation, and neuronal cell loss with immunohistochemistry. No difference in dimpling, penetration force, or the force after a 3-min rest period in the brain was observed. However, the force at the end of the insertion phase was significantly higher when inserting the probes at 100??m/s compared to 10??m/s. Furthermore, an expected tissue response was seen with an increase of glial and microglial reactivity around the probe. This reaction was similar along the entire length of the probe. However, evidence for a neuronal kill zone was observed only in the most superficial part of the implant. In this region, the lesion size was also greatest. Comparison of the tissue response between insertion speeds showed no differences. PMID:21896383

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

2011-09-01

353

Spontaneous Radiation Emission from Short, High Field Strength Insertion Devices  

SciTech Connect

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.

Geoffrey Krafft

2005-09-15

354

Wireless cardiac action potential transmission with ultrasonically inserted silicon microprobes  

NASA Astrophysics Data System (ADS)

This paper reports on the integration of ultrasonically inserted horn-shaped cardiac probes with wireless transmission of 3D cardiac action potential measurement for applications in ex vivo preparations such as monitoring the onset of ventricular fibrillation. Ultrasonically inserted silicon horn probes permit reduced penetration force during insertion, allowing silicon, a brittle material, to penetrate cardiac tissue. The probes also allow recording from multiple sites that are lithographically defined. An application-specific integrated circuit has been designed with a 40 dB amplifying stage and a frequency modulating oscillator at 95 MHz to wirelessly transmit the recorded action potentials. This ultrasonically inserted microprobe wireless system demonstrates the initial results in wireless monitoring of 3D action potential propagation, and the extraction of parameters of interest including the action potential duration and diastolic interval.

Shen, C. J.; Ramkumar, A.; Lal, A.; Gilmour, R. F., Jr.

2011-05-01

355

A new insertion sequence for incremental Delaunay triangulation  

NASA Astrophysics Data System (ADS)

Incremental algorithm is one of the most popular procedures for constructing Delaunay triangulations (DTs). However, the point insertion sequence has a great impact on the amount of work needed for the construction of DTs. It affects the time for both point location and structure update, and hence the overall computational time of the triangulation algorithm. In this paper, a simple deterministic insertion sequence is proposed based on the breadth-first-search on a Kd-tree with some minor modifications for better performance. Using parent nodes as search-hints, the proposed insertion sequence proves to be faster and more stable than the Hilbert curve order and biased randomized insertion order (BRIO), especially for non-uniform point distributions over a wide range of benchmark examples.

Liu, Jian-Fei; Yan, Jin-Hui; Lo, S. H.

2013-02-01

356

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

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

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

357

Pitch estimation of a deeply inserted cochlear implant electrode.  

PubMed

In this short communication, we evaluate the place-pitch relation of a newly designed, deeply inserted, cochlear implant electrode. The insertion depths ranged from 471 degrees to 662 degrees. Pitch perception was measured in eight subjects with monopolar stimulation on each electrode contact at intensities of 50% and 80% of the dynamic range. We observed a monotonic reduction of pitch estimate with insertion depth. For about half of the subjects, a flattening of the pitch estimate at the basal end of the electrode was seen, while for the other half, pitch continued to decrease monotonically up to the most apical part of the array. We conclude that deeper insertion could increase pitch range for at least some cochlear implant recipients, and could hence potentially increase group performance. PMID:15457819

Deman, Peter R; van Dijk, Bas; Offeciers, F Erwin; Govaerts, Paul J

2004-06-01

358

Measured and simulated reactivity insertion transients characteristics of NIRR-1  

Microsoft Academic Search

The power excursion characteristics of the Nigeria Research Reactor-1 (NIRR-1) under different reactivity insertion transients have been calculated using PARET\\/ANL 7.3 code. Experimental data of dynamic experiments performed in NIRR-1 facility during initial startup were used to benchmark the calculated data. For ramp insertion of total cold core excess reactivity of 3.77mK, the reactor quickly reaches a maximum power of

S. A. Jonah

2011-01-01

359

Laryngeal mask airway can be inserted with inhaled desflurane induction  

Microsoft Academic Search

Purpose. In this prospective, randomized, controlled trial, we investigated the reliability of laryngeal mask airway (LMA) insertion with inhaled desflurane. Methods. Eighty patients undergoing elective surgery were randomized into two groups to receive either 2.5 mg·kg 1 propofol (n 40) or tidal breath desflurane (n 40) induction followed by LMA insertion. All patients received fentanyl 1 µg·kg 1 2 min

Wai May Leong; Ee Lyn Ong

2005-01-01

360

Propofol or sevoflurane for laryngeal mask airway insertion  

Microsoft Academic Search

Purpose  Sevoflurane is a volatile anesthetic agent, which combines rapid, smooth inhalational induction of anesthesia with rapid recovery,\\u000a making it particularly suitable for day case anesthesia. The laryngeal mask airway is often also used in ambulatory anesthesia,\\u000a with intravenous propofol being the agent of choice for its insertion. Our objective was to compare the conditions for laryngeal\\u000a mask airway (LMA) insertion

Mary E. Molloy; Donal J. Buggy; Patrick Scanlon

1999-01-01

361

Simulation and optimization of the dual lithium ion insertion cell  

Microsoft Academic Search

The galvanostatic charge and discharge of a dual lithium ion insertion (rocking-chair) cell are modeled. Transport in the electrolyte is described with concentrated solution theory. Insertion of lithium into and out of the active electrode material is simulated using superposition, greatly simplifying the numerical calculations. Simulations results are presented for the LiâCâ{vert_bar}propylene carbonate + 1M LiClOâ{vert_bar}Li{sub y}MnâOâ cell, and these

Thomas F. Fuller; M. Doyle; J. Newman

1994-01-01

362

Gene transfer to the tendon-bone insertion site  

Microsoft Academic Search

This study investigated whether gene transfer to the tendon-bone insertion site is possible during early tendon-transplant healing using viral vectors. In addition, we evaluated the optimal gene delivery technique for an in vivo adenoviral gene transfer to a tendon-bone insertion site in a bone tunnel. Twenty-six rabbits underwent a bilateral transfer of the flexor digitorum longus tendon into a bone

Christian Lattermann; Boris A. Zelle; Janey D. Whalen; Axel W. A. Baltzer; Paul D. Robbins; Christopher Niyibizi; Christopher H. Evans; Freddie H. Fu

2004-01-01

363

Micro injection molding for mass production using LIGA mold inserts  

Microsoft Academic Search

Micro molding is one of key technologies for mass production of polymer micro parts and structures with high aspect ratios.\\u000a The authors developed a commercially available micro injection molding technology for high aspect ratio microstructures (HARMs)\\u000a with LIGA-made mold inserts and pressurized CO2 gasses. The test inserts made of nickel with the smallest surface details of 5 ?m with structural height

Takanori Katoh; Ryuichi Tokuno; Yanping Zhang; Masahiro Abe; Katsumi Akita; Masaharu Akamatsu

2008-01-01

364

Squeeze cast aluminium reinforced with mild steel inserts  

Microsoft Academic Search

The bonding of a mild steel insert to an Al-7Si alloy during squeeze casting has been studied for a range of processing conditions. Assessment of the mild steel\\/Al-7Si alloy interface shear strength has been made with a push-out test, and the results have been correlated with microstructural observations and residual stress calculations. Uncoated inserts do not exhibit any significant reaction

G. Durrant; M. Gallerneault; B. Cantor

1996-01-01

365

Stress generation in silicon particles during lithium insertion  

NASA Astrophysics Data System (ADS)

Using a fully-coupled diffusion-elasticity model with Butler-Volmer surface kinetics, we simulate the insertion of lithium into spherical silicon particles. Simulations predict the evolution of concentration, displacements, and stresses in the particles during the first insertion of Li. The particle response depends strongly on the reaction kinetics and the resulting stresses can be above the tensile failure stress of silicon depending on the particle size and discharge rate.

Golmon, Stephanie; Maute, Kurt; Lee, Se-Hee; Dunn, Martin L.

2010-07-01

366

Investigation of unsteady temperature fields of nonaxisymmetric nozzle inserts  

NASA Astrophysics Data System (ADS)

The problem of calculating unsteady heat-conduction in an eccentric nozzle insert subjected to intense heating is considered with reference to the thrust vector control of supersonic flight vehicles. The third boundary value problem of unsteady heat-conduction is solved analytically for an eccentric ring. Results of calculations are presented for a nozzle insert of small eccentricity, and a comparison is made with finite-difference results.

Makarenkova, L. G.

367

Enhanced EPR Sensitivity from a Ferroelectric Cavity Insert  

Microsoft Academic Search

We report the development of a simple ferroelectric cavity insert that increases the electron paramagnetic resonance (EPR) sensitivity by an order of magnitude when a sample is placed within it. The insert is a hollow cylinder (length 4.8 mm, outside diameter 1.7 mm, inside diameter 0.6 mm) made from a single crystal of KTaO3, which has a dielectric constant of

Yuri E. Nesmelov; Jack T. Surek; David D. Thomas

2001-01-01

368

Germ line insertion of mtDNA at the breakpoint junction of a reciprocal constitutional translocation  

Microsoft Academic Search

Constitutional chromosomal translocations are relatively common causes of human morbidity, yet the DNA double-strand break (DSB) repair mechanisms that generate them are incompletely understood. We cloned, sequenced and analyzed the breakpoint junctions of a familial constitutional reciprocal translocation t(9;11)(p24;q23). Within the 10-kb region flanking the breakpoints, chromosome 11 had 25% repeat elements, whereas chromosome 9 had 98% repeats, 95% of

Joan E. Willett-Brozick; Shazia A. Savul; Lauren E. Richey; Bora E. Baysal

2001-01-01

369

Hsp90 interacts with inducible NO synthase client protein in its heme-free state and then drives heme insertion by an ATP-dependent process.  

PubMed

Maturation of NOS enzymes requires that they incorporate heme to become active, but how this cellular process occurs is unclear. We investigated a role for chaperone heat shock protein 90 (hsp90) in enabling heme insertion into the cytokine-inducible mouse NOS. We used macrophage cell line RAW 264.7 and human embryonic kidney HEK293T cells and studied insertion of native heme during iNOS expression and insertion of exogenous heme into preformed apo-iNOS. Pulldown experiments showed that the hsp90-iNOS complex was present in cells, but the extent of their association was inversely related to iNOS heme content. Hsp90 was primarily associated with apo-iNOS monomer and was associated 11-fold less with heme-containing iNOS monomer or dimer in cells. Kinetic studies showed that hsp90 dissociation occurred coincident with cellular heme insertion into apo-iNOS (0.8 h(-1)). The hsp90 inhibitor radicicol or coexpression of an ATPase-defective hsp90 blocked heme insertion into apo-iNOS by 90 and 75%, respectively. The ATPase activity of hsp90 was not required for complex formation with iNOS but was essential for heme insertion to occur. We conclude that hsp90 plays a primary role in maturation of iNOS protein by interacting with the apoenzyme in cells and then driving heme insertion in an ATP-dependent manner. PMID:21357526

Ghosh, Arnab; Chawla-Sarkar, Mamta; Stuehr, Dennis J

2011-02-25

370

Exploring the nature of the translocon-assisted protein insertion.  

PubMed

The elucidation of the molecular nature of the translocon-assisted protein insertion is a challenging problem due to the complexity of this process. Furthermore, the limited availability of crucial structural information makes it hard to interpret the hints about the insertion mechanism provided by biochemical studies. At present, it is not practical to explore the insertion process by brute force simulation approaches due to the extremely lengthy process and very complex landscape. Thus, this work uses our previously developed coarse-grained model and explores the energetics of the membrane insertion and translocation paths. The trend in the calculated free-energy profiles is verified by evaluating the correlation between the calculated and observed effect of mutations as well as the effect of inverting the signal peptide that reflects the "positive-inside" rule. Furthermore, the effect of the tentative opening induced by the ribosome is found to reduce the kinetic barrier. Significantly, the trend of the forward and backward energy barriers provides a powerful way to analyze key energetics information. Thus, it is concluded that the insertion process is most likely a nonequilibrium process. Moreover, we provided a general formulation for the analysis of the elusive apparent membrane insertion energy, ?G(app), and conclude that this important parameter is unlikely to correspond to the free-energy difference between the translocon and membrane. Our formulation seems to resolve the controversy about ?G(app) for Arg. PMID:23269832

Rychkova, Anna; Warshel, Arieh

2012-12-26

371

Bits with diamond-coated inserts reduce gauge problems  

SciTech Connect

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.

Eckstrom, D. (Smith International Canada Ltd., Calgary (CA))

1991-06-17

372

Site-specific insertion of IS492 in Pseudoalteromonas atlantica.  

PubMed

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 x 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

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

2009-08-14

373

Site-Specific Insertion of IS492 in Pseudoalteromonas atlantica?  

PubMed Central

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.

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

2009-01-01

374

P-Element Insertion Alleles of Essential Genes on the Third Chromosome of Drosophila Melanogaster: Correlation of Physical and Cytogenetic Maps in Chromosomal Region 86e-87f  

PubMed Central

We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover ~25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty-one of these 38 complementation groups have at least one allele that has a P-element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven ``supercontigs'' that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the ?? element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms.

Deak, P.; Omar, M. M.; Saunders, RDC.; Pal, M.; Komonyi, O.; Szidonya, J.; Maroy, P.; Zhang, Y.; Ashburner, M.; Benos, P.; Savakis, C.; Siden-Kiamos, I.; Louis, C.; Bolshakov, V. N.; Kafatos, F. C.; Madueno, E.; Modolell, J.; Glover, D. M.

1997-01-01

375

A postimplantation lethal mutation induced by transgene insertion on mouse chromosome 8  

SciTech Connect

We have produced three lines of transgenic mice that contain additional copies of the mouse phosphoglycerate kinase 1 (Pgk1) gene. Two of these lines, 94-A and 94-K, which are descendants of a common founder, did not produce liveborn progeny carrying two copies of these transgenes (i.e., A/A, K/K, or A/K). Genotyping of midgestation embryos showed that A/K embryos are dead by Embryonic Day 10. Comparison of the level of transgene expression in the three transgenic lines ruled out PGK1 toxicity as the cause of death of A/A, A/K, and K/K embryos. The death of A/A, K/K, and A/K transgenic mice was therefore attributed to an insertional mutation disrupting a gene or genes essential for normal embryogenesis. Analysis of the structure of the 94-A and 94-K transgenes indicated that they differ in the number of tandem repeats and in the positions of the transgene-cellular DNA junctions. To determine if the two transgenes represent a single integration followed by a rearrangement or two independent integration events, we cloned the endogenous sequences surrounding the 94-A and 94-K transgene insertion sites. Restriction analysis of the isolated genomic clones indicated that the endogenous sequences abutting the 3{prime} ends of the 94-A and 94-K transgenes are separated by less than 20 kb, providing strong support for the single integration model. Further analysis indicated that the 94-A transgene is associated with a deletion of at least 18 kb and is located in the vicinity of a widely transcribed endogenous gene. Chromosomal mapping of the endogenous sequences flanking the 94-A and 94-K transgene insertions using mouse -hamster somatic cell hybrids and a (C57BL/6J X SPRET/Ei)F1 X SPRET/Ei backcross panel allowed us to assign the 94-A(K) transgene insertion to the subcentral region of mouse chromosome 8. 100 refs., 10 figs., 5 tabs.

Pravtcheva, D.D.; Wise, T.L. [Saint Louis Univ., MO (United States)

1995-12-10

376

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

Code of Federal Regulations, 2013 CFR

...false Posterior artificial tooth with a metal insert. 872.3900 Section 872...3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is a porcelain device with...

2013-04-01

377

Assembly Line  

NSDL National Science Digital Library

In this activity, learners work individually to assemble a product and then work in teams to design, construct, test, and redesign an assembly line process whose product must meet specific quality control criteria. Learners reflect and compare the two approaches. Note: Page 14 is not in the correct position. It should actually be between pages 2 and 3.

Ieee

2013-07-08

378

Cell lines  

US Patent & Trademark Office Database

Genomic instability in T-antigen expressing cells can be overcome by modifying the gene expressing T-antigen so that it lacks Bub1 binding. Stable cell lines can be produced by incorporation of the modified T-antigen gene, preferably together with the catalytic sub-unit of the telomerase construct.

2008-09-02

379

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

PubMed Central

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

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

2011-01-01

380

Timing of femoral prosthesis insertion during cemented arthroplasty: cement curing and static mechanical strength in an in vivo model  

PubMed Central

Background Modern cementing techniques aim to fix the implanted femoral prosthesis in the medullary cavity to minimize long-term complications such as aseptic loosening. The cure stage of bone cement into which the femoral component is being inserted is an important variable that is decided at the time of surgery. Late-cure cement is more viscous than early-cure cement and requires greater force on the part of the surgeon to insert the femoral prosthesis. We compared 2 cementing techniques, femoral component insertion into early-cure cement and insertion into late-cure cement, using an in vivo model to identify if cement cure stage affects the strength of the bone–cement interface. Methods We performed bilateral hemiarthroplasties using only the femoral component in vivo on paired porcine femora. The femora were harvested and cross-sectioned in preparation for strength testing. We measured bond strength by peak load required to push the femoral prosthesis and surrounding cement mantle free of the cancellous bone. Results All radiographs showed good cement interdigitation with no evidence of radiolucent lines at the bone–cement interface. We could not differentiate the early-cure and late-cure groups on postoperative radiographs. The mean failure load for the late-cure arthroplasties was 908 N (standard deviation [SD] 420 N), whereas the mean failure load for the conjugate early-cure arthroplasties was 503 N (SD 342 N). Conclusion Femoral component insertion into late-cure cement required significantly higher loads for push-out than femoral component insertion into early-cure cement.

Hunt, Stephen; Stone, Craig; Seal, Shane

2011-01-01

381

Lithium insertion in carbonaceous materials containing silicon  

NASA Astrophysics Data System (ADS)

Three different series of silicon-containing carbonaceous materials were synthesized for use as anodes in lithium ion cells. Disordered (or pregraphitic) carbons containing nanodispersed silicon were prepared by the chemical vapour deposition (CVD) of various chlorosilanes (SiClsb4, (CHsb3)sb2Clsb2Si, and (CHsb3)sb3ClSi) with benzene in two different apparatuses. Silicon oxycarbide glasses were synthesized by the pyrolysis of over 50 silicon-containing polymers at various temperatures, although the principal materials in the study were prepared at 1000sp°C. Finally, materials which we believe to be similar to disordered carbons containing nanodispersed silicon were prepared by the pyrolysis of various blends of pitches with polysilanes. Powder X-ray diffraction was used to learn about the structure of all the materials made. Thermal gravimetric analysis was used to determine the silicon content in the CVD materials and, when coupled to a residual gas analyzer, to study the decomposition process of the polymers. Near edge X-ray absorption spectroscopy measurements of the silicon L- and K-edges of CVD materials and the silicon K-edges of silicon oxycarbides were used to learn about local chemical environments of the silicon atoms. Lithium metal electrochemical test cells of the silicon-containing CVD materials showed larger capacities (up to 500 mAh/g) than pure carbons prepared in the same way (˜300 mAh/g). The additional capacity was observed to be centered near 0.4 V on charge, the average voltage observed for the removal of lithium from a silicon-lithium alloy. Chemical analysis showed that the stoichiometries of materials made by polymer pyrolysis were distributed over a well-defined region in the Si-O-C Gibbs phase diagram. An interesting series of materials is found near the line in the Si-O-C Gibbs triangle connecting carbon to SiOsb{1.3}. Lithium metal electrochemical test cells made using all the silicon oxycarbides synthesized showed that a stoichiometry of about Sisb{.25}Csb{.45}Osb{.30} gave the maximum reversible capacity (about 900 mAh/g). However, materials near this stoichiometry exhibit large irreversible capacities (>350 mAh/g) and significant hysteresis (the voltage difference between charge and discharge) in the voltage profile (˜0.8 V). In an attempt to reduce the oxygen content in one of the silicon oxycarbide glasses, a sample was washed in a dilute solution of hydrofluoric acid (HF) for times ranging from 2 minutes to 24 hours. The material lost, at most, 40 percent of its initial mass, although there was only a small change in its stoichiometry. In addition to the techniques mentioned above, small angle X-ray scattering and BET surface area measurements were used to study the microscopic pore network that was created by the HF washing. Lithium metal electrochemical test cells made using the product of pyrolysing pitch-polysilane blends showed that the capacity increased with silicon content from 340 mAh/g for pure carbon to a maximum of 600 mAh/g for samples with about 15 atomic % silicon (Sisb{.14}Osb{.09}Csb{.77}). The capacity then decreased to near zero as the composition approached SiC. These materials contain oxygen which is correlated to irreversible capacity loss. (Abstract shortened by UMI.)

Wilson, Alfred Macdonald

382

Transmission Lines  

NSDL National Science Digital Library

All About Circuits is a website that âÂÂprovides a series of online textbooks covering electricity and electronics.â Written by Tony R. Kuphaldt, the textbooks available here are wonderful resources for students, teachers, and anyone who is interested in learning more about electronics. This specific section, Transmission Lines, is the fourteenth chapter in Volume II â Alternating Current. A few of the topics covered in this chapter include: Circuits and the speed of light; Finite-length transmission lines; and Waveguides. Diagrams and detailed descriptions of concepts are included throughout the chapter to provide users with a comprehensive lesson. Visitors to the site are also encouraged to discuss concepts and topics using the All About Circuits discussion forums (registration with the site is required to post materials).

Kuphaldt, Tony R.

2008-07-03

383

Co-translational membrane insertion of mitochondrially encoded proteins.  

PubMed

The components of the mitochondrial proteome represent a mosaic of dual genetic origin: while most mitochondrial proteins are encoded by nuclear genes and imported into the organelle following synthesis in the cytosol, a small number of proteins is encoded by the mitochondrial genome. Though small in number, mitochondrial translation products are vital for cellular functionality as these proteins represent the core subunits of the respiratory chain and the ATPase which produce the vast majority of the cellular ATP. Mitochondrial translation products are almost exclusively highly hydrophobic polypeptides which are inserted into the inner membrane in the course of their synthesis. The machinery that mediates membrane insertion in mitochondria is deduced from that of their bacterial ancestors and hence shows profound similarities to the insertion machinery of prokaryotes. However, the specialization on the production of a very small set of translation products drove a severe reduction in the complexity of this system. The insertase Oxa1 forms the central component of the insertion machinery. Oxa1 directly binds to mitochondrial ribosomes and, together with the inner membrane protein Mba1, aligns the polypeptide exit tunnel of the ribosome with the insertion site at the inner membrane. The specific hallmarks and the critical components of this machinery are discussed in this review article. PMID:19962410

Ott, Martin; Herrmann, Johannes M

2009-12-02

384

Plasma cortisol concentrations after CIDR insertion in beef cows.  

PubMed

The objective of this study was to show plasma cortisol concentration after treatment with controlled internal drug release (CIDR) in non-suckling beef cows. On day 9 after oestrus, two cows were inserted with CIDR into the vagina for 24 h and the other two cows were treated as a control group. Four days later, the two control cows were treated with CIDR and the other two CIDR-treated cows were used as controls. Cortisol concentrations were determined by ELISA in plasma samples collected before, during and after insertion of CIDR. There was a significant increase in plasma cortisol concentrations (p<0.01) after insertion of CIDR. Mean (± SEM) plasma cortisol concentrations increased from 1.3 ± 0.4 to a peak of 8.8 ± 1.1 ng/ml at 5 h and then decreased to basal concentrations at 7 h after insertion of the device. In conclusion, the insertion of intra-vaginal device causes an increase in plasma cortisol concentrations in beef cows, although the pathophysiological significance of the elevation of cortisol is not known. PMID:20403126

Long, S T; Thinh, N C; Yusuf, M; Nakao, T

2011-02-01

385

Some physical and chemical indices of clique-inserted lattices  

NASA Astrophysics Data System (ADS)

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

Zhang, Zuhe

2013-10-01

386

Short indels are subject to insertion-biased gene conversion.  

PubMed

Recombination between homologous loci is accompanied by formation of heteroduplexes. Repairing mismatches in heteroduplexes often leads to single nucleotide substitutions in a process known as gene conversion. Gene conversion was shown to be GC-biased in different organisms; that is, a W(A or T)?S(G or C) substitution is more likely in this process than a S?W substitution. Here, we show that the insertion/deletion ratio for short noncoding indels that reach fixation between species is positively correlated with the recombination rate in Drosophila melanogaster, Homo sapiens, and Saccharomyces cerevisiae. This correlation is both due to an increase of the fixation rate of insertions and decrease of the fixation rate of deletions in regions of high recombination. Whole-genome data on indel polymorphism and divergence in D. melanogaster rule out mutation biases and selection as the cause of this trend, pointing to insertion-biased gene conversion as the most likely explanation. The bias toward insertions is the strongest for single-nucleotide indels, and decreases with indel length. In regions of high recombination rate this bias leads to an up to ?5-fold excess of fixed short insertions over deletions, and substantially affects the evolution of DNA segments. PMID:24033170

Leushkin, Evgeny V; Bazykin, Georgii A

2013-05-11

387

GAPDH regulates cellular heme insertion into inducible nitric oxide synthase.  

PubMed

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

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

2010-10-04

388

Anatomic safe zone of pin insertion point for distal clavicle fixation.  

PubMed

Clavicle fracture is the most common childhood fracture and one of the most common fractures in adults. Only some types of distal clavicular fractures, and dislocation of the acromioclavicular joint, require internal fixation. Many surgeons prefer closed pinning; however, the difficulty inserting many of the various kinds of pins from acromion into the medullary canal, of the distal clavicle, means the likelihood of iatrogenic complications from repeated drilling is heightened. The purpose of the present study was to establish what would be the optimum insertion point and direction for safe intramedullary pinning of the distal clavicle. Embalmed cadaveric shoulders (32) were studied. A bone window was created at the distal one-thirds of the clavicle, approximately 1.5 cm medial from the conoid tuberosity - as wide as could be freely, retrogradely drilled into the medullary canal of the distal clavicle. A 2.0-mm Kirschner wire was inserted until it penetrated the acromion. The point of emergence was recorded as ratio compared with the acromial width and length in coronal and sagittal planes, respectively. K-wire directions were measured as the angle between the K-wire and the reference line from the anterosuperior tubercle of the clavicle to the anterior angle of the acromion. The process was repeated until the acromion fractured 304 drillings were performed on 32 specimens. The length of the sagittal vs.coronal pin insertion point from the anterior vs. lateral borders of the acromion divided by its length vs. width averaged 0.325 +/- 0.04 and 0.397 +/- 0.09, respectively. The angle of the K-wire and the reference was 7.69 +/- 3.04 and 14.59 +/- 4.34 degrees in the coronal and horizontal planes, respectively. At 8 and 10 drillings survival was 0.72 (95%CI: 0.53-0.84) and 0.41 (95%CI: 0.24-0.57), respectively. The optimum pin inserting point for fixation of distal clavicle fracture and acromioclavicular joint dislocation is 32.5% and 39.7% of acromial length and width, respectively. If a 2.0-mm K-wire is used for fixation, drilling should not be repeated drilled more than 8 times to avoid sudden, high risk iatrogenic acromial fracture. PMID:16471101

Thumroj, Ekamol; Kosuwon, Weerachai; Kamanarong, Kimaporn

2005-11-01

389

Chromosomal Integration Pattern of a Helper-Dependent Minimal Adenovirus Vector with a Selectable Marker Inserted into a 27.4Kilobase Genomic Stuffer  

Microsoft Academic Search

vector with a neor gene inserted centrally into a noncoding 27.4-kb genomic stuffer element derived from the human X chromosome after infection of a sex chromosome aneuploid (X0) human glioblastoma cell line was studied. Our results indicate that even extensive homologies and abundant chromosomal repeat elements present in the vector did not lead to integration of the vector via homologous

MORITZ HILLGENBERG; HOLGER TONNIES; MICHAEL STRAUSS

2001-01-01

390

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

PubMed

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

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

1987-12-01

391

Refrigerated penetration insert for cryostat with axial thermal disconnect  

SciTech Connect

This patent describes an interface for a horizontal cryostat penetration insert for coupling a cryogenic refrigerator cold end to the cryostat penetration insert. The insert has an outer housing and a thermal radiation shield. The cryogenic refrigerator cold end has at least one stage. The interface comprises: an extension of the outer housing providing access to the interior of the housing; an extension of the thermal radiation shield into the housing extension, the shield having a flexible portion compressible in the axial direction, and a first thermal contact means affixed to the end of the thermal radiation shield extension. The first thermal contact means defines a central aperture and provides heat conduction from the shield extension to the first thermal contact means.

Miller, R.S.; Brzozowski, S.J.

1987-05-26

392

Parental perceptions of grommet insertion in children with cleft palate.  

PubMed

Otitis media with effusion is almost universal in children with cleft palate and can delay speech, language and educational development by causing hearing loss. Grommet insertion at the time of cleft palate repair is common. There is debate about whether the benefits of grommets outweigh the risk of complications. A postal questionnaire was used to investigate parental perceptions of middle-ear ventilation via grommet insertion in children attending the multidisciplinary cleft palate clinic. These children's case notes were reviewed. Many children had speech and language delay, but parents thought this improved after grommet insertion. Overall parents were pleased with the results. This confirms that grommets have an important part to play in the management of children with cleft palate. PMID:10767917

Greig, A V; Papesch, M E; Rowsell, A R

1999-12-01

393

Tool Life Detecting System Using Damage Sensor-Integrated Insert  

NASA Astrophysics Data System (ADS)

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.

Sekiya, Katsuhiko; Yamane, Yasuo; Torimoto, Ayumu

394

A Nasogastric Tube Inserted into the Gastrocutaneous Fistula  

PubMed Central

We reported a case in which a nasogastric tube was inserted into the gastrocutaneous fistula, diagnosed by abdominal computed tomography. A 78-year-old man with a history of recurrent cerebral hemorrhage had a percutaneous endoscopic gastrostomy tube due to dysphagia for 2 years. However, soft tissue infection at the gastrostomy site caused the removal of the tube. Immediately, antibiotic agents were infused. For appropriate hydration and medication, a nasogastric tube was inserted. However, there was no significant improvement of the soft tissue infection. Moreover, the amount of bloody exudate increased. Abdominal computed tomography revealed the nasogastric tube placed under the patient's skin via gastrocutaneous fistula. The nasogastric tube was removed, and an antibiotic agents were maintained. After 3 weeks, the signs of infection fully improved, and percutaneous endoscopic gastrostomy was performed again. This case shows necessities of an appropriate interval between removal of the gastrostomy tube and insertion of a nasogastric tube, and suspicion of existence of gastrocutaneous fistula.

Kim, Yang Soo; Kim, Joon Sung; Yu, In Hee; Jeong, Ji Young; Jung, Sung Hee; Jo, Yil Ryun

2011-01-01

395

Synchronized monochromator and insertion device energy scans at SLS  

SciTech Connect

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.

Krempasky, J.; Flechsig, U.; Korhonen, T.; Zimoch, D.; Quitmann, Ch.; Nolting, F. [Paul Scherrer Institut, Villigen, Swiss Light Source, 5235 Villigen PSI (Switzerland)

2010-06-23

396

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

PubMed

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

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

2012-04-26

397

Molecular archeology of L1 insertions in the human genome  

PubMed Central

Background As the rough draft of the human genome sequence nears a finished product and other genome-sequencing projects accumulate sequence data exponentially, bioinformatics is emerging as an important tool for studies of transposon biology. In particular, L1 elements exhibit a variety of sequence structures after insertion into the human genome that are amenable to computational analysis. We carried out a detailed analysis of the anatomy and distribution of L1 elements in the human genome using a new computer program, TSDfinder, designed to identify transposon boundaries precisely. Results Structural variants of L1 elements shared similar trends in the length and quality of their target site duplications (TSDs) and poly(A) tails. Furthermore, we found no correlation between the composition and genomic location of the pre-insertion locus and the resulting anatomy of the L1 insertion. We verified that L1 insertions with TSDs have the 5'-TTAAAA-3' cleavage site associated with L1 endonuclease activity. In addition, the second target DNA cut required for L1 insertion weakly matches the consensus pattern TTAAAA. On the other hand, the L1-internal breakpoints of deleted and inverted L1 elements do not resemble L1 endonuclease cleavage sites. Finally, the genome sequence data indicate that whereas singly inverted elements are common, doubly inverted elements are almost never found. Conclusions The sequence data give no indication that the creation of L1 structural variants depends on characteristics of the insertion locus. In addition, the formation of 5' truncated and 5' inverted L1s are probably not due to the action of the L1 endonuclease.

Szak, Suzanne T; Pickeral, Oxana K; Makalowski, Wojciech; Boguski, Mark S; Landsman, David; Boeke, Jef D

2002-01-01

398

Temporal relationship between Uniplant insertion and changes in cervical mucus.  

PubMed

This study was undertaken to determine the time required by a single implant containing nomegestrol acetate to affect cervical mucus production and sperm penetration in women. All subjects were investigated and, if necessary, treated for any kind of cervicitis or vaginitis prior to starting cervical mucus study. The subjects had not used hormonal contraception for at least three months prior to investigation. They were counseled to use condoms during this study and also to refrain from intercourse during the period of cervical mucus sampling. Follicular development and endometrial thickness were analyzed by transvaginal sonography. Cervical mucus examination, sperm penetration test, and transvaginal sonography were performed during the control cycle and during the first cycle of Uniplant use. Blood samples were taken for the measurement of estradiol, LH, and progesterone. Cervical mucus and sperm penetration tests were evaluated according to the World Health Organization (WHO) criteria. In the treated cycle, when cervical mucus reached a score of 8-10, Uniplant was inserted, independent of the day of the cycle. Cervical mucus was then collected at 0, 4, 8, 12, 24, 48, and 96 h later until a marked change in volume, consistency, ferning spinnbarkheit, and cellularity was observed. All samples were also used for sperm penetration test. Preovulatory estradiol and LH peak decreased significantly compared to pre-implant insertion. Progesterone levels were within the normal limit. Cervical mucus and sperm penetration tests were not affected by Uniplant in the first 12 h. Twenty-four hours after Uniplant insertion, cervical mucus and sperm penetration tests were affected in 70.6% of the women. Forty-eight hours after implant insertion, the women were affected. Follicular rupture occurred in the majority of the women 48 h after implant insertion. Based on these results, it is possible to conclude that Uniplant can affect estradiol and LH preovulatory peaks and disrupt the process of cervical mucus production and sperm penetration, but it was unable to prevent ovulation when inserted in the preovulatory phase. PMID:8922874

Barbosa, I C; Coutinho, E; Hirsch, C; Ladipo, O A; Olsson, S E; Ulmsten, U

1996-10-01

399

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

SciTech Connect

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.

Akker, Eric van den; Aarts, Lambertus H.J. [Department of Hematology, Erasmus University Medical Center, Dr. Molewaterplein 50, 3015 GE Rotterdam (Netherlands); Delwel, Ruud [Department of Hematology, Erasmus University Medical Center, Dr. Molewaterplein 50, 3015 GE Rotterdam (Netherlands)], E-mail: h.delwel@erasmusmc.nl

2007-09-30

400

Desflurane requirements for laryngeal mask airway insertion during inhalation induction  

Microsoft Academic Search

Purpose  We hypothesized that the simultaneous use of low concentrations (2O), and fentanyl would allow a laryngeal mask airway (LMA) to be inserted safely with inhalation induction of desflurane,\\u000a even in nonparalyzed patients. This prospective, observational study was performed to determine the 50% effective concentration\\u000a (EC50) of desflurane for LMA insertion in such patients.\\u000a \\u000a \\u000a \\u000a Methods  Twenty-two adult patients undergoing ambulatory surgical procedures

Hwa-Yong Shin; Jung-Ae Lim; Seong-Hyop Kim; Seung-Woo Baek; Duk-Kyung Kim

2009-01-01

401

Does notch size predict ACL insertion site size?  

Microsoft Academic Search

Purpose  The primary purpose of the current study is to identify a possible correlation between the femoral intercondylar notch size\\u000a and the ACL insertion site size. The secondary purpose is to determine if there is a difference between male and female notch\\u000a widths and insertion site sizes.\\u000a \\u000a \\u000a \\u000a \\u000a Methods  For this study, 82 patients (41 men and 41 women) with an average age

Femke Wolters; Sharon H. A. Vrooijink; Carola F. Van Eck; Freddie H. Fu

402

Rectal fist insertion. An unusual form of sexual behavior.  

PubMed

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

Shook, L L; Whittle, R; Rose, E F

1985-12-01

403

Factor Lines  

NSDL National Science Digital Library

This flash applet provides students with an activity to become more familiar with factors and multiples. The challenge is to arrange the four number cards (1, 2, 3 and 21) on a square of the grid to make as many different diagonal, vertical or horizontal lines as possible. The number card can be placed on a square of the grid if the square is the same number, a multiple of that number and or a factor of that number. Users have the ability to change the difficulty level. The Teachers' Notes page offers rationale, suggestions for implementation, key discussion questions, ideas for extension and support.

2009-10-01

404

Efficient simulation of chemical potentials and phase equilibria in associating fluids: monomer/dimer insertion versus gradual particle insertion in primitive water models  

NASA Astrophysics Data System (ADS)

Widom's test particle method for simulation of chemical potentials fails for associating (network forming) fluids. Therefore we study more sophisticated insertion methods, such as gradual particle insertion and the recent unbonded particle insertion of Tripathi and Chapman. We model strongly associating fluids by short-ranged primitive models (PMs) due to Nezbeda and co-workers. Recently, we showed that gradual insertion, in principle, is applicable to PM water. Here, we study systematically subcritical chemical potential isotherms, determine vapour-liquid coexistence densities and estimate the critical point density and temperature. For comparison we implement two variants of the Tripathi-Chapman algorithm. In the first case we use as unbonded test particles only particles which remain single molecules (monomers), when inserted and in the second case such test particles which remain as single molecules or form a two-particle cluster with an existing free molecule (monomer/dimer), when inserted. While monomer insertion improves Widom's method slightly, monomer/dimer insertion extends substantially the range of application to the subcritical temperatures studied by gradual insertion. But, for low temperatures monomer/dimer insertion requires extremely long Markov chains and significantly more computer time than gradual insertion. The Tripathi-Chapman algorithm - a direct extension of Widom's method - is much simpler than gradual insertion and therefore should be preferred at supercritical and critical conditions.

Vörtler, Horst L.; Kettler, Matthias

405

MR Inserts for Shock Wave Reduction in Warship Structures  

Microsoft Academic Search

The underwater explosion that has high energy brings about the shock wave and the pulsating gas bubble. In general, structural vibration from the shock wave is more serious than the pulsating gas bubble. This shock wave may damage the important fragile structures and equipment in the ship. This paper demonstrates the possibility of shock wave reduction in magneto-rheological (MR) inserts.

J. H. Kim; B. W. Kang; K. M. Park; S. B. Choi; K. S. Kim

2002-01-01

406

Hearing Aid Evaluation: Predicting Speech Gain from Insertion Gain.  

ERIC Educational Resources Information Center

This study, involving 11 older males with mild or moderate sensorineural loss, tested whether speech gain that results from a hearing aid fitting is predictable on the basis of unaided performance intensity curve, unaided narrow-band sound field thresholds, hearing aid insertion gain as a function of frequency, ambient noise, and internal hearing…

Dillon, Harvey

1993-01-01

407

A Comparison of Network Level Fault Injection with Code Insertion  

Microsoft Academic Search

This paper describes our research into the application of fault injection to Simple Object Access Protocol (SOAP) based Service Oriented-Architectures (SOA). We show that our previously devised WS-FIT method, when combined with parameter perturbation, gives comparable performance to Code Insertion techniques with the benefit that it is less invasive. Finally we demonstrate that this technique can be used to compliment

Nik Looker; Malcolm Munro; Jie Xu

2005-01-01

408

21 CFR 529.1940 - Progesterone intravaginal inserts.  

Code of Federal Regulations, 2013 CFR

...should be disposed after a single use. Administration of vaginal inserts for periods greater than 7 days may result in reduced fertility. Dinoprost solution provided by No. 000009 in § 510.600(c) of this chapter. (2) Ewes â(i) Amount ....

2013-04-01

409

Insert Technique for Constructing Artificial Red-Cockaded Woodpecker Cavities.  

National Technical Information Service (NTIS)

A complete guide is provided for excavating red-cockaded woodpecker (Picoides borealis) cavities. A hole 4 inches wide by 10 inches high by 6 inches deep is cut from a live pine (Pinus spp.) tree with a chainsaw, and a prefabricated cavity is inserted. Ca...

D. H. Allen

1991-01-01

410

Local effects in the vicinity of inserts in sandwich panels  

Microsoft Academic Search

The paper considers local bending effects induced in the vicinity of inserts in sandwich panels. Such local bending effects are associated with an increase of bending stresses in the sandwich faces and normal and shear stresses in the sandwich core. An earlier developed analytic model [Proceedings of the Sixth International Conference on Sandwich Structures (ICSS-6) (2002) 551] is adapted for

Elena Bozhevolnaya; Anders Lyckegaard; Ole. t. Thomsen; Vitaly Skvortsov

2004-01-01

411

Cylinder liner insert and method of making engine block therewith  

Microsoft Academic Search

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

N. L. Field; B. P. Winter

1990-01-01

412

Peripheral intravenous cannulation: safe insertion and removal technique.  

PubMed

The aim of this article is to reinforce good practice in insertion and removal techniques for peripheral intravenous cannulation. The article is intended as a practical guide. It is important that staff receive adequate education and training to undertake this skill competently, and also that they maintain competence in practice. PMID:17941430

Ingram, P; Lavery, I

413

33. INTERIOR VIEW OF FEEDER PREPARING TO INSERT THE FEEDER ...  

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

33. INTERIOR VIEW OF FEEDER PREPARING TO INSERT THE FEEDER ROD INTO ONE OF THE TWO LARGER NAIL OR SPIKE CUTTING MACHINES WHICH PRODUCE THE LARGER SIZE NAILS; NOTE THE OTHER SPIKE CUTTING MACHINE IN THE BACKGROUND - LaBelle Iron Works, Thirtieth & Wood Streets, Wheeling, Ohio County, WV

414

Planning of prosthetic ligament insertion in the knee  

Microsoft Academic Search

Prosthetic anterior cruciate ligaments (ACL) for the knee often suffer from pre-mature failure due to fatigue. Deformations of these ligaments are calculated as a function of their insertion orientation via a computer model. To help physically interpret these deformation values, the average flexion axis of the knee is calculated using a least-squares algorithm on experimental kinematic data. The results indicate

A. Richard; J. A. De Guise; S. Baldo; G. Dronin

1995-01-01

415

Observations on rotating needle insertions using a brachytherapy robot  

NASA Astrophysics Data System (ADS)

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.

Meltsner, M. A.; Ferrier, N. J.; Thomadsen, B. R.

2007-09-01

416

A Computer Based Simulator for Ultrasound Guided Needle Insertion Procedures  

Microsoft Academic Search

A system for the simulation of needle insertion procedures under ultrasound guidance is presented. This is a core skill in interventional radiology, currently acquired on an apprenticeship basis. The system is designed to improve the early stages of training, and detect people with poor natural ability. The simulator is based on a volumetric data set which is non-rigidly aligned to

D. Magee; D. Kessel

417

Insertion and deletion mutagenesis of the human cytomegalovirus genome  

SciTech Connect

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.

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

1987-10-01

418

Magnesium oxide inserts for the LECO Carbon Analyzer  

SciTech Connect

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.

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

1991-01-16

419

Rapid cloning of insect transposon insertion junctions using 'universal' PCR  

Microsoft Academic Search

Very highly degenerate primers with short specific 3' anchor sequences and 5' adaptors were used in con- junction with nested specific primers to amplify large numbers of unknown insertion junctions of the insect retrotransposon Woot, using genomic DNA as tem- plate for the polymerase chain reaction (PCR). This technique, sometimes referred to as universal PCR, is a powerful method for

R. W. Beeman; D. M. Stauth

1997-01-01

420

Bone registration method for robot assisted surgery: pedicle screw insertion  

Microsoft Academic Search

SUMMARY A registration method that identifies bone geometry with respect to a robotic manipulator arm is presented. Although the method is generally applicable to many orthopaedic internal fixation, it was only demonstrated for the insertion of pedicle screws in vertebral bodies for spine fixation. The method relies upon obtaining an impression of the vertebral bodies. Computed Tomography scans of both

K Abdel-Malek; D P McGowan; V K Goel; D Kowalski; A Hager

1997-01-01

421

Alu insertion polymorphisms in Native Americans and related Asian populations  

Microsoft Academic Search

Background: Alu insertions provide useful markers for the study of inter-population affinities and historical processes, but data on these systems are not numerous in Native Americans and related populations. Aim: The study aimed to answer the following questions: (a) do the population relationships found agree with ethnic, historical and geographical data? and (b) what can heterozygote levels and associated results

Jaqueline Battilana; Nelson J. R. Fagundes; Ana H. Heller; Angela Goldani; Loreta B. Freitas; Eduardo Tarazona-Santos; Batmunkh Munkhbat; Namid Munkhtuvshin; Mlu Krylov; Lidia Benevolenskaia; Frank C. Arnett; Mark A. Batzer; Prescott L. Deininger; Francisco M. Salzano; Sandro L. Bonatto

2006-01-01

422

Synchronised transmembrane insertion and glycosylation of a nascent membrane protein  

Microsoft Academic Search

Studies of the synthesis and incorporation of the vesicular stomatitis virus glycoprotein into membranes in a synchronised cell-free system demonstrate a tight coupling between polypeptide synthesis and membrane insertion, as a result of which the nascent chain crosses the membrane. The studies reveal a surprisingly precise sequence by which the nascent chain of this membrane glycoprotein is glycosylated in two

James E. Rothman; Harvey F. Lodish

1977-01-01

423

Insertion of an articulated human into a networked virtual environment  

Microsoft Academic Search

Most distributed interactive simulation (DIS) technology demonstrated in recent years has focused on vehicle interaction. The dismounted infantryman-the individual soldier-has been largely ignored or represented by static models. In six weeks of development, the Naval Postgraduate School, SARCOS Inc., and University of Pennsylvania, under Army Research Laboratory sponsorship, demonstrated the insertion of a fully articulated human figure into a DIS

David R. Pratt; Paul T. Barham; John Locke; Michael J. Zyda; Bryant Eastman; Timothy Moore; Klaus Biggers; Robert Douglass; Stephen Jacobsen; Michael Hollick; John Granieri; Hyeongseok Ko; Norman I. Badler

1994-01-01

424

Characteristics of joining inserts for composite sandwich panels  

Microsoft Academic Search

Composite sandwich constructions are widely employed in various light weight structures, because composite sandwich panels have high specific stiffness and high specific bending strength compared to solid panels. Since sandwich panels are basically unsuited to carry localized loads, the sandwich structure should provide joining inserts to transfer the localized loads to other structures.In this work, the load transfer characteristics of

Byoung Jung Kim; Dai Gil Lee

2008-01-01

425

Magnetic Ripple Correction in Tandem Mirrors by Ferromagnetic Inserts.  

National Technical Information Service (NTIS)

Magnetic ripple of 1% or more caused by discrete solenoid coils in the central cells of tandem mirrors may severely affect the MHD stability. The ripple amplitude can be reduced by an order of magnitude by ferromagnetic annuli inserted within the coils at...

G. W. Hamilton

1985-01-01

426

Direct observation of poloxamer 188 insertion into lipid monolayers.  

PubMed Central

P188, a triblock copolymer of the form poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) helps seal electroporated cell membranes, arresting the leakage of intracellular materials from the damaged cells. To explore the nature of the interaction between P188 and cell membranes, we have constructed a model system that assesses the ability of P188 to insert into lipid monolayers. Using concurrent Langmuir isotherm and fluorescence microscopy measurements, we find that P188 changes the phase behavior and morphology of the monolayers. P188 inserts into both dipalmitoylphosphatidlycholine and dipalmitoylphosphatidylglycerol monolayers at surface pressures equal to and lower than approximately 22 mN/m at 30 degrees C; this pressure corresponds to the maximal surface pressure attained by P188 on a pure water subphase. Similar results for the two phospholipids indicate that P188 insertion is not influenced by headgroup electrostatics. Because the equivalent surface pressure of a normal bilayer is on the order of 30 mN/m, the lack of P188 insertion above 22 mN/m further suggests the poloxamer selectively adsorbs into damaged portions of electroporated membranes, thereby localizing its effect. P188 is also found to be "squeezed out" of the monolayers at high surface pressures, suggesting a mechanism for the cell to be rid of the poloxamer when the membrane is restored.

Maskarinec, Stacey A; Hannig, Jurgen; Lee, Raphael C; Lee, Ka Yee C

2002-01-01

427

INSERTIONAL MUTAGENESIS SYSTEMS IN THE RED FLOUR BEETLE, TRIBOLIUM CASTANEUM  

Technology Transfer Automated Retrieval System (TEKTRAN)

The genome of Tribolium castaneum has been sequenced, and a preliminary assembly released (HGSC, Baylor College of Medicine). In order to progress toward a systematic, functional analysis of the genome we have developed a highly efficient, hybridization-based method of generating new insertions usi...

428

Laser Filter Inserts for Goggles, Sun, Wind and Dust.  

National Technical Information Service (NTIS)

In order to provide eye protection for military personnel, two types of laser-protective filter elements were developed for use as inserts in the goggles. The elements were designed to be snapped into the goggle frame in back of the standard ballistic len...

H. A. Smith M. J. Manning E. M. Healy

1994-01-01

429

PLANS FOR A 2ND INSERTION DEVICE IN CAMD  

Microsoft Academic Search

To allow the possible installation of a 2 nd Insertion Device in the CAMD Light Source, the lattice optic needs to be changed. The present configuration has a small vertical beta function in the long straight section containing the 7 Tesla wiggler. The new optic will give a small vertical beta at two long straight sections which are diametrically opposite.

M Fedurin; P Jines; T Miller; Y Wang

430

Ballistic Testing for Interceptor Body Armor Inserts Needs Improvement.  

National Technical Information Service (NTIS)

We are performing a series of Interceptor Body Armor audits in response to a congressional request. This audit evaluated product quality assurance for seven Army contracts, valued at $2.5 billion, for ballistic inserts awarded between 2004 and 2006. Speci...

2011-01-01

431

Embedded Micromirror Inserts for Optical Printed Circuit Boards  

Microsoft Academic Search

We present the use of an embedded 45deg micromirror, which is patterned in polymer photoresist using deep proton writing. The micromirror is metallized and inserted in a laser ablated cavity in the optical layer and in a next step covered with cladding material. Surface roughness measurements confirm the excellent quality of the mirror facet. Loss measurements have been carried out

Nina Hendrickx; JÜrgen Van Erps; Erwin Bosman; Christof Debaes; Hugo Thienpont; Peter Van Daele

2008-01-01

432

Palliation of proximal malignant biliary obstruction by endoscopic endoprosthesis insertion  

Microsoft Academic Search

For four years up to December 1987, 190 patients (median age 73 years) with proximal malignant biliary obstruction were treated by endoscopic endoprosthesis insertion. Altogether 101 had cholangiocarcinoma, 21 gall bladder carcinoma, 20 local spread of pancreatic carcinoma, and 48 metastatic malignancy. Fifty eight patients had type I, 54 type II, and 78 type III proximal biliary strictures (Bismuth classification).

A A Polydorou; S R Cairns; J F Dowsett; A R Hatfield; P R Salmon; P B Cotton; R C Russell

1991-01-01

433

Dynamic frequency scaling with buffer insertion for mixed workloads  

Microsoft Academic Search

This paper presents a method to reduce the energy of interactive systems for mixed workloads: multimedia applications that require constant output rates and sporadic jobs that need prompt responses. The authors' method divides multimedia programs into stages and inserts data buffers between them. Data buffering has three purposes: 1) to support constant output rates; 2) to allow frequency scaling for

Yung-hsiang Lu; Luca Benini; Giovanni De Micheli

2002-01-01

434

Inadvertent Insertion of Nasogastric Tube into the Brain  

Microsoft Academic Search

An unusual case of a nasogastric (Ryle's) tube inserted in the brain in a patient having severe head injury with skull base fractures is reported here along with a brief review of literature. A 35 years male was referred from a peripheral institute following head trauma with endotracheal tube and nasogastric tube in situ. A CT scan of the brain

AK Pandey; AK Sharma; BD Diyora; M Radhakrishnan

435

Scheduling with Inserted Idle Time: Problem Taxonomy and Literature Review  

Microsoft Academic Search

In the context of production scheduling, inserted idle time (IIT) occurs whenever a resource is deliberately kept idle in the face of waiting jobs. IIT schedules are particularly relevant in multimachine industrial situations where earliness costs and=or dynamically arriving jobs with due dates come into play. We provide a taxonomy of environments in which IIT scheduling is relevant, review the

John J. Kanet; V. Sridharan

2000-01-01

436

Bladder stone due to accidentally intravesically inserted intrauterine device.  

PubMed

Bladder stones occasionally develop due to foreign body in the bladder. A 45-year-old woman, who had had an intrauterine contraceptive device inserted 10 years earlier, presented with dysuria, frequency and voiding difficulty with suprapubic pain. The intrauterine device was found in the bladder with stone formation and was removed by cystoscopic procedure. PMID:22573181

Chae, Ji Yun; Kim, Jin Wook; Yoon, Cheol Yong; Park, Hong Seok; Moon, Du Geon; Oh, Mi Mi

2012-05-10

437

Dynamics of insertion sequence elements during experimental evolution of bacteria  

Microsoft Academic Search

We review the intersection between two areas of microbial evolution that were research foci of Michel Blot. One focus is the behavior of insertion sequence (IS) elements, including their role in promoting the evolutionary adaptation of their hosts. The other focus is experimental evolution, an approach that allows the dynamics of genomic and phenotypic change to be observed in the

Dominique Schneider; Richard E. Lenski

2004-01-01

438

Molecular archeology of L1 insertions in the human genome  

Microsoft Academic Search

BACKGROUND: As the rough draft of the human genome sequence nears a finished product and other genome-sequencing projects accumulate sequence data exponentially, bioinformatics is emerging as an important tool for studies of transposon biology. In particular, L1 elements exhibit a variety of sequence structures after insertion into the human genome that are amenable to computational analysis. We carried out a

Suzanne T Szak; Oxana K Pickeral; Wojciech Makalowski; Mark S Boguski; David Landsman; Jef D Boeke

2002-01-01

439

Insertion loss prediction of floating floors used in ship cabins  

Microsoft Academic Search

In this paper, vibration reduction in ship cabins by using floating floor is studied. Two theoretical models are developed and predicted insertion losses of floating floors are compared to experimental results, where measurements are done in the mock-up built for simulating typical ship cabins. The floating floor consists of upper board and mineral wool, which is in turn laid on

Sun-Il Cha; Ho-Hwan Chun

2008-01-01

440

Insert earphone modeling and measurement by IEC-60711 coupler.  

PubMed

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

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

2011-02-01

441

Discovering Sequence Motifs with Arbitrary Insertions and Deletions  

Microsoft Academic Search

Biology is encoded in molecular sequences: deciphering this encoding remains a grand scientific challenge. Functional regions of DNA, RNA, and protein sequences often exhibit characteristic but subtle motifs; thus, computational discovery of motifs in sequences is a fundamental and much-studied problem. However, most current algorithms do not allow for insertions or deletions (indels) within motifs, and the few that do

Martin C. Frith; Neil F. W. Saunders; Bostjan Kobe; Timothy L. Bailey

2008-01-01

442

Low-cost waveguide filters with printed-circuit inserts  

Microsoft Academic Search

An easy to manufacture waveguide filter is presented. This type of filter is based on printed-circuit-board inserts with resonator strips. The electromagnetic properties of the strips interacting with the electromagnetic field inside the waveguide are described by equivalent circuit elements. Filter synthesis and computer optimization is applied to design band-stop filters. Finally simulated results are compared with measurements

H. Schmiedel; M. Muller; A. Anufriev

1996-01-01

443

Pulmonary valve insertion late after repair of Fallot's tetralogy  

Microsoft Academic Search

Objectives: To analyze the results of pulmonary valve insertion late after initial repair of Fallot's tetralogy. Pulmonary insufficiency (PI) after correction of Fallot's tetralogy is usually well tolerated in the short term, but is associated with symptomatic right ventricular dilatation and an increased risk of ventricular arrhythmias over longer periods of time. Methods: From 1993 to July 2000, 51 patients

M. G. Hazekamp; M. M. J. Kurvers; P. H. Schoof; H. W. Vliegen; B. J. M. Mulder; A. A. W. Roest; J. Ottenkamp; R. A. E. Dion

2001-01-01

444

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

NASA Astrophysics Data System (ADS)

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 ?m2 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.

Contardo, L.; de Luca, M.; Biasotto, M.; Longo, R.; Olivo, A.; Pani, S.; di Lenarda, R.

2005-08-01

445

Germline Transgenesis and Insertional Mutagenesis in Schistosoma mansoni Mediated by Murine Leukemia Virus  

PubMed Central

Functional studies will facilitate characterization of role and essentiality of newly available genome sequences of the human schistosomes, Schistosoma mansoni, S. japonicum and S. haematobium. To develop transgenesis as a functional approach for these pathogens, we previously demonstrated that pseudotyped murine leukemia virus (MLV) can transduce schistosomes leading to chromosomal integration of reporter transgenes and short hairpin RNA cassettes. Here we investigated vertical transmission of transgenes through the developmental cycle of S. mansoni after introducing transgenes into eggs. Although MLV infection of schistosome eggs from mouse livers was efficient in terms of snail infectivity, >10-fold higher transgene copy numbers were detected in cercariae derived from in vitro laid eggs (IVLE). After infecting snails with miracidia from eggs transduced by MLV, sequencing of genomic DNA from cercariae released from the snails also revealed the presence of transgenes, demonstrating that transgenes had been transmitted through the asexual developmental cycle, and thereby confirming germline transgenesis. High-throughput sequencing of genomic DNA from schistosome populations exposed to MLV mapped widespread and random insertion of transgenes throughout the genome, along each of the autosomes and sex chromosomes, validating the utility of this approach for insertional mutagenesis. In addition, the germline-transmitted transgene encoding neomycin phosphotransferase rescued cultured schistosomules from toxicity of the antibiotic G418, and PCR analysis of eggs resulting from sexual reproduction of the transgenic worms in mice confirmed that retroviral transgenes were transmitted to the next (F1) generation. These findings provide the first description of wide-scale, random insertional mutagenesis of chromosomes and of germline transmission of a transgene in schistosomes. Transgenic lines of schistosomes expressing antibiotic resistance could advance functional genomics for these significant human pathogens. Database accession Sequence data from this study have been submitted to the European Nucleotide Archive (http://www.ebi.ac.uk/embl) under accession number ERP000379.

Suttiprapa, Sutas; Kines, Kristine J.; Tort, Jose F.; Mann, Victoria H.; Turner, Daniel J.; Berriman, Matthew; Brindley, Paul J.

2012-01-01

446

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

PubMed Central

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.

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

2010-01-01

447

Effectiveness of various drugs in controlling bleeding after IUD insertion.  

PubMed

The National Research Institute of Family Planning of Pakistan in April 1967 initiated a study to determine what drugs and other variables might aid in reducing IUD postinsertion bleeding. 768 women were admitted to the study after the insertion of a polyethylene plastic Lippes loop. Each woman was given 1 of the following medications (double-blind test) in a packet: 1) Vitamin-K (10 mg tid for 21 days), 2) Vitamin-C (50 mg tid for 21 days), 3) ferrous sulfate (.3 mg tid for 21 days), 4) Lyndiol (2.5 mg daily for 21 days), and 5) placebo (glucose-filled gelatin capsules, 1 daily for 21 days). 623 (81%) of the patients returned for the first-month follow-up. Second- and third-month follow-ups had far fewer returns; data analysis was thus restricted to the first month only. 4% of the loops were removed for bleeding. 91.5% reported some degree of postinsertion bleeding. Bleeding was reported as minimal or spotting and slight bleeding (less than normal menstrual flow), normal menstrual flow, or heavy bleeding and clots (more than normal menstrual flow). More of the young clients (ages 25-29) returned than expected (249 vs. 222.3); fewer of the older clients (ages 40-44) returned than expected (31 vs. 52.7). Ferrous sulfate was most effective in shifting the control pattern of bleeding to a pattern skewed to lesser degrees of bleeding. The least bleeding was reported for insertion 1-3 months postpartum. With regard to the time since the onset of the last menstrual period, insertion during lactation amenorrhea gave the best results, and insertion prior to Day 13 was much better than insertion during the last half of the cycle. PMID:12275143

Ali, R K; Satterthwaite, A P

1968-01-01

448

VME insertion device control at the Advanced Photon Source  

NASA Astrophysics Data System (ADS)

The Advanced Photon Source (APS) currently has 29 insertion devices (IDs) installed and operating. The need to remotely diagnose and correct problems has become increasingly important. This has been accomplished through the development of a new control system with greatly enhanced input/output (I/O) capabilities specifically targeted to this control task. The system features a custom VME control card and three rack-mounted interface chassis for ID control, encoder interface, and motor drive shutdown. The card provides device interlocks, limit switch logic, motor axis selection, digital I/O, and status feedback. This VME insertion device control was designed to operate with an eight-axis intelligent motor controller and a stepper-motor drive that accepts step and direction inputs. The front panel of the card has two connectors for all of the control signals for the stepper-motor drives. There is a third connector for the ID limit switch inputs and the emergency stop circuit, and a fourth connector provides 23 bits of digital outputs and 16 bits of digital inputs. Light-emitting diodes indicate which motions are inhibited by the limit switch logic. An experimental physics industrial control system (EPICS) (http://www.APS.ANL-GOV/EPICS) device driver was developed to access all the registers on the VME control card. Using standard EPICS records, the insertion device status can be viewed remotely. This minimizes downtime for APS ID beamline users by allowing faster resolution of any problems preventing a user from operating the insertion device. This new insertion device control has been in use at the APS since July of 1999. The design features of the control system and rationale for them will be presented, along with our experience in building, testing, installing, and operating the control system.

Smith, M.; Ramanathan, M.; Grimmer, J.; Merritt, M.

2002-03-01

449