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Arabidopsis T-DNA insertional lines for CDC25 are hypersensitive to hydroxyurea but not to zeocin or salt stress
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. PMID:20647223
Spadafora, Natasha D.; Doonan, John H.; Herbert, Robert J.; Bitonti, M. Beatrice; Wallace, Emily; Rogers, Hilary J.; Francis, Dennis
Implementing reverse genetics in Rosaceae: analysis of T-DNA flanking sequences of insertional mutant lines in the diploid strawberry, Fragaria vesca.
Reverse genetics is used for functional genomics research in model plants. To establish a model system for the systematic reverse genetics research in the Rosaceae family, we analyzed genomic DNA flanking the T-DNA insertions in 191 transgenic plants of the diploid strawberry, Fragaria vesca. One hundred and seventy-six T-DNA flanking sequences were amplified from the right border (RB) and 37 from the left border (LB) by thermal asymmetric interlaced PCR. Analysis of the T-DNA nick positions revealed that T-DNA was most frequently nicked at the cleavage sites. Analysis of 11 T-DNA integration sites indicated that T-DNA was integrated into the F. vesca genome by illegitimate recombination, as reported in other model plants: Arabidopsis, rice and tobacco. First, deletion of DNA was found at T-DNA integration target sites in all transgenic plants tested. Second, microsimilarities of a few base pairs between the left and/or right ends of the T-DNA and genomic sites were found in all transgenic plants tested. Finally, filler DNA was identified in four break-points. Out of 191 transgenic plants, T-DNA flanking sequences of 79 plants (41%) showed significant similarity to genes, elements or proteins of other plant species and 67 (35%) of the sequences are still unknown strawberry gene fragments. T-DNA flanking sequences of 126 plants (66%) showed homology to plant ESTs. This is the first report of T-DNA integration in a sizeable population of a rosaceous species. We have shown in this paper that T-DNA integration in strawberry is not random but directed by sequence microsimilarities in the host genome. PMID:20444194
Oosumi, Teruko; Ruiz-Rojas, Juan Jairo; Veilleux, Richard E; Dickerman, Allan; Shulaev, Vladimir
SNP discovery and genetic mapping of T-DNA insertional mutants in Fragaria vesca L
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
SNP discovery and genetic mapping of T-DNA insertional mutants in Fragaria vesca L.
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 sites of a population of 108 unique single copy mutants, we utilized thermal asymmetric interlaced PCR (hiTAIL-PCR) to amplify the flanking region surrounding either the left or right border of the T-DNA. Bioinformatics analysis of flanking sequences revealed little preference for insertion site with regard to G/C content; left borders tended to retain more of the plasmid backbone than right borders. Primers were developed from F. vesca flanking sequences to attempt to amplify products from both parents of the reference F. vesca 815 x F. bucharica 601 mapping population. Polymorphism occurred as: presence/absence of an amplification product for 16 primer pairs and different size products for 12 primer pairs, For 46 mutants, where polymorphism was not found by PCR, the amplification products were sequenced to reveal SNP polymorphism. A cleaved amplified polymorphic sequence/derived cleaved amplified polymorphism sequence (CAPS/dCAPS) strategy was then applied to find restriction endonuclease recognition sites in one of the parental lines to map the SNP position of 74 of the T-DNA insertion lines. BLAST search of flanking regions against GenBank revealed that 46 of 108 flanking sequences were close to presumed strawberry genes related to annotated genes from other plants. PMID:20349033
Ruiz-Rojas, J J; Sargent, D J; Shulaev, V; Dickerman, A W; Pattison, J; Holt, S H; Ciordia, A; Veilleux, Richard E
GABI-Kat SimpleSearch: an Arabidopsis thaliana T-DNA mutant database with detailed information for confirmed insertions.
Insertional mutagenesis approaches, especially by T-DNA, play important roles in gene function studies of the model plant Arabidopsis thaliana. GABI-Kat SimpleSearch (http://www.GABI-Kat.de) is a Flanking Sequence Tag (FST)-based database for T-DNA insertion mutants generated by the GABI-Kat project. Currently, the database contains >108,000 mapped FSTs from approximately 64,000 lines which cover 64% of all annotated A.thaliana protein-coding genes. The web interface allows searching for relevant insertions by gene code, keyword, line identifier, GenBank accession number of the FST, and also by BLAST. A graphic display of the genome region around the gene or the FST assists users to select insertion lines of their interests. About 3500 insertions were confirmed in the offspring of the plant from which the original FST was generated, and the seeds of these lines are available from the Nottingham Arabidopsis Stock Centre. The database now also contains additional information such as segregation data, gene-specific primers and confirmation sequences. This information not only helps users to evaluate the usefulness of the mutant lines, but also covers a big part of the molecular characterization of the insertion alleles. PMID:17062622
Li, Yong; Rosso, Mario G; Viehoever, Prisca; Weisshaar, Bernd
Bidirectional promoter trapping T-DNA for insertional mutagenesis in Verticillium dahliae.
Transfer DNA (T-DNA)-based random insertional mutagenesis is a universal forward genetic approach for gene identification and cloning in many phytopathogenic fungi. In a large number of randomly selected transformants, screening for mutants with a specific phenotype is laborious, especially for pathogenicity-defective mutants. To accelerate mutant screening and gene identification, a bidirectional promoter-trapping Ti binary vector, 1300-bisGFP-hyg, was constructed and deployed in this study. More than 6000 Verticillium dahliae transformants were obtained by the mediation of Agrobacterium tumefaciens carrying the vector. One thousand randomly selected transformants were cultured on Czapek-Dox and on Czapek-Dox plus cotton root extract media plates. The cultured transformants with green fluorescent protein (GFP) expression or changes in phenotype were selected and used in virulence or promoter-trapping assays. Based on the virulence assay of 60 transformants, the pathogenicity of 17 of these mutants was compromised. Ten pathogenicity-defective mutants were found with GFP expression, and 6 with expression in Czapek-Dox plus cotton root extract media specifically. Using TAIL-PCR (thermal asymmetric interlaced polymerase chain reaction), the T-DNA insertion sites were identified in 8 GFP-expressing transformants, including 5 pathogenicity-defective mutants and 3 unaffected transformants. Promoters of 6 genes were successfully trapped using the T-DNA method in this study. The nonpathogenic transformant 24C9 was the subject of additional investigation. It displayed strong GFP expression on water agar medium supplemented with cotton root extracts and on cotton seedling stems. The results obtained by Southern blot and quantitative real-time PCR confirmed that the transcription level of VdUGPU (encoding UTP-glucose-1-phosphate uridylyltransferase) was significantly reduced owing to T-DNA insertion in the gene promoter region. These results indicate that the bidirectional promoter-trapping Ti vector, combined with induction medium that contains root exudates, can be useful for identification of pathogenicity-related and functional genes in V. dahliae. PMID:24993736
Deng, Sheng; Wang, Cai-yue; Zhang, Xin; Lin, Ling
Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition.
The in vitro shoot culture of a T-DNA insertional mutant of Papaver somniferum L. established by the infection of Agrobacterium rhizogenes MAFF03-01724 accumulated thebaine instead of morphine as a major opium alkaloid. To develop a non-narcotic opium poppy and to gain insight into its genetic background, we have transplanted this mutant to soil, and analyzed its alkaloid content along with the manner of inheritance of T-DNA insertion loci among its selfed progenies. In the transplanted T0 primary mutant, the opium (latex) was found to be rich in thebaine (16.3% of dried opium) by HPLC analysis. The analyses on T-DNA insertion loci by inverse PCR, adaptor-ligation PCR, and quantitative real-time PCR revealed that as many as 18 copies of T-DNAs were integrated into a poppy genome in a highly complicated manner. The number of copies of T-DNAs was decreased to seven in the selected T3 progenies, in which the average thebaine content was 2.4-fold that of the wild type plant. This may indicate that the high thebaine phenotype was increasingly stabilized as the number of T-DNA copies was decreased. In addition, by reverse transcription PCR analysis on selected morphine biosynthetic genes, the expression of codeine 6-O-demethylase was clearly shown to be diminished in the T0 in vitro shoot culture, which can be considered as one of the key factors of altered alkaloid composition. PMID:24288085
Kawano, Noriaki; Kiuchi, Fumiyuki; Kawahara, Nobuo; Yoshimatsu, Kayo
The ben1-1 brassinosteroid-catabolism mutation is unstable due to epigenetic modifications of the intronic T-DNA insertion.
Loss-of-function genetic analysis plays a pivotal role in elucidating individual gene function as well as interactions among gene networks. The ease of gene tagging and cloning provided by transfer DNA (T-DNA) insertion mutants have led to their heavy use by the Arabidopsis research community. However, certain aspects of T-DNA alleles require caution, as highlighted in this study of an intronic insertion mutant (ben1-1) in the BEN1 (BRI1-5 ENHANCED 1) gene. As a part of our analysis of brassinosteroid catabolic enzymes, we generated a genetic triple-mutant from a cross between the bas1-2 sob7-1 double-null (T-DNA exonic insertion mutants of phyB-4 ACTIVATION TAGGED SUPPRESSOR 1 and SUPPRESSOR OF phyB-4 7) and ben1-1. As previously described, the single ben1-1 line behaves as a transcript null. However, in the triple-mutant background ben1-1 was reverted to a partial loss-of-function allele showing enhanced levels of the wild-type-spliced transcript. Interestingly, the enhanced expression of BEN1 remained stable when the ben1-1 single-mutant was reisolated from a cross with the wild type. In addition, the two genetically identical pretriple and posttriple ben1-1 mutants also differed phenotypically. The previously functional NPTII (NEOMYCIN PHOSPHOTRANSFERASE II) T-DNA marker gene (which encodes kanamycin resistance) was no longer functional in the recovered ben1-1 allele, though the length of the T-DNA insertion and the NPTII gene sequence did not change in the pretriple and posttriple ben1-1 mutants. Methylation analysis using both restriction endonuclease activity and bisulfite conversion followed by sequencing showed that the methylation status of the T-DNA is different between the original and the recovered ben1-1. These observations demonstrate that the recovered ben1-1 mutant is epigenetically different from the original ben1-1 allele. PMID:23893742
Sandhu, Kulbir Singh; Koirala, Pushpa Sharma; Neff, Michael M
Wide-scale screening of T-DNA lines for transcription factor genes affecting male gametophyte development in Arabidopsis.
Male gametophyte development leading to the formation of a mature pollen grain is precisely controlled at various levels, including transcriptional, post-transcriptional and post-translational, during its whole progression. Transcriptomic studies exploiting genome-wide microarray technologies revealed the uniqueness of pollen transcriptome and the dynamics of early and late successive global gene expression programs. However, the knowledge of transcription regulation is still very limited. In this study, we focused on the identification of pollen-expressed transcription factor (TF) genes involved in the regulation of male gametophyte development. To achieve this, the reverse genetic approach was used. Seventy-four T-DNA insertion lines were screened, representing 49 genes of 21 TF families active in either early or late pollen development. In the screen, ten phenotype categories were distinguished, affecting various structural or functional aspects, including pollen abortion, presence of inclusions, variable pollen grain size, disrupted cell wall structure, cell cycle defects, and male germ unit organization. Thirteen lines were not confirmed to contain the T-DNA insertion. Among 61 confirmed lines, about half (29 lines) showed strong phenotypic changes (i.e., ? 25% aberrant pollen) including four lines that produced a remarkably high proportion (70-100%) of disturbed pollen. However, the remaining 32 lines exhibited mild defects or resembled wild-type appearance. There was no significant bias toward any phenotype category among early and late TF genes, nor, interestingly, within individual TF families. Presented results have a potential to serve as a basal information resource for future research on the importance of respective TFs in male gametophyte development. PMID:22101548
Re?�k, David; Dupl'�kov�, Nikoleta; Honys, David
Changes in T-DNA methylation and expression are associated with phenotypic variation and plant regeneration in a crown gall tumor line
Phenotypic variation of an octopine-type crown gall tumor line resulting from changes in the pattern of T-DNA methylation and expression is described. Variants that grow as unorganized callus always express T-DNA transcripts 1 and 2. In shoot-forming variants (teratomas) only T-DNA transcript 4 is expressed. This line also regenerates normal-appearing, rooted plants in which all T-DNA expression is suppressed. Tissues
Richard M. Amasino; Ann L. T. Powell; Milton P. Gordon
Deep sequencing of the ancestral tobacco species Nicotiana tomentosiformis reveals multiple T-DNA inserts and a complex evolutionary history of natural transformation in the genus Nicotiana.
Nicotiana species carry cellular T-DNA sequences (cT-DNAs), acquired by Agrobacterium-mediated transformation. We characterized the cT-DNA sequences of the ancestral Nicotiana tabacum species Nicotiana tomentosiformis by deep sequencing. N. tomentosiformis contains four cT-DNA inserts derived from different Agrobacterium strains. Each has an incomplete inverted-repeat structure. TA is similar to part of the Agrobacterium rhizogenes 1724 mikimopine-type T-DNA, but has unusual orf14 and mis genes. TB carries a 1724 mikimopine-type orf14-mis fragment and a mannopine-agropine synthesis region (mas2-mas1-ags). The mas2' gene codes for an active enzyme. TC is similar to the left part of the A. rhizogenes A4 T-DNA, but also carries octopine synthase-like (ocl) and c-like genes normally found in A. tumefaciens. TD shows a complex rearrangement of T-DNA fragments similar to the right end of the A4 TL-DNA, and including an orf14-like gene and a gene with unknown function, orf511. The TA, TB, TC and TD insertion sites were identified by alignment with N. tabacum and Nicotiana sylvestris sequences. The divergence values for the TA, TB, TC and TD repeats provide an estimate for their relative introduction times. A large deletion has occurred in the central part of the N. tabacum cv. Basma/Xanthi TA region, and another deletion removed the complete TC region in N. tabacum. Nicotiana otophora lacks TA, TB and TD, but contains TC and another cT-DNA, TE. This analysis, together with that of Nicotiana glauca and other Nicotiana species, indicates multiple sequential insertions of cT-DNAs during the evolution of the genus Nicotiana. PMID:25219519
Chen, Ke; Dorlhac de Borne, Fran�ois; Szegedi, Ern�; Otten, L�on
www.mdpi.com/journal/pharmaceuticals Genetic and Phenotypic Analyses of a Papaver somniferum T-DNA Insertional Mutant with Altered Alkaloid Composition
Abstract: The in vitro shoot culture of a T-DNA insertional mutant of Papaver somniferum L. established by the infection of Agrobacterium rhizogenes MAFF03-01724 accumulated thebaine instead of morphine as a major opium alkaloid. To develop a non-narcotic opium poppy and to gain insight into its genetic background, we have transplanted this mutant to soil, and analyzed its alkaloid content along with the manner of inheritance of T-DNA insertion loci among its selfed progenies. In the transplanted T0 primary mutant, the opium (latex) was found to be rich in thebaine (16.3 % of dried opium) by HPLC analysis. The analyses on T-DNA insertion loci by inverse PCR, adaptor-ligation PCR, and quantitative real-time PCR revealed that as many as 18 copies of T-DNAs were integrated into a poppy genome in a highly complicated manner. The number of copies of T-DNAs was decreased to seven in the selected T3 progenies, in which the average thebaine content was 2.4-fold that of the wild type plant. This may indicate that the high thebaine phenotype was increasingly stabilized as the number of T-DNA copies was decreased. In addition, by reverse transcription PCR analysis on selected morphine biosynthetic genes, the expression of codeine 6-O-demethylase was clearly shown to be diminished in the T0 in vitro shoot
Noriaki Kawano; Fumiyuki Kiuchi; Nobuo Kawahara; Kayo Yoshimatsu
Oryza Tag Line, a phenotypic mutant database for the G�noplante rice insertion line library
To organize data resulting from the phenotypic characterization of a library of 30 000 T-DNA enhancer trap (ET) insertion lines of rice (Oryza sativa L cv. Nipponbare), we developed the Oryza Tag Line (OTL) database (http://urgi.versailles.inra.fr/OryzaTagLine/). OTL structure facilitates forward genetic search for specific phenotypes, putatively resulting from gene disruption, and/or for GUSA or GFP reporter gene expression patterns, reflecting ET-mediated endogenous gene detection. In the latest version, OTL gathers the detailed morpho-physiological alterations observed during field evaluation and specific screens in a first set of 13 928 lines. Detection of GUS or GFP activity in specific organ/tissues in a subset of the library is also provided. Search in OTL can be achieved through trait ontology category, organ and/or developmental stage, keywords, expression of reporter gene in specific organ/tissue as well as line identification number. OTL now contains the description of 9721 mutant phenotypic traits observed in 2636 lines and 1234 GUS or GFP expression patterns. Each insertion line is documented through a generic passport data including production records, seed stocks and FST information. 8004 and 6101 of the 13 928 lines are characterized by at least one T-DNA and one Tos17 FST, respectively that OTL links to the rice genome browser OryGenesDB. PMID:17947330
Larmande, Pierre; Gay, C�line; Lorieux, Mathias; P�rin, Christophe; Bouniol, Matthieu; Droc, Ga�tan; Sallaud, Christophe; Perez, Pascual; Barnola, Isabelle; Biderre-Petit, Corinne; Martin, J�r�me; Morel, Jean Beno�t; Johnson, Alexander A. T.; Bourgis, Fabienne; Ghesqui�re, Alain; Ruiz, Manuel; Courtois, Brigitte; Guiderdoni, Emmanuel
Genetics, structure, and prevalence of FP967 (CDC Triffid) T-DNA in flax.
The detection of T-DNA from a genetically modified flaxseed line (FP967, formally CDC Triffid) in a shipment of Canadian flaxseed exported to Europe resulted in a large decrease in the amount of flax planted in Canada. The Canadian flaxseed industry undertook major changes to ensure the removal of FP967 from the supply chain. This study aimed to resolve the genetics and structure of the FP967 transfer DNA (T-DNA). The FP967 T-DNA is thought to be inserted in at single genomic locus. The junction between the T-DNA and genomic DNA consisted of two inverted Right Borders with no Left Border (LB) flanking genomic DNA sequences recovered. This information was used to develop an event-specific quantitative PCR (qPCR) assay. This assay and an existing assay specific to the T-DNA construct were used to determine the genetics and prevalence of the FP967 T-DNA. These data supported the hypothesis that the T-DNA is present at a single location in the genome. The FP967 T-DNA is present at a low level (between 0.01 and 0.1%) in breeder seed lots from 2009 and 2010. None of the 11,000 and 16,000 lines selected for advancement through the Flax Breeding Program in 2010 and 2011, respectively, tested positive for the FP967 T-DNA, however. Most of the FP967 T-DNA sequence was resolved via PCR cloning and next generation sequencing. A 3,720�bp duplication of an internal portion of the T-DNA (including a Right Border) was discovered between the flanking genomic DNA and the LB. An event-specific assay, SAT2-LB, was developed for the junction between this repeat and the LB. PMID:25883881
Young, Lester; Hammerlindl, Joseph; Babic, Vivijan; McLeod, Jamille; Sharpe, Andrew; Matsalla, Chad; Bekkaoui, Faouzi; Marquess, Leigh; Booker, Helen M
Complementation of the embryo-lethal T-DNA insertion mutant of AUXIN-BINDING-PROTEIN 1 (ABP1) with abp1 point mutated versions reveals crosstalk of ABP1 and phytochromes
The function of the extracytoplasmic AUXIN-BINDING-PROTEIN1 (ABP1) is largely enigmatic. We complemented a homozygous T-DNA insertion null mutant of ABP1 in Arabidopsis thaliana Wassilewskia with three mutated and one wild-type (wt) ABP1 cDNA, all tagged C-terminally with a strepII�FLAG tag upstream the KDEL signal. Based on in silico modelling, the abp1 mutants were predicted to have altered geometries of the auxin binding pocket and calculated auxin binding energies lower than the wt. Phenotypes linked to auxin transport were compromised in these three complemented abp1 mutants. Red light effects, such as elongation of hypocotyls in constant red (R) and far-red (FR) light, in white light supplemented by FR light simulating shade, and inhibition of gravitropism by R or FR, were all compromised in the complemented lines. Using auxin- or light-induced expression of marker genes, we showed that auxin-induced expression was delayed already after 10min, and light-induced expression within 60min, even though TIR1/AFB or phyB are thought to act as receptors relevant for gene expression regulation. The expression of marker genes in seedlings responding to both auxin and shade showed that for both stimuli regulation of marker gene expression was altered after 10�20min in the wild type and phyB mutant. The rapidity of expression responses provides a framework for the mechanics of functional interaction of ABP1 and phyB to trigger interwoven signalling pathways. PMID:25392478
Effendi, Yunus; Ferro, Noel; Labusch, Corinna; Geisler, Markus; Scherer, G�nther F. E.
Complementation of the embryo-lethal T-DNA insertion mutant of AUXIN-BINDING-PROTEIN 1 (ABP1) with abp1 point mutated versions reveals crosstalk of ABP1 and phytochromes.
The function of the extracytoplasmic AUXIN-BINDING-PROTEIN1 (ABP1) is largely enigmatic. We complemented a homozygous T-DNA insertion null mutant of ABP1 in Arabidopsis thaliana Wassilewskia with three mutated and one wild-type (wt) ABP1 cDNA, all tagged C-terminally with a strepII-FLAG tag upstream the KDEL signal. Based on in silico modelling, the abp1 mutants were predicted to have altered geometries of the auxin binding pocket and calculated auxin binding energies lower than the wt. Phenotypes linked to auxin transport were compromised in these three complemented abp1 mutants. Red light effects, such as elongation of hypocotyls in constant red (R) and far-red (FR) light, in white light supplemented by FR light simulating shade, and inhibition of gravitropism by R or FR, were all compromised in the complemented lines. Using auxin- or light-induced expression of marker genes, we showed that auxin-induced expression was delayed already after 10 min, and light-induced expression within 60 min, even though TIR1/AFB or phyB are thought to act as receptors relevant for gene expression regulation. The expression of marker genes in seedlings responding to both auxin and shade showed that for both stimuli regulation of marker gene expression was altered after 10-20 min in the wild type and phyB mutant. The rapidity of expression responses provides a framework for the mechanics of functional interaction of ABP1 and phyB to trigger interwoven signalling pathways. PMID:25392478
Effendi, Yunus; Ferro, Noel; Labusch, Corinna; Geisler, Markus; Scherer, G�nther F E
Frequent spontaneous deletions of Ri T-DNA in Agrobacterium rhizogenes transformed potato roots and regenerated plants.
The presence of T-DNA was examined by Southern blot analysis in 16 regenerated shoot lines derived from 6 Agrobacterium rhizogenes-transformed root clones of Solanum tuberosum L. cv. Bintje. TR-DNA, present in regenerated shoot lines from 3 out of 6 root clones was correlated with the presence of opines. One root clone produced opines up to 2.5 years of subculture. However, plant regeneration from and prolonged subculturing of this root clone resulted in loss of opine synthesis, caused by deletion of TR-DNA. TL-DNA inserted at 1 to 5 independent loci was found in 14 of the 16 shoot lines. Surprisingly, 1 to 2 additional insertions next to similar insertions of TL-DNA were found in shoot lines from the same root clone (named 'sister' shoot lines) in 2 out of 4 root clones. Nevertheless, this did not result in gross phenotypic variation between sister shoot lines. Another root clone regenerated 1 shoot line with an Ri phenotype, containing 1 insertion of TL-DNA, and 2 shoot lines with a normal Bintje phenotype without TL-DNA. The 5th root clone showed no difference between sister shoot lines and the 6th root clone produced only 1 shoot line. We conclude that during prolonged root culture and during shoot regeneration from root clones deletion of TL- and TR-DNA insertions can occur. The significance of the frequency of deletion of T-DNA of the Ri plasmid is discussed. PMID:1966386
H�nisch ten Cate, C H; Loonen, A E; Ottaviani, M P; Ennik, L; van Eldik, G; Stiekema, W J
Peripherally inserted central line catheter infections in burn patients.
Peripherally inserted central line catheter (PICC) use has increased in burn patients. While the risks of central line associated blood stream infections (CLABSI) in burn patients have been well studied, the risks of PICC infections have yet to be fully elucidated. A retrospective chart review was performed of all burn patients with a PICC admitted to our regional burn center from 2006 to 2008. Information collected included: age, total body surface area (TBSA) burn injury, length of hospital stay, duration of PICC line insertion, hospital day of PICC line insertion, and development of infections. All mean values are presented as the mean � standard error. A total of 56 patients received a PICC during this time period. Eight patients developed PICC infections with an overall rate of 11.7/1000 catheter days. Burn patient CLABSI for the same time period was 12.9/1000 catheter days. Duration of PICC did not differ between infected and noninfected patients (13.3 � 6 vs. 12.1 � 9.6 days). Patients with PICC infections had a higher percentage of TBSA (42.7 � 3.2 vs. 20.6 � 2.4*), longer length of admission (72.4 � 6 vs. 35 � 3.2*), and later hospital day of PICC insertion (48 � 4.6 vs. 17.1 � 2.5*). On linear regression analysis, PICC infection was significantly associated with TBSA (B = 22.1*), length of admission (B = 37.4*), and hospital day of PICC insertion (B = 30.3*). On multivariate analysis, TBSA (Odds ratio [OR] = 1.05*), length of admission (OR = 1.04*), and hospital day of PICC insertion (OR = 1.05*) all independently increased the risk of developing a PICC infection. Patients with severe burn injury, long hospital admissions, and later hospital day of PICC insertions are at higher risk of developing PICC infections. This data suggests that protocols should be developed about routine PICC changes in this high-risk burn population. PMID:25055005
Barsun, Alura; Sen, Soman; Palmieri, Tina L; Greenhalgh, David G
The impact of central line insertion bundle on central line-associated bloodstream infection
Background Knowledge about the impact of each central line insertion bundle on central line-associated bloodstream infection (CLABSI) is limited. Methods A quality-improvement intervention, including education, central venous catheter (CVC) insertion bundle, process and outcome surveillance, have been introduced since March 2013. Outcome surveillances, including CLABSI per 1,000 catheter-days, CLABSI per 1,000 inpatient-days, and catheter utilization rates (days of catheter use divided by total inpatient-days), were measured. As a baseline measurement for a comparison, we retrospectively collected data from March 1, 2012 to December 31, 2012. Results During this 10-month period, there were a total of 687 CVC insertions, and 627 (91.2%) insertions were performed by intensivists. The rate of CLABSI significantly declined from 1.65 per 1000 catheter-day during the pre-intervention period to 0.65 per 1000 catheter-day post-intervention period (P?=?0.039). CLABSI more likely developed in subjects in which a maximal sterile barrier was not used compared with subjects in which it was used (P?=?0.03). Moreover, CVC inserted by non-intensivists were more likely to become infected than CVC inserted by intensivists (P?=?0.010). Conclusions This multidisciplinary infection control intervention, including a central line insertion care bundle, can effectively reduce the rate of CLABSI. The impact of different care bundle varies, and a maximal sterile barrier precaution during catheter insertion is an essential component of the care line insertion bundle. PMID:24985729
Prepackaged central line kits reduce procedural mistakes during central line insertion: a randomized controlled prospective trial
Background Central line catheter insertion is a complex procedure with a high cognitive load for novices. Providing a prepackaged all-inclusive kit is a simple measure that may reduce the cognitive load. We assessed whether the use of prepackaged all-inclusive central line insertion kits reduces procedural mistakes during central line catheter insertion by novices. Methods Thirty final year medical students and recently qualified physicians were randomized into two equal groups. One group used a prepackaged all-inclusive kit and the other used a standard kit containing only the central vein catheter and all other separately packaged components provided in a materials cart. The procedure was videotaped and analyzed by two blinded raters using a checklist. Both groups performed central line catheter insertion on a manikin, assisted by nursing students. Results The prepackaged kit group outperformed the standard kit group in four of the five quality indicators: procedure duration (26:26?�?3:50 min vs. 31:27?�?5:57 min, p?=?.01); major technical mistakes (3.1?�?1.4 vs. 4.8?�?2.6, p?=?.03); minor technical mistakes (5.2?�?1.7 vs. 8.0?�?3.2, p?=?.01); and correct steps (83?�?5% vs. 75?�?11%, p?=?.02). The difference for breaches of aseptic technique (1.2?�?0.8 vs. 3.0?�?3.6, p?=?.06) was not statistically significant. Conclusions Prepackaged all-inclusive kits for novices improved the procedure quality and saved staff time resources in a controlled simulation environment. Future studies are needed to address whether central line kits also improve patient safety in hospital settings. PMID:23631396
Genome-wide analysis of T-DNA integration into the chromosomes of Magnaporthe oryzae
Agrobacterium tumefaciens-mediated transformation (ATMT) has become a prevalent tool for functional genomics of fungi, but our understanding of T-DNA integration into the fungal genome remains limited relative to that in plants. Using a model plant-pathogenic fungus, Magnaporthe oryzae, here we report the most comprehensive analysis of T-DNA integration events in fungi and the development of an informatics infrastructure, termed a T-DNA analysis platform (TAP). We identified a total of 1110 T-DNA-tagged locations (TTLs) and processed the resulting data via TAP. Analysis of the TTLs showed that T-DNA integration was biased among chromosomes and preferred the promoter region of genes. In addition, irregular patterns of T-DNA integration, such as chromosomal rearrangement and readthrough of plasmid vectors, were also observed, showing that T-DNA integration patterns into the fungal genome are as diverse as those of their plant counterparts. However, overall the observed junction structures between T-DNA borders and flanking genomic DNA sequences revealed that T-DNA integration into the fungal genome was more canonical than those observed in plants. Our results support the potential of ATMT as a tool for functional genomics of fungi and show that the TAP is an effective informatics platform for handling data from large-scale insertional mutagenesis. PMID:17850257
Choi, Jaehyuk; Park, Jongsun; Jeon, Junhyun; Chi, Myoung-Hwan; Goh, Jaeduk; Yoo, Sung-Yong; Park, Jaejin; Jung, Kyongyong; Kim, Hyojeong; Park, Sook-Young; Rho, Hee-Sool; Kim, Soonok; Kim, Byeong Ryun; Han, Seong-Sook; Kang, Seogchan; Lee, Yong-Hwan
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 ...
Plenum window insertion loss in the presence of a line source--a scale model study.
The acoustical insertion losses of plenum windows installed on a building facade in the presence of a non-parallel line source are studied by using a 1:4 scaled down model in a semi-anechoic chamber in the present investigation. Two types of insertion losses, weighted by the normalized traffic noise spectrum (from the 100 Hz to 5000 Hz one-third octave bands), are defined with different references. The first one is for the case where the orientation of the building facade relative to the line source is fixed. The reference case is the opened window having the same orientation angle as the plenum window. The maximum and minimum insertion losses under this condition across the orientations tested are found to be around 14 dB and 5 dB, respectively. The other is the opposite situation where such orientation is allowed to change because of practical purposes and the reference for this condition is the opened window with its width span parallel to the line source. The corresponding maximum and minimum insertion losses are found to be around 18 dB and 8 dB, respectively. There are evidences showing that the lower order plenum acoustic modes are responsible for the relatively high low frequency insertion loss. PMID:23464017
Tong, Y G; Tang, S K
Paramutation-Like Interaction of T-DNA Loci in Arabidopsis
In paramutation, epigenetic information is transferred from one allele to another to create a gene expression state which is stably inherited over generations. Typically, paramutation describes a phenomenon where one allele of a gene down-regulates the expression of another allele. Paramutation has been described in several eukaryotes and is best understood in plants. Here we describe an unexpected paramutation-like trans SALK T-DNA interaction in Arabidopsis. Unlike most of the previously described paramutations, which led to gene silencing, the trans SALK T-DNA interaction caused an increase in the transcript levels of the endogenous gene (COBRA) where the T-DNA was inserted. This increased COBRA expression state was stably inherited for several generations and led to the partial suppression of the cobra phenotype. DNA methylation was implicated in this trans SALK T-DNA interaction since mutation of the DNA methyltransferase 1 in the suppressed cobra caused a reversal of the suppression. In addition, null mutants of the DNA demethylase ROS1 caused a similar COBRA transcript increase in the cobra SALK T-DNA mutant as the trans T-DNA interaction. Our results provide a new example of a paramutation-like trans T-DNA interaction in Arabidopsis, and establish a convenient hypocotyl elongation assay to study this phenomenon. The results also alert to the possibility of unexpected endogenous transcript increase when two T-DNAs are combined in the same genetic background. PMID:23272131
Xue, Weiya; Ruprecht, Colin; Street, Nathaniel; Hematy, Kian; Chang, Christine; Frommer, Wolf B.; Persson, Staffan; Niittyl�, Totte
Metal Insertion in a Microporous Metal-Organic Framework Lined with 2,2-Bipyridine
Metal Insertion in a Microporous Metal-Organic Framework Lined with 2,2-Bipyridine Eric D. Bloch, David Britt, Chain Lee, Christian J. Doonan,,� Fernando J. Uribe-Romo, Hiroyasu Furukawa, Jeffrey R), the first metal-organic framework with open 2,2-bipyridine (bpy) coordination sites. The material displays
High-frequency T-DNA-mediated gene tagging in plants.
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 PMID:2554318
Koncz, C; Martini, N; Mayerhofer, R; Koncz-Kalman, Z; K�rber, H; Redei, G P; Schell, J
High-frequency T-DNA-mediated gene tagging in plants.
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. PMID:2554318
Koncz, C; Martini, N; Mayerhofer, R; Koncz-Kalman, Z; K�rber, H; Redei, G P; Schell, J
Insertion device and beam line plans for the Advanced Photon Source: A report and recommendations by the Insertion Device and Beam Line Planning Committee
In the 7-GeV Advanced Photon Source (APS) Conceptual Design Report (CDR), fifteen complete experimental beam lines were specified in order to establish a representative technical and cost base for the components involved. In order to optimize the composition of the insertion devices and the beam line, these funds are considered a ''Trust Fund.'' The present report evaluates the optimization for the distribution of these funds so that the short- and long-term research programs will be most productive, making the facility more attractive from the user's point of view. It is recommended that part of the ''Trust Fund'' be used for the construction of the insertion devices, the front-end components, and the first-optics, minimizing the cost to potential users of completing a beam line. In addition, the possibility of cost savings resulting from replication and standardization of high multiplicity components (such as IDs, front ends, and first-optics instrumentation) is addressed. 2 refs., 5 tabs.
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Recessive Inheritance of Population-Specific Intronic LINE-1 Insertion Causes a Rotor Syndrome Phenotype.
Sequences of long-interspersed elements (LINE-1, L1) make up ?17% of the human genome. De novo insertions of retrotransposition-active L1s can result in genetic diseases. It has been recently shown that the homozygous inactivation of two adjacent genes SLCO1B1 and SLCO1B3 encoding organic anion transporting polypeptides OATP1B1 and OATP1B3 causes a benign recessive disease presenting with conjugated hyperbilirubinemia, Rotor syndrome. Here, we examined SLCO1B1 and SLCO1B3 genes in six Japanese diagnosed with Rotor syndrome on the basis of laboratory data and laparoscopy. All six Japanese patients were homozygous for the c.1738C>T nonsense mutation in SLCO1B1 and homozygous for the insertion of a ?6.1-kbp L1 retrotransposon in intron 5 of SLCO1B3, which altogether make up a Japanese-specific haplotype. RNA analysis revealed that the L1 insertion induced deleterious splicing resulting in SLCO1B3 transcripts lacking exon 5 or exons 5-7 and containing premature stop codons. The expression of OATP1B1 and OATP1B3 proteins was not detected in liver tissues. This is the first documented case of a population-specific polymorphic intronic L1 transposon insertion contributing to molecular etiology of recessive genetic disease. Since L1 activity in human genomes is currently seen as a major source of individual genetic variation, further investigations are warranted to determine whether this phenomenon results in other autosomal-recessive diseases. PMID:25546334
Kagawa, Tatehiro; Oka, Akira; Kobayashi, Yoshinao; Hiasa, Yoichi; Kitamura, Tsuneo; Sakugawa, Hiroshi; Adachi, Yukihiko; Anzai, Kazuya; Tsuruya, Kota; Arase, Yoshitaka; Hirose, Shunji; Shiraishi, Koichi; Shiina, Takashi; Sato, Tadayuki; Wang, Ting; Tanaka, Masayuki; Hayashi, Hideki; Kawabe, Noboru; Robinson, Peter N; Zemojtel, Tomasz; Mine, Tetsuya
Increased Polyamine Biosynthesis Enhances Stress Tolerance by Preventing the Accumulation of Reactive Oxygen Species: T-DNA Mutational Analysis of Oryza sativa Lysine Decarboxylase-like Protein 1
A highly oxidative stress-tolerant japonica rice line was isolated by T-DNA insertion mutation followed by screening in the presence of 50 mM H2O2. The T-DNA insertion was mapped to locus Os09g0547500, the gene product of which was annotated as lysine decarboxylase-like protein (GenBank accession No. AK062595). We termed this gene OsLDC-like 1, for Oryza sativa lysine decarboxylase-like 1. The insertion site was in the second exon and resulted in a 27 amino acid N-terminal deletion. Despite this defect in OsLDC-like 1, the mutant line exhibited enhanced accumulation of the polyamines (PAs) putrescine, spermidine, and spermine under conditions of oxidative stress. The generation of reactive oxygen species (ROS) in the mutant line was assessed by qRT-PCR analysis of NADPH oxidase (RbohD and RbohF), and by DCFH-DA staining. Cellular levels of ROS in osldc-like 1 leaves were significantly lower than those in the wild-type (WT) rice after exposure to oxidative, high salt and acid stresses. Exogenously-applied PAs such as spermidine and spermine significantly inhibited the stress-induced accumulation of ROS and cell damage in WT leaves. Additionally, the activities of ROS-detoxifying enzymes were increased in the homozygous mutant line in the presence or absence of H2O2. Thus, mutation of OsLDC-like 1 conferred an oxidative stress-tolerant phenotype. These results suggest that increased cellular PA levels have a physiological role in preventing stress-induced ROS and ethylene accumulation and the resultant cell damage. PMID:22965749
Jang, Su Jin; Wi, Soo Jin; Choi, Yoo Jin; An, Gynheung; Park, Ky Young
Inheritance and expression of a transgene insert in an aneuploid tobacco line.
A T-DNA locus comprising nptII, uidA and nos genes--all under the control of the nos promoter (this locus was designated K because it encodes resistance to Kanamycin)--was found to be inherited erratically in a transgenic tobacco line. This anomalous behavior was partially explained following a karyotype analysis of plants representing several generations: these plants were aneuploids, presumably for the K-containing chromosome. During four generations of sexual propagation, transgenic plants that were either trisomic or tetrasomic for the K-containing chromosome (i.e. 2n = 49 or 2n = 50, respectively) were obtained. The trisomic plants (2n = 48 + 1) were virtually indistinguishable phenotypically from normal euploids (2n = 4x = 48), whereas the tetrasomic plants (2n = 48 + 2) were smaller, had somewhat misshapen leaves and exhibited reduced fertility. Although the amount of NPTII protein in different trisomic (K--, KK-, KKK) and tetrasomic (KK--, KKK-) plants was generally consistent with a K dosage effect, the genetic behavior of each trisomic--with respect to segregation of KanR and marker gene activity in progeny--was unique and not completely explicable by invoking aneuploidy. Specifically, unexpected gains or losses of K could occur, suggesting the formation of double reductional gametes and/or frequent gene conversion at this locus. The susceptibility of K locus marker genes to trans-inactivation in the trisomic and tetrasomic lines was tested by crossing in partially homologous silencing loci. In all transgenotypes tested, the three K marker genes were sensitive to trans-silencing, which was accompanied by methylation in all copies of the nos promotor. In addition to this directed inactivation/methylation, the K locus could also undergo infrequent, spontaneous partial methylation, which produced stable epialleles. In most plants, however, the multiple copies of the nos promoter at this locus remained unmethylated and active through four generations in all transgenotypes examined. The significance of these results for irregular inheritance patterns, aneuploid syndromes and homology-dependent gene silencing is discussed. PMID:7808397
Matzke, M A; Moscone, E A; Park, Y D; Papp, I; Oberkofler, H; Neuhuber, F; Matzke, A J
Generation of Single-Copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-Mediated Resolution System1
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. PMID:17693537
De Buck, Sylvie; Peck, Ingrid; De Wilde, Chris; Marjanac, Gordana; Nolf, Jonah; De Paepe, Annelies; Depicker, Ann
Insertional Mutagenesis of Genes Required for Seed Development in Arabidopsis thaliana
The purpose of this project was to identify large numbers of Arabidopsis genes with essential functions during seed development. More than 120,000 T-DNA insertion lines were generated following Agrobacter- ium-mediated transformation. Transgenic plants were screened for defective seeds and putative mutants were subjected to detailed analysis in subsequent generations. Plasmid rescue and TAIL-PCR were used to recover plant sequences flanking
John McElver; Iris Tzafrir; George Aux; Rebecca Rogers; Carl Ashby; Kelsey Smith; Carla Thomas; Amy Schetter; Qing Zhou; Mary Ann Cushman; John Tossberg; Todd Nickle; Joshua Z. Levin; Marcus Law; David Meinke; David Patton
[Genetic mapping of T-DNA integration sites in Xa21 transgenic rice].
The transformation mediated by Agrobacterium has been successfully applied to rice in recent years. In the previous research we have transferred the Xa21 gene into five rice varieties of China, using Agrobacterium-mediated trasformation. In this study, T-DNA flanking sequences of Xa21 transgenic rice lines were obtained by using thermal asymmetric interlaced PCR (TAIL-PCR). The flanking sequences which are actual rice DNA were identified and located on molecular linkage map developed from a ZYQ8/JX17 double haploid (DH) population. A total of 22 T-DNA flanking rice sequences were isolated. Nineteen of them displayed RFLPs between the two parents, ZYQ8 and JX17, and were mapped on the rice chromosomes, 3, 4, 7, 9, 10, 11 and 12, respectively. The genetic mapping of T-DNA integration sites in Xa21 transgenic rice will benefit the study of position effect and stable inheritance of the transgene Xa21. PMID:12561472
Zhu, Xue-Feng; Chen, Xue-Wei; Li, Xiao-Bing; Qian, Qian; Huang, Da-Nian; Zhu, Li-Huang; Zhai, Wen-Xue
Generation of single-stranded T-DNA molecules during the initial stages of T-DNA transfer from Agrobacterium tumefaciens to plant cells
Activation of the T-DNA transfer process of Agrobacterium by the plant signal molecule acetosyringone generates a single-stranded, unipolar, linear T-DNA molecule (T-strand)-a potential conjugative intermediate in the transfer of the T-DNA to plant cells. Acetosyringone induction also leads to S1 nuclease-sensitive sites at the Ti plasmid T-DNA borders, and other molecular changes associated with the Ti plasmid T-DNA sequences, which
Scott E. Stachel; Benedikt Timmerman; Patricia Zambryski
Analysis of junctions between T-DNA and plant genome in transgenic Arabidopsis thaliana
To understand the mechanism forAgrobacterium- mediated transformation of plants, we analyzed the junctions between T-DNA and plant genome, using 12 individual transgenic lines\\u000a transformed with 7 different plant expression constructs. After performing TAIL-PCR, we sequenced 42 PCR products for analysis.\\u000a All of the RBs were nicked by VirD1\\/VirD2 proteins whereas only 62% of the LBs were. Additional deletions of the
Jeong Hoe Kim; Sangman Lee
SUMMARY Motivation: The aim of this work was to search for the nucleotide sequences in plant genome data banks similar to agrobacterial T-DNA fragments, in order to evaluate the role of naturally associated soilborne agrobacteria in plant evolution. Results: Depending on the variant and length of the T-DNA right-border fragment, we found from 2 to 115 nucleotide sequences within different
Analysis of T-DNA integration and generative segregation in transgenic winter triticale (x Triticosecale Wittmack)
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. PMID:23006412
Characterization at nucleotide resolution of the homogeneously staining region sites of insertion in two cancer cell lines
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
T-DNA from Agrobacterium Ti Plasmid is in the Nuclear DNA Fraction of Crown Gall Tumor Cells
The crown gall teratoma tumor line BT37, incited by Agrobacterium tumefaciens strain T37, has been found to contain part of the tumor-inducing plasmid, pTi T37, of the inciting strain. This foreign DNA segment, called T-DNA, is maintained at several copies per diploid tumor cell. We have examined subcellular DNA fractions from this tumor line in an effort to determine whether
Mary-Dell Chilton; Randall K. Saiki; Narendra Yadav; Milton P. Gordon; Francis Quetier
FISHES OF HAWAllAN ISLANDS. Page 197. After line 17 insert
together, the upper the larger, oval in shape; anal and dorsal fins similar, of equal height and ending posteriorly on a line; no detached rays; origin of anal over fifth dorsal ray; base of anal ain length of fish; anal a lighter bluish white, splotched anteriorly with dusky; axil and base of pectoral earth brown
Agrobacterium T-DNA integration: molecules and models
, NY 11794-5215, USA Genetic transformation mediated by Agrobacterium involves the transfer of a DNA-kingdom DNA transfer [1]. Although used mainly for plant genetic engineering [2], Agrobacterium can transform unexplored, although several different T-DNA integration mechanisms have been suggested. Recent genetic
Citovsky, Vitaly
Transcultural Diabetes Nutrition Algorithm (tDNA): Venezuelan Application
Medical nutrition therapy (MNT) is a necessary component of comprehensive type 2 diabetes (T2D) management, but optimal outcomes require culturally-sensitive implementation. Accordingly, international experts created an evidence-based transcultural diabetes nutrition algorithm (tDNA) to improve understanding of MNT and to foster portability of current guidelines to various dysglycemic populations worldwide. This report details the development of tDNA-Venezuelan via analysis of region-specific cardiovascular disease (CVD) risk factors, lifestyles, anthropometrics, and resultant tDNA algorithmic modifications. Specific recommendations include: screening for prediabetes (for biochemical monitoring and lifestyle counseling); detecting obesity using Latin American cutoffs for waist circumference and Venezuelan cutoffs for BMI; prescribing MNT to people with prediabetes, T2D, or high CVD risk; specifying control goals in prediabetes and T2D; and describing regional differences in prevalence of CVD risk and lifestyle. Venezuelan deliberations involved evaluating typical food-based eating patterns, correcting improper dietary habits through adaptation of the Mediterranean diet with local foods, developing local recommendations for physical activity, avoiding stigmatizing obesity as a cosmetic problem, avoiding misuse of insulin and metformin, circumscribing bariatric surgery to appropriate indications, and using integrated health service networks to implement tDNA. Finally, further research, national surveys, and validation protocols focusing on CVD risk reduction in Venezuelan populations are necessary. PMID:24699193
Nieto-Mart�nez, Ramfis; Hamdy, Osama; Marante, Daniel; In�s Marulanda, Mar�a; Marchetti, Albert; Hegazi, Refaat A.; Mechanick, Jeffrey I.
Murine interleukin 2 receptor beta chain: dysregulated gene expression in lymphoma line EL-4 caused by a promoter insertion.
The functional, high-affinity interleukin 2 receptor (IL-2R) consists of at least two receptor components, IL-2R alpha (p55) and IL-2R beta (p70-75). The cDNA encoding the murine IL-2R beta has been isolated by using the previously cloned cDNA for human IL-2R beta as a probe. Analysis of the cDNA revealed that the murine IL-2R beta shows a marked homology with the human IL-2R beta and that it is also structurally related to other cytokine receptors such as erythropoietin receptor. The cDNA-directed murine IL-2R beta formed high-affinity IL-2R in conjunction with the endogenous IL-2R alpha in a murine pro-B-cell line and could transduce IL-2-induced growth signal. In mouse lymphoma line EL-4, the IL-2R beta gene was found to be rearranged by the insertion of the long terminal repeat sequence of an intracisternal A particle, giving rise to constitutive expression of the IL-2R beta mRNA. Images PMID:2155425
Kono, T; Doi, T; Yamada, G; Hatakeyama, M; Minamoto, S; Tsudo, M; Miyasaka, M; Miyata, T; Taniguchi, T
Enhanced Expression of Bioactive Recombinant VEGF-111 with Insertion of Intronic Sequence in Mammalian Cell Lines.
Among all VEGF-A isoforms, VEGF-111 is particularly important in molecular biology research owing to its potent angiogenic properties and its remarkable resistance to proteolysis. These features make it a potential candidate for therapeutic use in ischemic diseases. VEGF-111 is not expressed in normal cells, but expression is induced by UV-B irradiation and exposure to genotoxic agents. Here, to increase expression at the transcriptional and translational levels, we synthesized and cloned recombinant VEGF-111 cDNA. Two fragments encoding exons 1-4 and intron 4/5 plus exon 8a were amplified and cloned into the pBud.CE4.1 vector using a class IIs restriction enzyme-based method. The expression of VEGF-111 in CHO-dhfr - and HEK 293 cell lines was evaluated with real-time PCR, dot blotting, and ELISA. VEGF expression was increased about 10- and 18-fold in transfected CHO-dhfr - and HEK 293 cells, respectively. Dot blotting and ELISA confirmed successful production of VEGF-111 in both cell lines. PMID:25820359
Hojati, Zohreh; Dehghanian, Fariba
Integration of Agrobacterium tumefaciens T-DNA in the Saccharomyces cerevisiae Genome by Illegitimate Recombination
Agrobacterium tumefaciens can transfer part of its Ti plasmid, the T-DNA, to plant cells where it integrates into the nuclear genome via illegitimate recombination. Integration of the T-DNA results in small deletions of the plant target DNA, and may lead to truncation of the T-DNA borders and the production of filler DNA. We showed previously that T-DNA can also be
Paul Bundock; Paul J. J. Hooykaas
Targeted Integration of T-DNA into the Tobacco Genome at Double-Stranded Breaks: New Insights on the Mechanism of T-DNA Integration
Agrobacterium tumefaciens T-DNA normally integrates into random sites in the plant genome. We have investigated targeting of T-DNA by nonhomologous end joining process to a specific double-stranded break created in the plant genome by I-CeuI endonuclease. Sequencing of genomic DNA\\/T-DNA junctions in targeted events revealed that genomic DNA at the cleavage sites was usually intact or nearly so, whereas donor
Mary-Dell M. Chilton; Qiudeng Que
T-DNA tagging of the translation initiation factor eIF-4A1 of Arabidopsis thaliana
A transgenic Arabidopsis thaliana line, generated using a T-DNA vector carrying a promoterless gus reporter gene, showed intense GUS expression in young leaves and rapidly growing stem tissues. The gus fusion has tagged the 3? region of the gene encoding the A. thaliana eukaryotic translation initiation factor eIF-4A1. Comparison of the genomic and cDNA sequence shows that the eIF-4A1 gene
Henri De Greve; Viet Khong Nguyen; Francine Deboeck; Lin Thia-Toong; Mansour Karimi; Jean-Pierre Hernalsteens
Application of Discrete Event Control to the Insertion Task of Electric Line Using 6-Link Electro-Hydraulic Manipulators with Dual Arm
NASA Astrophysics Data System (ADS)
Uninterrupted power supply has become indispensable during the maintenance task of active electric power lines as a result of today's highly information-oriented society and increasing demand of electric utilities. The maintenance task has the risk of electric shock and the danger of falling from high place. Therefore it is necessary to realize an autonomous robot system using electro-hydraulic manipulator because hydraulic manipulators have the advantage of electric insulation. Meanwhile it is relatively difficult to realize autonomous assembly tasks particularly in the case of manipulating flexible objects such as electric lines. In this report, a discrete event control system is introduced for automatic assembly task of electric lines into sleeves as one of a typical task of active electric power lines. In the implementation of a discrete event control system, LVQNN (learning vector quantization neural network) is applied to the insertion task of electric lines to sleeves. In order to apply these proposed control system to the unknown environment, virtual learning data for LVQNN was generated by fuzzy inference. By the experimental results of two types of electric lines and sleeves, these proposed discrete event control and neural network learning algorithm are confirmed very effective to the insertion tasks of electric lines to sleeves as a typical task of active electric power maintenance tasks.
Ahn, Kyoungkwan; Yokota, Shinichi
Agrobacterium-Mediated Gene Transfer Results Mainly in Transgenic Plants Transmitting T-DNA as a Single Mendelian Factor
Forty-four independent transformed tobacco plants were obtained from a cocultivation experiment with Agrobacterium tumefaciens strains carrying modified Ti-plasmids. The transformed plants were either self-fertilized or crossed with nontransformed plants or with other transformed plants. The segregation of a phenotypic marker (kanamycin resistance) in the progenies of these plants was determined. In 40 cases out of 44, the segregation of the kanamycin resistance marker is consistent with Mendelian genetics. Among these 40 clones, 35 contain a single kanamycin resistance locus. The five others segregate two independent resistance loci. In two of the single insert clones, the segregation ratio after selfing indicates that the T-DNA insertion may have caused a recessive lethal mutation. PMID:17246346
Budar, F.; Thia-Toong, L.; Van Montagu, M.; Hernalsteens, J.-P.
Generation and characterization of the Western Regional Research Center brachypodium T-DNA insertional mutant collection
Technology Transfer Automated Retrieval System (TEKTRAN)
The model grass Brachypodium distachyon (Brachypodium) is an excellent system for studying the basic biology underlying traits relevant to the use of grasses as food, forage and energy crops. To add to the growing collection of Brachypodium resources available to plant scientists, we further optim...
Published in: Plant Mol. Biol. (2005) 58:295-303 Relationship between allelic state of T-DNA and DNA methylation of
1 Published in: Plant Mol. Biol. (2005) 58:295-303 Relationship between allelic state of T that it is correlated with the allelic state of the T-DNA in parental lines. Analysis of the region between the T, and chromatin remodeling appeared to play a key function in epigenetic control in various organisms (Li et al
Paris-Sud XI, Université de
Central Line�Associated Bloodstream Infection in Hospitalized Children with Peripherally Inserted Central Venous Catheters: Extending Risk Analyses Outside the Intensive Care Unit
Background. Increasingly, peripherally inserted central venous catheters (PICCs) are placed for prolonged intravenous access. Few data exist regarding risk factors for central line�associated bloodstream infection (CLABSI) complicating PICCs in hospitalized children, especially children hospitalized outside the intensive care unit (ICU). Methods.?We identified all children with a PICC inserted at The Johns Hopkins Hospital (Baltimore, MD) from 1 January 2003 through 31 December 2009 and used Poisson regression models to identify risk factors for PICC-associated CLABSIs. Results.?A total of 2592 PICCs were placed in 1819 children. One hundred sixteen CLABSIs occurred over 44,972 catheter-days (incidence rate [IR], 2.58 cases per 1000 catheter-days; 95% confidence interval [CI], 2.07�3.00 cases per 1000 catheter-days). Independent predictors of CLABSI in the entire cohort included PICC dwell time of ?21 days (IR ratio [IRR], 1.53; 95% CI, 1.05�2.26), parenteral nutrition as indication for insertion (IRR, 2.24; 95% CI, 1.31�3.84), prior PICC-associated CLABSI (IRR, 2.48; 95% CI, 1.18�5.25), underlying metabolic condition (IRR, 2.07; 95% CI, 1.14�3.74), and pediatric ICU exposure during hospitalization (IRR, 1.80; 95% CI, 1.18�2.75). Risk factors for CLABSI in children without PICU exposure included younger age, underlying malignancy and metabolic conditions, PICCs inserted in the lower extremity, and a prior PICC-associated CLABSI. Conclusions.?Prolonged catheter dwell time, pediatric ICU exposure, and administration of parenteral nutrition as the indication for PICC insertion are important predictors of PICC-associated CLABSI in hospitalized children. A careful assessment of these risk factors may be important for future success in preventing CLABSIs in hospitalized children with PICCs. PMID:21454298
Advani, Sonali; Reich, Nicholas G.; Sengupta, Arnab; Gosey, Leslie
Characterization of a Chinese Hamster Ovary Cell Line Developed by Retroviral Insertional Mutagenesis That Is Resistant to Sindbis Virus Infection
The alphavirus Sindbis virus (SV) has a wide host range and infects many types of cultured cells in vitro. The outcome of infection is dependent on the strain of virus used for infection and the properties of the cells infected. To identify cellular determinants of susceptibility to SV infection we mutagenized Chinese hamster ovary (CHO) cells by retroviral insertion with
The maternal chromosome set is the target of the T-DNA in the in planta transformation of Arabidopsis thaliana.
In planta transformation methods are now commonly used to transform Arabidopsis thaliana by Agrobacterium tumefaciens. The origin of transformants obtained by these methods has been studied by inoculating different floral stages and examining gametophytic expression of an introduced beta-glucuronidase marker gene encoding GUS. We observed that transformation can still occur after treating flowers where embryo sacs have reached the stage of the third division. No GUS expression was observed in embryo sacs or pollen of plants infiltrated with an Agrobacterium strain bearing a GUS gene under the control of a gametophyte-specific promoter. To identify the genetic target we used an insertion mutant in which a gene essential for male gametophytic development has been disrupted by a T-DNA bearing a Basta resistance gene (B(R)). In this mutant the B(R) marker is transferred to the progeny only by the female gametes. This mutant was retransformed with a hygromycin resistance marker and doubly resistant plants were selected. The study of 193 progeny of these transformants revealed 25 plants in which the two resistance markers were linked in coupling and only one plant where they were linked in repulsion. These results point to the chromosome set of the female gametophyte as the main target for the T-DNA. PMID:10924482
Bechtold, N; Jaudeau, B; Jolivet, S; Maba, B; Vezon, D; Voisin, R; Pelletier, G
Insertional mutagenesis using Tnt1 retrotransposon in potato
Technology Transfer Automated Retrieval System (TEKTRAN)
Potato is the third most important food crop in the world. However, genetics and genomics research of potato has lagged behind many major crop species due to its autotetraploidy and a highly heterogeneous genome. Insertional mutagenesis using T-DNA or transposable elements, which is available in sev...
A LINE-1 insertion in DLX6 is responsible for cleft palate and mandibular abnormalities in a canine model of Pierre Robin sequence.
Cleft palate (CP) is one of the most commonly occurring craniofacial birth defects in humans. In order to study cleft palate in a naturally occurring model system, we utilized the Nova Scotia Duck Tolling Retriever (NSDTR) dog breed. Micro-computed tomography analysis of CP NSDTR craniofacial structures revealed that these dogs exhibit defects similar to those observed in a recognizable subgroup of humans with CP: Pierre Robin Sequence (PRS). We refer to this phenotype in NSDTRs as CP1. Individuals with PRS have a triad of birth defects: shortened mandible, posteriorly placed tongue, and cleft palate. A genome-wide association study in 14 CP NSDTRs and 72 unaffected NSDTRs identified a significantly associated region on canine chromosome 14 (24.2 Mb-29.3 Mb; p(raw?)=?4.64 � 10(-15)). Sequencing of two regional candidate homeobox genes in NSDTRs, distal-less homeobox 5 (DLX5) and distal-less homeobox 6 (DLX6), identified a 2.1 kb LINE-1 insertion within DLX6 in CP1 NSDTRs. The LINE-1 insertion is predicted to insert a premature stop codon within the homeodomain of DLX6. This prompted the sequencing of DLX5 and DLX6 in a human cohort with CP, where a missense mutation within the highly conserved DLX5 homeobox of a patient with PRS was identified. This suggests the involvement of DLX5 in the development of PRS. These results demonstrate the power of the canine animal model as a genetically tractable approach to understanding naturally occurring craniofacial birth defects in humans. PMID:24699068
Wolf, Zena T; Leslie, Elizabeth J; Arzi, Boaz; Jayashankar, Kartika; Karmi, Nili; Jia, Zhonglin; Rowland, Douglas J; Young, Amy; Safra, Noa; Sliskovic, Saundra; Murray, Jeffrey C; Wade, Claire M; Bannasch, Danika L
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
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
Copper Heat Exchanger for the External Auxiliary Bus-Bars Routing Line in the LHC Insertion Regions
The corrector magnets and the main quadrupoles of the LHC dispersion suppressors are powered by a special superconducting line (called auxiliary bus-bars line N), external to the cold mass and housed in a 50 mm diameter stainless steel tube fixed to the cold mass. As the line is periodically connected to the cold mass, the same gaseous and liquid helium cools both the magnets and the line. The final sub-cooling process (from around 4.5 K down to 1.9 K) consists in the phase transformation from liquid to superfluid helium. Heat is extracted from the line through the magnets via their point of junction. In dispersion suppressor zones, approximately 40 m long, the sub-cooling of the line is slightly delayed with respect to the magnets. This might have an impact on the readiness of the accelerator for operation. In order to accelerate the process, a special heat exchanger has been designed. It is located in the middle of the dispersion suppressor portion of the line. Its main function consists in providing a loca...
Garion, C; Seyvet, F; Sitko, M; Skoczen, B; Tock, J P
Shorter T-DNA or additional virulence genes improve Agrobactrium-mediated transformation
The effect of additional virulence (vir) genes and size of plasmid T-DNA in Agrobacterium tumefa- ciens was investigated for their impact on transformation efficiency. Transformation efficiency in tobacco, cotton, and rice was\\u000a increased when the T-DNA was 4.3 kb compared to 8.4 kb in size. However, when additional virG, virGN54D,virE, or virE\\/virG plasmids were included with the 8.4-kb T-DNA, transformation
S. H. Park; B.-M. Lee; M. G. Salas; M. Srivatanakul; R. H. Smith
Unregulated expression of the erythropoietin receptor gene caused by insertion of spleen focus-forming virus long terminal repeat in a murine erythroleukemia cell line.
A murine erythroleukemia (MEL) cell line, F5-5, expressed 10,000 binding sites for erythropoietin (EPO) per cell, 10-fold more than was expressed by other murine erythroleukemia cell lines and normal erythroid progenitors. Northern (RNA) and Southern blot analyses revealed overexpression of mRNA for the EPO receptor (EPOR) and rearrangement of one of the EPOR gene alleles in F5-5 cells, respectively. Molecular cloning of F5-5-derived cDNA encoding EPOR revealed that the 5' noncoding region of the EPOR cDNA corresponds to the 3' long terminal repeat sequence of the polycythemic strain of Friend spleen focus-forming virus (F-SFFVP). The aberrant EPOR transcripts containing the 3' long terminal repeat sequence were mainly expressed in F5-5 cells. The same integration upstream of the EPOR gene was also observed in other subclones and the parent cell line. It is possible that overexpression of EPOR by viral promoter insertion will confer growth advantage to an F-SFFVP-infected erythroid progenitor cell, leading to positive clonal selection through further leukemogenic steps. Images PMID:1656233
Hino, M; Tojo, A; Misawa, Y; Morii, H; Takaku, F; Shibuya, M
T-DNA integration and expression in a monocot crop plant after induction of Agrobacterium
Agrobacterium tumefaciens induces tumorous proliferations on dicotyledonous plants by transferring a specific region (T-DNA) of its tumour-inducing (Ti) plasmid into wound-activated plant cells, where it becomes integrated into nuclear DNA1 and expressed2. The vir region of the Ti plasmid is essential for T-DNA transfer3,4 and is induced by specific wound substances from plants5-9. Agrobacterium is therefore a powerful tool for
Willi Sch�fer; Andrea G�rz; G�nter Kahl
Arabidopsis genes essential for seedling viability: isolation of insertional mutants and molecular cloning.
We have undertaken a large-scale genetic screen to identify genes with a seedling-lethal mutant phenotype. From screening approximately 38,000 insertional mutant lines, we identified >500 seedling-lethal mutants, completed cosegregation analysis of the insertion and the lethal phenotype for >200 mutants, molecularly characterized 54 mutants, and provided a detailed description for 22 of them. Most of the seedling-lethal mutants seem to affect chloroplast function because they display altered pigmentation and affect genes encoding proteins predicted to have chloroplast localization. Although a high level of functional redundancy in Arabidopsis might be expected because 65% of genes are members of gene families, we found that 41% of the essential genes found in this study are members of Arabidopsis gene families. In addition, we isolated several interesting classes of mutants and genes. We found three mutants in the recently discovered nonmevalonate isoprenoid biosynthetic pathway and mutants disrupting genes similar to Tic40 and tatC, which are likely to be involved in chloroplast protein translocation. Finally, we directly compared T-DNA and Ac/Ds transposon mutagenesis methods in Arabidopsis on a genome scale. In each population, we found only about one-third of the insertion mutations cosegregated with a mutant phenotype. PMID:11779813
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
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.
Identification of pathogenicity-related genes in the rice pathogen Ustilaginoidea virens through random insertional mutagenesis.
Agrobacterium tumefaciens-mediated transformation (ATMT) is becoming a popular effective system as an insertional mutagenesis tool in filamentous fungi. To gain more insight into the cellular and molecular mechanisms in the pathogenesis of Ustilaginoidea virens, the causal agent of rice false smut disease, a T-DNA insertion mutant library of U. virens was established using ATMT. We optimized a range of conditions to improve the transformation efficiency. Transformants were most effectively obtained when the optimal co-cultivation time is 72h, with 50?M AS in medium and 100?l A. tumefaciens for co-cultivation, leading to the production of 160-185 hygromycin B resistant transformants per 1�10(5) conidia. Southern blot analysis indicated that 58.14% of transformants had a single T-DNA copy. Among 5600 transformants tested for virulence, 37 mutants with reproducible pathogenic defects were obtained. The flanking sequences of three avirulent tranformants (B20, B1015 and B1465) and two pathogenicity-reduced transformants (B726 and B785) were amplified by high-efficiency thermal asymmetric interlaced PCR. Sequence analyses revealed that single T-DNA insertion in mutant B20 targeted the coding region of a gene encoding a protein highly similar to SUN family protein, and in mutant B726 targeted upstream of a gene with unknown function. The two T-DNA insertion sites in mutant B785 were found in the coding region of a gene encoding C6 transcription factor, but failed in amplified flanking sequence of another T-DNA. Chromosomal rearrangement occurred in the genome of mutant B1016 and B1465 with single T-DNA insertion. Among avirulent mutants, B20 showed altered colony growth and pigmentation. The T-DNA insert in B20 was detected in the coding region of a gene named UvSUN2. Morphophysiological characterization analysis suggested that UvSUN2 might be a virulence factor, and possibly required for proper fungal growth, cell wall construction, and stress responses in U. virens. This study optimize and validate the transformation procedure, maximizing the number of single copy transformants and developing an efficient procedure for rescuing adjacent host sequences along with the inserted T-DNA. This is the first report of identification several candidate virulence factors and validated one in U. virens. Together with identification of avirulent mutants and their associated genes, results suggested that ATMT can effectively be used to identify genes associated with pathogenicity in U. virens, and provided novel insights into molecular mechanisms underlying virulence of this pathogen. PMID:25636735
Yu, Mina; Yu, Junjie; Hu, Jiankun; Huang, Lei; Wang, Yahui; Yin, Xiaole; Nie, Yafeng; Meng, Xiangkun; Wang, Weiduo; Liu, Yongfeng
Altered Growth and Wood Characteristics in Transgenic Hybrid Aspen Expressing Agrobacterium tumefaciens T-DNA Indoleacetic Acid-Biosynthetic Genes.
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. PMID:12228661
Tuominen, H.; Sitbon, F.; Jacobsson, C.; Sandberg, G.; Olsson, O.; Sundberg, B.
Genetic modification of potato development using Ri T-DNA
Forty-two potato plants were regenerated from a hairy-root line obtained after infection of a shoot of Solanum tuberosum cv �Desiree� with Agrobacterium rhizogenes strain LBA 9402 (pRil855). Transformed plants were uniform and had a distinct phenotype and development compared with untransformed controls. Their growth was vigorous, especially early in their development, their roots were abundant and showed reduced geotropism, their
G. Ooms; A. Karp; M. M. Burrell; D. Twell; J. Roberts
A plant transformation vector with a minimal T-DNA II. Irregular integration patterns of the T-DNA in the plant genome
A minimal T-DNA binary vector was used for Agrobacterium-mediated transfer of a chimeric T4 lysozyme gene located next to the left border, and transgenic potato plants which expressed T4 lysozyme protein were identified and further analysed. Frequent rearrangements of T4 lysozyme transgenes were detected. A vector derivative containing two matrix associated regions (MARs) flanking its multiple cloning site was constructed.
Petra Porsch; Astrid Jahnke
Protein apparatus for horizontal transfer of agrobacterial T-DNA to eukaryotic cells.
This review analyzes agrobacterial virulence proteins and recipient cell proteins involved in horizontal transfer of a T-DNA-protein complex. Specifically, it considers the early stages of the interactions of partners (signal exchange, attachment, close contact); T-DNA release from bacterial cells; channel formation for the transfer of ssDNA between the partners; transfer of agrobacterial T-DNA through the membrane, cytoplasm, and nuclear membrane of the recipient cell and its incorporation into the recipient cell genome. It further discusses possible pathways of agrobacterial ssDNA transfer to the recipient cells. In particular, the possible role of T-pili and VirE2 protein during conjugative transfer of agrobacterial ssDNA between donor and recipient cells is discussed. PMID:24460966
Chumakov, M I
Percutaneously inserted central catheter - infants
PICC - infants; PQC - infants; Pic line - infants; Per-Q cath - infants ... A percutaneously inserted central catheter (PICC) is a long, very thin, soft plastic tube that is put into a small blood vessel. This article addresses PICCs in ...
Peripherally inserted central catheter - insertion
PICC - insertion ... A PICC is a long, thin tube (called a catheter) that goes into a vein in your upper arm. ... in a large vein near your heart. The PICC helps carry nutrients and medicines into your body. ...
Involvement of KU80 in T-DNA integration in plant cells
27, 2005) In Agrobacterium-mediated genetic transformation of plant cells, the bacterium exports. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food. Following its nuclear import, the single-stranded T-DNA is stripped of its escorting proteins, most likely
Citovsky, Vitaly
T-DNA is organized predominantly in inverted repeat structures in plants transformed with Agrobacterium tumefaciens C58 derivatives
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
A simple and efficient method to identify replacements of P-lacZ by P-Gal4 lines allows obtaining Gal4 insertions in the bithorax complex of Drosophila.
The functional replacement of one gene product by another one is a powerful method to study specificity in development and evolution. In Drosophila, the Gal4/UAS method has been used to analyze in vivo such functional substitutions. To this aim, Gal4 lines that inactivate a gene and reproduce its expression pattern are required, and they can be frequently obtained by replacing pre-existing P-lacZ lines with such characteristics. We have devised a new method to quickly identify replacements of P-lacZ lines by P-Gal4 lines, and applied it successfully to obtain Gal4 insertions in the Ultrabithorax and Abdominal-B Hox genes. We have used these lines to study the functional replacement of a Hox gene by another one. Our experiments confirm that the abdominal-A gene can replace Ultrabithorax in haltere development but that it cannot substitute for Abdominal-B in the formation of the genitalia. PMID:16971094
de Navas, Luis; Foronda, David; Suzanne, Magali; S�nchez-Herrero, Ernesto
Temperature Affects the T-DNA Transfer Machinery of Agrobacterium tumefaciens
Early studies onAgrobacterium tumefaciensshowed that development of tumors on plants following infection byA. tumefacienswas optimal at temperatures around 22&C and did not occur at temperatures above 29&C. To assesswhetherthisinabilitytoinducetumorsisduetoadefectintheT-DNAtransfermachinery,mobilization of an incompatibility group Q (IncQ) plasmid by the T-DNA transfer machinery ofA. tumefacienswas tested at various temperatures. Optimal transfer occurred when matings were performed at 19&C, and transfer was not
A plant scaffold attached region detected close to a T-DNA integration site is active in mammalian cells.
Integration of foreign genes into plant genomes by the Agrobacterium T-DNA transfer system has been considered to occur at random. It has been speculated that the chromosomal structure of the integration site might affect the expression pattern of the introduced genes. To gain insight into the molecular structure of T-DNA integration sites and its possible impact on gene expression, we have examined plant DNA sequences in the vicinity of T-DNA borders. Analysis of a transgenic petunia plant containing a chloramphenicol acetyltransferase (CAT) gene regulated by the hemoglobin promoter (PAR) from Parasponia andersonii revealed a scaffold attachment region (SAR) close to one T-DNA end. In addition to having strong binding affinities for both animal and plant nuclear scaffolds this petunia SAR element is as active in mammalian cells as the authentic elements from mammalian sources. Images PMID:8052530
Dietz, A; Kay, V; Schlake, T; Landsmann, J; Bode, J
The Agrobacterium tumefaciens virC1 gene product binds to overdrive, a T-DNA transfer enhancer.
In Agrobacterium tumefaciens, a cis-active 24-base-pair sequence adjacent to the right border of the T-DNA, called overdrive, stimulates tumor formation by increasing the level of T-DNA processing. Recent results from our laboratory have suggested that the virC operon which enhances T-DNA processing probably does so because the VirC1 protein interacts with overdrive (N. Toro, A. Datta, M. Yanofsky, and E. W. Nester, Proc. Natl. Acad. Sci. USA 85:8558-8562, 1988). We report here the purification of the VirC1 protein from cells of Escherichia coli harboring a plasmid containing the coding sequences of the virC locus of the octopine Ti plasmid. By gel mobility shift and DNase I footprinting assays, we showed that this purified virC1 gene product binds to overdrive but not to the right border of T-DNA. Images PMID:2592351
Toro, N; Datta, A; Carmi, O A; Young, C; Prusti, R K; Nester, E W
Involvement of circular intermediates in the transfer of T-DNA from Agrobacterium tumefaciens to plant cells
Co-cultivation of Agrobacterium tumefaciens with plant cells leads to the induction of circular copies of the T-DNA segment of the large tumour-inducing (Ti) plasmid in the bacterial cells. These circular molecules are presumably intermediates in DNA transfer from the A. tumefaciens genome to the plant cells. In support of this suggestion, the junction of the T-DNA circles occurs precisely in
Zdena Koukol�kov�-Nicola; Raymond D. Shillito; Barbara Hohn; Kan Wang; Marc Van Montagu; Patricia Zembryski
Nucleotide sequence of the T-DNA region from the A grobacterium tumefaciens octopine Ti plasmid pTi15955
The complete nucleotide sequence of the transferred region (T-DNA) of an octopine tumor inducing (Ti) plasmid fromAgrobacterium tumefaciens (pTi15955) has been determined. A total of 24 595 nucleotides extending approximately 900 bases to either side of the outermost, T-DNA boundaries was sequenced. Computer analysis of the sequenced portion of the Ti plasmid revealed that recognition sites for 72 restriction endonucleases
R. F. Barker; K. B. Idler; D. V. Thompson; J. D. Kemp
Insertion sequences (ISs) constitute an important component of most bacterial genomes. Over 500 individual ISs have been described in the literature to date, and many more are being discovered in the ongoing prokaryotic and eukaryotic genome-sequencing projects. The last 10 years have also seen some striking advances in our understanding of the transposition process itself. Not least of these has been the development of various in vitro transposition systems for both prokaryotic and eukaryotic elements and, for several of these, a detailed understanding of the transposition process at the chemical level. This review presents a general overview of the organization and function of insertion sequences of eubacterial, archaebacterial, and eukaryotic origins with particular emphasis on bacterial elements and on different aspects of the transposition mechanism. It also attempts to provide a framework for classification of these elements by assigning them to various families or groups. A total of 443 members of the collection have been grouped in 17 families based on combinations of the following criteria: (i) similarities in genetic organization (arrangement of open reading frames); (ii) marked identities or similarities in the enzymes which mediate the transposition reactions, the recombinases/transposases (Tpases); (iii) similar features of their ends (terminal IRs); and (iv) fate of the nucleotide sequence of their target sites (generation of a direct target duplication of determined length). A brief description of the mechanism(s) involved in the mobility of individual ISs in each family and of the structure-function relationships of the individual Tpases is included where available. PMID:9729608
Mahillon, Jacques; Chandler, Michael
A pacemaker (PM) (or artificial PM, so as not to be confused with the heart's natural PM) is a medical device that uses electrical impulses, delivered by electrodes contracting the heart muscles, to regulate the beating of the heart. The primary purpose of this device is to maintain an adequate heart rate, either because the heart's natural PM is not fast enough, or there is a block in the heart's electrical conduction system. Modern PMs are externally programmable and allow the cardiologist to select the optimum pacing modes for individual patients. Some combine a PM and defibrillator in a single implantable device. PMs can be temporary or permanent. Temporary PMs are used to treat short-term heart problems, such as a slow heartbeat that's caused by a heart attack, heart surgery, or an overdose of medicine. Permanent PMs are used to control long-term heart rhythm problems. A PM can relieve some arrhythmia symptoms, such as fatigue and fainting. A PM also can help a person who has abnormal HRs resume a more active lifestyle. In the current mini review we will focus on the insertion of a PM and the possible pneumothorax that can be caused. PMID:25815303
Kotsakou, Maria; Kioumis, Ioannis; Lazaridis, George; Pitsiou, Georgia; Lampaki, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Tsakiridis, Kosmas; Katsikogiannis, Nikolaos; Karapantzos, Ilias; Karapantzou, Chrysanthi; Baka, Sofia; Mpoukovinas, Ioannis; Karavasilis, Vasilis; Rapti, Aggeliki; Trakada, Georgia; Zissimopoulos, Athanasios; Zarogoulidis, Konstantinos; Zarogoulidis, Paul
A pacemaker (PM) (or artificial PM, so as not to be confused with the heart�s natural PM) is a medical device that uses electrical impulses, delivered by electrodes contracting the heart muscles, to regulate the beating of the heart. The primary purpose of this device is to maintain an adequate heart rate, either because the heart�s natural PM is not fast enough, or there is a block in the heart�s electrical conduction system. Modern PMs are externally programmable and allow the cardiologist to select the optimum pacing modes for individual patients. Some combine a PM and defibrillator in a single implantable device. PMs can be temporary or permanent. Temporary PMs are used to treat short-term heart problems, such as a slow heartbeat that�s caused by a heart attack, heart surgery, or an overdose of medicine. Permanent PMs are used to control long-term heart rhythm problems. A PM can relieve some arrhythmia symptoms, such as fatigue and fainting. A PM also can help a person who has abnormal HRs resume a more active lifestyle. In the current mini review we will focus on the insertion of a PM and the possible pneumothorax that can be caused.
Kotsakou, Maria; Kioumis, Ioannis; Lazaridis, George; Pitsiou, Georgia; Lampaki, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Tsakiridis, Kosmas; Katsikogiannis, Nikolaos; Karapantzos, Ilias; Karapantzou, Chrysanthi; Baka, Sofia; Mpoukovinas, Ioannis; Karavasilis, Vasilis; Rapti, Aggeliki; Trakada, Georgia; Zissimopoulos, Athanasios; Zarogoulidis, Konstantinos
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
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
Differences in susceptibility of Arabidopsis ecotypes to crown gall disease may result from a deficiency in T-DNA integration.
We show that among ecotypes of Arabidopsis, there is considerable variation in their susceptibility to crown gall disease. Differences in susceptibility are heritable and, in one ecotype, segregate as a single major contributing locus. In several ecotypes, recalcitrance to tumorigenesis results from decreased binding of Agrobacterium to inoculated root explants. The recalcitrance of another ecotype occurs at a late step in T-DNA transfer. Transient expression of a T-DNA-encoded beta-glucuronidase gusA gene is efficient, but the ecotype is deficient in crown gall tumorigenesis, transformation to kanamycin resistance, and stable GUS expression. This ecotype is also more sensitive to gamma radiation than is a susceptible ecotype. DNA gel blot analysis showed that after infection by Agrobacterium, less T-DNA was integrated into the genome of the recalcitrant ecotype than was integrated into the genome of a highly susceptible ecotype. PMID:9090878
Nam, J; Matthysse, A G; Gelvin, S B
Application of Discrete Event Control to the Insertion Task of Electric Line Using 6Link Electro-Hydraulic Manipulators with Dual Arm
Uninterrupted power supply has become indispensable during the maintenance task of active electric power lines as a result of today's highly information-oriented society and increasing demand of electric utilities. The maintenance task has the risk of electric shock and the danger of falling from high place. Therefore it is necessary to realize an autonomous robot system using electro-hydraulic manipulator because
Kyoungkwan Ahn; Shinichi Yokota
Covalently bound VirD2 protein of Agrobacterium tumefaciens protects the T-DNA from exonucleolytic degradation.
We show that upon induction of Agrobacterium tumefaciens, free linear double-stranded T-DNA molecules as well as the previously described T-strands are generated from the Ti plasmid. A majority of these molecules are bound to a protein. We show that this protein is the product of the virulence gene virD2. This protein was found to be attached to the 5' terminus of processed T-DNA at the right border and to the rest of the Ti plasmid at the left border. The protein remnant after Pronase digestion rendered the right end of the double-stranded T-DNA resistant to 5'----3' exonucleolytic attack in vitro. The protein-DNA association was resistant to SDS, mercaptoethanol, mild alkali, piperidine, and hydroxylamine, indicating that it involves a covalent linkage. The possible involvement of this T-DNA-protein complex in replication, transduction to the plant, nuclear targeting, and integration into the plant nuclear DNA is discussed. Images PMID:2556703
D�rrenberger, F; Crameri, A; Hohn, B; Koukol�kov�-Nicola, Z
Core Vessel Insert Handling Robot for the Spallation Neutron Source
The Spallation Neutron Source provides the world's most intense pulsed neutron beams for scientific research and industrial development. Its eighteen neutron beam lines will eventually support up to twenty-four simultaneous experiments. Each beam line consists of various optical components which guide the neutrons to a particular instrument. The optical components nearest the neutron moderators are the core vessel inserts. Located approximately 9 m below the high bay floor, these inserts are bolted to the core vessel chamber and are part of the vacuum boundary. They are in a highly radioactive environment and must periodically be replaced. During initial SNS construction, four of the beam lines received Core Vessel Insert plugs rather than functional inserts. Remote replacement of the first Core Vessel Insert plug was recently completed using several pieces of custom-designed tooling, including a highly complicated Core Vessel Insert Robot. The design of this tool are discussed.
Graves, Van B [ORNL; Dayton, Michael J [ORNL
In planta transformation method for T-DNA transfer in orchids
NASA Astrophysics Data System (ADS)
Transgenic plant technology is an efficient tool to study the function of gene(s) in plant. The most popular and widely used technique is Agrobacterium-mediated transformation in which cocultivation was done by immersing the plant tissues/organ in overnight bacterial cultured for about 30 minutes to one hour under in vitro condition. In this experiment, we developed more easier technique that omitted the in vitro step during cocultivation with Agrobacterium, namely in planta transformation method. Pollinaria (compact pollen mass of orchid) of Phalaenopsis amabilis and Spathoglottis plicata orchids were used as target explants that were immersed into bacterial culture for 30 minutes, then dried up the pollinaria, the transformed pollinaria was used to pollinate orchid flowers. The T-DNA used for this experiments were Ubipro?PaFT/A. tumefaciens GV3101 for P. amabilis and MeEF1?2 pro?GUS/ A. tumefaciens LBA 4404 for S.plicata. Seeds that were produced from pollinated flowers were grown onto 10 mg/l hygromicin containing NP (New Phalaenopsis) medium. The existance of transgene in putative transformant protocorm (developing orchid embryo) genome was confirmed using PCR with specific primers of either PaFT or GUS genes. Histochemical GUS assay was also performed to the putative transformants. The result showed that transformation frequencies were 2.1 % in P. amabilis, and 0,53% in S. plicata. These results indicates that in planta transformation method could be used for Agrobacterium-mediated genetic transformation, with advantage easier and more secure work from contaminants than that of the in vitro method.
Semiarti, Endang; Purwantoro, Aziz; Mercuriani, Ixora S.; Anggriasari, Anida M.; Jang, Seonghoe; Suhandono, Sony; Machida, Yasunori; Machida, Chiyoko
Covalently Bound VirD2 Protein of Agrobacterium tumefaciens Protects the T-DNA from Exonucleolytic Degradation
We show that upon induction of Agrobacterium tumefaciens, free linear double-stranded T-DNA molecules as well as the previously described T-strands are generated from the Ti plasmid. A majority of these molecules are bound to a protein. We show that this protein is the product of the virulence gene virD2. This protein was found to be attached to the 5' terminus
Franz Durrenberger; Andreas Crameri; Barbara Hohn; Zdena Koukolikova-Nicola
Axenic mycorrhization of wild type and transgenic hybrid aspen expressing T-DNA indoleacetic acid-biosynthetic genes
We describe and document the in vitro synthesis of ectomycorrhiza between roots of wild type and transgenic aspen (Populus tremula � P. tremuloides), expressing Agrobacterium tumefaciens T-DNA indoleacetic acid (IAA)-biosynthetic genes, and Amanita muscaria. Plantlets were raised from tissue culture. The root system of approximately 4-week-old plantlets was transferred to Petri\\u000a dishes and incubated together with fungal mycelia under sterile
R. Hampp; M. Ecke; C. Schaeffer; T. Wallenda; A. Wingler; I. Kottke; B. Sundberg
T-DNA gene 5 of Agrobacterium modulates auxin response by autoregulated synthesis of a growth hormone antagonist in plants.
Oncogenes carried by the transferred DNA (T-DNA) of Agrobacterium Ti plasmids encode the synthesis of plant growth factors, auxin and cytokinin, and induce tumour development in plants. Other T-DNA genes regulate the tumorous growth in ways that are not yet understood. To determine the function of T-DNA gene 5, its coding region was expressed in Escherichia coli. Synthesis of the gene 5 encoded protein (26 kDa) correlated with a 28-fold increase in conversion of tryptophan to indole-3-lactate (ILA), an auxin analogue. Expression of chimeric gene 5 constructs in transgenic tobacco resulted in overproduction of ILA that enhanced shoot formation in undifferentiated tissues and increased the tolerance of germinating seedlings to the inhibitory effect of externally supplied auxin. Promoter analysis of gene 5 in plants revealed that its expression was inducible by auxin and confined to the vascular phloem cells. cis-regulatory elements required for auxin regulation and phloem specific expression of gene 5 were mapped to a 90 bp promoter region that carried DNA sequence motifs common to several auxin induced plant promoters, as well as a binding site for a nuclear factor, Ax-1. ILA was found to inhibit the auxin induction of the gene 5 promoter and to compete with indole-3-acetic acid (IAA) for in vitro binding to purified cellular auxin binding proteins. It is suggested therefore that ILA autoregulates its own synthesis and thereby modulates a number of auxin responses in plants. Images PMID:1756712
K�rber, H; Strizhov, N; Staiger, D; Feldwisch, J; Olsson, O; Sandberg, G; Palme, K; Schell, J; Koncz, C
T-DNA and opine synthetic loci in tumors incited by Agrobacterium tumefaciens A281 on soybean and alfalfa plants
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.
Characterization of Arabidopsis thaliana lines with altered seed storage protein profiles using synchrotron-powered FT-IR spectromicroscopy.
Arabidopsis thaliana lines expressing only one cruciferin subunit type (double-knockout; CRUAbc, CRUaBc, or CRUabC) or devoid of cruciferin (triple-knockout; CRU-) or napin (napin-RNAi) were generated using combined T-DNA insertions or RNA interference approaches. Seeds of double-knockout lines accumulated homohexameric cruciferin and contained similar protein levels as the wild type (WT). Chemical imaging of WT and double-knockout seeds using synchrotron FT-IR spectromicroscopy (amide I band, 1650 cm(-1), ?C?O) showed that proteins were concentrated in the cell center and protein storage vacuoles. Protein secondary structure features of the homohexameric cruciferin lines showed predominant ?-sheet content. The napin-RNAi line had lower ?-helix content than the WT. Lines entirely devoid of cruciferin had high ?-helix and low ?-sheet levels, indicating that structurally different proteins compensate for the loss of cruciferin. Lines producing homohexameric CRUC showed minimal changes in protein secondary structure after pepsin treatment, indicating low enzyme accessibility. The Synchrotron FT-IR technique provides information on protein secondary structure and changes to the structure within the cell. PMID:23298281
Withana-Gamage, Thushan S; Hegedus, Dwayne D; Qiu, Xiao; Yu, Peiqiang; May, Tim; Lydiate, Derek; Wanasundara, Janitha P D
Identification of tyrosine 706 in the kinase insert as the major colony-stimulating factor 1 (CSF-1)-stimulated autophosphorylation site in the CSF-1 receptor in a murine macrophage cell line.
The receptor for colony-stimulating factor 1 (CSF-1) is a ligand-activated protein-tyrosine kinase. It has been shown previously that the CSF-1 receptor is phosphorylated on serine in vivo and that phosphorylation on tyrosine can be induced by stimulation with CSF-1. We studied the phosphorylation of the CSF-1 receptor by using the BAC1.2F5 murine macrophage cell line, which naturally expresses CSF-1 receptors. Two-dimensional tryptic phosphopeptide mapping showed that the CSF-1 receptor is phosphorylated on several different serine residues in vivo. Stimulation with CSF-1 at 37 degrees C resulted in rapid phosphorylation on tyrosine at one major site and one or two minor sites. We identified the major site as Tyr-706. The identity of Tyr-706 was confirmed by mutagenesis. This residue is located within the kinase insert domain. There was no evidence that Tyr-973 (equivalent to Tyr-969 in the human CSF-1 receptor) was phosphorylated following CSF-1 stimulation. When cells were stimulated with CSF-1 at 4 degrees C, additional phosphotyrosine-containing phosphopeptides were detected and the level of phosphorylation of the individual phosphotyrosine-containing phosphopeptides was substantially increased. In addition, we show that CSF-1 receptors are capable of autophosphorylation at six to eight major sites in vitro. Images PMID:2160591
van der Geer, P; Hunter, T
Regulation of oncogene expression in T-DNA-transformed host plant cells.
Virulent Agrobacterium tumefaciens strains integrate their T-DNA into the plant genome where the encoded agrobacterial oncogenes are expressed and cause crown gall disease. Essential for crown gall development are IaaH (indole-3-acetamide hydrolase), IaaM (tryptophan monooxygenase) and Ipt (isopentenyl transferase), which encode enzymes for the biosynthesis of auxin (IaaH, IaaM) and cytokinin (Ipt). Although these oncogenes are well studied as the tumor-inducing principle, nothing is known about the regulation of oncogene expression in plant cells. Our studies show that the intergenic regions (IGRs) between the coding sequences (CDS) of the three oncogenes function as promoters in plant cells. These promoters possess a eukaryotic sequence organization and cis-regulatory elements for the binding of plant transcription factors. WRKY18, WRKY40, WRKY60 and ARF5 were identified as activators of the Ipt promoter whereas IaaH and IaaM is constitutively expressed and no transcription factor further activates their promoters. Consistent with these results, the wrky triple mutant plants in particular, develops smaller crown galls than wild-type and exhibits a reduced Ipt transcription, despite the presence of an intact ARF5 gene. WRKY40 and WRKY60 gene expression is induced by A. tumefaciens within a few hours whereas the ARF5 gene is transcribed later during crown gall development. The WRKY proteins interact with ARF5 in the plant nucleus, but only WRKY40 together with ARF5 synergistically boosts the activation of the Ipt promoter in an auxin-dependent manner. From our data, we propose that A. tumefaciens initially induces WRKY40 gene expression as a pathogen defense response of the host cell. The WRKY protein is recruited to induce Ipt expression, which initiates cytokinin-dependent host cell division. With increasing auxin levels triggered by ubiquitous expression of IaaH and IaaM, ARF5 is activated and interacts with WRKY40 to potentiate Ipt expression and balance cytokinin and auxin levels for further cell proliferation. PMID:25615824
Zhang, Yi; Lee, Chil-Woo; Wehner, Nora; Imdahl, Fabian; Svetlana, Veselova; Weiste, Christoph; Dr�ge-Laser, Wolfgang; Deeken, Rosalia
Regulation of Oncogene Expression in T-DNA-Transformed Host Plant Cells
Virulent Agrobacterium tumefaciens strains integrate their T-DNA into the plant genome where the encoded agrobacterial oncogenes are expressed and cause crown gall disease. Essential for crown gall development are IaaH (indole-3-acetamide hydrolase), IaaM (tryptophan monooxygenase) and Ipt (isopentenyl transferase), which encode enzymes for the biosynthesis of auxin (IaaH, IaaM) and cytokinin (Ipt). Although these oncogenes are well studied as the tumor-inducing principle, nothing is known about the regulation of oncogene expression in plant cells. Our studies show that the intergenic regions (IGRs) between the coding sequences (CDS) of the three oncogenes function as promoters in plant cells. These promoters possess a eukaryotic sequence organization and cis-regulatory elements for the binding of plant transcription factors. WRKY18, WRKY40, WRKY60 and ARF5 were identified as activators of the Ipt promoter whereas IaaH and IaaM is constitutively expressed and no transcription factor further activates their promoters. Consistent with these results, the wrky triple mutant plants in particular, develops smaller crown galls than wild-type and exhibits a reduced Ipt transcription, despite the presence of an intact ARF5 gene. WRKY40 and WRKY60 gene expression is induced by A. tumefaciens within a few hours whereas the ARF5 gene is transcribed later during crown gall development. The WRKY proteins interact with ARF5 in the plant nucleus, but only WRKY40 together with ARF5 synergistically boosts the activation of the Ipt promoter in an auxin-dependent manner. From our data, we propose that A. tumefaciens initially induces WRKY40 gene expression as a pathogen defense response of the host cell. The WRKY protein is recruited to induce Ipt expression, which initiates cytokinin-dependent host cell division. With increasing auxin levels triggered by ubiquitous expression of IaaH and IaaM, ARF5 is activated and interacts with WRKY40 to potentiate Ipt expression and balance cytokinin and auxin levels for further cell proliferation. PMID:25615824
Zhang, Yi; Lee, Chil-Woo; Wehner, Nora; Imdahl, Fabian; Svetlana, Veselova; Weiste, Christoph; Dr�ge-Laser, Wolfgang; Deeken, Rosalia
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
Genetic analysis of the individual T-DNA genes of Agrobacterium tumefaciens ; further evidence that two genes are involved in indole-3-acetic acid synthesis
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. Inz�; A. Follin; M. Van Lijsebettens; C. Simoens; C. Genetello; M. Van Montagu; J. Schell
Effects of P Element Insertions on Quantitative Traits in Drosophila Melanogaster
P element mutagenesis was used to construct 94 third chromosome lines of Drosophila melanogaster which contained on average 3.1 stable P element inserts, in an inbred host strain background previously free of P elements. The homozygous and heterozygous effects of the inserts on viability and abdominal and sternopleural bristle number were ascertained by comparing the chromosome lines with inserts to insert-free control lines of the inbred host strain. P elements reduced average homozygous viability by 12.2% per insert and average heterozygous viability by 5.5% per insert, and induced recessive lethal mutations at a rate of 3.8% per insert. Mutational variation for the bristle traits averaged over both sexes was 0.03V(e) per homozygous P insert and 0.003V(e) per heterozygous P insert, where V(e) is the environmental variance. Mutational variation was greater for the sexes considered separately because inserts had large pleiotropic effects on sex dimorphism of bristle characters. The distributions of homozygous effects of inserts on the bristle traits were asymmetrical, with the largest effects in the direction of reducing bristle number; and highly leptokurtic, with most of the increase in variance contributed by a few lines with large effects. The inserts had partially recessive effects on the bristle traits. Insert lines with extreme bristle effects had on average greatly reduced viability. PMID:1311697
Mackay, TFC.; Lyman, R. F.; Jackson, M. S.
Oxalate-minus mutants of Sclerotinia sclerotiorum via T-DNA insertion accumulate fumarate in culture and retain pathogenicity on plants
Technology Transfer Automated Retrieval System (TEKTRAN)
Sclerotinia sclerotiorum is a ubiquitous necrotrophic pathogen capable of infecting over 400 plant species including many economically important crops. Oxalic acid production has been shown in numerous studies to be a pathogenicity factor for S. sclerotiorum through several mechanisms. During our ra...
Insight into the molecular requirements for pathogenicity of Fusarium oxysporum f. sp. lycopersici through large-scale insertional mutagenesis
Background Fusarium oxysporum f. sp. lycopersici is the causal agent of vascular wilt disease in tomato. In order to gain more insight into the molecular processes in F. oxysporum necessary for pathogenesis and to uncover the genes involved, we used Agrobacterium-mediated insertional mutagenesis to generate 10,290 transformants and screened the transformants for loss or reduction of pathogenicity. Results This led to the identification of 106 pathogenicity mutants. Southern analysis revealed that the average T-DNA insertion is 1.4 and that 66% of the mutants carry a single T-DNA. Using TAIL-PCR, chromosomal T-DNA flanking regions were isolated and 111 potential pathogenicity genes were identified. Conclusions Functional categorization of the potential pathogenicity genes indicates that certain cellular processes, such as amino acid and lipid metabolism, cell wall remodeling, protein translocation and protein degradation, seem to be important for full pathogenicity of F. oxysporum. Several known pathogenicity genes were identified, such as those encoding chitin synthase V, developmental regulator FlbA and phosphomannose isomerase. In addition, complementation and gene knock-out experiments confirmed that a glycosylphosphatidylinositol-anchored protein, thought to be involved in cell wall integrity, a transcriptional regulator, a protein with unknown function and peroxisome biogenesis are required for full pathogenicity of F. oxysporum. PMID:19134172
Michielse, Caroline B; van Wijk, Ringo; Reijnen, Linda; Cornelissen, Ben JC; Rep, Martijn
Pearls Educator's Guide Insert
NSDL National Science Digital Library
This four-page PDF insert takes an informative look at pearls, oysters, and mollusks. Useful to both educators and students, it includes information and images from the Pearls exhibition at the American Museum of Natural History.
Agrobacterium T-DNA integration into the plant genome can occur without the activity of key non-homologous end-joining proteins.
Non-homologous end joining (NHEJ) is the major model proposed for Agrobacterium T-DNA integration into the plant genome. In animal cells, several proteins, including KU70, KU80, ARTEMIS, DNA-PKcs, DNA ligase IV (LIG4), Ataxia telangiectasia mutated (ATM), and ATM- and Rad3-related (ATR), play an important role in 'classical' (c)NHEJ. Other proteins, including histone H1 (HON1), XRCC1, and PARP1, participate in a 'backup' (b)NHEJ process. We examined transient and stable transformation frequencies of Arabidopsis thaliana roots mutant for numerous NHEJ and other related genes. Mutants of KU70, KU80, and the plant-specific DNA LIGASE VI (LIG6) showed increased stable transformation susceptibility. However, these mutants showed transient transformation susceptibility similar to that of wild-type plants, suggesting enhanced T-DNA integration in these mutants. These results were confirmed using a promoter-trap transformation vector that requires T-DNA integration into the plant genome to activate a promoterless gusA (uidA) gene, by virus-induced gene silencing (VIGS) of Nicotiana benthamiana NHEJ genes, and by biochemical assays for T-DNA integration. No alteration in transient or stable transformation frequencies was detected with atm, atr, lig4, xrcc1, or parp1 mutants. However, mutation of parp1 caused high levels of T-DNA integration and transgene methylation. A double mutant (ku80/parp1), knocking out components of both NHEJ pathways, did not show any decrease in stable transformation or T-DNA integration. Thus, T-DNA integration does not require known NHEJ proteins, suggesting an alternative route for integration. PMID:25641249
Park, So-Yon; Vaghchhipawala, Zarir; Vasudevan, Balaji; Lee, Lan-Ying; Shen, Yunjia; Singer, Kamy; Waterworth, Wanda M; Zhang, Zhanyuan J; West, Christopher E; Mysore, Kirankumar S; Gelvin, Stanton B
Efforts to deregulate Rainbow papaya in Japan: Molecular Characterization of Transgene and Vector Inserts
Technology Transfer Automated Retrieval System (TEKTRAN)
Transformation plasmid-derived insert number and insert site sequence in 55-1 line papaya derivatives Rainbow and SunUp was determined as part of a larger petition to allow its import into Japan (Suzuki, et al., 2007, 2008). Three insertions were detected by Southern analysis and their correspondin...
The Advanced Light Source (ALS), the first US third generation synchrotron radiation source, is currently under construction at the Lawrence Berkeley Laboratory. The low-emittance, 1.5 GeV electron storage ring and the insertion devices are specifically designed to produce high brightness beams in the UV to soft X-Ray range. The planned initial complement of insertion devices includes four 4.6 m long undulators, with period lengths of 3.9 cm, 5.0 cm (2) and 8.0 cm, and a 2.9 m long wiggler of 16 cm period length. Undulator design is well advanced and fabrication has begun on the 5.0 cm and 8.0 cm period length undulators. This paper discusses ALS insertion device requirements; general design philosophy; and design of the magnetic structure, support structure/drive systems, control system and vacuum system. 18 refs., 9 figs., 5 tabs.
Hoyer, E.; Chin, J.; Halbach, K.; Hassenzahl, W.V.; Humphries, D.; Kincaid, B.; Lancaster, H.; Plate, D.
Cloning of Insertion Site Flanking Sequence and Construction of Transfer DNA Insert Mutant Library in Stylosanthes Colletotrichum
Stylosanthes sp. is the most important forage legume in tropical areas worldwide. Stylosanthes anthracnose, which is mainly caused by Colletotrichum gloeosporioides, is a globally severe disease in stylo production. Little progress has been made in anthracnose molecular pathogenesis research. In this study, Agrobacterium tumefaciens-mediated transformation was used to transform Stylosanthes colletotrichum strain CH008. The major factors of the genetic transformation system of S. colletotrichum were optimized as follows: A. tumefaciens� AGL-1 concentration (OD600), 0.8; concentration of Colletotrichum conidium, 1�106 conidia/mL; acetosyringone concentration, 100 mmol/L; induction time, 6 h; co-culture temperature, 25�C; and co-culture time, 3 d. Thus, the transformation efficiency was increased to 300�400 transformants per 106 conidia. Based on the optimized system, a mutant library containing 4616 mutants was constructed, from which some mutants were randomly selected for analysis. Results show that the mutants were single copies that could be stably inherited. The growth rate, spore amount, spore germination rate, and appressorium formation rate in some mutants were significantly different from those in the wild-type strain. We then selected the most appropriate method for the preliminary screening and re-screening of each mutant�s pathogenic defects. We selected 1230 transformants, and obtained 23 strains with pathogenic defects, namely, 18 strains with reduced pathogenicity and five strains with lost pathogenicity. Thermal asymmetric interlaced PCR was used to identify the transfer DNA (T-DNA) integration site in the mutant that was coded 2430, and a sequence of 476 bp was obtained. The flanking sequence of T-DNA was compared with the Colletotrichum genome by BLAST, and a sequence of 401 bp was found in Contig464 of the Colletotrichum genome. By predicting the function of the flanking sequence, we discovered that T-DNA insertion in the promoter region of the putative gene had 79% homology with the aspartate aminotransferase gene in Magnaporthe oryzae (XP_003719674.1). PMID:25361073
Huang, Guixiu; Gao, Jianming; Zhang, Shiqing; Zheng, Jinlong; Liu, Qiaolian; Xi, Jingen
Sequence alignment in bioinformatics Dynamic programming activity Every living organism has DNA, a sequence of four nucleotides A, C, G, and T. DNA is copied and transmitted
Sequence alignment in bioinformatics Dynamic programming activity Every living organism has DNA, a sequence of four nucleotides A, C, G, and T. DNA is copied and transmitted from parent to child, but from, organisms with a common ancestor can end up with fairly different DNA sequences. Now that it is relatively
Zirbel, Craig L.
Characterization of pre-insertion loci of de novo L1 insertions
The human Long Interspersed Element-1 (LINE-1) and the Short Interspersed Element (SINE) Alu comprise 28% of the human genome. They share the same L1-encoded endonuclease for insertion, which recognizes an A+T-rich sequence. Under a simple model of insertion distribution, this nucleotide preference would lead to the prediction that the populations of both elements would be biased towards A+T-rich regions. Genomic L1 elements do show an A+T-rich bias. In contrast, Alu is biased towards G+C-rich regions when compared to the genome average. Several analyses have demonstrated that relatively recent insertions of both elements show less G+C content bias relative to older elements. We have analyzed the repetitive element and G+C composition of more than 100 pre-insertion loci derived from de novo L1 insertions in cultured human cancer cells, which should represent an evolutionarily unbiased set of insertions. An A+T-rich bias is observed in the 50 bp flanking the endonuclease target site, consistent with the known target site for the L1 endonuclease. The L1, Alu, and G+C content of 20 kb of the de novo pre-insertion loci show a different set of biases than those observed for fixed L1s in the human genome. In contrast to the insertion sites of genomic L1s, the de novo L1 pre-insertion loci are relatively L1-poor, Alu-rich and G+C-neutral. Finally, a statistically significant cluster of de novo L1 insertions was localized in the vicinity of the c-myc gene. These results suggest that the initial insertion preference of L1, while A+T-rich in the initial vicinity of the break site, can be influenced by the broader content of the flanking genomic region and have implications for understanding the dynamics of L1 and Alu distributions in the human genome. PMID:17067767
Gasior, Stephen L.; Preston, Graeme; Hedges, Dale J.; Gilbert, Nicolas; Moran, John V.; Deininger, Prescott L.
Remote Control Inserting the batteries
Top View Rear View Inserting the batteries 1 3Press in on the arrow mark and slide in the direction of the arrow to remove the battery cover. 2 Insert two AA size batteries, making sure their polarities match the and marks inside the battery compartment. Insert the side tabs of the battery cover into their slots
Kostic, Milivoje M.
Effect of medium on charge transport properties of modified poly(dA)-poly(dT) DNA molecular wire
NASA Astrophysics Data System (ADS)
By using tight binding Hamiltonian approach, the medium's effect on charge transport properties of modified poly(dA)-poly(dT) DNA molecular wire is studied. The DNA sequences used are modified poly(dA)-poly(dT) sequence by replacing 50 base pairs randomly with GC or CG base pairs. The DNA is contacted by metallic leads at both ends. The medium's effect on DNA is modeled as backbone onsite energy disorder. The DNA is taken to be at room temperature. The charge transport properties of the two sequences are studied by calculating the transmission probabilities of the charges using the scattering and transfer matrix methods simultaneously. Then the current-voltage (I-V) characteristics are calculated from the transmission probabilities using the Landauer-B�ttiker Formalism. The theoretical differential conductance curve is then calculated from the I-V characteristic curve. The I-V results show that as the backbone disorder strength increases, the maximum current decreases for both sequences. However, the threshold voltage can increase or decrease with increasing backbone disorder depending on the sequence. The transmission results show that the position of transmission peaks changed with backbone disorder strength. Comparison with previous published results using poly(dG)-poly(dC) sequence shows that in contrast to that sequence, backbone disorder can decrease the threshold voltage in one of the sequences studied. This may point to a new understanding of the interplay between environmental and sequence disorder effect on charge transport in DNA.
Suhendro, D. K.; Yudiarsah, E.; Saleh, R.
Impedance calculation for ferrite inserts
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
An essential virulence protein of Agrobacterium tumefaciens , VirB4, requires an intact mononucleotide binding domain to function in transfer of T-DNA
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
Conserved function in Nicotiana tabacum of a single Drosophila hsp70 promoter heat shock element when fused to a minimal T-DNA promoter
To demonstrate the extent of evolutionary conservation in the mechanism of induction of heat shock genes between plants and animals, the minimal sequence from the Drosophila hsp70 promoter sufficient to confer heat shock inducible transcription in tobacco was determined. Segments of the hsp70 promoter were fused to a minimal promoter of the T-DNA indole-3-acetamide hydrolase (iaaH) gene, in a chimaeric
David Wing; Csaba Koncz; Jeff Schell
Effect of loss of T-DNA genes on MIA biosynthetic pathway gene regulation and alkaloid accumulation in Catharanthus roseus hairy roots
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
Detection of transformed cells in crown gall tumors using the GUS reporter gene and correlation of GUS stained cells with T-DNA gene activity
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))
Existence of a bovine LINE repetitive insert that appears in the cDNA of bovine protein BCNT in ruminant, but not in human, genomes 1 A portion of these findings was presented in a preliminary report at the 13th Annual meeting on oncogenes (18�21 June 1997, Frederick). The nucleotide sequences studied in this paper will appear in the DDBJ\\/EMBL\\/GenBank Databases with the following Accession Nos: bovine bcnt; AB005652 (genomic); human BCNT AB009285(EST1020018), AB004907 (genomic), AB009269 (genomic), AB009270 (genomic), AB009271(genomic), and mouse deer bcnt; AB005651 (genomic). 1
A novel protein, BCNT, originally isolated from bovine brain and named after Bucentaur, contains an internal portion that is translated from part of bovine LINE repetitive sequence (Bov-B LINE). Human cDNA highly homologous to the bovine bcnt (bbcnt) cDNA has been isolated but does not contain a sequence similar to the Bov-B LINE insert (Nobukuni, T., Kobayashi, M., Omori, A.,
Ichiro Takahashi; Takahiro Nobukuni; Haruo Ohmori; Mariko Kobayashi; Shoji Tanaka; Kazuhiko Ohshima; Norihiro Okada; Tohru Masui; Katsuyuki Hashimoto; Shintaro Iwashita
A role for the anaphase promoting complex in hormone regulation
To increase our knowledge of anaphase promoting complex (APC\\/C) function during plant development, we characterized an Arabidopsis thaliana T-DNA-insertion line where the T-DNA fell within the 5? regulatory region of the APC10 gene. The insert disrupted endogenous expression, resulting in overexpression of APC10 mRNA from the T-DNA- internal CaMV 35S promoter, and increased APC10 protein. Overexpression of APC10 produced phenotypes
Donna L. Lindsay; Peta C. Bonham-Smith; Spike Postnikoff; Gordon R. Gray; Troy A. A. Harkness
Two independent retrotransposon insertions at the same site within the coding region of BTK.
Insertion of endogenous retrotransposon sequences accounts for approximately 0.2% of disease causing mutations. These insertions are mediated by the reverse transcriptase and endonuclease activity of long interspersed nucleotide (LINE-1) elements. The factors that control the target site selection in insertional mutagenesis are not well understood. In our analysis of 199 unrelated families with proven mutations in BTK, the gene responsible for X-linked agammaglobulinemia, we identified two families with retrotransposon insertions at exactly the same nucleotide within the coding region of BTK. Both insertions, an SVA element and an AluY sequence, occurred 12 bp before the end of exon 9. Both had the typical hallmarks of a retrotransposon insertion including target site duplication and a long poly A tail. The insertion site is flanked by AluSx sequences 1 kb upstream and 1 kb downstream and an unusual 60 bp sequence consisting of only As and Ts is located in intron 9, 60 bp downstream of the insertion. The occurrence of two retrotransposon sequences at exactly the same site suggests that this site is vulnerable to insertional mutagenesis. A better understanding of the factors that make this site vulnerable will shed light on the mechanisms of LINE-1 mediated insertional mutagenesis. PMID:15712380
Conley, Mary Ellen; Partain, Julie D; Norland, Shannon M; Shurtleff, Sheila A; Kazazian, Haig H
A High-Throughput Arabidopsis Reverse Genetics System
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
The favorable transfer free energy for a transmembrane (TM) ?-helix between the aqueous phase and lipid bilayer underlies the stability of membrane proteins. However, the connection between the energetics and process of membrane protein assembly by the Sec61/SecY translocon complex in vivo is not clear. Here, we directly determine the partitioning free energies of a family of designed peptides using three independent approaches: an experimental microsomal Sec61 translocon assay, a biophysical (spectroscopic) characterization of peptide insertion into hydrated planar lipid bilayer arrays, and an unbiased atomic-detail equilibrium folding-partitioning molecular dynamics simulation. Remarkably, the measured free energies of insertion are quantitatively similar for all three approaches. The molecular dynamics simulations show that TM helix insertion involves equilibrium with the membrane interface, suggesting that the interface may play a role in translocon-guided insertion. PMID:25204588
Ulmschneider, Martin B.; Ulmschneider, Jakob P.; Schiller, Nina; Wallace, B. A.; von Heijne, Gunnar; White, Stephen H.
Variational theory of insertion devices
NASA Astrophysics Data System (ADS)
Using constant-action constraints, a variational approach to isolating optimum particle trajectories in insertion devices is presented which is based on assigning performance functions to insertion devices which are functionals of the particle trajectory, and then extremizing the performance functions by varying the particle trajectory. Results from the application of the approach to a concrete example are given. The analysis suggests that the restricted on-axis brightness-enhancing undulator is helical, and it provides a numerical value for the best brightness. The constraints may be of a dynamical or geometrical nature, each leading to the definition of a new type of insertion device.
Tatchyn, R.
Insertional Gene Activation by Lentiviral and Gammaretroviral Vectors? �
Gammaretroviral and lentiviral vectors are promising tools for gene therapy, but they can be oncogenic. The development of safer vectors depends on a quantitative assay for insertional mutagenesis. Here we report a rapid, inexpensive, and reproducible assay which uses a murine cell line to measure the frequency of interleukin-3 (IL-3)-independent mutants. Lentiviral and gammaretroviral vectors cause insertional mutagenesis at similar frequencies; however, they use different mechanisms. Human immunodeficiency virus (HIV)-based vectors generate mutants by insertion only into the growth hormone receptor (Ghr) locus. The HIV enhancer/promoter is active in the absence of the HIV Tat protein in this locus, and an HIV/Ghr spliced transcript expresses GHR and cells respond to GH. Deletion of the enhancer/promoter in a self-inactivating HIV-based vector prevents this mechanism of insertional mutagenesis. In contrast, gammaretroviral vectors insert into other loci, including IL-3 and genes identified as common insertion sites in the Retroviral Tagged Cancer Gene Database (RTCGD). PMID:18945765
Bokhoven, Marieke; Stephen, Sam L.; Knight, Sean; Gevers, Evelien F.; Robinson, Iain C.; Takeuchi, Yasuhiro; Collins, Mary K.
Supplies-(tools such as oral syringes, disposable spoon, cup, gloves, pad or paper towels, etc. needed to measure or prepare the drug.)List preparation instructionsInsert administration instruction (e.g., with or without food).
Peripherally inserted central catheters revisited
Background: This study compares central venous catheters (CVC) and peripherally inserted central catheters (PICC) for indications for insertion, complications, and economic impact.Methods: A retrospective review of 838 (283 CVC, 555 PICC) consecutively placed venous catheters reflected 49,365 CVC and 11,814 PICC days.Results: There were 57 (20%) complications in the CVC group, 197 (35%) complications in the PICC group. PICC were
Jeffrey R. Smith; Mark L. Friedell; Michael L. Cheatham; Samuel P. Martin; Michael J. Cohen; John D. Horowitz
Development of control system to automate the PCB pin insertion process
This thesis describes the development of the control system that runs the automated pin insertion machine in a surface mount technology assembly line. The control system is divided into 2 subsystems viz. pin sorting and ...
Isaac, Rejin
Quantum Diffusion in Polaron Model of poly(dG)-poly(dC) and poly(dA)-poly(dT) DNA polymers
We numerically investigate quantum diffusion of an electron in a model of poly(dG)-poly(dC) and poly(dA)-poly(dT) DNA polymers with fluctuation of the parameters due to the impact of colored noise. The randomness is introduced by fluctuations of distance between two consecutive bases along the stacked base pairs. We demonstrate that in the model the decay time of the correlation can control the spread of the electronic wavepacket. Furthermore it is shown that in a motional narrowing regime the averaging over fluctuation causes ballistic propagation of the wavepacket, and in the adiabatic regime the electronic states are affected by localization.
H. Yamada; E. B. Starikov; D. Hennig
independence will be a challenge. Another possible solution which avoids this problem is to use TEX to create a graphics image or table. This has can be done through the use of macros, LaTEX's picture environment, table macros, or by using Pi... work has been done at describing a minimum set of macros to perform standard graphics functions [12]. PiCTEX was designed to create mathematical figures (lines, curves, plots, etc. . . ) [15). In many respects, it resembles LaTEX's picture...
Kattawar, Gregory Williford
Transposon insertion mapping with PIMMS � Pragmatic Insertional Mutation Mapping System
The PIMMS (Pragmatic Insertional Mutation Mapping System) pipeline has been developed for simple conditionally essential genome discovery experiments in bacteria. Capable of using raw sequence data files alongside a FASTA sequence of the reference genome and GFF file, PIMMS will generate a tabulated output of each coding sequence with corresponding mapped insertions accompanied with normalized results enabling streamlined analysis. This allows for a quick assay of the genome to identify conditionally essential genes on a standard desktop computer prioritizing results for further investigation. Availability: The PIMMS script, manual and accompanying test data is freely available at https://github.com/ADAC-UoN/PIMMS
Blanchard, Adam M.; Leigh, James A.; Egan, Sharon A.; Emes, Richard D.
Gene Insertion Patterns and Sites
NASA Astrophysics Data System (ADS)
During the past 25 years, the molecular analysis of transgene insertion patterns and sites in plants has greatly contributed to our understanding of the mechanisms underlying transgene integration, expression, and stability in the nuclear genome. Molecular characterization is also an essential step in the safety assessment of genetically modified crops. This chapter describes the standard experimental procedures used to analyze transgene insertion patterns and loci in cereals and grasses transformed using Agrobacterium tumefaciens or direct transfer of DNA. Methods and protocols enabling the determination of the number and configuration of transgenic loci via a combination of inheritance studies, polymerase chain reaction, and Southern analyses are presented. The complete characterization of transgenic inserts in plants is, however, a holistic process relying on a wide variety of experimental approaches. In this chapter, these additional approaches are not detailed but references to relevant bibliographic records are provided.
Vain, Philippe; Thole, Vera
Insertional Mutagenesis Using Tnt1 Retrotransposon in Potato1[OPEN
Insertional mutagenesis using transfer DNA or transposable elements, which is an important tool in functional genomics and is well established in several crops, has not been developed in potato (Solanum tuberosum). Here, we report the application of the tobacco (Nicotiana tabacum) Tnt1 retrotransposon as an insertional mutagen in potato. The Tnt1 retrotransposon was introduced into a highly homozygous and self-compatible clone, 523-3, of the diploid wild potato species Solanum chacoense. Transposition of the Tnt1 elements introduced into 523-3 can be efficiently induced by tissue culture. Tnt1 preferentially inserted into genic regions in the potato genome and the insertions were stable during sexual reproduction, making Tnt1 an ideal mutagen in potato. Several distinct phenotypes associated with plant stature and leaf morphology were discovered in mutation screening from a total of 38 families derived from Tnt1-containing lines. We demonstrate that the insertional mutagenesis system based on Tnt1 and the 523-3 clone can be expanded to the genome-wide level to potentially tag every gene in the potato genome. PMID:23898040
Duangpan, Saowapa; Zhang, Wenli; Wu, Yufang; Jansky, Shelley H.; Jiang, Jiming
Pneumothorax as a complication of central venous catheter insertion.
The central venous catheter (CVC) is a catheter placed into a large vein in the neck [internal jugular vein (IJV)], chest (subclavian vein or axillary vein) or groin (femoral vein). There are several situations that require the insertion of a CVC mainly to administer medications or fluids, obtain blood tests (specifically the "central venous oxygen saturation"), and measure central venous pressure. CVC usually remain in place for a longer period of time than other venous access devices. There are situations according to the drug administration or length of stay of the catheter that specific systems are indicated such as; a Hickman line, a peripherally inserted central catheter (PICC) line or a Port-a-Cath may be considered because of their smaller infection risk. Sterile technique is highly important here, as a line may serve as a port of entry for pathogenic organisms, and the line itself may become infected with organisms such as Staphylococcus aureus and coagulase-negative Staphylococci. In the current review we will present the complication of pneumothorax after CVC insertion. PMID:25815301
Tsotsolis, Nikolaos; Tsirgogianni, Katerina; Kioumis, Ioannis; Pitsiou, Georgia; Baka, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Rapti, Aggeliki; Trakada, Georgia; Katsikogiannis, Nikolaos; Tsakiridis, Kosmas; Karapantzos, Ilias; Karapantzou, Chrysanthi; Barbetakis, Nikos; Zissimopoulos, Athanasios; Kuhajda, Ivan; Andjelkovic, Dejan; Zarogoulidis, Konstantinos; Zarogoulidis, Paul
Pneumothorax as a complication of central venous catheter insertion
The central venous catheter (CVC) is a catheter placed into a large vein in the neck [internal jugular vein (IJV)], chest (subclavian vein or axillary vein) or groin (femoral vein). There are several situations that require the insertion of a CVC mainly to administer medications or fluids, obtain blood tests (specifically the �central venous oxygen saturation�), and measure central venous pressure. CVC usually remain in place for a longer period of time than other venous access devices. There are situations according to the drug administration or length of stay of the catheter that specific systems are indicated such as; a Hickman line, a peripherally inserted central catheter (PICC) line or a Port-a-Cath may be considered because of their smaller infection risk. Sterile technique is highly important here, as a line may serve as a port of entry for pathogenic organisms, and the line itself may become infected with organisms such as Staphylococcus aureus and coagulase-negative Staphylococci. In the current review we will present the complication of pneumothorax after CVC insertion.
Tsotsolis, Nikolaos; Tsirgogianni, Katerina; Kioumis, Ioannis; Pitsiou, Georgia; Baka, Sofia; Papaiwannou, Antonis; Karavergou, Anastasia; Rapti, Aggeliki; Trakada, Georgia; Katsikogiannis, Nikolaos; Tsakiridis, Kosmas; Karapantzos, Ilias; Karapantzou, Chrysanthi; Barbetakis, Nikos; Zissimopoulos, Athanasios; Kuhajda, Ivan; Andjelkovic, Dejan; Zarogoulidis, Konstantinos
Electromagnetically tracked placement of a peripherally inserted central catheter
NASA Astrophysics Data System (ADS)
This paper describes a computer program to utilize electromagnetic tracking guidance during insertion of peripherally inserted central catheters. Placement of a Peripherally Inserted Central Catheter (PICC) line is a relatively simple, routine procedure in which a catheter is inserted into the veins of the lower arm and threaded up the arm to the vena cava to sit just above the heart. However, the procedure requires x-ray verification of the catheter position and is usually done under continuous fluoroscopic guidance. The computer program is designed to replace fluoroscopic guidance in this procedure and make PICC line placement a bedside procedure. This would greatly reduce the time and resources dedicated to this procedure. The physician first goes through a quick registration procedure to register the patient space with the computer screen coordinates. Once registration is completed, the program provides a continuous, real-time display of the position of the catheter tip overlaid on an x-ray image of the patient on an adjacent computer screen. Both the position and orientation of the catheter tip is shown. The display is very similar to that shown when using fluoroscopy.
Sacolick, Laura; Patel, Neilesh; Tang, Jonathan; Levy, Elliot; Cleary, Kevin R.
Logo insertion in MPEG transcoder
Various studies show that a basic architecture of an MPEG-2 transcoder consists of a cascaded decoder\\/encoder. Such a transcoder re-uses the motion vectors and minimizes the changes of macroblock mode. One interesting feature that can be added on this type of transcoder is to insert a translucent logo into the transcoded bit-stream
K. Panusopone; Xuemin Chen; Fan Ling
Formation of nanofilms on cell surfaces to improve the insertion efficiency of a nanoneedle into cells
Highlights: Black-Right-Pointing-Pointer We examined the insertion efficiency of nanoneedles into fibroblast and neural cells. Black-Right-Pointing-Pointer Nanofilms formed on cell surfaces improved the insertion efficiency of nanoneedles. Black-Right-Pointing-Pointer Nanofilms improved the insertion efficiency even in Y27632-treated cells. -- Abstract: A nanoneedle, an atomic force microscope (AFM) tip etched to 200 nm in diameter and 10 {mu}m in length, can be inserted into cells with the aid of an AFM and has been used to introduce functional molecules into cells and to analyze intracellular information with minimal cell damage. However, some cell lines have shown low insertion efficiency of the nanoneedle. Improvement in the insertion efficiency of a nanoneedle into such cells is a significant issue for nanoneedle-based cell manipulation and analysis. Here, we have formed nanofilms composed of extracellular matrix molecules on cell surfaces and found that the formation of the nanofilms improved insertion efficiency of a nanoneedle into fibroblast and neural cells. The nanofilms were shown to improve insertion efficiency even in cells in which the formation of actin stress fibers was inhibited by the ROCK inhibitor Y27632, suggesting that the nanofilms with the mesh structure directly contributed to the improved insertion efficiency of a nanoneedle.
Amemiya, Yosuke [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan)] [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan); Kawano, Keiko [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-26 Naka-cho, Koganei, Tokyo 184-8588 (Japan)] [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-26 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Matsusaki, Michiya; Akashi, Mitsuru [Department of Applied Chemistry, Graduate School of Engineering Science, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan)] [Department of Applied Chemistry, Graduate School of Engineering Science, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871 (Japan); Nakamura, Noriyuki [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-26 Naka-cho, Koganei, Tokyo 184-8588 (Japan)] [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-26 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Nakamura, Chikashi, E-mail: firstname.lastname@example.com [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan) [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan); Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-26 Naka-cho, Koganei, Tokyo 184-8588 (Japan)
Photoemission study of poly(dA)-poly(dT) DNA : Experimental and theoretical approach to the electronic density of states
We present results of an ultraviolet photoemission spectroscopy study of artificially synthesized poly(dA)-poly(dT) DNA molecules on $p$-type Si substrates. For comparison, we also present the electronic density of states (DOS) calculated using an \\emph{ab initio} tight-binding method based on density-functional theory (DFT). Good agreement was obtained between experiment and theory. The spectra of DNA networks on the Si substrate showed that the Fermi level of the substrate is located in the middle of the band gap of DNA. The spectra of thick ($\\sim 70$ nm) DNA films showed a downward shift of $\\sim 2$ eV compared to the network samples.
Hiroki Wadati; Kozo Okazaki; Yasuhiro Niimi; Atsushi Fujimori; Hitoshi Tabata; Jared Pikus; James P. Lewis
Focus on peripherally inserted central catheters in critically ill patients.
Venous access devices are of pivotal importance for an increasing number of critically ill patients in a variety of disease states and in a variety of clinical settings (emergency, intensive care, surgery) and for different purposes (fluids or drugs infusions, parenteral nutrition, antibiotic therapy, hemodynamic monitoring, procedures of dialysis/apheresis). However, healthcare professionals are commonly worried about the possible consequences that may result using a central venous access device (CVAD) (mainly, bloodstream infections and thrombosis), both peripherally inserted central catheters (PICCs) and centrally inserted central catheters (CICCs). This review aims to discuss indications, insertion techniques, and care of PICCs in critically ill patients. PICCs have many advantages over standard CICCs. First of all, their insertion is easy and safe -due to their placement into peripheral veins of the arm- and the advantage of a central location of catheter tip suitable for all osmolarity and pH solutions. Using the ultrasound-guidance for the PICC insertion, the risk of hemothorax and pneumothorax can be avoided, as well as the possibility of primary malposition is very low. PICC placement is also appropriate to avoid post-procedural hemorrhage in patients with an abnormal coagulative state who need a CVAD. Some limits previously ascribed to PICCs (i.e., low flow rates, difficult central venous pressure monitoring, lack of safety for radio-diagnostic procedures, single-lumen) have delayed their start up in the intensive care units as common practice. Though, the recent development of power-injectable PICCs overcomes these technical limitations and PICCs have started to spread in critical care settings. Two important take-home messages may be drawn from this review. First, the incidence of complications varies depending on venous accesses and healthcare professionals should be aware of the different clinical performance as well as of the different risks associated with each type of CVAD (CICCs or PICCs). Second, an inappropriate CVAD choice and, particularly, an inadequate insertion technique are relevant-and often not recognized-potential risk factors for complications in critically ill patients. We strongly believe that all healthcare professionals involved in the choice, insertion or management of CVADs in critically ill patients should know all potential risk factors of complications. This knowledge may minimize complications and guarantee longevity to the CVAD optimizing the risk/benefit ratio of CVAD insertion and use. Proper management of CVADs in critical care saves lines and lives. Much evidence from the medical literature and from the clinical practice supports our belief that, compared to CICCs, the so-called power-injectable peripherally inserted central catheters are a good alternative choice in critical care. PMID:25374804
Cotogni, Paolo; Pittiruti, Mauro
An Inverse Pcr Screen for the Detection of P Element Insertions in Cloned Genomic Intervals in Drosophila Melanogaster
We developed a screening approach that utilizes an inverse polymerase chain reaction (PCR) to detect P element insertions in or near previously cloned genes in Drosophila melanogaster. We used this approach in a large scale genetic screen in which P elements were mobilized from sites on the X chromosome to new autosomal locations. Mutagenized flies were combined in pools, and our screening approach was used to generate probes corresponding to the sequences flanking each site of insertion. These probes then were used for hybridization to cloned genomic intervals, allowing individuals carrying insertions in them to be detected. We used the same approach to perform repeated rounds of sib-selection to generate stable insertion lines. We screened 16,100 insert bearing individuals and recovered 11 insertions in five intervals containing genes encoding members of the kinesin superfamily in Drosophila melanogaster. In addition, we recovered an insertion in the region including the Larval Serum Protein-2 gene. Examination by Southern hybridization confirms that the lines we recovered represent genuine insertions in the corresponding genomic intervals. Our data indicates that this approach will be very efficient both for P element mutagenesis of new genomic regions and for detection and recovery of ``local'' P element transposition events. In addition, our data constitutes a survey of preferred P element insertion sites in the Drosophila genome and suggests that insertion sites that are mutable at a rate of ~10(-4) are distributed every 40-50 kb. PMID:7713430
Dalby, B.; Pereira, A. J.; Goldstein, LSB.
Field errors in hybrid insertion devices
Hybrid magnet theory as applied to the error analyses used in the design of Advanced Light Source (ALS) insertion devices is reviewed. Sources of field errors in hybrid insertion devices are discussed.
Schlueter, R.D. [Lawrence Berkeley Lab., CA (United States)
A Case of Anterior Interosseous Nerve Syndrome After Peripherally Inserted Central Catheter (PICC) Line Insertion
Palsies involving the anterior interosseous nerve comprise less than 1% of all upper extremity nerve palsies. Patients often present initially with acute pain in the proximal forearm, lasting several hours to days. The pain subsides, to be followed by paresis or total paralysis of the pronator quadratus, flexor pollicis longus and the radial half of the flexor profundus, either individually
M E Puhaindran; H P Wong
Insert metering plates for gas turbine nozzles
The invention comprises a metering plate which is assembled to an impingement insert for use in the nozzle of a gas turbine. The metering plate can have one or more metering holes and is used to balance the cooling flow within the nozzle. A metering plate with multiple holes reduces static pressure variations which result from the cooling airflow through the metering plate. The metering plate can be assembled to the insert before or after the insert is inserted into the nozzle.
Burdgick, Steven S.; Itzel, Gary; Chopra, Sanjay; Abuaf, Nesim; Correia, Victor H.
Genic DNA methylation changes during in vitro organogenesis: organ specificity and conservation between parental lines of epialleles.
During differentiation, in vitro organogenesis calls for the adjustment of the gene expression program toward a new fate. The role of epigenetic mechanisms including DNA methylation is suggested but little is known about the loci affected by DNA methylation changes, particularly in agronomic plants for witch in vitro technologies are useful such as sugar beet. Here, three pairs of organogenic and non-organogenic in vitro cell lines originating from different sugar beet (Beta vulgaris altissima) cultivars were used to assess the dynamics of DNA methylation at the global or genic levels during shoot or root regeneration. The restriction landmark genome scanning for methylation approach was applied to provide a direct quantitative epigenetic assessment of several CG methylated genes without prior knowledge of gene sequence that is particularly adapted for studies on crop plants without a fully sequenced genome. The cloned sequences had putative roles in cell proliferation, differentiation or unknown functions and displayed organ-specific DNA polymorphism for methylation and changes in expression during in vitro organogenesis. Among them, a potential ubiquitin extension protein 6 (UBI6) was shown, in different cultivars, to exhibit repeatable variations of DNA methylation and gene expression during shoot regeneration. In addition, abnormal development and callogenesis were observed in a T-DNA insertion mutant (ubi6) for a homologous sequence in Arabidopsis. Our data showed that DNA methylation is changed in an organ-specific way for genes exhibiting variations of expression and playing potential role during organogenesis. These epialleles could be conserved between parental lines opening perspectives for molecular markers. PMID:22486767
Maury, St�phane; Trap-Gentil, Marie-V�ronique; H�brard, Claire; Weyens, Guy; Delaunay, Alain; Barnes, Steve; Lefebvre, Marc; Joseph, Claude
JT/LJT connector insert material evaluation
Different insert (insulator) materials are undergoing evaluation to replace the Fiberite E-3938 BE96 material currently used. Also being evaluated is the reconfiguration of the insert and metal shell-edge geometries for the purpose of reducing the alleged interference principally responsible for insert damage.
Baca, J.R.F.
Insertion device calculations with mathematica
The design of accelerator insertion devices such as wigglers and undulators has usually been aided by numerical modeling on digital computers, using code in high level languages like Fortran. In the present era, there are higher level programming environments like IDL{reg_sign}, MatLab{reg_sign}, and Mathematica{reg_sign} in which these calculations may be performed by writing much less code, and in which standard mathematical techniques are very easily used. The authors present a suite of standard insertion device modeling routines in Mathematica to illustrate the new techniques. These routines include a simple way to generate magnetic fields using blocks of CSEM materials, trajectory solutions from the Lorentz force equations for given magnetic fields, Bessel function calculations of radiation for wigglers and undulators and general radiation calculations for undulators.
Carr, R. [Stanford Synchrotron Radiation Lab., CA (United States); Lidia, S. [Univ. of California, Davis, CA (United States)
Keyhole mandrel with insert pocket
A sidepocket mandrel having a one piece body section with a keyhole cross section extending from one end of the body to the other end of the body forms an open bore and an offset bore. A one piece integral insert having a locking shoulder and a sidepocket includes an external shape to mate in the offset bore in the lower portion of the body allowing the upper portion of the offset bore to function as a discriminator.
Johnston, R. A.
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
Analysis of inserts in prokaryote genomes
NASA Astrophysics Data System (ADS)
Nucleotide genomic signals satisfy regularities that reveal restrictions in the distribution of nucleotides and pairs of nucleotides along DNA sequences. Structurally, a chromosome appears to be more than a plain text, by satisfying symmetry constrains that evoke the rhythm and rhyme in poems. These regularities make it easy to identify exogenous inserts in the genomes of prokaryotes, because such inserts obey different regularities than the background sequence. The paper presents instances of inserts found in the genomes of Bacillus subtilis, Mycobacterium tuberculosis and other prokaryotes. Inserts of exogenous material are frequently accompanied by complementary inserts tending to restore the original constrains.
Cristea, Paul Dan; Tuduce, Rodica Aurora
Microneedle insertion force reduction using vibratory actuation.
The effect of vibratory actuation on microneedle insertion force was investigated. Hollow micro hypodermic injection needles were fabricated by a two-wafer polysilicon micromolding process. A vibratory actuator operating in the kHz range was coupled with the hypodermic microneedles. The force to insert microneedles into excized animal tissue was measured with a load cell. Results showed a greater than 70% reduction in microneedle insertion force by using vibratory actuation. The application of vibratory actuation provides a promising method to precisely control the microneedle insertion forces to overcome microneedle structural material limitations, minimize insertion pain, and enhance the efficiency of drug delivery. PMID:15377826
Yang, Ming; Zahn, Jeffrey D
Effects of Single P-Element Insertions on Bristle Number and Viability in Drosophila Melanogaster
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. PMID:8722781
Lyman, R. F.; Lawrence, F.; Nuzhdin, S. V.; Mackay, TFC.
Endonuclease-independent insertion provides an alternative pathway for L1 retrotransposition in the human genome
LINE-1 elements (L1s) are a family of highly suc- cessful retrotransposons comprising �17% of the human genome, the majority of which have inserted through an endonuclease-dependent mechanism termed target-primed reverse transcription. Recent in vitro analyses suggest that in the absence of non- homologous end joining proteins, L1 elements may utilize an alternative, endonuclease-independent pathway for insertion. However, it remains unknown
Shurjo K. Sen; Charles T. Huang; Kyudong Han; Mark A. Batzer
Summary of experimental insertions workshop
The last ISABELLE workshop of the summer 1976 series, which was held at Brookhaven, August 16-20, focused on the design and utilization of the experimental insertions. The goals of the workshop, which were somewhat more general than might be suggested by the title, are: (1) review the ISABELLE proposal from the point of view of experimental use; (2) contribute useful information on the ''open questions'' in the ISABELLE design; (3) develop data for experimental equipment and operating cost estimates; and (4) project a first approximation to ISABELLE operating modes.
Sandweiss, J.; Month, M.
Insertional mutagenesis by transposable elements in the mammalian genome.
Several mammalian repetitive transposable genetic elements were characterized in recent years, and their role in mutagenesis is delineated in this review. Two main groups have been described: elements with symmetrical termini such as the murine IAP sequences and the human THE 1 elements and elements characterized by a poly-A rich tail at the 3' end such as the SINE and LINE sequences. The characteristic property of such mobile elements to spread and integrate in the host genome leads to insertional mutagenesis. Both germline and somatic mutations have been documented resulting from the insertion of the various types of mammalian repetitive transposable genetic elements. As foreseen by Barbara McClintock, such genetic events can cause either the activation or the inactivation of specific genes, resulting in their identification via an altered phenotype. Several disease states, such as hemophilia and cancer, are the result of this apparent aspect of genome instability. PMID:8385004
Amariglio, N; Rechavi, G
Localization properties of electronic states in a polaron model of poly(dG)-poly(dC) and poly(dA)-poly(dT) DNA polymers.
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
Automatic Steering of Manually Inserted Needles
Bevel-tipped flexible needles can be robotically steered to reach clinical targets along curvilinear paths in 3D. Manual needle insertion allows the clinician to control the insertion speed, ensuring patient safety. This paper presents a control law for automatic 3D steering of manually inserted flexible needles, enabling path-following control. A look-ahead proportional controller for position and orientation is presented. The look-ahead distance is a linear function of insertion speed. Simulations in a 3D brain-like environment demonstrate the performance of the proposed controller. Experimental results also show the feasibility of this technique in 2D and 3D environments. PMID:24752485
Wu, Guofan; Li, Xiao; Lehocky, Craig A.; Riviere, Cameron N.
Needle insertion into soft tissue: a survey.
Needle insertion in soft tissue has attracted considerable attention in recent years due to its application in minimally invasive percutaneous procedures such as biopsies and brachytherapy. This paper presents a survey of the current state of research on needle insertion in soft tissue. It examines the topic from several aspects, e.g. modeling needle insertion forces, modeling tissue deformation and needle deflection during insertion, robot-assisted needle insertion, and the effect of different trajectories on tissue deformation. All studies show that the axial force of a needle during insertion in soft tissue is the summation of different forces distributed along the needle shaft such as stiffness force, frictional force and cutting force. Some studies have modeled these forces. The force data in some procedures is used for identifying tissue layers as the needle is inserted or for path planning. Needle deflection and tissue deformation are major problems for accurate needle insertion and attempts have been made to model them. Using current models several insertion techniques have been developed which are briefly reviewed in this paper. PMID:16938481
Abolhassani, Niki; Patel, Rajni; Moallem, Mehrdad
Nozzle insert for mixed mode fuel injector
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)
Target Site Specificity of the Tos17 Retrotransposon Shows a Preference for Insertion within Genes and against Insertion in Retrotransposon-Rich Regions of the Genome
Because retrotransposons are the major component of plant genomes, analysis of the target site selection of retrotransposons is important for understanding the structure and evolution of plant genomes. Here, we examined the target site specificity of the rice retrotransposon Tos17, which can be activated by tissue culture. We have produced 47,196 Tos17-induced insertion mutants of rice. This mutant population carries ?500,000 insertions. We analyzed >42,000 flanking sequences of newly transposed Tos17 copies from 4316 mutant lines. More than 20,000 unique loci were assigned on the rice genomic sequence. Analysis of these sequences showed that insertion events are three times more frequent in genic regions than in intergenic regions. Consistent with this result, Tos17 was shown to prefer gene-dense regions over centromeric heterochromatin regions. Analysis of insertion target sequences revealed a palindromic consensus sequence, ANGTT-TSD-AACNT, flanking the 5-bp target site duplication. Although insertion targets are distributed throughout the chromosomes, they tend to cluster, and 76% of the clusters are located in genic regions. The mechanisms of target site selection by Tos17, the utility of the mutant lines, and the knockout gene database are discussed. PMID:12897251
Miyao, Akio; Tanaka, Katsuyuki; Murata, Kazumasa; Sawaki, Hiromichi; Takeda, Shin; Abe, Kiyomi; Shinozuka, Yoriko; Onosato, Katsura; Hirochika, Hirohiko
Insertion slanting strabismus surgical procedures.
Insertion slanting recessions or biased resections have been reported to be useful for treating A- and V-pattern strabismus, convergence insufficiency, and convergence excess esotropia. Paradoxically, good results have been reported with methods that are opposite in nature. For example, some researchers would recess the medial rectus muscles and slant the superior pole of each muscle back farther than the inferior pole (Simonsz/von Graefe method) for a V-pattern esotropia, and others would slant the inferior poles back farther (Bietti method). The Simonsz/von Graefe method seems to be based on sound concepts of oculomotor mechanics. The Bietti method has been justified based on a misquoting and misinterpretation of previous work by Alan Scott, MD. Probably neither method contributes substantially to the outcome of strabismus surgery because sarcomere remodeling should rapidly negate the effect of the slanting. Most likely it is the recession or resection itself that affects the outcome. PMID:22159685
Kushner, Burton J
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Cho, Sung-Bae
Effects of insertion conditions on tissue strain and vascular damage during neuroprosthetic device insertion
NASA Astrophysics Data System (ADS)
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 assessments of damage were made for insertions using devices with different tip shapes inserted at different speeds. Direct damage to the vasculature included severing, rupturing and dragging, and was often observed several hundred micrometers from the insertion site. Slower insertions generally resulted in more vascular damage. Cortical surface features greatly affected insertion success; insertions attempted through pial blood vessels resulted in severe tissue compression. Automated image analysis techniques were developed to quantify tissue deformation and calculate mean effective strain. Quantitative measures demonstrated that, within the range of experimental conditions studied, faster insertion of sharp devices resulted in lower mean effective strain. Variability within each insertion condition indicates that multiple biological factors may influence insertion success. Multiple biological factors may contribute to tissue distortion, thus a wide variability was observed among insertions made under the same conditions.
Bjornsson, C. S.; Oh, S. J.; Al-Kofahi, Y. A.; Lim, Y. J.; Smith, K. L.; Turner, J. N.; De, S.; Roysam, B.; Shain, W.; Kim, S. J.
Active interlock for storage ring insertion devices
The insertion devices in the NSLS (National Synchrotron Light Source) X-ray ring produce such intense photon beams that passive measures alone are not adequate to protect the vacuum chamber. An active interlock is used to prevent thermal damage from improperly steered electron beams. The interlock system measures the position of the electron beam on both sides of the insertion devices
J. L. Rothman; R. J. Nawrocky
Coaxial needle insertion assistant for epidural puncture
Detection of internal puncture during manual needle insertion is challenging due to the large friction force between a needle shaft and surrounding tissue, which masks small changes in force at the needle tip. A novel robotic coaxial needle insertion assistant was developed to enhance operator perception during epidural puncture. The coaxial needle separates the cutting force at the needle tip
Yoshihiko Koseki; Danilo De Lorenzo; Kiyoyuki Chinzei; Allison M. Okamura
Insert presenter logo here on slide master.
University of Auckland PKI: Lemon Markets and Lemonade Session ID: STAR-304 Session Classification: Intermediate Insert presenter logo here on slide master. See hidden slide 2 for directions Agenda 2 Lemon master. See hidden slide 2 for directions3 Lemon Markets / PKI Markets Insert presenter logo here
Gutmann, Peter
Inserting random and site-specific changes into the genome of chickens.
During the past decade, modifications to the chicken genome have evolved from random insertions of small transgenes using viral vectors to site-specific deletions using homologous recombination vectors and nontargeted insertions of large transgenes using phi-31 integrase. Primordial germ cells (PGC) and gonocytes are the germline-competent cell lines in which targeted modifications and large transgenes are inserted into the genome. After extended periods of in vitro culture, PGC retain their capacity to form functional gametes when reintroduced in vivo. Rates of stable germline modification vary from 1 � 10(-5) for nontargeted insertions to 1 � 10(-8) for targeted insertions. Following transfection, clonally derived cell lines are expanded, injected into Stage 13-15 Hamburger and Hamilton embryos, and putative chimeras are incubated to term in surrogate shells. Green fluorescent protein (GFP) is incorporated into transgenes to reveal the presence of genetically modified PGC in culture and the extent of colonization of the gonad during the first week posthatch. If the extent of colonization is adequate, cohorts of putative chimeras are reared to sexual maturity. Semen is collected and the contribution from donor PGC is estimated by evaluating GFP expression using flow cytometry and PCR. The most promising candidates are selected for breeding to obtain G1 heterozygote offspring. To date, this protocol has been used to (1) knockout the immunoglobulin heavy and light chain genes and produce chickens lacking humoral immunity, (2) insert human V genes and arrays of pseudo V genes into the heavy and light immunoglobulin loci to produce chickens making antibodies with human V regions, (3) insert GFP into nontargeted locations within the genome to produce chickens expressing GFP, and (4) insert Cre recombinase into the genome to produce chickens that excise sequences of DNA flanked by loxP sites. PMID:25828572
Collarini, Ellen; Leighton, Philip; Pedersen, Darlene; Harriman, Bill; Jacob, Roy; Mettler-Izquierdo, Shelley; Yi, Henry; van de Lavoir, Marie-Cecile; Etches, Robert J
Human Lamina Cribrosa Insertion and Age
Purpose. To test the hypothesis that in healthy human eyes the lamina cribrosa (LC) insertion into the pia mater increases with age. Methods. The optic nerve heads (ONHs) of donor eyes fixed at either 5 or 50 mm Hg of IOP were sectioned, stained, and imaged under bright- and dark-field conditions. A 3-dimensional (3D) model of each ONH was reconstructed. From the 3D models we measured the area of LC insertion into the peripapillary scleral flange and into the pia, and computed the total area of insertion and fraction of LC inserting into the pia. Linear mixed effect models were used to determine if the measurements were associated with age or IOP. Results. We analyzed 21 eyes from 11 individuals between 47 and 91 years old. The LC inserted into the pia in all eyes. The fraction of LC inserting into the pia (2.2%�29.6%) had a significant decrease with age (P = 0.049), which resulted from a nonsignificant increase in the total area of LC insertion (P = 0.41) and a nonsignificant decrease in the area of LC insertion into the pia (P = 0.55). None of the measures was associated with fixation IOP (P values 0.44�0.81). Differences between fellow eyes were smaller than differences between unrelated eyes. Conclusions. The LC insertion into the pia mater is common in middle-aged and older eyes, and does not increase with age. The biomechanical and vascular implications of the LC insertion into the pia mater are not well understood and should be investigated further. PMID:22956611
Sigal, Ian A.; Flanagan, John G.; Lathrop, Kira L.; Tertinegg, Inka; Bilonick, Richard
Clonal analysis of heterogeneous crown gall tumor tissues induced by wild-type and shooter mutant strains of Agrobacterium tumefaciens -expression of T-DNA genes
Summary Tumors were induced by anAgrobacterium tumefaciens strain with a wild-type octopine Ti plasmid and by shooter mutants with a transposon insertion in the auxin-locus of the T-region. Cloning of isolated axenic tumor tissues revealed that in all cases they consisted of tumor cells (10�26%) next to a majority of normal cells. The tumor clones that had been induced by
G. M. S. van Slogteren; J. H. C. Hoge; P. J. J. Hooykaas; R. A. Schilperoort
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2010 CFR
...2010-04-01 2010-04-01 false Patient package inserts for estrogens. 310...Specific New Drugs or Devices � 310.515 Patient package inserts for estrogens. (a) Requirement for a patient package insert. FDA concludes...
Insertion and removal of Implanon: practical considerations.
Implanon, a novel contraceptive implant containing the progestogen, etonogestrel (68 mg), provides highly effective, well-tolerated contraception for up to 3 years. Its single-rod design, semi-rigid polymer base and convenient pre-loaded presentation mean that Implanon has potential insertion and removal advantages over the six-capsule Norplant implant. Insertion of Implanon is simple: the applicator needle is positioned subdermally and the cannula is withdrawn, leaving the implant rod in place. Implanon is removed using the 'pop out' technique, involving a 2-mm incision. A meta-analysis of data from seven open-label, randomized studies in 1,378 women compared the ease of insertion and removal of the Implanon and Norplant implants and the frequency of associated complications. It was approximately four times quicker to insert and remove Implanon than Norplant (mean insertion times 1.1 vs. 4.3 min, respectively; mean removal times 2.6 vs.10.2 min, respectively). Insertion complications were very rare with both Implanon (0.3%) and Norplant (0.0%). However, Implanon was associated with a significantly lower frequency of removal complications (0.2 vs. 4.8% with Norplant; p < 0.001). Implant site abnormalities were uncommon. The simplicity of the insertion and removal procedures with Implanon may have significant implications for contraceptive implant training programs and user counselling. PMID:11246605
Mascarenhas, L
Insertion sequence elements in Lactococcus garvieae.
Insertion sequences are the simplest intracellular Mobile Genetic Elements which can occur in very high numbers in prokaryotic genomes, where they play an important evolutionary role by promoting genome plasticity. As such, the studies on the diversity and distribution of insertion sequences in genomes not yet investigated can contribute to improve the knowledge on a bacterial species and to identify new transposable elements. The present work describes the occurrence of insertion sequences in Lactococcus garvieae, an opportunistic emerging zoonotic and human pathogen, also associated with different food matrices. To date, no insertion elements have been described for L. garvieae in the IS element database. The analysis of the twelve published L. garvieae genomes identified 15 distinct insertion sequences that are members of the IS3, IS982, IS6, IS21 and IS256 families, including five new elements. Most of the insertion sequences in L. garvieae show substantial homology to the Lactococcus lactis elements, suggesting the movement of IS between these two species phylogenetically closely related. ISLL6 elements belonging to IS3 family were most abundant, with several copies distributed in 9 of the 12 genomes analyzed. An alignment analysis of two complete genomes carrying multi-copies of this insertion sequence indicates a possible involvement of ISLL6 in chromosomal rearrangement. PMID:25447909
Eraclio, Giovanni; Ricci, Giovanni; Fortina, Maria Grazia
Insertion of an SVA element, a nonautonomous retrotransposon, in PMS2 intron 7 as a novel cause of Lynch syndrome.
Heterozygous germline mutations in the mismatch repair gene PMS2 predispose carriers for Lynch syndrome, an autosomal dominant predisposition to cancer. Here, we present a LINE-1-mediated retrotranspositional insertion in PMS2 as a novel mutation type for Lynch syndrome. This insertion, detected with Southern blot analysis in the genomic DNA of the patient, is characterized as a 2.2 kb long 5' truncated SVA_F element. The insertion is not detectable by current diagnostic testing limited to MLPA and direct Sanger sequencing on genomic DNA. The molecular nature of this insertion could only be resolved in RNA from cultured lymphocytes in which nonsense-mediated RNA decay was inhibited. Our report illustrates the technical problems encountered in the detection of this mutation type. Especially large heterozygous insertions will remain unnoticed because of preferential amplification of the smaller wild-type allele in genomic DNA, and are probably underreported in the mutation spectra of autosomal dominant disorders. PMID:22461402
van der Klift, Heleen M; Tops, Carli M; Hes, Frederik J; Devilee, Peter; Wijnen, Juul T
The BDGP gene disruption project: Single transposon insertions associated with 40 percent of Drosophila genes
The Berkeley Drosophila Genome Project (BDGP) strives to disrupt each Drosophila gene by the insertion of a single transposable element. As part of this effort, transposons in more than 30,000 fly strains were localized and analyzed relative to predicted Drosophila gene structures. Approximately 6,300 lines that maximize genomic coverage were selected to be sent to the Bloomington Stock Center for public distribution, bringing the size of the BDGP gene disruption collection to 7,140 lines. It now includes individual lines predicted to disrupt 5,362 of the 13,666 currently annotated Drosophila genes (39 percent). Other lines contain an insertion at least 2 kb from others in the collection and likely mutate additional incompletely annotated or uncharacterized genes and chromosomal regulatory elements. The remaining strains contain insertions likely to disrupt alternative gene promoters or to allow gene mis-expression. The expanded BDGP gene disruption collection provides a public resource that will facilitate the application of Drosophila genetics to diverse biological problems. Finally, the project reveals new insight into how transposons interact with a eukaryotic genome and helps define optimal strategies for using insertional mutagenesis as a genomic tool.
Bellen, Hugo J.; Levis, Robert W.; Liao, Guochun; He, Yuchun; Carlson, Joseph W.; Tsang, Garson; Evans-Holm, Martha; Hiesinger, P. Robin; Schulze, Karen L.; Rubin, Gerald M.; Hoskins, Roger A.; Spradling, Allan C.
Upper extremity deep vein thrombosis: a complication of an indwelling peripherally inserted central venous catheter
Key Clinical Message We report a case of peripherally inserted central venous catheter (PICC)-associated deep vein thrombosis (DVT). Ultrasound images and video of subclavian thrombus are presented. PICC line-associated DVT, particularly in cancer patients is not uncommon. Point-of-care Emergency Department ultrasound can readily diagnose this complication and device removal is not always necessary.
Carr, Peter J; Rippey, James C R
Experience with Phase II insertion devices in the x-ray ring
The installation of Phase II insertion devices in the NSLS x-ray ring was completed in February 1989, when the hybrid wiggler magnet (HBW) for beam line X21 and the superconducting wiggler (SCW) for beam line X17 were installed. Prior to this time, the soft x-ray undulator (SXU) and X1, another hybrid wiggler at X25, and the ten period mini-undulator at X13 had been installed. 8 refs., 4 figs., 2 tabs.
Decker, G.; Galayda, J.; Krinsky, S.; Solomon, L.
FOA Lecture 16: Insertion Loss Testing
NSDL National Science Digital Library
This is Lecture 16 in the FOA series on fiber optics. This lecture is about insertion loss testing. Insertion loss testing tests an installed cable plant with a light source and power meter in a method that is similar to how a communications system transmits data over the cable plant. Insertion loss testing is used to test the cable plant and compare it to the loss budget calculated in the design phase to ensure the planned communications systems will work over the cable plant. Running time for the lecture is 15:50. Flash is required to view the video.
Elliptically polarizing adjustable phase insertion device
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)
euL1db: the European database of L1HS retrotransposon insertions in humans
Retrotransposons account for almost half of our genome. They are mobile genetics elements�also known as jumping genes�but only the L1HS subfamily of Long Interspersed Nuclear Elements (LINEs) has retained the ability to jump autonomously in modern humans. Their mobilization in germline�but also some somatic tissues�contributes to human genetic diversity and to diseases, such as cancer. Here, we present euL1db, the European database of L1HS retrotransposon insertions in humans (available at http://euL1db.unice.fr). euL1db provides a curated and comprehensive summary of L1HS insertion polymorphisms identified in healthy or pathological human samples and published in peer-reviewed journals. A key feature of euL1db is its sample-wise organization. Hence L1HS insertion polymorphisms are connected to samples, individuals, families and clinical conditions. The current version of euL1db centralizes results obtained in 32 studies. It contains >900 samples, >140 000 sample-wise insertions and almost 9000 distinct merged insertions. euL1db will help understanding the link between L1 retrotransposon insertion polymorphisms and phenotype or disease. PMID:25352549
Mir, Ashfaq A.; Philippe, Claude; Cristofari, Ga�l
Single connector provides safety fuses for multiple lines
NASA Technical Reports Server (NTRS)
Fuse-bearing sleeve which is inserted between the male and female members of a multiple-line connector contains a safety fuse for each pin of the connector assembly. The sleeve is easily and quickly opened for fuse replacement.
Weber, G. J.
Reading between the LINEs: Human Genomic Variation Induced by LINE1 Retrotransposition
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
Peripherally inserted central catheter - dressing change
PICC - dressing change ... You have a peripherally inserted central catheter (PICC). This is a tube that goes into a vein in your arm. It carries nutrients and medicines into your body. It is also ...
Insertable fluid flow passage bridgepiece and method
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)
Shrink-Fit Solderable Inserts Seal Hermetically
NASA Technical Reports Server (NTRS)
Shrink-fit stainless-steel insert in aluminum equipment housing allows electrical connectors to be replaced by soldering, without degrading hermeticity of housing or connector. Welding could destroy electrostatic-sensitive components and harm housing and internal cables. Steel insert avoids problems because connector soldered directly to it rather than welded to housing. Seals between flange and housing, and between connector and flange resistant to leaks, even after mechanical overloading and thermal shocking.
Croucher, William C.
Electrochemical insertion of sodium into carbon
Electrochemical insertion of sodium ions into carbon using solid polymer electrolytes or organic liquid electrolytes is described. Cells with the configuration Na P(EO)(sub *)NaCF3SO3 CP(EO) = polyethylene oxide or Na\\/liquid electrolyte\\/C were galvanostatically discharged, charged, and cycled. The extent of insertion into C (i.e., x in Na(+)C(sub x -)) was found to be a strong function of the type and
Marca M. Doeff; Yanping Ma; Steven J. Visco; Lutgard C. Dejonghe
Compliant Control of Threaded Fastener Insertion
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
Gluteus minimus: observations on its insertion.
In 17 adult and 3 full term fetuses the hip joints were dissected to expose the insertion of gluteus minimus. An attachment of the deep surface of the tendon as it passes over the hip joint capsule was found in every case. Histological examination of this insertion confirmed the presence of short intramuscular tendons firmly anchoring the tendon to the capsule. It is suggested that this attachment retracts the capsule during hip joint motion, thereby preventing capsular entrapment. PMID:11273048
Walters, J; Solomons, M; Davies, J
Bioinformatics of High-Throughput Insertional Mutagenesis
\\u000a Bioinformatics plays critical roles to handle large amount of sequence data from insertional mutagenesis. First, computational\\u000a approaches are used to develop rapid sequence analysis pipelines and biological databases. Millions of reads from an insertion\\u000a mutagenesis screening are mapped to genomic locations and be annotated to their target genes rapidly by pipeline, and such\\u000a sequence-based data is stored and managed in
Keiko Akagi; Ming Yi; Jean Roayaei; Robert M. Stephens
Efficient instruction sequencing with Inline Target Insertion
NASA Technical Reports Server (NTRS)
Inline target insertion, a specific compiler and pipeline implementation method for delayed branches with squashing, is defined. The method is shown to offer two important features not discovered in previous studies. First, branches inserted into branch slots are correctly executed. Second, the execution returns correctly from interrupts or exceptions with only one program counter. These two features result in better performance and less software/hardware complexity than conventional delayed branching mechanisms.
Hwu, Wen-Mei W.; Chang, Pohua P.
Rectal suppository: commonsense and mode of insertion.
Rectal suppository is a well-known form of medication and its use is increasing. The commonest shape is one with an apex (pointed end) tapering to a base (blunt end). Because of a general lack of information about mode of insertion, we asked 360 lay subjects (Egyptians and non-Egyptians) and 260 medical personnel (physicians, pharmacists, and nurses) by questionnaire which end they inserted foremost. Apart from 2 individuals, all subjects suggested insertion with the apex foremost. Commonsense was the most frequent basis for this practice (86.9% of lay subjects and 84.6% of medical personnel) followed by information from a relative, a friend, or medical personnel, or from study at medical school. Suppository insertion with the base or apex foremost was compared in 100 subjects (60 adults, 40 infants and children). Retention with the former method was more easily achieved in 98% of the cases, with no need to introduce a finger in the anal canal (1% vs 83%), and lower expulsion rate (0% vs 3%). The designer of the "torpedo-shaped" suppository suggested its insertion with apex foremost. Our data suggest that a suppository is better inserted with the base foremost. Reversed vermicular contractions or pressure gradient of the anal canal might press it inwards. PMID:1681170
Abd-el-Maeboud, K H; el-Naggar, T; el-Hawi, E M; Mahmoud, S A; Abd-el-Hay, S
Central Solenoid Insert Technical Specification
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
A large-scale collection of phenotypic data describing an insertional mutant population to facilitate functional analysis of rice genes
In order to facilitate the functional analysis of rice genes, we produced about 50,000 insertion lines with the endogenous\\u000a 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\\u000a more than one mutant phenotype. The most frequently observed phenotype was low fertility,
Akio Miyao; Yukimoto Iwasaki; Hidemi Kitano; Jun-Ichi Itoh; Masahiko Maekawa; Kazumasa Murata; Osamu Yatou; Yasuo Nagato; Hirohiko Hirochika
Nozzle cavity impingement/area reduction insert
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)
We review the properties and uses of cell lines in Drosophila research, emphasizing the variety of lines, the large body of genomic and transcriptional data available for many of the lines, and the variety of ways the lines have been used to provide tools for and insights into the developmental, molecular, and cell biology of Drosophila and mammals. PMID:24434506
Cherbas, Lucy; Gong, Lei
Toolbox for mobile-element insertion detection on cancer genomes.
Mobile elements constitute greater than 45% of the human genome as a result of repeated insertion events during human genome evolution. Although most of mobile elements are fixed within the human population, some elements (including ALU, long interspersed elements (LINE) 1 (L1), and SVA) are still actively duplicating and may result in life-threatening human diseases such as cancer, motivating the need for accurate mobile-element insertion (MEI) detection tools. We developed a software package, TANGRAM, for MEI detection in next-generation sequencing data, currently serving as the primary MEI detection tool in the 1000 Genomes Project. TANGRAM takes advantage of valuable mapping information provided by our own MOSAIK mapper, and until recently required MOSAIK mappings as its input. In this study, we report a new feature that enables TANGRAM to be used on alignments generated by any mainstream short-read mapper, making it accessible for many genomic users. To demonstrate its utility for cancer genome analysis, we have applied TANGRAM to the TCGA (The Cancer Genome Atlas) mutation calling benchmark 4 dataset. TANGRAM is fast, accurate, easy to use, and open source on https://github.com/jiantao/Tangram. PMID:25452688
Lee, Wan-Ping; Wu, Jiantao; Marth, Gabor T
Toolbox for Mobile-Element Insertion Detection on Cancer Genomes
Mobile elements constitute greater than 45% of the human genome as a result of repeated insertion events during human genome evolution. Although most of mobile elements are fixed within the human population, some elements (including ALU, long interspersed elements (LINE) 1 (L1), and SVA) are still actively duplicating and may result in life-threatening human diseases such as cancer, motivating the need for accurate mobile-element insertion (MEI) detection tools. We developed a software package, TANGRAM, for MEI detection in next-generation sequencing data, currently serving as the primary MEI detection tool in the 1000 Genomes Project. TANGRAM takes advantage of valuable mapping information provided by our own MOSAIK mapper, and until recently required MOSAIK mappings as its input. In this study, we report a new feature that enables TANGRAM to be used on alignments generated by any mainstream short-read mapper, making it accessible for many genomic users. To demonstrate its utility for cancer genome analysis, we have applied TANGRAM to the TCGA (The Cancer Genome Atlas) mutation calling benchmark 4 dataset. TANGRAM is fast, accurate, easy to use, and open source on https://github.com/jiantao/Tangram. PMID:25452688
Lee, Wan-Ping; Wu, Jiantao; Marth, Gabor T
Mosaic retroposon insertion patterns in placental mammals.
One and a half centuries after Charles Darwin and Alfred Russel Wallace outlined our current understanding of evolution, a new scientific era is dawning that enables direct observations of genetic variation. However, pure sequence-based molecular attempts to resolve the basal origin of placental mammals have so far resulted only in apparently conflicting hypotheses. By contrast, in the mammalian genomes where they were highly active, the insertion of retroelements and their comparative insertion patterns constitute a neutral, virtually homoplasy-free archive of evolutionary histories. The "presence" of a retroelement at an orthologous genomic position in two species indicates their common ancestry in contrast to its "absence" in more distant species. To resolve the placental origin controversy we extracted approximately 2 million potentially phylogenetically informative, retroposon-containing loci from representatives of the major placental mammalian lineages and found highly significant evidence challenging all current single hypotheses of their basal origin. The Exafroplacentalia hypothesis (Afrotheria as the sister group to all remaining placentals) is significantly supported by five retroposon insertions, the Epitheria hypothesis (Xenarthra as the sister group to all remaining placentals) by nine insertion patterns, and the Atlantogenata hypothesis (a monophyletic clade comprising Xenarthra and Afrotheria as the sister group to Boreotheria comprising all remaining placentals) by eight insertion patterns. These findings provide significant support for a "soft" polytomy of the major mammalian clades. Ancestral successive hybridization events and/or incomplete lineage sorting associated with short speciation intervals are viable explanations for the mosaic retroposon insertion patterns of recent placental mammals and for the futile search for a clear root dichotomy. PMID:19261842
Churakov, Gennady; Kriegs, Jan Ole; Baertsch, Robert; Zemann, Anja; Brosius, J�rgen; Schmitz, J�rgen
RERTR-12 Insertion 2 Irradiation Summary Report
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
Processed pseudogene insertions in somatic cells
Processed pseudogenes are copies of messenger RNAs that have been reverse transcribed into DNA and inserted into the genome using the enzymatic activities of active L1 elements. Processed pseudogenes generally lack introns, end in a 3� poly A, and are flanked by target site duplications. Until recently, very few polymorphic processed pseudogenes had been discovered in mammalian genomes. Now several studies have found a number of polymorphic processed pseudogenes in humans. Moreover, processed pseudogenes can occur in somatic cells, including in various cancers and in early fetal development. One recent somatic insertion of a processed pseudogene has caused a Mendelian X-linked disease, chronic granulomatous disease. PMID:25184004
Rare complication of a dialysis catheter insertion
Insertion of a dialysis catheter is a very common procedure in renal medicine, which is associated with one or more complications in ?15% of patients. Central venous catheter vascular erosion is a rare but serious complication, the incidence appears to be between 0.4 and 1% which can be difficult to diagnose, leading to increased morbidity and mortality. We report a case of a persistent left-sided superior vena cava (PLSVC) and a possible central venous catheter vascular erosion following insertion of a left-sided internal jugular dialysis catheter. We discuss the mechanisms, diagnosis and measures to limit vascular erosion and also briefly discuss PLSVC.
Balasubramanian, Santhanakrishnan; Gupta, Sanjay; Nicholls, Marcus; Laboi, Paul
Circular permutant GFP insertion folding reporters
Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.
Waldo, Geoffrey S. (Santa Fe, NM); Cabantous, Stephanie (Los Alamos, NM)
Circular permutant GFP insertion folding reporters
Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.
Waldo, Geoffrey S.; Cabantous, Stephanie
Circular permutant GFP insertion folding reporters
Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.
Waldo, Geoffrey S. (Santa Fe, NM); Cabantous, Stephanie (Los Alamos, NM)
Circular permutant GFP insertion folding reporters
Provided are methods of assaying and improving protein folding using circular permutants of fluorescent proteins, including circular permutants of GFP variants and combinations thereof. The invention further provides various nucleic acid molecules and vectors incorporating such nucleic acid molecules, comprising polynucleotides encoding fluorescent protein circular permutants derived from superfolder GFP, which polynucleotides include an internal cloning site into which a heterologous polynucleotide may be inserted in-frame with the circular permutant coding sequence, and which when expressed are capable of reporting on the degree to which a polypeptide encoded by such an inserted heterologous polynucleotide is correctly folded by correlation with the degree of fluorescence exhibited.
Waldo, Geoffrey S; Cabantous, Stephanie
ALS insertion device block measurement and inspection
The performance specifications for ALS insertion devices require detailed knowledge and strict control of the Nd-Fe-B permanent magnet blocks incorporated in these devices. This paper describes the measurement and inspection apparatus and the procedures designed to qualify and characterize these blocks. A detailed description of a new, automated Helmholtz coil facility for measurement of the three components of magnetic moment is included. Physical block inspection and magnetic moment measurement procedures are described. Together they provide a basis for qualifying blocks and for specifying placement of blocks within an insertion devices' magnetic structures. 1 ref., 4 figs.
Marks, S.; Carrieri, J.; Cook, C.; Hassenzahl, W.V.; Hoyer, E.; Plate, D.
Neutron absorber inserts for 55-gal drums
Transport and temporary storage of more than 200 g of fissile material in 55-gal drums at the Rocky Flats Environmental Technology Site (RFETS) have received significant attention during the cleanup mission. This paper discusses successful applications and results of extensive computer studies. Interim storage and movement of fissile material in excess of standard drum limits (200 g) in a safe configuration have been accomplished using special drum inserts. Such inserts have constrained the contents of a drum to two 4-{ell} bottles. The content of the bottles was limited to 600 g Pu or U in solution or a total of 1200 g for the entire drum. The inserts were a simple design constructed of stainless steel, forming a vertical cylindrical pipe into which two bottles, one on top of the other, could be centered in the drum. The remaining drum volume was configured to preclude any additional bottle placement external to the vertical cylinder. Such inserts in drums were successfully used in moving high-concentration solution from one building to another for chemical processing. Concern about the knowledge of fissile material concentration in bottles prompted another study for drum inserts. The past practice had been to load up to fourteen 4-{ell} bottles into 55-gal drums, provided the fissile material concentration was <6 g fissile/{ell}, and the total drum contents of 200 g fissile was not exceeded. Only one determination of the solution concentration was needed. An extensive safety analysis concluded that a single measurement of bottle content could not ensure compliance with double-contingency-criterion requirements. A second determination of the bottle contents was required before bottles could be placed in a 55-gal drum. Al alternative to a dual-measurement protocol, which is for bolstering administrative control, was to develop an engineered safety feature that would eliminate expensive tests and administrative decisions. A drum insert design was evaluated that would ensure subcriticality in a drum even if the concentration of fissile material solution exceeded 6 g/{ell}. The challenge posed to the study was to maximize the number of bottles with a concentration up to 150 g/{ell} that could be safely placed into a drum. A concentration of 150 g of fissile material per liter is the maximum expected solution concentration in bottles at RFETS. A series of computer analyses were undertaken to design special drum inserts. A design was completed for a 55-gal drum insert to hold up to ten bottles containing 600 g fissile material in solution per bottle or 6 kg per drum. This insert would protect against accidental criticality caused by misloading high-concentration-solution bottles into a drum designated for low-concentration-bottle transport. The drum with the inserts could be used for transport or temporary storage of high-concentration solutions in 4-{ell} bottles. Conceptually, the drum with its inserts could also be employed for interim storage and transport of fissile material in forms other than solutions in 4-{ell} bottles.
Wilson, R.E.; Kim, Y.S.; Toffer, H.
77 GHz VCO Design Using a Coupled-Line Inductor with Spiral-Shaped EBG Structure
area [1] or to design a slow-wave microstrip line with low insertion loss and wide stop-band [277 GHz VCO Design Using a Coupled-Line Inductor with Spiral-Shaped EBG Structure Jae-Hoon Song1-line inductor with spiral-shaped electromagnetic band- gap (EBG) structure. The coupled-line inductor enhances Q
Nam, Sangwook
Immediate postabortal insertion of intrauterine devices.
Background The use of an effective contraceptive may be necessary after an abortion. Insertion of an intrauterine device (IUD) may be done the same day or later. Immediate IUD insertion is an option since the woman is not pregnant, pain of insertion is less because the cervical os is open, and her motivation to use contraception may be high. However, insertion of an IUD immediately after a pregnancy ends carries risks, such as spontaneous expulsion.Objectives To assess the safety and efficacy of IUD insertion immediately after spontaneous or induced abortion.Search methods We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, POPLINE, ClinicalTrials.gov,and ICTRP in January 27, 2014. We also contacted investigators to identify other trials.Selection criteria We sought all randomised controlled trials (RCTs) with at least one treatment arm that involved IUD insertion immediately after an induced abortion or after curettage for spontaneous abortion.Data collection and analysis We evaluated the methodological quality of each report and abstracted the data. We focused on discontinuation rates for accidental pregnancy, perforation, expulsion, and pelvic inflammatory disease.We computed the weighted average of the rate ratios.We compute drisk ratios (RRs) with 95% Confidence Intervals (CIs).We performed an intention-to-treat (ITT) analysis by including all randomised participants in the analysis according to the Cochrane Handbook for Systematic Reviews of Interventions.Main results We identified 12 trials most of which are of moderate risk of bias involving 7,119 participants which described random assignment.Five trials randomised to either immediate or delayed insertion of IUD. One of them randomised to immediate versus delayed insertion of Copper 7 showed immediate insertion of the Copper 7 was associated with a higher risk of expulsion than was delayed insertion(RR 11.98, 95% CI 1.61 to 89.35,1 study, 259 participants); the quality of evidence was moderate. Moderate quality of evidence also suggests that use and expulsion of levonorgestrel-releasing intrauterine system or CuT380A was more likely for immediate compared to delayed insertion risk ratio (RR) 1.40 (95% CI 1.24 to 1.58; 3 studies; 878 participants) and RR 2.64 ( 95% CI 1.16 to 6.00; 3 studies; 878 participants) respectively. Another trial randomised to the levonorgestrel IUD or Nova T showed discontinuation rates due to pregnancy were likely to be higher for women in the Nova T group. (MD 8.70, 95% CI 3.92 to 13.48;1 study; 438 participants);moderate quality evidence.Seven trials examined immediate insertion of IUD only. From meta-analysis of two multicentre trials, pregnancy was less likely for the TCu 220C versus the Lippes Loop (RR 0.43, 95% CI 0.24 to 0.75; 2 studies; 2257 participants ) as was expulsion (RR 0.61, 95% CI0.46 to 0.81; 2 studies; 2257 participants). Estimates for the TCu 220 versus the Copper 7 were RR 0.42 ( 95% CI 0.23 to 0.77; 2 studies, 2,274 participants) and RR 0.68, (95% CI 0.51 to 0.91); 2 studies, 2,274 participants), respectively. In other work, adding copper sleeves to the Lippes Loop improved efficacy (RR 3.40, 95% CI 1.28 to 9.04, 1 study, 400 participants) and reduced expulsion(RR 3.00, 95% CI 1.51 to 5.97; 1 study, 400 participants).Authors' conclusions Moderate quality evidence shows that insertion of an IUD immediately after abortion is safe and practical. IUD expulsion rates appear higher immediately after abortions compared to delayed insertions. However, at six months postabortion, IUD use is higher following immediate insertion compared to delayed insertion. PMID:25101364
Okusanya, Babasola O; Oduwole, Olabisi; Effa, Emmanuel E
Insertion Loss of Personal Protective Clothing
'The use of personal protective clothing that covers the head is a common practice in many industries. Such personal protective clothing will impact the sound pressure level and the frequency content of sounds to which the wearer will be exposed. The use of such clothing, then, may impact speech and alarm audibility. A measure of the impact of such clothing is its insertion loss. Insertion loss measurements were performed on four types of personal protective clothing in use by Westinghouse Savannah River Company personnel which utilize cloth and plastic hood configurations to protect the head. All clothing configurations tested at least partially cover the ears. The measurements revealed that insertion loss of the items tested was notable at frequencies above 1000 Hz only and was a function of material stiffness and acoustic flanking paths to the ear. Further, an estimate of the clothing''s noise reduction rating reveals poor performance in that regard, even though the insertion loss of the test articles was significant at frequencies at and above 1000 Hz.'
Shull D.J.; Biesel, V.B.; Cunefare, K.A.
Embedded Multiprocessor Technology for VHSIC Insertion
NASA Technical Reports Server (NTRS)
Viewgraphs on embedded multiprocessor technology for VHSIC insertion are presented. The objective was to develop multiprocessor system technology providing user-selectable fault tolerance, increased throughput, and ease of application representation for concurrent operation. The approach was to develop graph management mapping theory for proper performance, model multiprocessor performance, and demonstrate performance in selected hardware systems.
Hayes, Paul J.
Inserting new technology into small missions
NASA Technical Reports Server (NTRS)
Part of what makes small missions small is that they have less money. Executing missions at low cost implies extensive use of cost sharing with other missions or use of existing solutions. Luckily, there are methods for creating new technology and inserting it into faster-better-cheaper missions.
Deutsch, L. J.
Lymphatic Leak Complicating Central Venous Catheter Insertion
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: email@example.com; Kleidon, Tricia M. [Great Ormond Street Hospital for Children, Department of Radiology (United Kingdom)
Online Advertising Insertion Order Form Company name
Online Advertising Insertion Order Form Advertiser Company name: Address: City: State: Zip: Contact: Billing Instructions Gross Amount : $ Net Amount: $ Please refer to 2014 Advertising Rate Sheet Less 15% agency discount or other applicable discount (please specify below): Send invoices to: Advertiser
Butler, Laurie J.
Insert your NHS Logo Here Homeopathic Services
Insert your NHS Logo Here 1 Homeopathic Services 1 Summary This paper outlines the rationale for an approach to the commissioning of Homeopathic services by [NHS commissioning organisation]. The majority to commission new referrals to homeopathic services, and provides for six months treatment for those who
Colquhoun, David
Hall of Ocean Life Educator's Guide Insert
NSDL National Science Digital Library
The museum's Milstein Family Hall of Ocean Life explores the diverse, complex web of life supported by the ocean and the vital inter-relationships between human and aquatic systems. This insert to the hall guide is designed to help you maximize your trip to the museum.
ANL/APS/TB53 Insertion Devices
is the calibration system [4]. This system consists of the calibration electromagnet and a set of Nuclear Magnetic moments of individual permanent magnets [5]. ID Magnetic Specifications An electron beam passing throughANL/APS/TB53 Insertion Devices: Magnetic Measurements and Tuning I. Vasserman, E. Gluskin, J
Kemner, Ken
Lymphatic Leak Complicating Central Venous Catheter Insertion
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
Frequency, Gradience, and Variation in Consonant Insertion
ERIC Educational Resources Information Center
This dissertation addresses the extent to which linguistic behavior can be described in terms of the projection of patterns from existing lexical items, through an investigation of Korean reduplication. Korean has a productive pattern of reduplication in which a consonant is inserted in a vowel-initial base, illustrated by forms such as "alok"--"t�
An, Young-ran
Insert or gene Laboratory Gene transfer vectora
Insert or gene Laboratory Gene transfer vectora Host rangeb functionc containment leveld MMLV based, CC BSL-2 including neurons TX BSL-2+/BSL-3 Poxvirus based- Broad host range S, E, M, T, DR, MP BSL-2 cells. c General categories of cellular genes and functions: S, structural proteins: actin, myosin, etc
Kay, Mark A.
Systematic reverse genetics of transfer-DNA-tagged lines of Arabidopsis
The authors have developed an efficient reverse-genetics protocol that uses expedient pooling and hybridization strategies to identify individual transfer-DNA insertion lines from a collection of 6,000 independently transformed lines in as few as 36 polymerase chain reactions. The authors have used this protocol to systematically isolate Arabidopsis lines containing insertional mutations in individual cytochrome P450 genes. In higher plants P450
Rodney G. Winkler; Michael R. Frank; David W. Galbraith; R. Feyereisen; K. A. Feldmann
Technology assessment for CVD-diamond-coated cutting tool inserts
This study assesses the potential for manufacturing diamond-coated, cutting tool inserts by plasma-enhanced chemical vapor deposition (PA-CVD), an emerging technology expected to compete with the static, high-pressure sintering method presently used for the commercial production of polycrystalline diamond composite (PDC) coatings on tool inserts. Using a 4-step manufacturing model based upon microwave PA-CVD for diamond growth, insert manufacturing costs were estimated for three technology scenarios, representing the present (1988, 1st generation) near-term (1990-1999, 2nd generation), and possible long-term (2,000+, 3rd generation) development of diamond PA-CVD technology. Model calculations were then analyzed to provide research guidance on insert manufacturing characteristics, insert performance, and insert life required for the development of markets for thin (1-25 micron) and thick (500-1,000 micron) CVD-diamond-coated inserts. Analysis shows that present technology requires a prohibitively large investment in multiple reactors to grow thin-diamond-coated inserts in commercial quantities. However, technology now under development shows promise for the economical manufacture of thin-diamond-coated inserts, provided tough, adherent coatings can be achieved at high yield. The US market for thin-diamond-coated WC-Co inserts could approach 2M inserts ($25M) by the year 2000. Thick, CVD-diamond-coated inserts (500-1,000 microns), necessary for the most demanding cutting applications, will require development of new technology to be competitive with sintered PDC inserts.
Walton, K.R.; Dismukes, J.P. (Exxon Research and Engineering Co., Annandale, NJ (United States)); Krueger, R.A.; Field, F.R. III; Clark, J.P. (Massachusetts Inst. of Tech., Cambridge (United States))
Effects of insertion conditions on tissue strain and vascular damage during neuroprosthetic device insertion
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
BREEDING & GENETICS Linkage Analysis of Transgenes Inserted into Cotton via Agrobacterium tumefaciens Transformation
The location of transgenes inserted into a genome are important in genetic studies and breeding programs. We conducted linkage analysis between 2,4-D resistant transgenes and 14 morphological marker genes in upland cotton (Gossypium hirsutum L.). Two separate germlines that exhibited monogenic dominance for resistance to 2,4-D were selected for linkage analysis. Multiple marker lines T582 and T586 were crossed with
Russell J. Kohel; Jerry E. Quisenberry; Greg Cartwright; John Yu
A New Ray-Tracing Program Capable of Simulating Insertion-Device Synchrotron Radiation Sources
A new ray-tracing program was developed to facilitate optical design of synchrotron radiation beam lines and monochromators. The program uses Monte Carlo modelling to simulate characteristics of insertion-device sources such as undulators and wigglers as well as those of normal bending magnets. The program is successfully applied to the optical design of monochromator systems for a new multipole wiggler\\/undulator beam
Yasuji Muramatsu; Yasuo Ohishi; Hideki Maezawa
Asymptotically good codes correcting insertions, deletions, and transpositions
We present simple, polynomial-time encodable and decodable codes which are asymptotically good for channels allowing insertions, deletions and transpositions. As a corollary, they achieve exponential error probability in a stochastic model of insertion-deletion.
Schulman, L.J. [Georgia Institute Technology, Atlanta, GA (United States); Zuckerman, D. [Univ. of Texas, Austin, TX (United States)
A large-scale collection of phenotypic data describing an insertional mutant population to facilitate functional analysis of rice genes
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. PMID:17180734
Iwasaki, Yukimoto; Kitano, Hidemi; Itoh, Jun-Ichi; Maekawa, Masahiko; Murata, Kazumasa; Yatou, Osamu; Nagato, Yasuo; Hirochika, Hirohiko
Mutual Inductance Modeling for Multiple RLC Interconnects with Application to Shield Insertion Junmou Zhang and Eby G. Friedman
Mutual Inductance Modeling for Multiple RLC Interconnects with Application to Shield Insertion of Rochester Rochester, New York 146270231 Abstract-- An effective mutual inductance is proposed in this paper lines into only two coupled RLC interconnects. The concept of an effective mutual inductance is further
Friedman, Eby G.
A dilution refrigerator insert for standard ILL cryostats K. Neumaier
773 A dilution refrigerator insert for standard ILL cryostats K. Neumaier WMI Garching, F.R.G. A-1.2 K) the sample stick was replaced by a dilution refrigerator insert with a minimum no temperatures, we replaced the sample stick by a dilution refrigerator insert (Fig. 1). The large cooling power
Paris-Sud XI, Universit� de
A fast algorithm for context-aware buffer insertion
We study the problem of performing buffer insertion in the context of a given layout in a practical situation, there are restrictions on where buffers may be inserted while routing over such regions may be possible (e.g., due to the presence of macro cells). As a result, it is desirable to perform route planning and biffer insertion simultaneously. Further it
Ashok Jagannathan; Sung-Woo Hur; John Lillis
A fast algorithm for context-aware buffer insertion
We study the problem of performing buffer insertion in the context of a given layout. In a practical situation, there are restrictions on where buffers may be inserted; for instance, it may be possible to route wires over a preplaced macro cell, but may not be possible to insert buffers in that region. As a result, it is desirable to
Ashok Jagannathan; Sung-Woo Hur; John Lillis
Membrane Fission Is Promoted by Insertion of Amphipathic Helices and
Membrane Fission Is Promoted by Insertion of Amphipathic Helices and Is Restricted by Crescent BAR membrane curva- ture. Here, we investigate membrane fission by shallow hydrophobic insertions- brane fission driven by hydrophobic insertions. A quantitative assay for vesiculation reveals an antag
McMahon, Harvey
Electrochemical insertion of sodium into carbon
NASA Astrophysics Data System (ADS)
Electrochemical insertion of sodium ions into carbon using solid polymer electrolytes or organic liquid electrolytes is described. Cells with the configuration Na P(EO)(sub *)NaCF3SO3 CP(EO) = polyethylene oxide or Na/liquid electrolyte/C were galvanostatically discharged, charged, and cycled. The extent of insertion into C (i.e., x in Na(+)C(sub x -)) was found to be a strong function of the type and particle size of the carbon used, and the reversibility of the process was highly dependent upon the type of electrolyte used. The possibility of designing sodium ion rocking chair cells is discussed, and a first-generation example, using a petroleum coke anode, polymer electrolyte, and sodium cobalt bronze cathode is described.
Doeff, Marca M.; Ma, Yanping; Visco, Steven J.; Dejonghe, Lutgard C.
Innovations in central vascular access device insertion.
The new millennium has brought with it many technological advances that have affected every industry, especially healthcare and nursing. The key to success will be the ability of nurses to acclimate to these changes. To accomplish this, nurses need to advance their individual practice and skills, and to pursue and maintain an education in their respective fields. At no other time in healthcare history have so many changes taken place in such a short time, and nurses must remain abreast of changes to ensure career securement, advancement, and the safety of their patients. This article focuses on peripherally inserted central catheters and the changing technology for their insertion, maintenance, and training requirements. PMID:15965380
Pieger-Mooney, Sandi
Comparison of insertional RNA editing in Myxomycetes.
RNA editing describes the process in which individual or short stretches of nucleotides in a messenger or structural RNA are inserted, deleted, or substituted. A high level of RNA editing has been observed in the mitochondrial genome of Physarum polycephalum. The most frequent editing type in Physarum is the insertion of individual Cs. RNA editing is extremely accurate in Physarum; however, little is known about its mechanism. Here, we demonstrate how analyzing two organisms from the Myxomycetes, namely Physarum polycephalum and Didymium iridis, allows us to test hypotheses about the editing mechanism that can not be tested from a single organism alone. First, we show that using the recently determined full transcriptome information of Physarum dramatically improves the accuracy of computational editing site prediction in Didymium. We use this approach to predict genes in the mitochondrial genome of Didymium and identify six new edited genes as well as one new gene that appears unedited. Next we investigate sequence conservation in the vicinity of editing sites between the two organisms in order to identify sites that harbor the information for the location of editing sites based on increased conservation. Our results imply that the information contained within only nine or ten nucleotides on either side of the editing site (a distance previously suggested through experiments) is not enough to locate the editing sites. Finally, we show that the codon position bias in C insertional RNA editing of these two organisms is correlated with the selection pressure on the respective genes thereby directly testing an evolutionary theory on the origin of this codon bias. Beyond revealing interesting properties of insertional RNA editing in Myxomycetes, our work suggests possible approaches to be used when finding sequence motifs for any biological process fails. PMID:22383871
Chen, Cai; Frankhouser, David; Bundschuh, Ralf
Thermally Activated Retainers For Insertion In Gaps
NASA Technical Reports Server (NTRS)
Mechanical retainers of new type for use with gap filler easy to install and to attach themselves securely. Concept based on shape-memory properties of metal alloy Nitinol, alloy of nickel and titanium. Retainers conceived for use with Space Shuttle insulating tiles but used on other assemblies of blocks or tiles configured similarly. Tabs bent outward made flush by cooling below memory transition temperature. After insertion in gap and reheating, tabs spring outward.
Grimaldi, Margaret E.; Hartz, Leslie S.
Needle Insertion Test by Porcine Ligamentum Flavum
\\u000a Needles for the epidural anesthesia are designed, as a curved tip allows direction of the catheter in the epidural space.\\u000a Purpose of the current study was to check the effect of the curved tip and slower insertion speed on puncturing dynamics for\\u000a the ligamentum flavum. Porcine spines from a slaughterhouse were employed. Two Tuhoy needles fabricated by different manufacture\\u000a and
K. Naemura; H. Saito
Inserting new technology into small missions
NASA Technical Reports Server (NTRS)
Part of what makes small missions small is that they have less money. Executing missions at low cost implies extensive use of cost sharing with other missions or use of existing solutions. However, in order to create many small missions, new technology must be developed, applied, and assimilated. Luckily, there are methods for creating new technology and inserting it into faster-better-cheaper (FBC) missions.
Deutsch, L. J.
Insertional Polymorphisms of Endogenous Feline Leukemia Viruses
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
[Historical evolution of package inserts in Brazil].
In Brazil, package inserts provide key information on pharmaceuticals. The current study analyzes the evolution of package inserts and the impact on this process by scientific research and development, globalization of information, and various health policies. The study began with a retrospective review of Brazilian health legislation until 1920, the year when the National Public Health Department was created. The analysis of documents on the evolution of health regulation in Brazil began with the Brazilian Pharmaceutical Collection-Health Rulings. The second stage of the study involved a search of standards and norms in VISALEGIS: Health Surveillance Legislation, Portal for Legislation from the National Congressional Information System and the Health Legislation System. Package inserts became an important vehicle for information in the country and underwent important regulatory changes in the latter half of the 20th century. From 1946 to 2006, the number of mandatory items increased, with more in-depth description. However, the standardization of information for medicines with the same active ingredient failed to materialize, despite its importance and the various legal initiatives in this direction. PMID:18392350
Caldeira, Telma Rodrigues; Neves, Eug�nio Rodrigo Zimmer; Perini, Edson
Database documentation of retrotransposon insertion polymorphisms.
Retrotransposons constitute more than 40 percent of the human genome with L1, Alu, SVA, and HERVs known to remain active in transposition. Retrotransposition contribute to genetic diversity in the form of retrotransposon insertion polymorphism (RIP) that is defined as the presence or absence of a retrotransposon insertion among human populations at a specific genomic location. So far close to 5000 cases of RIPs have been identified with more than 50 cases associated with disease. A large number of new RIPs are being and to be identified from newly available personal genomes data, making RIPs an important source of genetic variations/mutations that deserve proper documentation. In this review, we discuss the special characteristics of RIPs and the challenges in their compiling and annotating, and we examine the current status of database documentation of RIPs and describe in details the design, data schema, and utilities of dbRIP, which is currently the only database dedicated to the documentation of retrotransposon insertion polymorphism. Some future perspectives and outstanding issues associated with documentation of RIPs are also presented. PMID:22201974
Liang, Ping; Tang, Wanxiangfu
Optimization of {beta} functions through insertion devices
It is generally noted that at an undulator straight section, the horizontal beta function is made to be large while the vertical beta function is relatively small. On the other hand, at a wiggler straight section, both horizontal and vertical beta functions are made to be small. In this note we describe a procedure with which optimum settings of the beta functions in the insertion straight section are to be determined. For this we consider separately for the undulator radiation and the radiation from the wiggler device. Since the brilliance of radiation is a canonically conserved quantity, we use the brilliance as a figure of merit for the consideration. Then the optimization process is to find a set of horizontal and vertical beta functions which would give the maximum brilliance when the natural emittance, emittance coupling constant, length of insertion device and the photon energy to which the insertion device is optimized are specified. The next step of the study is to find sensitivities of the beta functions to the brilliance.
Cho, Y.
Effect of Insertion Speed on Tissue Response and Insertion Mechanics of a Chronically Implanted Silicon-Based Neural Probe
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
Vector�Hexamer PCR Isolation of All Insert Ends from a YAC Contig of the Mouse Igh?Locus
We have developed a simple PCR strategy, termed vector�hexamer PCR, that is unique in its ability to easily recover every insert end from large insert clones in YAC and BAC vectors. We used this method to amplify and isolate all insert ends from a YAC contig covering the mouse Igh locus. Seventy-seven ends were amplified and sequenced from 36 YAC clones from four libraries in the pYAC4 vector. Unexpectedly, 40% of the insert ends of these YACs were LINE1 repeats. Nonrepetitive ends were suitable for use as probes on Southern blots of digested YACs to identify overlaps and construct a contig. The same strategy was used successfully to amplify insert ends from YACs in the pRML vector from the Whitehead Institute/MIT-820 mouse YAC library and from BACs in pBeloBAC11. The simplicity of this technique and its ability to isolate every end from large insert clones are of great utility in genomic investigation. [The nucleotide sequence data reported in this paper are accessible in GenBank under accession nos. B07512�B07598.] PMID:9647641
Herring, Christopher D.; Chevillard, Christophe; Johnston, Sean L.; Wettstein, Peter J.; Riblet, Roy
Recessed impingement insert metering plate for gas turbine nozzles
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)
Insertion of coherence requests for debugging a multiprocessor
A method and system are disclosed to insert coherence events in a multiprocessor computer system, and to present those coherence events to the processors of the multiprocessor computer system for analysis and debugging purposes. The coherence events are inserted in the computer system by adding one or more special insert registers. By writing into the insert registers, coherence events are inserted in the multiprocessor system as if they were generated by the normal coherence protocol. Once these coherence events are processed, the processing of coherence events can continue in the normal operation mode.
Blumrich, Matthias A. (Ridgefield, CT); Salapura, Valentina (Chappaqua, NY)
Modal Analysis of Deepwater Mooring Lines Based on a Variational Formulation
is expected more damping is obtained. Two realistic designs of mooring lines were compared: one made up with a main insert of steel rope, called �Steel System�, and one composed by a main insert of polyester, named �Polyester System�. Comparing the natural...
Martinez Farfan, Jose Alberto
Assembly and Insertion of a Self-Fitting Hearing Aid
A self-fitting hearing aid has been proposed as a viable option to meet the need for rehabilitation in areas where audiology services are unreliable. A successful outcome with a self-fitting hearing aid pivots in part on the clarity of the instructions accompanying the device. The aims of this article are (a) to review the literature to determine features that should be incorporated into written health-care materials and factors to consider in the design process when developing written instructions for a target audience of older adults and (b) to apply this information to the development of a set of written instructions as the first step in self-fitting of a hearing aid, assembling four parts and inserting the aid into the ear. The method involved a literature review of published peer reviewed research. The literature revealed four steps in the development of written health-care materials: planning, design, assessment of suitability, and pilot testing. Best practice design principles for each step were applied in the development of instructions for how to assemble and insert a hearing aid. Separate booklets were developed for the left and right aids and the content of each consisted of simple line drawings accompanied by captions. The reading level was Grade 3.5 equivalent and the Flesch Reading Ease Score was 91.1 indicating that the materials were �very easy� to read. It is essential to follow best practice design principles when developing written health-care materials to motivate the reader, maximize comprehension, and increase the likelihood of successful application of the content. PMID:22389434
Hickson, Louise; Meyer, Carly
A piezoelectric vibration-based syringe for reducing insertion force
NASA Astrophysics Data System (ADS)
Puncturing of the human skin with a needle is perhaps the most common invasive medical procedure. Clinical studies have revealed that tissue deformation and needle deflection are the primary problem for needle misplacement in percutaneous procedures. To avoid this, various techniques for reducing insertion forces during needle insertion have been considered. This paper presents a piezoelectric vibration-based syringe to reduce insertion force. AC power was applied to the piezoelectric elements to vibrate the needle with high frequency and thereby reduce the friction and cutting forces between the needle and tissue. Vibration mode shapes of the needle were observed by finite element analysis and verified by experimental results. Effects of reducing insertion force via the vibrating needle were also confirmed by inserting the needle into the porcine tissues. The proposed syringe, which minimizes the insertion force and overcomes limitations of needle materials, can be widely utilized in robot-assisted needle insertion systems.
Huang, Y. C.; Tsai, M. C.; Lin, C. H.
Meta-analysis of gross insertions causing human genetic disease: novel mutational mechanisms and the role of replication slippage.
Although gross insertions (>20 bp) comprise <1% of disease-causing mutations, they nevertheless represent an important category of pathological lesion. In an attempt to study these insertions in a systematic way, 158 gross insertions ranging in size between 21 bp and approximately 10 kb were identified using the Human Gene Mutation Database (www.hgmd.org). A careful meta-analytical study revealed extensive diversity in terms of the nature of the inserted DNA sequence and has provided new insights into the underlying mutational mechanisms. Some 70% of gross insertions were found to represent sequence duplications of different types (tandem, partial tandem, or complex). Although most of the tandem duplications were explicable by simple replication slippage, the three complex duplications appear to result from multiple slippage events. Some 11% of gross insertions were attributable to nonpolyglutamine repeat expansions (including octapeptide repeat expansions in the prion protein gene [PRNP] and polyalanine tract expansions) and evidence is presented to support the contention that these mutations are also caused by replication slippage rather than by unequal crossing over. Some 17% of gross insertions, all >or=276 bp in length, were found to be due to LINE-1 (L1) retrotransposition involving different types of element (L1 trans-driven Alu, L1 direct, and L1 trans-driven SVA). A second example of pathological mitochondrial-nuclear sequence transfer was identified in the USH1C gene but appears to arise via a novel mechanism, trans-replication slippage. Finally, evidence for another novel mechanism of human genetic disease, involving the possible capture of DNA oligonucleotides, is presented in the context of a 26-bp insertion into the ERCC6 gene. PMID:15643617
Chen, Jian-Min; Chuzhanova, Nadia; Stenson, Peter D; F�rec, Claude; Cooper, David N
Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) furnace development
NASA Astrophysics Data System (ADS)
The Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) are microgravity furnaces under development at Marshall Space Flight Center. The furnaces are being developed for the first Materials Science Research Rack (MSRR-1) of the Materials Science Research Facility (MSRF), one of the first International Space Station (ISS) scientific payloads. QMI is a Bridgman furnace with quench capability for studying interface behavior during directional solidification of metallic and alloy materials. DMI will be a Bridgman-Stockbarger furnace to study diffusion processes in semiconductors. The design for each insert, both QMI and DMI, is driven by specific science, operations and safety requirements, as well as by constraints arising from resource limitations, such as volume, mass and power. Preliminary QMI analysis and testing indicates that the design meets these requirements. .
Crouch, Myscha R.; Carswell, William E.; Farmer, Jeff; Rose, Fred; Tidwell, Paul H.
Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) Furnace Development
NASA Technical Reports Server (NTRS)
The Quench Module Insert (QMI) and the Diffusion Module Insert (DMI) are microgravity furnaces under development at Marshall Space Flight Center. The furnaces are being developed for the first Materials Science Research Rack (MSRR-1) of the Materials Science Research Facility (MSRF), one of the first International Space Station (ISS) scientific payloads. QMI is a Bridgman furnace with quench capability for studying interface behavior during directional solidification of metallic and alloy materials. DMI will be a Bridgman-Stockbarger furnace to study diffusion processes in semiconductors. The design for each insert, both QMI and DMI, is driven by specific science, operations and safety requirements, as well as by constraints arising from resource limitations, such as volume, mass and power. Preliminary QMI analysis and testing indicates that the design meets these requirements.
Crouch, Myscha R.; Carswell, William E.; Farmer, Jeff; Rose, Fred; Tidwell, Paul H., II
NSDL National Science Digital Library
In this design challenge activity, learners build a device to carry a Ping-Pong ball from the top of a zip line to the bottom in four seconds (or less!). Learners follow the design process to build the carrier that slides quickly down a zip line string, figure out how to keep something balanced, and identify ways to reduce friction.
Insertion torque versus mechanical resistance of mini-implants inserted in different cortical thickness
Objective This study aimed to measure insertion torque, tip mechanical resistance to fracture and transmucosal neck of mini-implants (MI) (Conex�o Sistemas de Pr�teseT), as well as to analyze surface morphology. Methods Mechanical tests were carried out to measure the insertion torque of MIs in different cortical thicknesses, and tip mechanical resistance to fracture as well as transmucosal neck of MIs. Surface morphology was assessed by scanning electron microscopy (SEM) before and after the mechanical tests. Results Values of mechanical resistance to fracture (22.14 N.cm and 54.95 N.cm) were higher and statistically different (P < 0.05) from values of insertion torque for 1-mm (7.60 N.cm) and 2-mm (13.27 N.cm) cortical thicknesses. Insertion torque was statistically similar (P > 0.05) to torsional fracture in the tip of MI (22.14 N.cm) when 3 mm cortical thickness (16.11 N.cm) and dense bone (23.95 N.cm) were used. Torsional fracture of the transmucosal neck (54.95 N.cm) was higher and statistically different (P < 0.05) from insertion torsional strength in all tested situations. SEM analysis showed that the MIs had the same smooth surface when received from the manufacturer and after the mechanical tests were performed. Additionally, no significant marks resulting from the manufacturing process were observed. Conclusion All mini-implants tested presented adequate surface morphology. The resistance of mini-implants to fracture safely allows placement in 1 and 2-mm cortical thickness. However, in 3-mm cortical thickness and dense bones, pre-drilling with a bur is recommended before insertion. PMID:25162571
Santos, Renata de Faria; Ruellas, Antonio Carlos de Oliveira; Fernandes, Daniel Jogaib; Elias, Carlos Nelson
Effect of the Starting Point of Half-Pin Insertion on the Insertional Torque of the Pin at the Tibia
Purpose The authors have observed a failure to achieve secure fixation in elderly patients when inserting a half-pin at the anteromedial surface of the tibia. The purpose of this study was to compare two methods for inserting a half-pin at tibia diaphysis in elderly patients. Materials and Methods Twenty cadaveric tibias were divided into Group C or V. A half-pin was inserted into the tibias of Group C via the conventional method, from the anteromedial surface to the interosseous border of the tibia diaphysis, and into the tibias of Group V via the vertical method, from the anterior border to the posterior surface at the same level. The maximum insertion torque was measured during the bicortical insertion with a torque driver. The thickness of the cortex was measured by micro-computed tomography. The relationship between the thickness of the cortex engaged and the insertion torque was investigated. Results The maximum insertion torque and the thickness of the cortex were significantly higher in Group V than Group C. Both groups exhibited a statistically significant linear correlation between torque and thickness by Spearman's rank correlation analysis. Conclusion Half-pins inserted by the vertical method achieved purchase of more cortex than those inserted by the conventional method. Considering that cortical thickness and insertion torque in Group V were significantly greater than those in Group C, we suggest that the vertical method of half-pin insertion may be an alternative to the conventional method in elderly patients. PMID:25510759
Kim, Sung-Jae; Kim, Sung-Hwan; Kim, Young-Hwan
Autosoliton propagation and mapping problem in optical fiber lines with lumped nonlinear devices.
A theoretical model is developed to describe the propagation of ultrashort optical pulses in fiber transmission systems in the quasilinear regime, with periodically inserted in-line nonlinear optical devices. PMID:16090101
Boscolo, S; Derevyanko, S A; Turitsyn, S K; Kovalev, A S; Bogdan, M M
Commercial Generic Bioprocessing Apparatus Science Insert - 03
NASA Technical Reports Server (NTRS)
Commercial Generic Bioprocessing Apparatus Science Insert - 03 (CSI-03) is the third set of investigations in the CSI program series. The CSI program provides the K-12 community opportunities to utilize the unique microgravity environment of the International Space Station as part of the regular classroom to encourage learning and interest in science, technology, engineering and math. CSI-03 will examine the complete life cycle of the painted lady butterfly and the ability of an orb weaving spider to spin a web, eat and remain healthy in space.
Moreno, Nancy; Stodieck, Louis; Cushing, Paula; Stowe, Mark; Hamilton, Mary Ann; Werner, Ken
Analytic closed orbit analysis for RHIC insertion
Analytic closed orbit analysis is performed to evaluate the tolerance of quadrupole misalignment and dipole errors (b{sub 0},a{sub 0}) in the RHIC insertion. Sensitivity coefficients of these errors are tabulated for different {beta}{sup 0} values. Using these sensitivity tables, we found that the power supplies ripple of 10{sup {minus}4} can cause closed orbit motion of 0.05 mm at the IP in comparison with the rms beam size of 0.3 mm. It is desirable to have the power supply ripple less than 10{sup {minus}5}. 2 refs., 1 fig., 2 tabs.
Lee, S.Y. (Indiana Univ., Bloomington, IN (USA). Dept. of Physics); Tepikian, S. (Brookhaven National Lab., Upton, NY (USA))
STRUCTURAL BIOLOGY: Membrane Protein Insertion and Stability
NSDL National Science Digital Library
Access to the article is free, however registration and sign-in are required. The translocon is an endoplasmic reticulum membrane protein that has the dual ability to insert a newly synthesized polypeptide chain into a cell membrane or secrete it across a cell membrane. This process has been exploited to assess the energetics of transmembrane -helices, as described in two recent studies by the von Heijne and White groups (published here and elsewhere). In a Perspective, MacKinnon discusses the implications of this work for membrane protein structure and function.
Roderick MacKinnon (Howard Hughes Medical Institute, Rockefeller University; )
Transcatheter aortic valve insertion (TAVI): a review
The introduction of transcatheter aortic valve insertion (TAVI) has transformed the care provided for patients with severe aortic stenosis. The uptake of this procedure is increasing rapidly, and clinicians from all disciplines are likely to increasingly encounter patients being assessed for or having undergone this intervention. Successful TAVI heavily relies on careful and comprehensive imaging assessment, before, during and after the procedure, using a range of modalities. This review outlines the background and development of TAVI, describes the nature of the procedure and considers the contribution of imaging techniques, both to successful intervention and to potential complications. PMID:24258463
Morgan-Hughes, G; Roobottom, C
Artists's Conception of Cassini Saturn Orbit Insertion
NASA Technical Reports Server (NTRS)This is an artists concept of Cassini during the Saturn Orbit Insertion (SOI) maneuver, just after the main engine has begun firing. The spacecraft is moving out of the plane of the page and to the right (firing to reduce its spacecraft velocity with respect to Saturn) and has just crossed the ring plane.The SOI maneuver, which is approximately 90 minutes long, will allow Cassini to be captured by Saturn's gravity into a five-month orbit.Cassini's close proximity to the planet after the maneuver offers a unique opportunity to observe Saturn and its rings at extremely high resolution.
Stress-inducible OsP5CS2 gene is essential for salt and cold tolerance in rice
We isolated a rice T-DNA tagging line, in which T-DNA was inserted into the sixth intron of OsP5CS2. This gene encodes for a protein that is highly homologous to ?1-pyrroline-5-carboxylate synthetase (P5CS), a proline biosynthesis enzyme. The T-DNA contained the promoterless gus gene, allowing generation of a gene fusion between OsP5CS2 and gus. Therefore, the expression pattern of OsP5CS2 could
Junghe Hur; Ki-Hong Jung; Choon-Hwan Lee; Gynheung An
Stress-inducible OsP5CS2gene is essential for salt and cold tolerance in rice
We isolated a rice T-DNA tagging line, in which T-DNA was inserted into the sixth intron of OsP5CS2. This gene encodes for a protein that is highly homologous to ? 1-pyrroline-5-carboxylate synthetase (P5CS), a proline biosynthesis enzyme. The T-DNA contained the promoterless gus gene, allowing generation of a gene fusion between OsP5CS2 and gus. Therefore, the expression pattern of OsP5CS2
Junghe Hur; Ki-Hong Jung; Choon-Hwan Leeb; Gynheung Ana
Peripherally inserted central venous catheter safety in burn care: a single-center retrospective cohort review.
The use of peripherally inserted central catheter (PICC) line for central venous access in thermally injured patients has increased in recent years despite a lack of evidence regarding safety in this patient population. A recent survey of invasive catheter practices among 44 burn centers in the United States found that 37% of burn units use PICC lines as part of their treatment protocol. The goal of this study was to compare PICC-associated complication rates with the existing literature in both the critical care and burn settings. The methodology involved is a single institution retrospective cohort review of patients who received a PICC line during admission to a regional burn unit between 2008 and 2013. Fifty-three patients were identified with a total of seventy-three PICC lines. The primary outcome measurement for this study was indication for PICC line discontinuation. The most common reason for PICC line discontinuation was that the line was no longer indicated (45.2%). Four cases of symptomatic upper extremity deep vein thrombosis (5.5%) and three cases of central line-associated bloodstream infection (4.3%, 2.72 infections per 1000 line days) were identified. PICC lines were in situ an average of 15 days (range 1 to 49 days). We suggest that PICC line-associated complication rates are similar to those published in the critical care literature. Though these rates are higher than those published in the burn literature, they are similar to central venous catheter-associated complication rates. While PICC lines can be a useful resource in the treatment of the thermally injured patient, they are associated with significant and potentially fatal risks. PMID:25501778
Austin, Ryan E; Shahrokhi, Shahriar; Bolourani, Siavash; Jeschke, Marc G
[Ceramic acetabulum cup inserts for hip endoprostheses].
The articulating components of a total hip prosthesis are the spherical femoral head and the acetabular cup. Particularly high rates of wear are seen with cups made of polyethylene, and the abrasion particles cause osteolysis, which often makes surgical revision necessary. Despite considerable progress in the development of total hip replacement systems over the last 20 years, there is still a need to eliminate or further minimize the problem of osteolysis, which is also referred to as polyethylene disease. Clinical experience over the last 20 years has shown that by using Biolox (medical grade alumina) femoral heads and acetabular cups it is possible to achieve very low wear rates, which histological studies have shown to be readily tolerated. The secondary conditions needed to develop modern modular ceramic acetabular cup inserts are discussed. For the fixation of cup inserts, the well known taper fixation concept has proved to be of value. A concept taking into account the needs of the surgeon and the manufacturing constraints applying to ceramic materials is proposed. PMID:8679913
Willmann, G; K�lberer, H; Pfaff, H G
Radiological Insertion and Management of Peritoneovenous Shunt
The purpose of the study was to report our experience of the management of complications following the insertion of a peritoneovenous shunt for intractable malignant ascites. From June 1999 to January 2006, 26 patients underwent insertion of a peritoneovenous shunt for ascites by interventional radiologists. We have used ultrasound and shuntography to assist in the diagnosis of the cause of shunt blockage. Successful techniques for the restoration of the shunt function include port- pumping, stripping of any fibrin sheath, and revision of either the venous or peritoneal catheter. The procedure was initially successful in all patients with continued patency until death in 17. A further four patients are still alive with a functioning shunt. There was one rapid postprocedure death resulting from pulmonary edema. Two patients developed pneumothorax, managed successfully with either a chest drain or aspiration. Shunt dysfunction occurred eight times in seven patients. There were five successful revisions in four patients. Overall, shunt patency has been maintained in 80.1% of patients. Shunt dysfunction is seen in a significant number of patients, but successful revision of the shunt can be achieved in the majority.
Bratby, M. J. [St George's Hospital, Radiology Department (United Kingdom); Hussain, F. F., E-mail: email@example.com; Lopez, A. J. [Royal Surrey County Hospital and St. Luke's Cancer Centre, Radiology Department (United Kingdom)
Black hole on film following catheter insertion.
Vascular accesses both permanent and temporary play an essential role in the procedure of hemodialysis (HD). The double lumen catheter (DLC) is one form of temporal vascular access which may produce different complications based on where the DLC is inserted. Here, we report a case of a cirrhotic patient receiving HD. The DLC was inserted through the left femoral vein. Later on, an unusual catheter position, mimicking a perforation in the iliac vessel, was noted in the incidental image finding. After venography, we concluded that this DLC had entered the dilated left gonadal vein, and was working well. This condition may be explained by portal hypertension leading to increased frequency of collateral branches and easy dilatation of the left gonadal vein. This condition may develop in a cirrhotic patient with portal hypertension, but there is no harm to a patient. However, this condition leads to an interesting computed tomography image, which may result in misinterpretation of the imaging reports and a clinical misdiagnosis. PMID:22304472
Chen, Hsuan-Wei; Chen, Jin-Shuen
Phoenix Conductivity Probe Inserted in Martian Soil
NASA Technical Reports Server (NTRS)This series of six images from the Robotic Arm Camera on NASA's Phoenix Mars Lander records the first time that the four spikes of the lander's thermal and electrical conductivity probe were inserted into Martian soil. The images were taken on July 8, 2008, during the Phoenix mission's 43rd Martian day, or sol, since landing. The insertion visible from the shadows cast on the ground on that sol was a validation test of the procedure. The spikes on the probe are about 1.5 centimeters or half an inch long. The science team will use the probe tool to assess how easily heat and electricity move through the soil from one spike to another. Such measurements can provide information about frozen or unfrozen water in the soil. The probe is mounted on the 'knuckle' of Phoenix's Robotic Arm. It has already been used for assessing water vapor in the atmosphere when it is held above the ground. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is led by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.
Lithium Insertion Chemistry of Some Iron Vanadates
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.
NSDL National Science Digital Library
This iOS app helps students to visualize number sentences and create models for addition, subtractions, multiplication, and division. The number line can be adjusted to represent multiples of numbers from one to one hundred.
Clarity Innovations, Inc.
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.
NSDL National Science Digital Library
In this brief article the numerous uses of the number line are detailed: counting, measurement, addition, subtraction, decimals, and fractions. The article contains visual representations of the some of the concepts and links to related topics.
... A A A Beau's lines (crosswise nail plate grooves) are due to severe illness or certain medications. ... occur along the fingernails and the toenails. The grooves extending across the nail plate often span its ...
Design and evaluation of moxifloxacin hydrochloride ocular inserts.
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
Lithium insertion mechanism in tin-based spinel sulfides
Cu�MSn�S� (M = Fe, Co) spinel compounds have been studied as cathode materials in Li\\/LiPF�(1M)-PC-EC\\/spinel sulfide cells. To understand the lithium insertion mechanism, both pristine compounds and chemically inserted products were characterized by X-ray Powder Diffraction (XPD), ¹¹⁹Sn Moessbauer Spectroscopy (MS), and X-ray Absorption Spectroscopy (XAS) at the sulfur K-edge. To accommodate the inserted lithium ions, two steps of reduction
C. Branci; J. Sarradin; J. Olivier-Fourcade; J. C. Jumas
Optimal and efficient buffer insertion and wire sizing
We present optimal solutions to the following problems: (1) post-layout buffer insertion, (2) wire-sizing and (3) simultaneous buffer insertion and wire-sizing. We optimize a practical objective function: required arrival time. To the best of our knowledge, this work represents the first sub-exponential algorithms for these problems. In experiments, we observe substantial improvements over previous results for buffer insertion, and up
John Lillis; Chung-Kuan Cheng; Ting-Ting Y. Lin
Relaxation phenomena in lithium-ion-insertion cells
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
Intrauterine device insertion in the postpartum period: A systematic review.
Abstract Objectives Given new research on postpartum placement of levonorgestrel and copper intrauterine devices (IUDs), our objective was to update a prior systematic review of the safety and expulsion rates of postpartum IUDs. Methods We searched MEDLINE, CENTRAL, LILACS, POPLINE, Web of Science, and ClinicalTrials.gov databases for articles between the database inception until July 2013. We included studies that compared IUD insertion time intervals and routes during the postpartum period. We used standard abstract forms and the United States Preventive Services Task Force grading system to summarise and assess the quality of the evidence. Results We included 18 articles. New evidence suggests that a levonorgestrel releasing-intrauterine system (LNG-IUS) insertion within 48 hours of delivery is safe. Postplacental insertion and insertion between 10 minutes and 48 hours after delivery result in higher expulsion rates than insertion 4 to 6 weeks postpartum, or non-postpartum insertion. Insertion at the time of caesarean section is associated with lower expulsion rates than postplacental insertion at the time of vaginal delivery. Conclusions This review supports the evidence that insertion of an intrauterine contraceptive within the first 48 hours of vaginal or caesarean delivery is safe. Expulsion rates should be further studied in larger randomised controlled trials. PMID:25397890
Sonalkar, Sarita; Kapp, Nathalie
Wind tunnel supplementary Mach number minimum section insert
NASA Technical Reports Server (NTRS)
A device is described which changes the Mach number capability of a wind tunnel without permanently altering the existing nozzle of the tunnel. An insert is removably attached to the wall of the existing nozzle expansion area thereby creating a second minimum section upstream of the model test section. The added insert may be removed without complicated and expensive changes to the basic wind tunnel. In one embodiment, a removable insert is disposed within wind tunnel nozzle walls with a portion of the flow boundary layer being bled off from the tunnel via passageway and tunnel exit to reduce the extent of separated flow normally occuring upstream of the insert contraction section.
Couch, L. M. (inventor)
Contraceptive sponge patient insert warns of TSS.
The Today contraceptive sponge patient insert is being revised to reflect concerns about the possible relationship between toxic shock syndrome (TSS) and the vaginal contraceptive sponge. The sponge was first marketed over-the-counter in June 1983. It is made of polyurethane impregnated with the spermicide nonoxynol-9 and is intended to provide 24 hours of contraception. At the time of approval, the Food and Drug Administration (FDA) required a patient insert containing a warning that clinical trials had not been large enough to assess the risk of TSS, and that users should seek medical care if they developed the listed symptoms of TSS. In February 1984, 4 cases of confirmed TSS were reported to be associated with sponge use. Of these, 2 occurred in women who had difficulty removing the product, 1 occurred in a woman 37 days postpartum, and 1 occurred in a woman who had left the sponge in place for 5 days. All were nonmenstruating white women, 20-29 years of age. As of July 1, 1984 there had been 4 additional confirmed nonmenstruating cases of TSS with the contraceptive sponge. The manufacturer of the sponge, VLI Corporation, added a warning in February on the outside packaging and in the patient insert. It advises the user to consult her physician or clinic immediately if she experiences 2 or more of the warning signs of TSS, to remove the sponge within the specified time limit, and to consult one's physician or clinic before using this product after childbirth, miscarriage, or abortion. The data should be regarded as preliminary, but the rate of TSS in nonmenstruating women using the contraceptive sponge is estimated to be less than that in menstruating women using tampons but more than the background rate for nonmenstruating women. TSS patients should be managed as those suffering from other forms of shock. Large volumes of fluid and supportive care should be used as needed. A vaginal examination should be performed with removal of any tampon or vaginal contraceptive. There is no evidence that antibiotics are effective in treating active TSS cases, but beta-lactamase-resistant antistaphylococcal antibiotics seem to be effective in preventing recurrences. PMID:6479516
Method of joining a vane cavity insert to a nozzle segment of a gas turbine
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)
Impact of Orthodontic Decompensation on Bone Insertion
There has always been concern in determining the relationship between orthodontic tooth movement and the consequent biological costs to the periodontium and tooth root. The possibility of evaluating the tooth and bone morphology by CBCT allows more accurate analysis of qualitative and quantitative aspects of these processes. This paper presents a case report of a 20-year-old male patient with Class III malocclusion and hyperdivergent facial pattern, who was surgically treated. A significant amount of labial movement of mandibular incisors was performed during orthodontic treatment before surgery. CBCT was used for evaluation of buccal and lingual bone plates before and after tooth decompensation. The changes in the bone insertion level of maxillary and mandibular incisors in the present case encourage a reflection on the treatment protocol in individuals with dentoskeletal discrepancies. PMID:25436157
Guedes, Fabio Pinto; Capelozza Filho, Leopoldino; Garib, Daniela Gamba; Nary Filho, Hugo; Borgo, Evandro Jos�; Cardoso, Mauricio de Almeida
Increasing prion propensity by hydrophobic insertion.
Prion formation involves the conversion of proteins from a soluble form into an infectious amyloid form. Most yeast prion proteins contain glutamine/asparagine-rich regions that are responsible for prion aggregation. Prion formation by these domains is driven primarily by amino acid composition, not primary sequence, yet there is a surprising disconnect between the amino acids thought to have the highest aggregation propensity and those that are actually found in yeast prion domains. Specifically, a recent mutagenic screen suggested that both aromatic and non-aromatic hydrophobic residues strongly promote prion formation. However, while aromatic residues are common in yeast prion domains, non-aromatic hydrophobic residues are strongly under-represented. Here, we directly test the effects of hydrophobic and aromatic residues on prion formation. Remarkably, we found that insertion of as few as two hydrophobic residues resulted in a multiple orders-of-magnitude increase in prion formation, and significant acceleration of in vitro amyloid formation. Thus, insertion or deletion of hydrophobic residues provides a simple tool to control the prion activity of a protein. These data, combined with bioinformatics analysis, suggest a limit on the number of strongly prion-promoting residues tolerated in glutamine/asparagine-rich domains. This limit may explain the under-representation of non-aromatic hydrophobic residues in yeast prion domains. Prion activity requires not only that a protein be able to form prion fibers, but also that these fibers be cleaved to generate new independently-segregating aggregates to offset dilution by cell division. Recent studies suggest that aromatic residues, but not non-aromatic hydrophobic residues, support the fiber cleavage step. Therefore, we propose that while both aromatic and non-aromatic hydrophobic residues promote prion formation, aromatic residues are favored in yeast prion domains because they serve a dual function, promoting both prion formation and chaperone-dependent prion propagation. PMID:24586661
Gonzalez Nelson, Aaron C; Paul, Kacy R; Petri, Michelina; Flores, Noe; Rogge, Ryan A; Cascarina, Sean M; Ross, Eric D
Increasing Prion Propensity by Hydrophobic Insertion
Prion formation involves the conversion of proteins from a soluble form into an infectious amyloid form. Most yeast prion proteins contain glutamine/asparagine-rich regions that are responsible for prion aggregation. Prion formation by these domains is driven primarily by amino acid composition, not primary sequence, yet there is a surprising disconnect between the amino acids thought to have the highest aggregation propensity and those that are actually found in yeast prion domains. Specifically, a recent mutagenic screen suggested that both aromatic and non-aromatic hydrophobic residues strongly promote prion formation. However, while aromatic residues are common in yeast prion domains, non-aromatic hydrophobic residues are strongly under-represented. Here, we directly test the effects of hydrophobic and aromatic residues on prion formation. Remarkably, we found that insertion of as few as two hydrophobic residues resulted in a multiple orders-of-magnitude increase in prion formation, and significant acceleration of in vitro amyloid formation. Thus, insertion or deletion of hydrophobic residues provides a simple tool to control the prion activity of a protein. These data, combined with bioinformatics analysis, suggest a limit on the number of strongly prion-promoting residues tolerated in glutamine/asparagine-rich domains. This limit may explain the under-representation of non-aromatic hydrophobic residues in yeast prion domains. Prion activity requires not only that a protein be able to form prion fibers, but also that these fibers be cleaved to generate new independently-segregating aggregates to offset dilution by cell division. Recent studies suggest that aromatic residues, but not non-aromatic hydrophobic residues, support the fiber cleavage step. Therefore, we propose that while both aromatic and non-aromatic hydrophobic residues promote prion formation, aromatic residues are favored in yeast prion domains because they serve a dual function, promoting both prion formation and chaperone-dependent prion propagation. PMID:24586661
Petri, Michelina; Flores, Noe; Rogge, Ryan A.; Cascarina, Sean M.; Ross, Eric D.
A translocation, insertion and deletion distance formula for sorting genomes
Sorting genomes by translocation, insertion and deletions has already been researched for years such as in. However, the authors in did not consider the case that a gene in the target genome does not appear in the source genome. Translocation is a reciprocal operation in sorting genomes, and deletion and insertion are reciprocal to each other. In this paper, we
Hao Fanchang; Luan Junfeng; Zhu Daming; Feng Haodi
An algorithm for progressive multiple alignment of sequences with insertions.
Dynamic programming algorithms guarantee to find the optimal alignment between two sequences. For more than a few sequences, exact algorithms become computationally impractical, and progressive algorithms iterating pairwise alignments are widely used. These heuristic methods have a serious drawback because pairwise algorithms do not differentiate insertions from deletions and end up penalizing single insertion events multiple times. Such an unrealistically high penalty for insertions typically results in overmatching of sequences and an underestimation of the number of insertion events. We describe a modification of the traditional alignment algorithm that can distinguish insertion from deletion and avoid repeated penalization of insertions and illustrate this method with a pair hidden Markov model that uses an evolutionary scoring function. In comparison with a traditional progressive alignment method, our algorithm infers a greater number of insertion events and creates gaps that are phylogenetically consistent but spatially less concentrated. Our results suggest that some insertion/deletion "hot spots" may actually be artifacts of traditional alignment algorithms. PMID:16000407
L�ytynoja, Ari; Goldman, Nick
Load beam unit replaceable inserts for dry coal extrusion pumps
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.
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Efficient Processing of Exact Top-k Queries be inserted later Abstract The top-k query is employed in a wide range of applications to generate a ranked in two phases find a cut-off threshold for the top-k result scores, then evaluate all the records
Ding, Xuhua
Hysteresis during lithium insertion in hydrogen-containing carbons
The authors studied lithium insertion in hydrogen-containing carbons heated at temperatures near 700 C. High capacities with large hysteresis (lithium insertion into these carbons at nearly 0 V and removal at nearly 1 V) were shown to be proportional to the hydrogen content of the samples. It is believed that the lithium atoms may bind on hydrogen-terminated edges of hexagonal
Tao Zheng; J. R. Dahn; W. R. McKinnon
Stochastic Models of Sequence Evolution including Insertion-Deletion events
Comparison of sequences that have descended from a common an- cestor based on an explicit stochastic model of substitutions, insertions an d dele- tions has risen to prominence in the last decade. Making statements about the positions of insertions-deletions (abbr. indels) is central in sequence an d genome analysis and is called alignment. This statistical approach is harder concep tually
Istv�n Mikl�s; �d�m Nov�k; Rahul Satija; Rune Lyngs�; Jotun Hein
Peripherally Inserted Central Catheter Placement Using the Sonic Flashlight
involving physicians and nurses. J Vasc Interv Radiol 2009; 20:13801383 Abbreviation: PICC peripherally inserted central catheter PERIPHERALLY inserted central cathe- ters (PICCs) are increasingly being used with peripheral intravenous catheters. This makes PICCs the preferred access method for many uses, including long
Stetten, George
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
A Study of Package Inserts in Southern India
Introduction: Package insert is an officially approved document that accompanies a drug. It is intended to provide information for the safe and effective use of a drug and contains information based on regulatory guidelines. Sometimes, information provided in the package inserts is suboptimal which can led to medication errors. This study was undertaken to assess the presentation and completeness of clinical information provided in the currently available package inserts for anti-diabetic, anti-hypertensive and hypolipedemic drugs in India. Material and Methods: Around 130 package inserts were collected from pharmacies located at different areas of Bangalore. They were analyzed based on criteria mentioned in Schedule D of Drug and Cosmetic act 1945. Results and Observations: Out of 134 package inserts, 64 were anti-diabetics, 40 anti-hypertensives, and 30 hypolipedemics. Out of them, 31 (23.14%) belonged to Grade �A� (including all injectable preparations) and remaining 76.86% to Grade �B�. None of the PIs belonged to Grade �C�. The inserts were inadequate in many aspects; for example, they had unclear instructions about generic name of other ingredients used, about handling, undesirable effects, pediatric and geriatric use, and guidelines for use of the drugs. Conclusion: This study indicated that information relevant to the safe and effective use of medication was not mentioned in the analyzed package inserts. It is, therefore, recommended to update the existing package inserts based on criteria mentioned in the Schedule D of Drug and Cosmetic Act, 1945. PMID:24392376
Ramdas, Deepak; Chakraborty, Ananya; HS, Swaroop; Faizan, Syed; Kumar V, Praveen; BN, Srinivas
Deep venous thrombosis after percutaneous insertion of vena caval filters
Purpose: A large multicenter study has recently questioned the overall clinical efficacy of vena caval filters, especially when inserted prophylactically, because of the subsequent development of deep venous thrombosis (DVT) at the insertion site. We examined the incidence of this complication with newer, smaller diameter percutaneous devices. Methods: We reviewed our vascular surgery and interventional radiology clinical registries to identify
John Blebea; Ryan Wilson; Peter Waybill; Marsha M. Neumyer; Judy S. Blebea; Karla M. Anderson; Robert G. Atnip
Lithium Insertion In Silicon Nanowires: An ab Initio Study
Lithium Insertion In Silicon Nanowires: An ab Initio Study Qianfan Zhang, Wenxing Zhang, Wenhui Wan, and � School of Physics, Peking University, Beijing 100871, China ABSTRACT The ultrahigh specific lithium ion opportunities for energy storage. However, a systematic theoretical study on lithium insertion in SiNWs remains
Cui, Yi
Combined plasma and thermal hollow cathode insert model
NASA Technical Reports Server (NTRS)
In this paper, we present the first results from a Hollow Cathode Thermal (HCThermal) model that uses the spatially distributed plasma fluxes calculated by the InsertRegion of an Orificed Cathode (IROrCa2D) code as the heat source to predict the hollow cathode and insert temperatures.
Katz, Ira; Polk, James E.; Mikellides, Ionnis G.; Goebel, Dan m.; Hornbeck, Sarah E.
Roll casting of wire inserted aluminum alloy strip
Casting of wire inserted composite strip was tried using a downward melt drag twin roll caster. Casting and insert of the wire was operated by one process. Therefore, many steps to make metal matrix composite alloy can be saved by the process of the present study. The effects of the important casting conditions, which are roll speed and nozzle slit,
T. Haga; K. Takahashi; H. Inui; H. Sakaguchi; H. Watari; S. Kumai
Computational Mechanics manuscript No. (will be inserted by the editor)
Computational Mechanics manuscript No. (will be inserted by the editor) Nonlinear fluid- tural and finite volume code OpenFOAM for fluid mechanics. This is achieved by using the Component the date of receipt and acceptance should be inserted later Abstract The main focus of the present article
Paris-Sud XI, Universit� de
Orbit Insertion by Mars Reconnaissance Orbiter (Artist's Concept)
NASA Technical Reports Server (NTRS)This is an artist's concept of NASA's Mars Reconnaissance Orbiter during the critical process of Mars orbit insertion. In order to be captured into orbit around Mars, the spacecraft must conduct a 25-minute rocket burn when it is just shy of reaching the planet. As pictured, it will pass under the red planet's southern hemisphere as it begins the insertion burn.
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.
Gas turbine nozzle vane insert and methods of installation
A pair of hollow elongated insert bodies are disposed in one or more of the nozzle vane cavities of a nozzle stage of a gas turbine. Each insert body has an outer wall portion with apertures for impingement-cooling of nozzle wall portions in registration with the outer wall portion. The insert bodies are installed into the cavity separately and spreaders flex the bodies toward and to engage standoffs against wall portions of the nozzle whereby the designed impingement gap between the outer wall portions of the insert bodies and the nozzle wall portions is achieved. The spreaders are secured to the inner wall portions of the insert bodies and the bodies are secured to one another and to the nozzle vane by welding or brazing.
Miller, William John (Simpsonville, SC); Predmore, Daniel Ross (Clifton Park, NY); Placko, James Michael (West Chester, OH)
Detection for processing history of seam insertion and contrast enhancement
NASA Astrophysics Data System (ADS)
With the development of manipulations techniques of digital images, digital image forensic technology is becoming more and more necessary. However, the determination of processing history of multi-operation is still a challenge problem. In this paper, we improve the traditional seam insertion algorithm, and propose corresponding detection method. Then an algorithm that focuses on detecting the processing history of seam insertion and contrast enhancement is proposed, which can be widely used in practical image forgery. Based on comprehensive analysis, we have discovered the inherent relationship between seam insertion and contrast enhancement. Different orders of processing make different impacts on images. By using the newly proposed algorithm, both contrast enhancement followed by seam insertion and seam insertion followed by contrast enhancement can be detected correctly. Plenty of experiments have been implemented to prove the accuracy.
Li, Jianwei; Zhao, Yao; Ni, Rongrong
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.
Image reconstruction algorithm for a special geometry of the PET-insert system
We are developing a PET-insert system with a half-ring insert that can not only image breast at a high resolution as other dedicated mammography systems but also image the chest and ancillary wall at a better resolution than the existing PET systems. Three different types of coincidences are acquired; coincidences between insert and insert (type II), coincidences between insert and
Debashish Pal; Daniel Keesing; Joseph A. O'Sullivan; Sergey Komarov; Yuan-Chuan Tai
Mining for Single Nucleotide Polymorphisms and Insertions/Deletions in Maize Expressed Sequence Tag Data1
We have developed a computer based method to identify candidate single nucleotide polymorphisms (SNPs) and small insertions/deletions from expressed sequence tag data. Using a redundancy-based approach, valid SNPs are distinguished from erroneous sequence by their representation multiple times in an alignment of sequence reads. A second measure of validity was also calculated based on the cosegregation of the SNP pattern between multiple SNP loci in an alignment. The utility of this method was demonstrated by applying it to 102,551 maize (Zea mays) expressed sequence tag sequences. A total of 14,832 candidate polymorphisms were identified with an SNP redundancy score of two or greater. Segregation of these SNPs with haplotype indicates that candidate SNPs with high redundancy and cosegregation confidence scores are likely to represent true SNPs. This was confirmed by validation of 264 candidate SNPs from 27 loci, with a range of redundancy and cosegregation scores, in four inbred maize lines. The SNP transition/transversion ratio and insertion/deletion size frequencies correspond to those observed by direct sequencing methods of SNP discovery and suggest that the majority of predicted SNPs and insertion/deletions identified using this approach represent true genetic variation in maize. PMID:12746514
Batley, Jacqueline; Barker, Gary; O'Sullivan, Helen; Edwards, Keith J.; Edwards, David
[Advantages and disadvantages of IUD insertion immediately after abortion].
In 672 women the interval insertion of intrauterine contraception devices (IUD) was applied: Lippes loop D in 138 women. In 530 women the insertion was applied immediately after artificial abortion: Lippes loop C in 390 and Lippes loop D in 140 cases. Data on the effectiveness of intrauterine contraception were biostatistically treated. One- and two-year gross cumulative rates of pregnancy were 4.4 and 6.4, respectively 3.7 and 6.3 for the interval insertion of Lippes loop C and D, and 2.1 and 4.5, respectively 4.2 and 11.2 for the postabortion insertion of the same devices. Two-year cumulative rates of spontaneous expulsions of 18.5 and 8.1 for interval insertions were significantly higher than in the group of women in whom the insertion of the same devices was performed immediately after abortion and in whom these rates were 11.1 and 5.9. The removal of intrauterine contraception devices because of bleeding and pain, or because of both, with the rates of 6.4 and 3.2 in 100 women after a two-year use, was less frequent in the group of women in whom interval insertion was performed than in the group of postabortion insertions with the corresponding rates of 7.3 and 9.9, respectively. To prevent repeated abortions, a frequent use of postabortion IUD insertion is recommended. Only when an intensified bleeding is expected, it is advisable to postpone insertion at least until the first menstruation. PMID:7132427
Matrljan, I; Randi?, L
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
The DrosDel Collection: A Set of P-Element Insertions for Generating Custom Chromosomal Aberrations in Drosophila melanogaster
We describe a collection of P-element insertions that have considerable utility for generating custom chromosomal aberrations in Drosophila melanogaster. We have mobilized a pair of engineered P elements, p{RS3} and p{RS5}, to collect 3243 lines unambiguously mapped to the Drosophila genome sequence. The collection contains, on average, an element every 35 kb. We demonstrate the utility of the collection for
Edward Ryder; Fiona Blows; Michael Ashburner; Rosa Bautista-Llacer; Jenny Drummond; Jane Webster; David Gubb; Nicola Gunton; Glynnis Johnson; Cahir J; David Huen; Punita Sharma; Zoltan Asztalos; Heiko Baisch; Janet Schulze; Maria Kube; Kathrin Kittlaus; Gunter Reuter; Peter Maroy; Janos Szidonya; Asa Rasmuson-Lestander; Karin Ekstr�m; Barry Dickson; Christoph Hugentobler; Hugo Stocker; Ernst Hafen; Jean Antoine Lepesant; Gert Pflugfelder; Bernard Mechler; Florenci Serras; Montserrat Corominas; Stephan Schneuwly; Thomas Preat; John Roote; Steven Russell
Adaptive Evolution Involving Gene Duplication and Insertion of a Novel Ty1 \\/ copia Like Retrotransposon in Soybean
Gene duplication is a major force for generating evolutionary novelties that lead to adaptations to environments. We previously\\u000a identified two paralogs encoding phytochrome A (phyA), GmphyA1 and GmphyA2, in soybean, a paleopolyploid species. GmphyA2 is encoded by the E4 locus responsible for photoperiod sensitivity. In photoperiod insensitive lines, GmphyA2 is inactivated by the insertion of a retrotransposon in exon 1.
Akira Kanazawa; Baohui Liu; Fanjiang Kong; Sachiko Arase; Jun Abe
We review the long wavelength description of three-dimensional liquids of entangled lines with an average orientation along the z-axis. Originally developed for polymer nematics, a closely related approach is applicable to the vortex liquid state of high temperature superconductors, and to ferro- and electrorheological fluids. The theory predicts unusual behavior of the three-dimensional structure function near the origin of reciprocal
David R. Nelson
Similar integration but different stability of Alus and LINEs in the human genome
Alus and LINEs (LINE1) are widespread classes of repeats that are very unevenly distributed in the human genome. The majority of GC-poor LINEs reside in the GC-poor isochores whereas GC-rich Alus are mostly present in GC-rich isochores. The discovery that LINES and Alus share similar target site duplication and a common AT-rich insertion site specificity raised the question as to
Adam Pavl???ek; Kamel Jabbari; Jan Pa?es; V�clav Pa?es; Ji??? Hejnar; Giorgio Bernardi
AKATSUKI status after the Venus orbit insertion
NASA Astrophysics Data System (ADS)
Japanese Venus Climate Orbiter 'AKATSUKI' (PLANET-C) was successfully launched at 06:58:22JST on May 21, by H-IIA F17. After the separation from H-IIA, the telemetry from AKATSUKI was normally detected by DSN Goldstone station (10:00JST) and the solar cell paddles' expansion was confirmed. AKATSUKI was put into the 3-axis stabilized mode in the initial operation from Uchinoura station and the critical operation was finished at 20:00JST on the same day. AKATSUKI will arrive at Venus on Dec 7, 2010. From the orbit around Venus, it will take pictures of the distribution of clouds and minor gas components at different wavelength by 4 cameras. These data will be analyzed to study the meteorology on Venus, especially the mechanism of the super-rotation of Venusian atmosphere which is still the top mystery of the Venus science. Simultaneous observation with ESA's Venus Express is highly expected to study the Venusian atmosphere. We will present the present status of AKATSUKI after the Venus orbit insertion with the latest results from the cameras.
Nakamura, M.; Ishii, N.; Imamura, T.; Ueno, M.; Yamazaki, A.; Satoh, T.; Suzuki, M.; Iwagami, N.; Taguchi, M.; Watanabe, S.; Takahashi, Y.; Fukuhara, T.; Ohtsuki, S.; PLANET-C Project Team
IYA2009 newspaper insert in your community
NASA Astrophysics Data System (ADS)
The Gemini PIO (Public Information Officer) offers suggestions on how to approach your local newspaper about a newspaper insert for your community being published during IYA2009. Local government support, articles by astronomers, advertisers, and appointing someone within your organisation to manage the content will be discussed. We will explain the timeline required, the number of personnel hours required, developmental stages and income a local newspaper would have to generate to produce a quality, table-top supplement. In 2003, over 30,000 copies of Stars over Mauna Kea, a special supplement in tabloid format were produced and distributed in the local newspapers in Hilo, Hawaii. The publication, 48 pages in total, featured profiles of the observatories on Mauna Kea, stories about the geology and legends of Mauna Kea, and historical information about the evolution of astronomy in Hawaii. In additionthe publication included a series of essays titled In their own words. These articles were written by key members of the astronomy community. Sixty thousand copies of Stars over Mauna Kea II were printed as a follow-up to the first edition in 2005. This second edition included an article on the `Imiloa Astronomy Education Center, explanations about the types of telescopes on Mauna Kea and columns written by scientists about the fascinating and significant discoveries made on Hawaii. Personal stories about careers in astronomy were also highlighted. In Chile, a similar eight-page supplement, featuring Gemini, CTIO and SOAR telescopes, was published in 2005 and 5000 copies were distributed throughout the country.
Harvey, J.
Insertion sequence IST3091 of Thiobacillus ferrooxidans.
An insertion sequence, designated as IST3091, was located adjacent to the putative origin of replication region of plasmid pTFI91 of Thiobacillus ferrooxidans TFI-91. The DNA sequence of the transposase gene of IST3091 revealed similarity with that of IS30, IS1086, IS4351, and the integrase gene of SpV1-R8A2 B (a bacteriophage of Spiroplasma citri). The sequence of IST3091 is 1063 bp long with partially matched 30-bp terminal inverted repeats. Several restriction fragments of plasmid pTFI91 of T. ferrooxidans containing the IST3091 element were cloned into the vector pHSG398. The hybrid plasmids (pBTL) were transformed into Escherichia coli NK7379 containing a miniF plasmid, which was devoid of transposable elements. The transposition function of the IST3091 element was confirmed by mobilizing hybrid plasmids via conjugation from transformed E. coli NK7379 (donor) to E. coli M8820 (recipient). The presence of the transposed element in transconjugants was detected by polymerase chain reaction amplification. PMID:9226869
Chakravarty, L; Kittle, J D; Tuovinen, O H
Phoenix Conductivity Probe Inserted into Martian Soil
NASA Technical Reports Server (NTRS)NASA's Phoenix Mars Lander inserted the four needles of its thermal and conductivity probe into Martian soil during the 98th Martian day, or sol, of the mission and left it in place until Sol 99 (Sept. 4, 2008). The Robotic Arm Camera on Phoenix took this image on the morning of Sol 99 while the probe's needles were in the ground. The science team informally named this soil target 'Gandalf.' The thermal and conductivity probe measures how fast heat and electricity move from one needle to an adjacent one through the soil or air between the needles. Conductivity readings can be indicators about water vapor, water ice and liquid water. The probe is part of Phoenix's Microscopy, Electrochemistry and Conductivity suite of instruments. The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.
The ATLAS Insertable B-Layer Project
The ATLAS experiment will upgrade its Pixel Detector with the installation of a new pixel layer in 2013-14. The new sub-detector, named Insertable B-layer (IBL), will be installed between the existing Pixel Detector and a new smaller radius beam-pipe at a radius of 3.3 cm. To cope with the high radiation and pixel occupancy due to the proximity to the interaction point, a new read-out chip and two different silicon sensor technologies (planar and 3D) have been de- veloped. Furthermore, the physics performance should be improved through the reduction of pixel size while a low material budget should be imposed. A new mechanical support using lightweight staves and a CO2 based cooling system is used. An overview of the IBL project and the status of the production of staves and the qualification of the assembly procedure, the loaded module electrical integrity and the read-out chain will be presented.
Miucci, A; The ATLAS collaboration
Structural basis for outer membrane lipopolysaccharide insertion.
Lipopolysaccharide (LPS) is essential for most Gram-negative bacteria and has crucial roles in protection of the bacteria from harsh environments and toxic compounds, including antibiotics. Seven LPS transport proteins (that is, LptA-LptG) form a trans-envelope protein complex responsible for the transport of LPS from the inner membrane to the outer membrane, the mechanism for which is poorly understood. Here we report the first crystal structure of the unique integral membrane LPS translocon LptD-LptE complex. LptD forms a novel 26-stranded ?-barrel, which is to our knowledge the largest ?-barrel reported so far. LptE adopts a roll-like structure located inside the barrel of LptD to form an unprecedented two-protein 'barrel and plug' architecture. The structure, molecular dynamics simulations and functional assays suggest that the hydrophilic O-antigen and the core oligosaccharide of the LPS may pass through the barrel and the lipid A of the LPS may be inserted into the outer leaflet of the outer membrane through a lateral opening between strands ?1 and ?26 of LptD. These findings not only help us to understand important aspects of bacterial outer membrane biogenesis, but also have significant potential for the development of novel drugs against multi-drug resistant pathogenic bacteria. PMID:24990744
Dong, Haohao; Xiang, Quanju; Gu, Yinghong; Wang, Zhongshan; Paterson, Neil G; Stansfeld, Phillip J; He, Chuan; Zhang, Yizheng; Wang, Wenjian; Dong, Changjiang
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...
21 CFR 872.3900 - Posterior artificial tooth with a metal insert.
Code of Federal Regulations, 2012 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...
21 CFR 872.3900 - Posterior artificial tooth with a metal insert.
Code of Federal Regulations, 2011 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...
21 CFR 310.501 - Patient package inserts for oral contraceptives.
Code of Federal Regulations, 2010 CFR
...2010-04-01 2010-04-01 false Patient package inserts for oral contraceptives...New Drugs or Devices � 310.501 Patient package inserts for oral contraceptives. (a) Requirement for a patient package insert. The safe and...
21 CFR 872.3900 - Posterior artificial tooth with a metal insert.
Code of Federal Regulations, 2010 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...
Isthmus-guided cortical bone trajectory for pedicle screw insertion.
Herein is described cortical bone trajectory (CBT), a new path for pedicle screw insertion for lumbar vertebral fusion. Because the points of insertion are under the end of the inferior articular process, and because the screws are inserted toward the lateral side, there is less soft tissue development than with the conventional technique; the CBT technique therefore enables less invasive surgery than the conventional technique. However, it has some drawbacks. For example, in the original CBT approach, the points of insertion are in the vicinity of the end of the inferior articular process. Because this joint has been destroyed in many patients who have indications for intervertebral fusion surgery, it is sometimes difficult to use it as a reference point for screw insertion location. With severe lateral slippage, the screw insertion site can become significantly dislocated sideways, with possible resultant damaging to the spinal canal and/or nerve root. The CBT technique here involved inserting the screws while keeping clear of the intervertebral foramen with the assistance of side view X-ray fluoroscopy and using the end of the inferior articular process and the isthmus as points of reference for screw location. PMID:25179360
Iwatsuki, Koichi; Yoshimine, Toshiki; Ohnishi, Yu-ichiro; Ninomiya, Kosi; Ohkawa, Toshika
The genomic landscape of polymorphic human nuclear mitochondrial insertions
The transfer of mitochondrial genetic material into the nuclear genomes of eukaryotes is a well-established phenomenon that has been previously limited to the study of static reference genomes. The recent advancement of high throughput sequencing has enabled an expanded exploration into the diversity of polymorphic nuclear mitochondrial insertions (NumtS) within human populations. We have developed an approach to discover and genotype novel Numt insertions using whole genome, paired-end sequencing data. We have applied this method to a thousand individuals in 20 populations from the 1000 Genomes Project and other datasets and identified 141 new sites of Numt insertions, extending our current knowledge of existing NumtS by almost 20%. We find that recent Numt insertions are derived from throughout the mitochondrial genome, including the D-loop, and have integration biases that differ in some respects from previous studies on older, fixed NumtS in the reference genome. We determined the complete inserted sequence for a subset of these events and have identified a number of nearly full-length mitochondrial genome insertions into nuclear chromosomes. We further define their age and origin of insertion and present an analysis of their potential impact to ongoing studies of mitochondrial heteroplasmy and disease. PMID:25348406
Dayama, Gargi; Emery, Sarah B.; Kidd, Jeffrey M.; Mills, Ryan E.
Estimating the fitness effect of an insertion sequence.
Since its discovery, mobile DNA has fascinated researchers. In particular, many researchers have debated why insertion sequences persist in prokaryote genomes and populations. While some authors think that insertion sequences persist only because of occasional beneficial effects they have on their hosts, others argue that horizontal gene transfer is strong enough to overcome their generally detrimental effects. In this study, we model the long-term fate of a prokaryote cell population, of which a small proportion of cells has been infected with one insertion sequence per cell. Based on our model and the distribution of IS5, an insertion sequence for which sufficient data is available in 525 fully sequenced proteobacterial genomes, we show that the fitness cost of insertion sequences is so small that they are effectively neutral or only slightly detrimental. We also show that an insertion sequence infection can persist and reach the empirically observed distribution if the rate of horizontal gene transfer is at least as large as the fitness cost, and that this rate is well within the rates of horizontal gene transfer observed in nature. In addition, we show that the time needed to reach the observed prevalence of IS5 is unrealistically long for the fitness cost and horizontal gene transfer rate that we computed. Occasional beneficial effects may thus have played an important role in the fast spreading of insertion sequences like IS5. PMID:22252506
Bichsel, Manuel; Barbour, A D; Wagner, Andreas
Insertional tagging of at least two loci associated with resistance to adenine arabinoside in Toxoplasma gondii, and cloning of the adenosine kinase locus
A genetic approach has been exploited to investivate adenylate salvage pathways in the protozoan parasite Toxoplasma gondii, a purine auxotroph. Using a new insertional mutagenesis vector designed to facilitate the rescue of tagged loci even when multiple plasmids integrate as a tandem array, 15 independent clonal lines resistant to the toxic nucleoside analog adenine arabinoside (AraA) were generated. Approximately two-thirds
William J. Sullivan; Chi-Wu Chiang; Craig M. Wilson; Fardos N. M. Naguib; Mahmoud H. el Kouni; Robert G. K. Donald; David S. Roos
Copy-number dependent transpositions and excisions of the mdg-1 mobile element in inbred lines of Drosophila melanogaster
The chromosomal location of the mobile element mdg-1 was studied in 17 highly-inbred lines of Drosophila melanogaster. Although some lines were stable for their pattern of insertion sites from the 15th to the 27th and 35th brother�sister generations, others showed a high rate of gain of new insertion sites or a high rate of excision (loss of elements). In one
C Bi�mont; A Aouar
International Workshop on Magnetic Measurements of Insertion Devices
The International Workshop on Magnetic Measurements of Insertion Devices was held at the Advanced Photon Source, Argonne National Laboratory, on September 28--29, 1993. The workshop brought together scientists and engineers from Europe, Japan, and the United States to discuss the following topics: Special techniques for magnetic measurements of insertion devices, magnetic tolerances of the insertion devices for third generation synchrotron radiation sources, methods for and accuracy of the multipole moments measurements, magnetic sensors, among other topics. The workshop included thirteen presentations that are collected in this volume.
Not Available
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.
Isolation and molecular characterization of a Spotted leaf 18 mutant by modified activation-tagging in rice
A lesion mimic mutant that we designated Spotted leaf 18 (Spl18) was isolated from 13,000 activation-tagging lines of rice produced by our modified activation-tagging vector and further\\u000a characterized. Spl18 was dominant and its phenotype was linked to the T-DNA insertion. An ORF was located about 500�bp downstream of the inserted\\u000a T-DNA, and the deduced protein, designated OsAT1, showed sequence similarity
Masaki Mori; Chikako Tomita; Kazuhiko Sugimoto; Morifumi Hasegawa; Nagao Hayashi; Joseph G. Dubouzet; Hirokazu Ochiai; Hitoshi Sekimoto; Hirohiko Hirochika; Shoshi Kikuchi
Error compensation in insertion-region magnets
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
Assessment of genetic variation for the LINE1 retrotransposon from next generation sequence data
BACKGROUND: In humans, copies of the Long Interspersed Nuclear Element 1 (LINE-1) retrotransposon comprise 21% of the reference genome, and have been shown to modulate expression and produce novel splice isoforms of transcripts from genes that span or neighbor the LINE-1 insertion site. RESULTS: In this work, newly released pilot data from the 1000 Genomes Project is analyzed to detect
Eric Rouchka; Diego E Montoya-Durango; Vilius Stribinskis; Kenneth Ramos; Ted Kalbfleisch
Myositis ossificans of the humeral insertion of pectoralis major
We report on a rare case of myositis ossificans of the humeral insertion of pectoralis major muscle following a single episode of trauma which, to our knowledge, has not previously been documented. PMID:25114419
Mogere, Viga; Thyagarajan, David S. K.; Fairbairn, K. Julia; Wallace, Angus
Pressure molding of powdered materials improved by rubber mold insert
NASA Technical Reports Server (NTRS)
Pressure molding tungsten microspheres is accomplished by applying hydraulic pressure to a silicone rubber mold insert with several barrel shaped chambers which is placed in a steel die cavity. This technique eliminates castings containing shear fractures.
Computational Prediction of Insertional RNA Editing Ralf Bundschuh
it is translated · Example: mitochondrion of Physarum polycephalum most prevalent editing event: C insertion e as possible · Situation for mitochondrion of Physarum polycephalum: six protein coding genes
Bundschuh, Ralf
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2013 CFR
...estrogens requires that patients be fully informed of the benefits and risks involved in...patients with a patient package insert...drug's benefits and risks. An estrogen...Instructions on how a patient may reduce the risks of estrogen...
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2014 CFR
...estrogens requires that patients be fully informed of the benefits and risks involved in...patients with a patient package insert...drug's benefits and risks. An estrogen...Instructions on how a patient may reduce the risks of estrogen...
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2012 CFR
...estrogens requires that patients be fully informed of the benefits and risks involved in...patients with a patient package insert...drug's benefits and risks. An estrogen...Instructions on how a patient may reduce the risks of estrogen...
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2011 CFR
...estrogens requires that patients be fully informed of the benefits and risks involved in...patients with a patient package insert...drug's benefits and risks. An estrogen...Instructions on how a patient may reduce the risks of estrogen...
The use of dimorphic Alu insertions in human DNA fingerprinting
We have characterized certain Human Specific Alu Insertions as either dimorphic (TPA25, PV92, APO), sightly dimorphic (C2N4 and C4N4) or monomorphic (C3N1, C4N6, C4N2, C4N5, C4N8), based on studies of Caucasian, Asian, American Black and African Black populations. Our approach is based upon: (1) PCR amplification using primers directed to the sequences that flank the site of insertion of the different Alu elements studied; (2) gel electrophoresis and scoring according to the presence or absence of an Alu insertion in one or both homologous chromosomes; (3) allelic frequencies calculated and compared according to Hardy-Weinberg equilibrium. Our DNA fingerprinting procedure using PCR amplification of dimorphic Human Specific Alu insertions, is stable enough to be used not only as a tool for genetic mapping but also to characterize populations, study migrational patterns and track the inheritance of human genetic disorders.
Novick, G.E.; Gonzalez, T.; Garrison, J.; Novick, C.C.; Herrera, R.J. [Florida International Univ., Miami, FL (United States). Dept. of Biological Sciences; Batzer, M.A. [Lawrence Livermore National Lab., CA (United States); Deininger, P.L. [Louisiana State Univ., New Orleans, LA (United States). Medical Center
External-memory search trees with fast insertions
This thesis provides both experimental and theoretical contributions regarding external-memory dynamic search trees with fast insertions. The first contribution is the implementation of the buffered repository B-tree, a ...
Nelson, Jelani (Jelani Osei)
Effectiveness Review Analysis (Insert Effectiveness Review Name) Root Cause
Effectiveness Review Analysis � (Insert Effectiveness Review Name) 1 of 1 Root Cause: Corrective action address the root cause? 2. Does the corrective action prevent recurrence of similar conditions due
Management of insertional Achilles tendinopathy through a Cincinnati incision
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 and debridement of the peritendinous and tendon tissue with osteotomy of the calcaneum through a transverse skin incision at the level of the Achilles insertion. Results This method has been used since 2002 on over 40 patients for exposure of the Achilles tendon insertion and the distal Achilles tendon. Conclusion The Cincinnati incision allows adequate exposure, has minimal risk of symptomatic iatrogenic nerve injury, and has minimal problems related to the scar. PMID:17697370
Carmont, Michael R; Maffulli, Nicola
LS-283, Rev.1 Radiation Dose Measurements of the Insertion
Nd-Fe-B permanent magnets are also used. Growing concern for the lifetime of APS insertion devices,15]. The change in the color intensity, or optical density, is measured using a radiachromic reader, or simple
Kemner, Ken
Master Thesis: Simulation of plastic deformation in cemented carbide inserts
in cemented carbide, high-speed steel and other hard materials such as diamond, cubic boron nitride.g. turning or milling, the inserts are exposed to large forces at high temperatures. This will result
Haviland, David
The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes.
A fundamental goal of genetics and functional genomics is to identify and mutate every gene in model organisms such as Drosophila melanogaster. The Berkeley Drosophila Genome Project (BDGP) gene disruption project generates single P-element insertion strains that each mutate unique genomic open reading frames. Such strains strongly facilitate further genetic and molecular studies of the disrupted loci, but it has remained unclear if P elements can be used to mutate all Drosophila genes. We now report that the primary collection has grown to contain 1045 strains that disrupt more than 25% of the estimated 3600 Drosophila genes that are essential for adult viability. Of these P insertions, 67% have been verified by genetic tests to cause the associated recessive mutant phenotypes, and the validity of most of the remaining lines is predicted on statistical grounds. Sequences flanking >920 insertions have been determined to exactly position them in the genome and to identify 376 potentially affected transcripts from collections of EST sequences. Strains in the BDGP collection are available from the Bloomington Stock Center and have already assisted the research community in characterizing >250 Drosophila genes. The likely identity of 131 additional genes in the collection is reported here. Our results show that Drosophila genes have a wide range of sensitivity to inactivation by P elements, and provide a rationale for greatly expanding the BDGP primary collection based entirely on insertion site sequencing. We predict that this approach can bring >85% of all Drosophila open reading frames under experimental control. PMID:10471706
Spradling, A C; Stern, D; Beaton, A; Rhem, E J; Laverty, T; Mozden, N; Misra, S; Rubin, G M
A natural allele of Nxf1 suppresses retrovirus insertional mutations
Endogenous retroviruses have shaped the evolution of mammalian genomes. Host genes that control the effects of retrovirus insertions are therefore of great interest. The modifier-of-vibrator-1 locus (Mvb1) controls levels of correctly processed mRNA from genes mutated by endogenous retrovirus insertions into introns, including the Pitpnvb tremor mutation and the Eya1BOR model of human branchiootorenal syndrome. Positional complementation cloning identifies Mvb1
Jennifer A Floyd; David A Gold; Dorothy Concepcion; Tiffany H Poon; Xiaobo Wang; Elizabeth Keithley; Dan Chen; Erica J Ward; Steven B Chinn; Rick A Friedman; Hon-Tsen Yu; Kazuo Moriwaki; Toshihiko Shiroishi; Bruce A Hamilton
Production Recesses for Replaceable Cutting Inserts of Milling Tools
NASA Astrophysics Data System (ADS)
This article describes the production of recesses for replaceable cutting inserts of milling tools. The recess manufacture is exemplified on a D20 shanktype milling cutter with three replaceable cutting inserts. A new production technology and its operating cycle are demonstrated. A com-parison of the old and the new technologies shows the percentage saving of change cycles and used tools, including overall evaluation of the tech-nology.
Kousal, Luk�; Sad�lek, Marek
Impediments to rapid insertion of innovative displays and peripherals
NASA Astrophysics Data System (ADS)
In order to optimize system performance and minimize cost for a system to fill capability gaps, an improvement to rapid insertion of innovative display and peripheral technology is required to take advantage of human-machine intersections. Current approaches to testing and integration impedes successful rapid insertion of innovative technology for new systems and incremental upgrades. Considerations to innovative displays and peripherals must occur further to the left of the lifecycle to be successful and key integration areas must be address for success.
Nicholson, Gail
Evaluation of preferable insertion routes for esophagogastroduodenoscopy using ultrathin endoscopes
AIM: To evaluate the discomfort associated with esophagogastroduodenoscopy (EGD) using an ultrathin endoscope through different insertion routes. METHODS: This study (January 2012-March 2013) included 1971 consecutive patients [male/female (M/F), 1158/813, 57.5 � 11.9 years] who visited a single institute for annual health checkups. Transnasal EGD was performed in 1394 patients and transoral EGD in 577. EGD-associated discomfort was assessed using a visual analog scale score (VAS score: 0-10). RESULTS: Multivariate analysis revealed gender (M vs F: 4.02 � 2.15 vs 5.06 � 2.43) as the only independent predictor of the VAS score in 180 patients who underwent EGD for the first time; whereas it revealed gender (M vs F 3.60 � 2.20 vs 4.84 � 2.37), operator, age group (A: < 39 years; B: 40-49 years; C: 50-59 years; D: 60-69 years; E: > 70 years; A/B/C/D/E: 4.99 � 2.32/4.34 � 2.49/4.19 � 2.31/3.99 � 2.27/3.63 � 2.31), and type of insertion as independent predictors in the remaining patients. Subanalysis for gender, age group, and insertion route revealed that the VAS score decreased with age regardless of gender and insertion route, was high in female patients regardless of age and insertion route, and was low in males aged over 60 years who underwent transoral insertion. CONCLUSION: Although comprehensive analysis revealed that the insertion route may not be an independent predictor of the VAS score, transoral insertion may reduce EGD-associated discomfort in elderly patients. PMID:24803817
Ono, Satoshi; Niimi, Keiko; Fujishiro, Mitsuhiro; Takahashi, Yu; Sakaguchi, Yoshiki; Nakayama, Chiemi; Minatsuki, Chihiro; Matsuda, Rie; Hirayama-Asada, Itsuko; Tsuji, Yosuke; Mochizuki, Satoshi; Kodashima, Shinya; Yamamichi, Nobutake; Ozeki, Atsuko; Matsumoto, Lumine; Ohike, Yumiko; Yamazaki, Tsutomu; Koike, Kazuhiko
Evolution models with base substitutions, insertions, deletions, and selection
NASA Astrophysics Data System (ADS)
The evolution model with parallel mutation-selection scheme is solved for the case when selection is accompanied by base substitutions, insertions, and deletions. The fitness is assumed to be either a single-peak function (i.e., having one finite discontinuity) or a smooth function of the Hamming distance from the reference sequence. The mean fitness is calculated exactly in large-genome limit. In the case of insertions and deletions the evolution characteristics depend on the choice of reference sequence.
Saakian, D. B.
Simulation and optimization of the dual lithium ion insertion cell
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 Li�C�{vert_bar}propylene carbonate + 1M LiClO�{vert_bar}Li{sub y}Mn�O� cell, and these
Thomas F. Fuller; M. Doyle; J. Newman
Roll Casting of Net Inserted Aluminum Alloy Strip
The net-inserted metal matrix composite strip could be cast by a twin roll caster at only one process. The caster for composite\\u000a strip was assembled by combination of the open-top melt drag type and the downward melt drag type caster. The matrix was Al-12mass%Si,\\u000a and inserted net was made from SUS304 stainless steel. The diameter of the wire of the
Toshio Haga; Kenta Takahashi; Ryoji Nakamura; Hisaki Watari
In vivo evaluation of nicotine lyophilised nasal insert in sheep.
The nasal route offers an attractive means of delivering a drug directly to the systemic circulation and avoiding hepatic first-pass metabolism, although rapid mucociliary clearance can be detrimental to nasal absorption. The in vitro and in vivo characteristics of a nasal insert formulation prepared by lyophilisation of a viscous HPMC gel solution designed to overcome this problem were studied. In vitro release of nicotine from the lyophilised insert was compared with powder and spray formulations. Stability and characterisation studies were carried out using dynamic vapour sorption, scanning electron microscopy and HPLC analysis. Nicotine formulations were administered to eight wether sheep in a randomised four-way cross-over study, and plasma nicotine assessed comparing the nasal insert formulation with conventional nasal powder, nasal spray and IV doses. In vitro release studies demonstrated prolonged nicotine release from the nasal insert formulation compared to a powder and liquid. In vivo plasma profiles appeared to show prolonged plasma nicotine levels compared to the conventional formulations, although T(max), C(max) and AUC parameters for the insert were not significantly different due to high variability in the pharmacokinetic data. In conclusion, the nasal insert displayed a promising prolonged plasma profile, which must be investigated further to provide statistical significance to prove the effect. PMID:16183221
McInnes, Fiona J; Thapa, Panna; Baillie, Alan J; Welling, Peter G; Watson, David G; Gibson, Ian; Nolan, Andrea; Stevens, Howard N E
Sensorless Motion Planning for Medical Needle Insertion in Deformable Tissues
Minimally invasive medical procedures such as biopsies, anesthesia drug injections, and brachytherapy cancer treatments require inserting a needle to a specific target inside soft tissues. This is difficult because needle insertion displaces and deforms the surrounding soft tissues causing the target to move during the procedure. To facilitate physician training and preoperative planning for these procedures, we develop a needle insertion motion planning system based on an interactive simulation of needle insertion in deformable tissues and numerical optimization to reduce placement error. We describe a 2-D physically based, dynamic simulation of needle insertion that uses a finite-element model of deformable soft tissues and models needle cutting and frictional forces along the needle shaft. The simulation offers guarantees on simulation stability for mesh modications and achieves interactive, real-time performance on a standard PC. Using texture mapping, the simulation provides visualization comparable to ultrasound images that the physician would see during the procedure. We use the simulation as a component of a sensorless planning algorithm that uses numerical optimization to compute needle insertion offsets that compensate for tissue deformations. We apply the method to radioactive seed implantation during permanent seed prostate brachytherapy to minimize seed placement error. PMID:19126473
Alterovitz, Ron; Goldberg, Kenneth Y.; Pouliot, Jean; Hsu, I-Chow
Bits with diamond-coated inserts reduce gauge problems
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))
Gain of function mutation in tobacco MADS box promoter switch on the expression of flowering class B genes converting sepals to petals.
One mutant transgenic line displaying homeotic conversion of sepals to petals with other phenotypic aberrations was selected and characterized at molecular level. The increased transcript level of gene encoding anthocyanidin synthase and petal specific class B genes, GLOBOSA and DEFECIENS in sepals of mutant line may be responsible for its homeotic conversion to petaloid organs. While characterizing this mutant line for locus identification, T-DNA was found to be inserted in 3' untranslated region of promoter of class B MADS box gene, GLOBOSA. Here, CaMV 35S promoter of T-DNA might be deriving the expression of class B genes. PMID:24362510
Mahajan, Monika; Yadav, Sudesh Kumar
Metal Insertion in a Microporous Metal-Organic Framework Lined with 2,2 '-Bipyridine
Reaction of AlCl{sub 3}�6H{sub 2}O with 2,2?-bipyridine-5,5?-dicarboxylic acid (H{sub 2}bpydc) affords Al(OH)(bpydc) (1, MOF-253), the first metal?organic framework with open 2,2?-bipyridine (bpy) coordination sites. The material displays a BET surface area of 2160 m{sup 2}/g and readily complexes metals to afford, for example, 1�xPdCl{sub 2} (x = 0.08, 0.83) and 1�0.97Cu(BF{sub 4}){sub 2}. EXAFS spectroscopy performed on 1�0.83PdCl{sub 2} reveals the expected square planar coordination geometry, matching the structure of the model complex (bpy)PdCl{sub 2}. Significantly, the selectivity factor for binding CO{sub 2} over N{sub 2} under typical flue gas conditions is observed to increase from 2.8 in 1 to 12 in 1�0.97Cu(BF{sub 4}){sub 2}.
Bloch, Eric D; Britt, David; Lee, Chain; Doonan, Christian J; Uribe-Romo, Fernando J; Furukawa, Hiroyasu; Long, Jeffrey R; Yaghi, Omar M
The nature of the transition mismatches with Watson-Crick architecture: the G*�T or G�T* DNA base mispair or both? A QM/QTAIM perspective for the biological problem.
This study provides the first accurate investigation of the tautomerization of the biologically important guanine*�thymine (G*�T) DNA base mispair with Watson-Crick geometry, involving the enol mutagenic tautomer of the G and the keto tautomer of the T, into the G�T* mispair (?G?=?.99?kcal?mol(-1), population?=?15.8% obtained at the MP2 level of quantum-mechanical theory in the continuum with ??=?4), formed by the keto tautomer of the G and the enol mutagenic tautomer of the T base, using DFT and MP2 methods in vacuum and in the weakly polar medium (??=?4), characteristic for the hydrophobic interfaces of specific protein-nucleic acid interactions. We were first able to show that the G*�T?G�T* tautomerization occurs through the asynchronous concerted double proton transfer along two antiparallel O6H���O4 and N1���HN3 H-bonds and is assisted by the third N2H���O2 H-bond, that exists along the entire reaction pathway. The obtained results indicate that the G�T* base mispair is stable from the thermodynamic point of view complex, while it is dynamically unstable structure in vacuum and dynamically stable structure in the continuum with ??=?4 with lifetime of 6.4�10(-12)?s, that, on the one side, makes it possible to develop all six low-frequency intermolecular vibrations, but, on the other side, it is by three orders less than the time (several ns) required for the replication machinery to forcibly dissociate a base pair into the monomers during DNA replication. One of the more significant findings to emerge from this study is that the short-lived G�T* base mispair, which electronic interaction energy between the bases (-23.76?kcal?mol(-1)) exceeds the analogical value for the G�C Watson-Crick nucleobase pair (-20.38?kcal?mol(-1)), "escapes from the hands" of the DNA replication machinery by fast transforming into the G*�T mismatch playing an indirect role of its supplier during the DNA replication. So, exactly the G*�T mismatch was established to play the crucial role in the spontaneous point mutagenesis. PMID:24842163
Brovarets', Ol'ha O; Hovorun, Dmytro M
Technology Transfer Automated Retrieval System (TEKTRAN)
Arabidopsis UDP-sugar pyrophosphorylase (AtUSP) is a broad substrate enzyme that catalyzes the synthesis of UDP-sugars. We identified two knockout lines (usp-1 and usp-2) with T-DNA insertional events within the AtUSP coding region. From selfed heterozygous individuals, only wild-type and heterozygo...
LETTER TO THE EDITOR F-box protein AFB4 plays a crucial role in plant growth,
of physiological, molecular, and genetic approaches to characterize a T-DNA insertion line (GABI-KAT ac- cession no confers defects in many aspects of the plant life cycle including lateral root development, hypocotyl elon- gation, leaf organogenesis, flowering time, seed forma- tion, and disease resistance to specific
Palva, Tapio
Small bowel injury after suprapubic catheter insertion presenting 3 years after initial insertion.
A 77-year-old woman was referred to urology with blockages of her suprapubic catheter (SPC). The catheter was replaced easily in the emergency department, however, no urine was draining, only a cloudy green fluid was visible. On cystoscopy bilious material was identified in the bladder. There was no catheter visible. There seemed to be a fistulous tract entering the bladder at the left dome. The urethra was dilated, a urethral catheter was placed and the SPC was removed. A CT demonstrated that the SPC tract transfixed a loop of pelvic small bowel and entered the bladder with no intraperitoneal contrast leak. The patient recovered well and did not require laparotomy. This case emphasises that bowel perforation, although rare, must be considered as a complication of SPC placement even years after initial insertion when catheter problems arise. Unusually, we learn that this complication may not present with abdominal pain or peritonism. PMID:24326435
Gallagher, Kevin M; Good, Daniel W; Brush, John P; Alhasso, Ammar; Al-hasso, Ammar; Stewart, Grant D
Definition of Metrics to Evaluate Cochlear Array Insertion Forces Performed with Forceps, Insertion Tool, or Motorized Tool in Temporal Bone Specimens
Introduction. In order to achieve a minimal trauma to the inner ear structures during array insertion, it would be suitable to control insertion forces. The aim of this work was to compare the insertion forces of an array insertion into anatomical specimens with three different insertion techniques: with forceps, with a commercial tool, and with a motorized tool. Materials and Methods. Temporal bones have been mounted on a 6-axis force sensor to record insertion forces. Each temporal bone has been inserted, with a lateral wall electrode array, in random order, with each of the 3 techniques. Results. Forceps manual and commercial tool insertions generated multiple jerks during whole length insertion related to fits and starts. On the contrary, insertion force with the motorized tool only rose at the end of the insertion. Overall force momentum was 1.16 � 0.505?N (mean � SD, n = 10), 1.337 � 0.408?N (n = 8), and 1.573 � 0.764?N (n = 8) for manual insertion with forceps and commercial and motorized tools, respectively. Conclusion. Considering force momentum, no difference between the three techniques was observed. Nevertheless, a more predictable force profile could be observed with the motorized tool with a smoother rise of insertion forces. PMID:25126565
Ferrary, Evelyne; Sterkers, Olivier
The Insertion Green Monster (iGM) Method for Expression of Multiple Exogenous Genes in Yeast
1 The Insertion Green Monster (iGM) Method for Expression of Multiple Exogenous Genes in Yeast;2 Running title: The Insertion Green Monster Method Key words: synthetic biology, multi-gene insertions Green Monster' (iGM) set of expression vectors that enable precise insertion of many heterologous genes
Roth, Frederick
Diamond studded insert drag bit with strategically located hydraulic passages for mud motors
A diamond studded insert drag bit is disclosed having a multiplicity of individual diamond insert cutter blanks inserted in the face of the bit. The diamond insert blanks are so positioned to maximize penetration of the bit in a borehole. The bit further includes fluid passages strategically located in the bit face to provide uniform flow, cooling and continuous cleaning
Management of pain associated with the insertion of intrauterine contraceptives
BACKGROUND Most intrauterine contraception (IUC) placements do not require pain relief. However, small proportions of nulliparous (?17%) and parous (?11%) women experience substantial pain that needs to be proactively managed. This review critically evaluates the evidence for pain management strategies, formulates evidence-based recommendations and identifies data gaps and areas for further research. METHODS A PubMed literature search was undertaken. Relevant articles on management of pain associated with IUC insertion, published in English between 1980 and November 2012, were identified using the following search terms: �intrauterine contraception�, �insertion� and �pain�. RCTs were included; further relevant articles were also identified and included as appropriate. RESULTS Seventeen studies were identified and included: 12 RCTs and one non-randomized study of pre-insertion oral analgesia, cervical priming and local anaesthesia; one systematic review and one RCT on post-insertion analgesia and two non-randomized studies on non-pharmacological interventions. There was no conclusive evidence that any prophylactic pharmacological intervention reduces pain associated with IUC insertion. However, most of the regimens studied were adopted from hysteroscopy or abortion and effectiveness in specific subsets of women has not been studied adequately. A systematic review found non-steroidal anti-inflammatory agents (NSAID) to be effective in reactively treating post-insertion pain, but no benefit was found with prophylactic use. CONCLUSIONS No prophylactic pharmacological intervention has been adequately evaluated to support routine use for pain reduction during or after IUC insertion. Women's anxiety about the procedure may contribute to higher levels of perceived pain, which highlights the importance of counselling, and creating a trustworthy, unhurried and professional atmosphere in which the experience of the provider also has a major role; a situation frequently referred to as �verbal anaesthesia�. PMID:23670222
Gemzell-Danielsson, K.; Mansour, D.; Fiala, C.; Kaunitz, A.M.; Bahamondes, L.
Palladium- and Nickel-Catalyzed Carbon�Carbon Bond Insertion Reactions with Alkylidenesilacyclopropanes
Palladium and nickel catalysts promoted highly selective carbon�carbon bond insertion reactions with di-tert-butyl-alkylidenesilacyclopropanes. Pd(PPh3)4 was demonstrated to be the optimal catalyst, allowing for a variety of carbon�carbon ?-bond insertion reactions. Depending on the nature of the carbon�carbon ? bond, the insertion reaction proceeded with either direct insertion into the carbon(sp2)�silicon bond or with allylic transposition. Ring-substituted alkylidenesilacyclopropanes required a nickel catalyst to afford insertion products. Using Ni(cod)2 as the carbon�carbon bond insertion catalyst, new double alkyne insertion products and alkene isomerization products were observed. PMID:20419110
Buchner, Kay M.; Woerpel, K. A.
Optical Performance of the GM/CA-CAT Canted Undulator Beam lines for Protein Crystallography
A new macromolecular crystallographic facility developed by GM/CA-CAT is operational at the Advanced Photon Source (APS). The facility consists of three beamlines: two lines based on the first 'hard' dual canted undulators and one bending magnet beamline. The ID lines are operational, and the BM line is being commissioned. Both insertion device (ID) beamlines are independently tunable over a wide energy range. The inboard ID lines have been upgraded with a new insertion device to provide enhanced performance for MAD phasing experiments near the selenium and bromine K-edges. The ID line monochromators' crystals are indirectly, cryogenically cooled for improved performance and reliability. Focusing is achieved by long bimorph mirrors in a Kirkpatrick-Baez geometry. This paper describes the design of the beam lines and the optical characterization of the mirrors and monochromators.
Fischetti, Robert F.; Yoder, Derek W.; Xu Shenglan; Stepanov, Sergey; Makarov, Oleg; Benn, Richard; Corcoran, Stephen [GM/CA-CAT, Biosciences Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Diete, Wolfgang; Schwoerer-Boehing, Markus; Signorato, Riccardo; Schroeder, Leif [ACCEL Instruments GmbH, Friedrich-Ebert Strasse 1, D-51429 Bergisch Gladbach (Germany); Berman, Lonny [National Synchrotron Light Source, Brookhaven National Laboratory, Upton, NY 11973 (United States); Viccaro, P. James [University of Chicago, CARS-CAT, Argonne National Laboratory, Argonne, IL 60439 (United States); Smith, Janet L. [GM/CA-CAT, Biosciences Division, Argonne National Laboratory, Argonne, IL 60439 (United States); Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109 (United States)
A Medicago truncatula Tobacco Retrotransposon Insertion Mutant Collection with Defects in Nodule Development and Symbiotic Nitrogen Fixation1[W][OA
A Tnt1-insertion mutant population of Medicago truncatula ecotype R108 was screened for defects in nodulation and symbiotic nitrogen fixation. Primary screening of 9,300 mutant lines yielded 317 lines with putative defects in nodule development and/or nitrogen fixation. Of these, 230 lines were rescreened, and 156 lines were confirmed with defective symbiotic nitrogen fixation. Mutants were sorted into six distinct phenotypic categories: 72 nonnodulating mutants (Nod?), 51 mutants with totally ineffective nodules (Nod+ Fix?), 17 mutants with partially ineffective nodules (Nod+ Fix+/?), 27 mutants defective in nodule emergence, elongation, and nitrogen fixation (Nod+/? Fix?), one mutant with delayed and reduced nodulation but effective in nitrogen fixation (dNod+/? Fix+), and 11 supernodulating mutants (Nod++Fix+/?). A total of 2,801 flanking sequence tags were generated from the 156 symbiotic mutant lines. Analysis of flanking sequence tags revealed 14 insertion alleles of the following known symbiotic genes: NODULE INCEPTION (NIN), DOESN�T MAKE INFECTIONS3 (DMI3/CCaMK), ERF REQUIRED FOR NODULATION, and SUPERNUMERARY NODULES (SUNN). In parallel, a polymerase chain reaction-based strategy was used to identify Tnt1 insertions in known symbiotic genes, which revealed 25 additional insertion alleles in the following genes: DMI1, DMI2, DMI3, NIN, NODULATION SIGNALING PATHWAY1 (NSP1), NSP2, SUNN, and SICKLE. Thirty-nine Nod? lines were also screened for arbuscular mycorrhizal symbiosis phenotypes, and 30 mutants exhibited defects in arbuscular mycorrhizal symbiosis. Morphological and developmental features of several new symbiotic mutants are reported. The collection of mutants described here is a source of novel alleles of known symbiotic genes and a resource for cloning novel symbiotic genes via Tnt1 tagging. PMID:22679222
Pislariu, Catalina I.; D. Murray, Jeremy; Wen, JiangQi; Cosson, Viviane; Muni, RajaSekhara Reddy Duvvuru; Wang, Mingyi; A. Benedito, Vagner; Andriankaja, Andry; Cheng, Xiaofei; Jerez, Ivone Torres; Mondy, Samuel; Zhang, Shulan; Taylor, Mark E.; Tadege, Million; Ratet, Pascal; Mysore, Kirankumar S.; Chen, Rujin; Udvardi, Michael K.
Mechanisms of syn-insertion of alkynes and allenes into gold-silicon bonds: a comprehensive experimental/theoretical study.
A detailed mechanistic study is reported for the syn-insertion of alkynes and allenes in the Au-Si bonds of complexes (R3P)Au-SiR'Ph2 (R = Ph, Me and R' = t-Bu, Ph). Kinetic experiments indicate that (i) the reaction is first-order in alkyne and gold silyl complex and (ii) it requires a rather low enthalpy of activation and a relatively large negative entropy of activation [?H(�) = 13.7 (�1.6) kcal�mol(-1) and ?S(�) = -32.0 (�5.0) cal�mol(-1)�K(-1) for the reaction of (Ph3P)Au-Si(t-Bu)Ph2 with methyl propiolate], in line with a bimolecular associative transformation. The different mechanistic pathways have been explored by DFT calculations. Accordingly, the reaction is found to proceed via a two-step inner-sphere mechanism: (i) first, the alkyne coordinates to the gold silyl complex to form a ?-complex; (ii) the subsequent migratory insertion step is rate determining and occurs in a concerted manner. Provided dispersion effects are taken into account (B97D functional), the enthalpy of activation estimated theoretically [?H(�) = 11.5 kcal�mol(-1)] is in good agreement with that measured experimentally. The influence of the ? substrate (methyl propiolate, dimethyl acetylene dicarboxylate, phenyl acetylene, ethyl 2,3-butadienoate) has been analyzed theoretically, and the regioselectivity of the insertion has been rationalized. In particular, the unexpected selectivity observed experimentally with the allene is shown to result from the insertion of the terminal nonactivated C?C double bond into the Au-Si bond of (Ph3P)Au-SiPh3, followed by an original isomerization (Au/Si exchange process). This study provides unambiguous evidence for coordination-insertion at gold and thereby strongly supports the possible occurrence of inner-sphere mechanisms during the functionalization of alkynes and allenes. PMID:25000559
Joost, Maximilian; Estevez, Laura; Mallet-Ladeira, Sonia; Miqueu, Karinne; Amgoune, Abderrahmane; Bourissou, Didier
The Smooth Muscle Myosin Seven Amino Acid Heavy Chain Insert's Kinetic Role in the Crossbridge Cycle for Mouse Bladder
The seven amino acid insert in the smooth muscle myosin heavy chain is thought to regulate the kinetics of contraction, contributing to the differences between fast and slow smooth muscle. The effects of this insert on force and stiffness were determined in bladder tissue of a transgenic mouse line expressing the insert SMB at one of three levels: an SMB wild type (+/+), an SMA homozygous type (?/?) and a heterozygous type (+/?). For skinned muscle, an increase in MgADP or inorganic phosphate (Pi) should shift the distribution of crossbridges in the actomyosin ATPase (AMATPase) to increase the relative population of the crossbridge state prior to ADP release and Pi release, respectively. Exogenous ADP increased force and stiffness in a manner consistent with increasing the Ca2+ concentration in both the +/+ and +/? mouse types. However, the ?/? type showed a significantly greater increase in force than in stiffness suggesting that immediately prior to ADP release, the AMATPase either has an additional force producing isomerization state or a slower ADP dissociation rate for the ?/? type compared to the +/+ or +/? types. Exogenous Pi led to a significantly greater decrease in stiffness than in force for all three mouse types suggesting that there is a force producing state prior to Pi release. In addition, the increase in Pi showed similar changes in the +/+ and ?/? types whereas in the +/? type the decreases in both force and stiffness were greater than the other two mouse types indicating that the insert can affect the cooperativity between myosin heads. In conclusion, the seven amino acid insert modulates the kinetics and/or states of the AMATPase, which could lead to differences in the kinetics of contraction between fast and slow smooth muscle. PMID:12562924
Karagiannis, Peter; Babu, Gopal J; Periasamy, Muthu; Brozovich, Frank V
Aberrant splicing and transcription termination caused by P element insertion into the intron of a Drosophila gene
Insertional mutagenesis screens using the P[lacZ, rosy{sup +}] (PZ) transposable element have provided thousands of mutant lines for analyzing genes of varied function in the fruitfly, Drosophila melanogaster. As has been observed with other P elements, many of the PZ-induced mutations result from insertion of the P element into the promoter or 5{prime} untranslated regions of the affected gene. We document here a novel mechanism for mutagenesis by this element. We show that sequences present within the element direct aberrant splicing and termination events that produce an mRNA composed of 5{prime} sequences from the mutated gene (in this case, pipsqueak) and 3{prime} sequences from within the P[lacZ, rosy{sup +}] element. These truncated RNAs could yield proteins with dominant mutant effects. 43 refs., 4 figs.
Horowitz, H.; Berg, C.A. [Univ. of Washington, Seattle, WA (United States)
Insertion and Excision of the Transposable Element Mariner in Drosophila
The transposable element mariner is active in both germline and somatic cells of Drosophila mauritiana. Activity of the element is greatly enhanced in the presence of Mos1, a genetic factor identified as an autonomous copy of mariner. A strain of D. mauritiana containing Mos1 and other copies of mariner was used to initiate a screen for visible mutations. More than 20 mutations were obtained, including alleles of white, yellow and vermilion. Six alleles were characterized at the molecular level, and all were found to contain a mariner element inserted into the affected gene. Four insertions into the white locus were sequenced to determine the exact site of insertion of mariner. There appears to be little sequence specificity requirement for mariner insertion, other than an absolute requirement for the dinucleotide TA, which is duplicated upon insertion. Sequences of phenotypically wild-type germline and somatic revertants obtained from various white alleles, including the previously isolated w(pch) allele, were obtained using the polymerase chain reaction. Mariner excision is imprecise in both germline and soma, and the most frequent excision events are the same in the two tissues. Mutant derivatives of w(pch) were also studied, and were found to exhibit a wide range of molecular structures and phenotypes. PMID:2160399
Bryan, G.; Garza, D.; Hartl, D.
Lithium insertion mechanism in tin-based spinel sulfides
Cu{sub 2}MSn{sub 3}S{sub 8} (M = Fe, Co) spinel compounds have been studied as cathode materials in Li/LiPF{sub 6}(1M)-PC-EC/spinel sulfide cells. To understand the lithium insertion mechanism, both pristine compounds and chemically inserted products were characterized by X-ray Powder Diffraction (XPD), {sup 119}Sn Moessbauer Spectroscopy (MS), and X-ray Absorption Spectroscopy (XAS) at the sulfur K-edge. To accommodate the inserted lithium ions, two steps of reduction have been observed. First, for a low lithium content (up to 2 Li per formula), a copper reduction is occurring. Moreover, for the iron-based compound the copper atoms extraction is associated with a migration of iron atoms from the 16c octahedral sites toward the 8a tetrahedral sites. Second, for a high lithium content (above 2 Li per formula) a complex and simultaneous reduction of tin(IV) toward tin(II) and tin(0) has been clearly observed as well as a distortion of sulfur coordination polyhedra during the insertion process. From an electrochemical point of view, these spinels are rather interesting as model compounds regarding the lithium insertion study.
Branci, C.; Sarradin, J.; Olivier-Fourcade, J.; Jumas, J.C.
Mapping Insertions, Deletions and SNPs on Venter's Chromosomes
Background The very recent availability of fully sequenced individual human genomes is a major revolution in biology which is certainly going to provide new insights into genetic diseases and genomic rearrangements. Results We mapped the insertions, deletions and SNPs (single nucleotide polymorphisms) that are present in Craig Venter's genome, more precisely on chromosomes 17 to 22, and compared them with the human reference genome hg17. Our results show that insertions and deletions are almost absent in L1 and generally scarce in L2 isochore families (GC-poor L1+L2 isochores represent slightly over half of the human genome), whereas they increase in GC-rich isochores, largely paralleling the densities of genes, retroviral integrations and Alu sequences. The distributions of insertions/deletions are in striking contrast with those of SNPs which exhibit almost the same density across all isochore families with, however, a trend for lower concentrations in gene-rich regions. Conclusions Our study strongly suggests that the distribution of insertions/deletions is due to the structure of chromatin which is mostly open in gene-rich, GC-rich isochores, and largely closed in gene-poor, GC-poor isochores. The different distributions of insertions/deletions and SNPs are clearly related to the two different responsible mechanisms, namely recombination and point mutations. PMID:19543403
Costantini, Maria; Bernardi, Giorgio
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 ?m 2 was used for the acquisition process. Seven hundred and twenty projections were performed over 180� range using a high-resolution rotator. The projections were reconstructed using standard algorithms for tomographic reconstruction. The apical infiltration was evaluated by verifying if black spots were detectable on the images. The specimens of groups 3 and 4 showed a better apical seal than the ones of groups 1 and 2. Post insertion, when a ZOE-based sealer is used, increases the apical gap even if it does not seem to be clinically relevant and sufficient to be a cause of endodontic failure. The new method for analysis appeared to be effective to detect small defects in endodontic obturation, analysing guttapercha-sealer and sealer-dentin interfaces.
Contardo, L.; De Luca, M.; Biasotto, M.; Longo, R.; Olivo, A.; Pani, S.; Di Lenarda, R.
Germline Transgenesis and Insertional Mutagenesis in Schistosoma mansoni Mediated by Murine Leukemia Virus
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. PMID:22911241
Suttiprapa, Sutas; Kines, Kristine J.; Tort, Jos� F.; Mann, Victoria H.; Turner, Daniel J.; Berriman, Matthew; Brindley, Paul J.
Functional importance of the NH2-terminal insertion sequence of lung surfactant protein B
Lung surfactant protein B (SP-B) is required for proper surface activity of pulmonary surfactant. In model lung surfactant lipid systems composed of saturated and unsaturated lipids, the unsaturated lipids are removed from the film at high compression. It is thought that SP-B helps anchor these lipids closely to the monolayer in three-dimensional cylindrical structures termed �nanosilos� seen by atomic force microscopy imaging of deposited monolayers at high surface pressures. Here we explore the role of the SP-B NH2 terminus in the formation and stability of these cylindrical structures, specifically the distribution of lipid stack height, width, and density with four SP-B truncation peptides: SP-B 1�25, SP-B 9�25, SP-B 11�25, and SP-B 1�25Nflex (prolines 2 and 4 substituted with alanine). The first nine amino acids, termed the insertion sequence and the interface seeking tryptophan residue 9, are shown to stabilize the formation of nanosilos while an increase in the insertion sequence flexibility (SP-B 1�25Nflex) may improve peptide functionality. This provides a functional understanding of the insertion sequence beyond anchoring the protein to the two-dimensional membrane lining the lung, as it also stabilizes formation of nanosilos, creating reversible repositories for fluid lipids at high compression. In lavaged, surfactant-deficient rats, instillation of a mixture of SP-B 1�25 (as a monomer or dimer) and synthetic lung lavage lipids quickly improved oxygenation and dynamic compliance, whereas SP-B 11�25 surfactants showed oxygenation and dynamic compliance values similar to that of lipids alone, demonstrating a positive correlation between formation of stable, but reversible, nanosilos and in vivo efficacy. PMID:20023175
Frey, Shelli L.; Pocivavsek, Luka; Waring, Alan J.; Walther, Frans J.; Hernandez-Juviel, Jose M.; Ruchala, Piotr
Novel low-loss delay line for broadband phased antenna array applications
The microwave propagation velocity along a nonlinear transmission line is a function of dc bias, hence, a nonlinear transmission line (NLTL) can be utilized as a broadband delay line. A hybrid NLTL has been fabricated in a proof-of-principle experimental concept test where a 1.1-ns true time delay with <4-dB insertion loss has been measured in good agreement with theory. A
W.-M. Zhaug; R. P. Hsia; C. Liang; G. Song; C. W. Domier
A Theory and Design of an Insulating Insert in a Cathodically Protected Underground Water Pipeline. II. The Corrosion Rate of Anodic Zones and Effectiveness of the Insert
As a continuation of the previous study [1], the current flowing through an insulating tubular insert is calculated for a cathodically protected, underground water pipeline. The solution obtained allows calculating the anodic corrosion rate inside a pipe adjacent to the insert, as well as the insert's effectiveness. The main regularities in changing in the above parameters, as well as influencing
L. I. Freiman
Synchronized monochromator and insertion device energy scans at SLS
NASA Astrophysics Data System (ADS)
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.
Krempask�, J.; Flechsig, U.; Korhonen, T.; Zimoch, D.; Quitmann, Ch.; Nolting, F.
Synchronized monochromator and insertion device energy scans at SLS
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)
Soluble gentamicin ophthalmic inserts as a drug delivery system.
A comparison was made of soluble 14C-gentamicin ophthalmic inserts with drop, ointment, and the subconjunctival routes of administration. The insert is a solid, solubilizable collagen polymer containing 14C-gentamicin. We compared the levels of 14C-gentamicin in the rabbit tear film and in multiple corneal and scleral biopsies to determine which route of administration gave the best results. The wafer route of administration gave the highest tear film and tissue concentration of drug. The tear film concentration by subconjunctival injection was surprisingly low. Soluble collagen inserts offer a new method of delivering high doses of gentamicin in infected corneal tissue in a convenient and atraumatic fashion. PMID:655927
Bloomfield, S E; Miyata, T; Dunn, M W; Bueser, N; Stenzel, K H; Rubin, A L
A Nasogastric Tube Inserted into the Gastrocutaneous Fistula
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. PMID:22506228
Kim, Yang Soo; Kim, Joon Sung; Yu, In Hee; Jeong, Ji Young; Jung, Sung Hee; Jo, Yil Ryun
Electrochemical insertion of lithium into polyacrylonitrile-based disordered carbons
Electrochemical lithium insertion into polyacrylonitrile (PAN)-based disordered carbons was studied using the techniques of discharge/charge tests, cyclic voltammetry, and {sup 7}Li nuclear magnetic resonance (NMR) spectroscopy. The PAN-based carbons were prepared by vacuum pyrolysis of PAN at 500, 800, and 1,000 C. They showed charge capacities between 254 and 380 mAh/g in the first cycle. {sup 7}Li NMR spectra showed two kinds of lithium insertion sites in the PAN-based carbons: a reversible site where lithium is removed in the subsequent charge process and an irreversible site where lithium remains intact. The NMR results suggest that lithium in fully Li-inserted PAN-based carbons has an ionic character, and reversible site lithium resides between negatively charged carbon layers.
Jung, Y.; Suh, M.C.; Lee, S.I.; Shim, S.C.; Kwak, J. [Korea Advanced Inst. of Science and Technology, Taejon (Korea, Republic of). Dept. of Chemistry; Lee, H.; Kim, M. [Korea Research Inst. of Standards and Science, Taejon (Korea, Republic of). Div. of Chemistry and Radiation
Variation in the insertion of the palmaris longus tendon.
The palmaris longus is harvested as a tendon graft in various surgical procedures. We herein report the variations in the insertion of the palmaris longus tendon. During a routine dissection, a rare variation in the insertion of the palmaris longus tendon was observed. In the left forearm, the palmaris longus tendon bifurcated, while in the right forearm, the palmaris longus tendon trifurcated, giving rise to an accessory muscle, which passed superficial to the ulnar artery and ulnar nerve. The accessory muscle was supplied by a deep branch of the ulnar nerve, and the ulnar artery was observed to be tortuous. During reconstructive surgeries, surgeons should bear in mind the accessory muscle. Also, since the palmaris longus muscle provides a very useful graft in tendon surgery, every surgeon should be aware of the variations in the insertion of the palmaris longus tendon. PMID:25640108
Sunil, Vinutha; Rajanna, Shubha; Gitanjali; Kadaba, Jayanthi
Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
Gene targeting refers to the precise modification of a genetic locus using homologous recombination. The generation of novel cell lines and transgenic mouse models using this method necessitates the construction of a �targeting� vector, which contains homologous DNA sequences to the target gene, and has for many years been a limiting step in the process. Vector construction can be performed in vivo in Escherichia coli cells using homologous recombination mediated by phage recombinases using a technique termed recombineering. Recombineering is the preferred technique to subclone the long homology sequences (>4kb) and various targeting elements including selection markers that are required to mediate efficient allelic exchange between a targeting vector and its cognate genomic locus. Typical recombineering protocols follow an iterative scheme of step-wise integration of the targeting elements and require intermediate purification and transformation steps. Here, we present a novel recombineering methodology of vector assembly using a multiplex approach. Plasmid gap repair is performed by the simultaneous capture of genomic sequence from mouse Bacterial Artificial Chromosome libraries and the insertion of dual bacterial and mammalian selection markers. This subcloning plus insertion method is highly efficient and yields a majority of correct recombinants. We present data for the construction of different types of conditional gene knockout, or knock-in, vectors and BAC reporter vectors that have been constructed using this method. SPI vector construction greatly extends the repertoire of the recombineering toolbox and provides a simple, rapid and cost-effective method of constructing these highly complex vectors. PMID:25590226
Reddy, Thimma R.; Kelsall, Emma J.; Fevat, L�na M.S.
Analysis of transposon insertion mutants highlights the diversity of mechanisms underlying male progamic development in Arabidopsis.
To identify genes with essential roles in male gametophytic development, including postpollination (progamic) events, we have undertaken a genetic screen based on segregation ratio distortion of a transposon-borne kanamycin-resistance marker. In a population of 3359 Arabidopsis Ds transposon insertion lines, we identified 20 mutants with stably reduced segregation ratios arising from reduced gametophytic transmission. All 20 mutants showed strict cosegregation of Ds and the reduced gametophytic transmission phenotype. Among these, 10 mutants affected both male and female transmission and 10 mutants showed male-specific transmission defects. Four male and female (ungud) mutants and 1 male-specific mutant showed cellular defects in microspores and/or in developing pollen. The 6 remaining ungud mutants and 9 male-specific (seth) mutants affected pollen functions during progamic development. In vitro and in vivo analyses are reported for 5 seth mutants. seth6 completely blocked pollen germination, while seth7 strongly reduced pollen germination efficiency and tube growth. In contrast, seth8, seth9, or seth10 pollen showed reduced competitive ability that was linked to slower rates of pollen tube growth. Gene sequences disrupted in seth insertions suggest essential functions for putative SETH proteins in diverse processes including protein anchoring, cell wall biosynthesis, signaling, and metabolism. PMID:15342534
Lalanne, Eric; Michaelidis, Christos; Moore, James M; Gagliano, Wendy; Johnson, Andrew; Patel, Ramesh; Howden, Ross; Vielle-Calzada, Jean-Phillippe; Grossniklaus, Ueli; Twell, David
RJPrimers: unique transposable element insertion junction discovery and PCR primer design for marker development
Transposable elements (TE) exist in the genomes of nearly all eukaryotes. TE mobilization through �cut-and-paste� or �copy-and-paste� mechanisms causes their insertions into other repetitive sequences, gene loci and other DNA. An insertion of a TE commonly creates a unique TE junction in the genome. TE junctions are also randomly distributed along chromosomes and therefore useful for genome-wide marker development. Several TE-based marker systems have been developed and applied to genetic diversity assays, and to genetic and physical mapping. A software tool �RJPrimers� reported here allows for accurate identification of unique repeat junctions using BLASTN against annotated repeat databases and a repeat junction finding algorithm, and then for fully automated high-throughput repeat junction-based primer design using Primer3 and BatchPrimer3. The software was tested using the rice genome and genomic sequences of Aegilops tauschii. Over 90% of repeat junction primers designed by RJPrimers were unique. At least one RJM marker per 10 Kb sequence of A. tauschii was expected with an estimate of over 0.45 million such markers in a genome of 4.02 Gb, providing an almost unlimited source of molecular markers for mapping large and complex genomes. A web-based server and a command line-based pipeline for RJPrimers are both available at http://wheat.pw.usda.gov/demos/RJPrimers/. PMID:20497996
You, Frank M.; Wanjugi, Humphrey; Huo, Naxin; Lazo, Gerard R.; Luo, Ming-Cheng; Anderson, Olin D.; Dvorak, Jan; Gu, Yong Q.
Ka-band full-360� analog phase shifter with low insertion loss
NASA Astrophysics Data System (ADS)
A new reflection-type wideband 360� monolithic-microwave integrated-circuit (MMIC) analog phase shifter at the Ka-band is proposed. The phase shifter is designed based on the principle of vector synthesis. Three Lange couplers are employed in the phase shifter, which is fabricated by the standard 0.25-?m GaAs process. We use four 4 � 40 ?m GaAs HEMTs as the reflection loads. A microstrip line in parallel with the device is used as an inductance to counteract the parasitic capacitance of the device so that the reflection load performs like a pure resistance and the insertion loss can be decreased. In this phase shifter, a folded Lange coupler is utilized to reduce the size of the chip. The size of the proposed MMIC phase shifter is only 2.0 � 1.2 mm2. The measurement results show that the insertion loss is 5.0 � 0.8 dB and a 360� continuously tunable range across 27�32 GHz is obtained with miniscule DC power consumption.
Mengyi, Cao; Yang, Lu; Jiaxing, Wei; Jiaxin, Zheng; Xiaohua, Ma; Yue, Hao
Neutron reflectometry of supported hybrid bilayers with inserted peptide
The insertion of a synthetic amphiphilic, ?-helical peptide into a supported hybrid bilayer membrane (HBM) was studied by neutron reflectometry to elucidate the resulting nanostructure. The HBM consisted of a self-assembled monolayer of perdeuterated octadecanethiol on gold and an overlying leaflet of acyl-deuterated phosphatidylcholine (d-DMPC). Using contrast variation, several reflectivity spectra were recorded for each step of film fabrication, and simultaneously modeled. This analysis indicated that peptide insertion into the DMPC lipid leaflet is the likeliest mode of incorporation. PMID:21274414
Smith, Matthew B.; McGillivray, Duncan J.; Genzer, Jan; L�sche, Mathias; Kilpatrick, Peter K.
An innovation in insert visual reinforcement audiometry in children.
Visual reinforcement audiometry (VRA) is an important behavioural test of hearing in young children. The use of insert earphones during VRA provides a reliable method of obtaining ear- and frequency-specific data. Two testers are usually required for this procedure. The first tester operates the audiometer and other controls while the second tester distracts the child with various toys. The paper describes a technique of providing a visual cue to the second tester when a sound stimulus is introduced through the insert earphones. PMID:15829066
Jayarajan, V; Nandi, R; Caldicott, B
Method of modifying a volume mesh using sheet insertion
A method and machine-readable medium provide a technique to modify a hexahedral finite element volume mesh using dual generation and sheet insertion. After generating a dual of a volume stack (mesh), a predetermined algorithm may be followed to modify (refine) the volume mesh of hexahedral elements. The predetermined algorithm may include the steps of locating a sheet of hexahedral mesh elements, determining a plurality of hexahedral elements within the sheet to refine, shrinking the plurality of elements, and inserting a new sheet of hexahedral elements adjacently to modify the volume mesh. Additionally, another predetermined algorithm using mesh cutting may be followed to modify a volume mesh.
Borden, Michael J. (Albuquerque, NM); Shepherd, Jason F. (Albuquerque, NM)
Tail-anchored membrane protein insertion into the endoplasmic reticulum
Membrane proteins are inserted into the endoplasmic reticulum by two highly conserved parallel pathways. The well-studied co-translational pathway uses the signal recognition particle and its receptor for targeting, and the Sec61 translocon for membrane integration. A recently discovered post-translational pathway uses an entirely different set of factors involving transmembrane domain-selective cytosolic chaperones and an accompanying receptor at the endoplasmic reticulum. Elucidation of the structural and mechanistic basis of this post-translational membrane protein insertion pathway highlights general principles shared between the two pathways and key distinctions unique to each. PMID:22086371
Hegde, Ramanujan S.; Keenan, Robert J.
New chicane magnet design for insertion device straights at the Advanced Light Source
A chicane magnet incorporating counter-rotating permanent magnet pairs together with trim coils has been designed for use in the Advanced Light Source (ALS) straights in conjunction with two insertion devices. In particular, this design is being developed for use in the existing beam line (BL) 4 elliptically polarizing undulator (EPU) straight and in the BL11 EPU straight, currently under design and construction. The purpose of the chicane is to provide a fixed angular separation between two successive EPU photon fans, and to correct steering perturbations resulting from EPU polarization state changes. Polarization changes occur on the time scale of one second; associated steering corrections must be accomplished in less than a second. Hysteresis associated with conventional iron core electromagnets prevents fast steering correction to the required precision. This consideration motivated the iron-free design presented here.
Marks, Steve; Schlueter, Ross; Anderson, David; Gath, William; Jung, Jin-Young; Robin, David; Steier, Christoph; Stevens, Troy
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Stateless Multicounter 5 3 Watson-Crick machines and special two-head Watson-Crick automata (WK-auto- mata). These biologically motivated machines. Keywords Watson-Crick automata, counter machine, DNA computing, membrane computing, reversal bounded
Egecioglu, �?mer
Achilles tendon rupture following coblation for insertional Achilles tendinosis.
Radiofrequency microdebridement for Achilles tendinosis is a relatively new technique. We report a case of Achilles tendon rupture in a patient eight weeks after coblation for his right insertional Achilles tendinosis. We believe that this is the first reported case of Achilles tendon rupture following this new treatment of radiofrequency microdebridement for chronic Achilles tendinosis. PMID:20307450
Akhtar, M A; Montgomery, H; Shenolikar, A
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Permanence criteria for semi-free profinite the condition of a profinite group being semi-free, which is more general than being free and more restrictive than being quasi-free. In particular, every projective semi-free profinite group is free. We prove
Harbater, David
Fast, Completely Reversible Li Insertion in Vanadium Pentoxide Nanoribbons
ABSTRACT Layered-structure nanoribbons with efficient electron transport and short lithium ion insertion level the chemical, structural, and electrical transformations of V2O5 nanoribbons. We found that transformation of V2O5 into the -Li3V2O5 phase depends not only on the width but also the thickness
Cui, Yi
Basis of Identities of the Algebra of Simplified Insertions
for words of length ,k k . 1. Introduction Given a free associative algebra [ ]Ass X on a set of generators in [ ]Ass X , and if s sb u= , where s F and su are monomials in [ ]Ass X , then ( )s sb a u (see [2, 3]). Let's call the new algebra [ ], ,Ass X + the algebra of simplified insertions on the set
Adhesion degradation of rubber and steel insert for conveyor belts
The problem of degradation of rubber in conveyor belts is very important because they incorporate a lot of rubber and have a high value. In the paper we have analyzed the situation of rubber conveyor belts reinforced with metal insertions because it was found that they can sometimes be obsolete pretty fast due to the degradation of the connection between
Dan Dobrot?
Alu insertion polymorphisms in Native Americans and related Asian populations
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
Amyloid protein unfolding and insertion kinetics on neuronal membrane mimics
Atomistic details of beta-amyloid (Abeta ) protein unfolding and lipid interaction kinetics mediated by the neuronal membrane surface are important for developing new therapeutic strategies to prevent and cure Alzheimer's disease. Using all-atom MD simulations, we explored the early unfolding and insertion kinetics of 40 and 42 residue long Abeta in binary lipid mixtures with and without cholesterol that mimic
Liming Qiu; Creighton Buie; Mark Vaughn; Kwan Cheng
Pulmonary valve insertion late after repair of Fallot's tetralogy
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
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Meshless Methods for Physics-Based Modeling the challenges by proposing a meshless computational frame- work for dynamic modeling and simulation of solids and thin-shells represented as point samples. Our meshless framework can directly compute the elastic
Qin, Hong
EPJ manuscript No. (will be inserted by the editor)
EPJ manuscript No. (will be inserted by the editor) Simulations of two-dimensional foam rheology of bubbles with fixed areas surrounded #12;2 A. Wyn et al.: 2D Foam Rheology by films that are circular arcs to slide past each other in plastic events known as T1 topological changes (figure 1) [25]. These occur
Cox, Simon
Noname manuscript No. (will be inserted by the editor)
that it is able to control the variance of extrapolation, while maintaining, and often improving, predictive of accomplishing favor- able (and stable) predictive performance. A general overview of model regularizationNoname manuscript No. (will be inserted by the editor) Prediction-Based Regularization Using Data
Keinan, Alon
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Continuous Blooming of Convex Polyhedra Erik We construct the first two continuous bloomings of all convex polyhedra. First, the source unfolding can be continuously bloomed. Second, any unfolding of a convex polyhedron can be refined (further cut
Demaine, Erik
EPJ manuscript No. (will be inserted by the editor)
EPJ manuscript No. (will be inserted by the editor) Configurational statistics of densely and fully packed loops in the negative-weight percolation model O. Melchert and A. K. Hartmann Institut f¨ur Physik threshold, where random-walk behavior is obtained for d 6. 1 Introduction The statistical properties
Peinke, Joachim
Isocyanide insertion: de novo synthesis of trifluoromethylated phenanthridine derivatives.
A mechanistically new strategy has been described for the simple, practical, and environmentally friendly preparation of 6-(trifluoromethyl)phenanthridine derivatives using ionic isocyanide insertion from biphenyl isocyanide derivatives and Umemoto's reagent. These reactions were promoted only by inorganic base in good-to-excellent chemical yields without any external stoichiometric oxidants and radical initiators. PMID:24180686
Cheng, Yuanzheng; Jiang, Heng; Zhang, Yan; Yu, Shouyun
Computational Mechanics manuscript No. (will be inserted by the editor)
for Porous Media Models in Soil Mechanics Christian Wieners 1 , Martin Ammann 2 , Stefan Diebels 2 , Wolfgang for porous media models in soil mechanics are a di�cult task for the engineering modelling as wellComputational Mechanics manuscript No. (will be inserted by the editor) Parallel 3-d simulations
Wieners, Christian
21 CFR 529.1940 - Progesterone intravaginal inserts.
Code of Federal Regulations, 2011 CFR
...insert contains: (1) 1.38 grams (g) progesterone in molded silicone over a nylon spine. (2) 0.3 g progesterone in molded silicone over a flexible nylon spine. (b) Sponsor. See No. 000009 in � 510.600(c) of this...
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Multi-Dimensional Top-k Dominating Queries Man Lung Yiu · Nikos Mamoulis Abstract The top-k dominating query returns k data ob- jects which, it combines the advantages of top-k and skyline queries without sharing their disadvan- tages: (i) the output
Mamoulis, Nikos
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Distributed Top-k Query Processing, such as top-k, that enable users to retrieve only a limited set of the most interesting data objects. As dataTo, for efficient top-k processing over multiple servers, where each server stores autonomously a fraction
Nørvåg, Kjetil
INSERT YOUR BUILDING NAME HERE: BUILDING EMERGENCY PLAN Date Adopted: 6/4/13 Date Revised: 6/4/13 Prepared By: Tracey Simmerman #12;PURDUE UNIVERSITY BUILDING EMERGENCY PLAN VERSION 3 2 Table of Contents Suspension or Campus Closure SECTION 3: BUILDING INFORMATION 3.1 Building Deputy/Alternate Building Deputy
Physical Defect Modeling for Fault Insertion in System Reliability Test
Hardware fault-insertion test (FIT) is a promising method for system reliability test and diagnosis coverage measurement. It improves the speed of releasing a quality diagnostic program before manufacturing and provides feedbacks of fault tolerance of a very complicated large system. Certain level insufficient fault tolerance can be fixed in the current system but others may require ASIC or overall system
Zhaobo Zhang; Zhanglei Wang; Xinli Gu; Krishnendu Chakrabarty
Physical defect modeling for fault insertion in system reliability test
Hardware fault-insertion test (FIT) is a promising method for system reliability test and diagnosis coverage measurement. It improves the speed of releasing a quality diagnostic program before manufacturing and provides feedbacks of fault tolerance of a very complicated large system. Certain level insufficient fault tolerance can be fixed in the current system but others may require ASIC or overall system
Zhaobo Zhang; Zhanglei Wang; Xinli Gu; Krishnendu Chakrabarty
Simple method of insertion of Xomed one piece septal button.
Silicone septal buttons have been available to obturate septal perforations since the 1970s. Various methods of insertion have been suggested but the procedure is not always easy. We have devised an alternative method which has proved quick and simple to perform. PMID:1613352
al-Khabori, M J
Computer Modeling of Protocellular Functions: Peptide Insertion in Membranes
NASA Technical Reports Server (NTRS)
Lipid vesicles became the precursors to protocells by acquiring the capabilities needed to survive and reproduce. These include transport of ions, nutrients and waste products across cell walls and capture of energy and its conversion into a chemically usable form. In modem organisms these functions are carried out by membrane-bound proteins (about 30% of the genome codes for this kind of proteins). A number of properties of alpha-helical peptides suggest that their associations are excellent candidates for protobiological precursors of proteins. In particular, some simple a-helical peptides can aggregate spontaneously and form functional channels. This process can be described conceptually by a three-step thermodynamic cycle: 1 - folding of helices at the water-membrane interface, 2 - helix insertion into the lipid bilayer and 3 - specific interactions of these helices that result in functional tertiary structures. Although a crucial step, helix insertion has not been adequately studied because of the insolubility and aggregation of hydrophobic peptides. In this work, we use computer simulation methods (Molecular Dynamics) to characterize the energetics of helix insertion and we discuss its importance in an evolutionary context. Specifically, helices could self-assemble only if their interactions were sufficiently strong to compensate the unfavorable Free Energy of insertion of individual helices into membranes, providing a selection mechanism for protobiological evolution.
Rodriquez-Gomez, D.; Darve, E.; Pohorille, A.
Combined Transistor Sizing with Bu er Insertion for Timing Optimization
Combined Transistor Sizing with Bu er Insertion for Timing Optimization Yanbin Jiang y Sachin S are signi - cantly better than the results given by merely using a TILOS-like transistor sizing algorithm alone. 1 Introduction The transistor sizing problem 1, 2, 3 is often formulated as Minimize Area subject
Sapatnekar, Sachin
EPJ manuscript No. (will be inserted by the editor)
EPJ manuscript No. (will be inserted by the editor) Proposal for a Raman X-ray Free Electron Laser-ray free electron laser is proposed, based on a Raman process occurring during the interaction between- and gamma-ray laser 41.60.Cr Free-electron lasers 42.65.Dr Stimulated Raman Scattering 1 Introduction
Boyer, Edmond
People, events, and technology insertion in the Ada program
There are three components of the Ada program that I want to mention in this panel - dedicated people, major milestone events, and technology insertion.Ada is succeeding because of the participation, hard work, and enthusiastic support by many hundreds of dedicated professionals. During the time I was associated with the AJPO, I had the very good fortune to work with
Robert Mathis
Starting up 1. Insert the diskette in the floppy drive
Starting up 1. Insert the diskette in the floppy drive 2. Double�click the My Computer icon 3. Double�click the Floppy (A:) icon to open the diskette 4. Double�click on the file Sample to launch Excel. A formula stars with the = character 3. Some built-in formulas are: SUM AVERAGE COUNT SMALL 4. In the cell
Van Cleave, Nancy K.
Characterization of an insertion sequence, IS12528, from Gluconobacter suboxydans.
A novel insertion sequence element, IS12528, was found to be associated with inactivation of the alcohol dehydrogenase by insertion in the adhA gene, which encodes the primary dehydrogenase subunit of the three-component membrane-bound alcohol dehydrogenase complex in Gluconobacter suboxydans. Cloning and sequencing analyses revealed that IS12528 was 905 bp in length and had a terminal inverted repeat of 18 bp. In addition, IS12528 was found to generate a 3-bp duplication (TMA, where M represents C or A) at the inserted site upon transposition. IS12528 encoded one long product of 274 amino acids that was rich in basic amino acids. This protein showed significant homology with putative transposases of the IS1031 family isolated from Acetobacter xylinum, which belongs to another genus of acetic acid bacteria. IS12528-like sequences were distributed in a wide variety of acetic acid bacteria, as determined by Southern hybridization and PCR. These observations suggest that IS12528 is one of the insertion sequences that are responsible for genetic instability leading to deficiencies in various physiological properties in a variety of acetic acid bacteria. PMID:9055428
Kondo, K; Horinouchi, S
Acceptance of Advertising A written insertion order is
Acceptance of Advertising A written insertion order is required for all advertisements. All advertisements are subject to approval. The University of Chicago Press reserves the right to reject advertising for any reason. The advertiser and/or its advertising agency are entirely responsible for the lawfulness
McReynolds, Ben
Insertion and deletion mutagenesis of the human cytomegalovirus genome
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.
Genetic Instability in Drosophila melanogaster: Deletion Induction by Insertion Sequences
Females of Drosophila melanogaster heteroallelic or homoallelic for X chromosome giant (gt) mutants generate deletions involving the wild-type alleles at two X chromosome gene loci: yellow body color (y) and white eye color (w). The deletions, bidirectional in the case of y and with fixed endpoint in the case of w, are associated with particular X chromosomes. Distinctive insertion sequences,
M. M. Green
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Mathematical modeling of respiratory system of the cardio-respiratory system and its regulation, the analysis of normal respiratory mechanics, as well of the respiratory system [31] [38]. Although several models of the respiratory mechanics have been proposed
Paris-Sud XI, Universit� de
Astronomy & Astrophysics manuscript no. (will be inserted by hand later)
be attributed to various hydrodynamic and hydromagnetic instabilities that can arise in differentially rotatingAstronomy & Astrophysics manuscript no. (will be inserted by hand later) The stability of Physics and Technology, 194021 St. Petersburg, Russia January 28, 2005 Abstract. The stability properties
External Tank (ET) Bipod Fitting Bolted Attachment Locking Insert Performance
NASA Technical Reports Server (NTRS)
Following STS-107, the External Tank (ET) Project implemented corrective actions and configuration changes at the ET bipod fitting. Among the corrective actions, the existing bolt lock wire which provided resistance to potential bolt rotation was removed. The lock wire removal was because of concerns with creating voids during foam application and potential for lock wire to become debris. The bolts had been previously lubricated to facilitate assembly but, because of elimination of the lock wire, the ET Project wanted to enable the locking feature of the insert. Thus, the lubrication was removed from bolt threads and instead applied to the washer under the bolt head. Lubrication is necessary to maximize joint pre-load while remaining within the bolt torque specification. The locking feature is implemented by thread crimping in at four places in the insert. As the bolt is torqued into the insert the bolt threads its way past the crimped parts of the insert. This provides the locking of the bolt, as torque is required to loosen the joint after clamping.
Larsen, Curtis E.; Wilson, Tim R.; Elliott, Kenny B.; Raju, Ivatury S.; McManamen, John
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Sparse Deconvolution Methods for Ultrasonic the sparsity assumption about the measured ultrasonic signal as prior knowl- edge. The MP algorithm is used. For blind deconvolution, where the transducer impulse response is unknown, we offer a general Gaussian echo
Plonka, Gerlind
Solving Witten's SFT by insertion of operators on projectors
Following Okawa, we insert operators at the boundary of regulated star algebra projectors to construct the leading order tachyon vacuum solution of open string field theory. We also calculate the energy density of the solution and the ratio between the kinetic and the cubic terms. A universal relationship between these two quantities is found. We show that for any twist
Haitang Yang
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Estimating Deep Web Data Source Size the size of a deep web data source that is accessible by queries only. Since most deep web data sources. The results show that our method has small bias and standard deviation. Keywords Deep web, estimators, capture
Lu, Jianguo
Palliation of proximal malignant biliary obstruction by endoscopic endoprosthesis insertion
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
A Two-Level Inspection Model With Technological Insertions
This paper presents a model for optimal asset maintenance inspection services. The model is designed to support through-life service in the form of multiple nested inspections and maintenance to meet defined asset availability and capability requirements, as well as achieving successful through-life technology insertions. The inspections and maintenance activities are assumed to be performed at more than one level, but
Wenbin Wang; Matthew J. Carr; Tommy W. S. Chow; Michael G. Pecht
Peripherally Inserted Central Catheters by Nurses using the Sonic Flashlight
Peripherally Inserted Central Catheters by Nurses using the Sonic Flashlight David Wang1,5, Nikhil functions by fixing the relative geometry of the ultrasound transducer, the display, and a half-silvered mirror to produce a virtual image at the scanned anatomy within the body. Through the half- silvered
Stetten, George
Simultaneous Routing and Buffer Insertion for High Performance Interconnect
We present an algorithm for simultaneously finding a rectilinear Steiner tree T and buffer insertion points into T. The objective of the algorithm is to minimize a cost function (e.g., total area or power) subject to given timing constraints on the sinks of the net. An interesting side-effect of our approach is that we are able to derive an entire
John Lillis; Chung-kuan Cheng; Ting-ting Y. Lin
PATAT 2012 manuscript No. (will be inserted by the editor)
PATAT 2012 manuscript No. (will be inserted by the editor) Self-Rostering applied to case studies and discuss its application to a number of practical case studies. Keywords Personnel Rostering · Shift defines the staffing demand, i.e., specifies, per day, the number of employees that need to perform
Al Hanbali, Ahmad
Direct observation of poloxamer 188 insertion into lipid monolayers.
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. PMID:11867460
Maskarinec, Stacey A; Hannig, J�rgen; Lee, Raphael C; Lee, Ka Yee C
Endometrial Adenocarcinoma Following the Insertion of a Mirena IUCD
Background. The case histories of two patients who developed endometrial adenocarcinoma with a Mirena intrauterine system (IUS) in place are reported.Case. Two patients had a Mirena IUS inserted to treat abnormal vaginal bleeding. After 12 and 36 months with the device in place they presented with a recurrence of their heavy, irregular menstrual bleeding. Endometrial biopsies confirmed the presence of
Kevin Jones; Mel Georgiou; Derek Hyatt; Tim Spencer; Hilary Thomas
Analysis of Early-Insertion Standard Coalesced Hashing
This paper analyzes the early-insertion standard coalesced hashing method (EISCH), which is a variant of the standard coalesced hashing algorithm (SCH) described in [Knu73], [Vit80] and [Vit82b]. The analysis answers the open problem posed in [Vit80...
Chen, Wen-Chin; Vitter, Jeffrey Scott
Polyglutamine expansion in huntingtin increases its insertion into lipid bilayers
An expanded polyglutamine (Q) tract (>37Q) in huntingtin (htt) causes Huntington disease. Htt associates with membranes and polyglutamine expansion in htt may alter membrane function in Huntington disease through a mechanism that is not known. Here we used differential scanning calorimetry to examine the effects of polyQ expansion in htt on its insertion into lipid bilayers. We prepared synthetic lipid
Kimberly B. Kegel; Vitali Schewkunow; Ellen Sapp; Nicholas Masso; Erich E. Wanker; Marian DiFiglia; Wolfgang H. Goldmann
Molecular archeology of L1 insertions in the human genome
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
Noname manuscript No. (will be inserted by the editor)
words music video generation, multi-media analysis, music and video synchronization 1 Introduction analysis module and a matching module. In the analysis module, we segment the music and video using flowNoname manuscript No. (will be inserted by the editor) Automated Music Video Generation Using Multi
Lee, In-Kwon
Noname manuscript No. (will be inserted by the editor)
-truth. This technique is similar to external evaluation of attribute-based clustering methods. The other two wellNoname manuscript No. (will be inserted by the editor) Communities Validity: Methodical Evaluation to statistically validate the clustering result, and relative evaluation to compare alternative clustering results
Zaiane, Osmar R.
Mechanism of fluid infusion during microneedle insertion and retraction
Previous work has shown that infusion flow rates can be increased by an order of magnitude by partially retracting microneedles after insertion into the skin. This study sought to determine the mechanism by which retraction increases fluid infusion by piercing human cadaver skin with single microneedles, fixing the skin after retracting microneedles to different distances, and examining skin microstructure by
Wijaya Martanto; Jason S. Moore; Tracey Couse; Mark R. Prausnitz
Teaching oddity and insertion to Head Start children
Two interventions were offered to Head Start preschoolers. Half of the children played numerous learning set games designed to teach the children (1) the oddity principle and (2) how to insert objects into a series. Mastery of these concepts is theoretically an important aspect of school readiness. The other half of the children engaged in the I Can Problem Solve
Dennis Ciancio; A. Cristina Rojas; Kelly McMahon; Robert Pasnak
Noname manuscript No. (will be inserted by the editor)
· Gyroscope · Hierarchical Linear Model 1 Introduction Regular physical activity plays an important role and acceptance should be inserted later Abstract Walking is a commonly available activity to maintain a email@example.com B. Adar Emken · Donna Spruijt-Metz Dept. of Preventive Medicine, University of Southern California
Sukhatme, Gaurav S.
Quality & Quantity manuscript No. (will be inserted by the editor)
under the scope of empirical investigation. Keywords Qualitative research · Measurement · FalsifiabilityQuality & Quantity manuscript No. (will be inserted by the editor) How to State General Qualitative Facts in Psychology? Stéphane Vautier Received: date / Accepted: date Abstract In what form should
Paris-Sud XI, Université de
Networking Science manuscript No. (will be inserted by the editor)
Networking Science manuscript No. (will be inserted by the editor) Diffusion of Real-Time Information in Overlaying Social-Physical Networks: Network Coupling and Clique Structure Dajun Qian · Osman the diffusion behavior of real-time in- formation in an overlaying social-physical network. Typi- cally, real
Yagan, Osman
Noname manuscript No. (will be inserted by the editor)
of spanning trees for a given set of traffic demand matrices. Solving the TE problem in large EthernetNoname manuscript No. (will be inserted by the editor) Multi-objective Traffic Engineering for Data Center Networks Trong-Viet Ho � Yves Deville � Olivier Bonaventure Received: date / Accepted: date
Deville, Yves
EPJ manuscript No. (will be inserted by the editor)
@esstin.uhp-nancy.fr, firstname.lastname@example.com the date of receipt and acceptance should be inserted later Abstract the redefinition of the whole traffic control system. Cur- rently, the basic device of traffic control is the track of localized cracks as well as large shellings. The two supply frequencies make the sensor sensitive
Paris-Sud XI, Universit� de
Queueing Systems manuscript No. (will be inserted by the editor)
as n , by virtue of the law of large numbers. Consequently, for any fixed traffic intensity Queueing Systems manuscript No. (will be inserted by the editor) Heavy-Traffic Limits for Nearly Deterministic Queues: Stationary Distributions Karl Sigman � Ward Whitt Received: date / Accepted: date Abstract
Whitt, Ward
Insights on human evolution: an analysis of Alu insertion polymorphisms
We analyzed the genetic profile of 563 individuals from 12 geographically targeted human populations from Europe, Asia and Africa using 27 human-specific polymorphic Alu insertions. Phylogenetic analyses indicated a clear correspondence between genetic profiles and historical patterns of gene flow and genetic drift. Sub-Saharan African populations (Benin, Cameroon, Kenya and Rwanda) formed a visibly differentiated cluster, indicating the role of
Maria C Terreros; Miguel A Alfonso-S�nchez; Gabriel E Novick; Javier R Luis; Harlette Lacau; Robert K Lowery; Maria Regueiro; Rene J Herrera
Autonomous Robots manuscript No. (will be inserted by the editor)
Autonomous Robots manuscript No. (will be inserted by the editor) Recognition, Prediction: The approach of inferring user's intended task and optimizing low-level robot motions has promise for mak- ing robot teleoperation interfaces more intuitive and respon- sive. But most existing methods assume
Indiana University
Mathematische Zeitschrift manuscript No. (will be inserted by the editor)
Mathematische Zeitschrift manuscript No. (will be inserted by the editor) Leonid V. Kovalev Diego Investigator award under grant DMS 0601926. J.-M.W. was supported by the NSF grant DMS 0400810. L. V. Kovalev West Green Street, Urbana, IL 61801, USA. E-mail: email@example.com #12;2 L.V. Kovalev, D. Maldonado, J
Wu, Jang-Mei
Stability analysis of chickpea large genomic DNA inserts in Agrobacterium.
Technology Transfer Automated Retrieval System (TEKTRAN)
Agrobacterium tumefaciens-mediated transformation of large DNA inserts directly into plants facilitates the transfer of gene clusters and flanking regulatory elements. It is recommended that the integrity of large genomic fragments in Agrobacterium be verified prior to plant transformation. In this ...
Tungsten trioxide films during cation insertion: in situ optical characterization
At the moment WO3 is the most suitable material as cathodic film for electrochromic devices, mainly because of its good mechanical and physical properties and efficiency in coloring and bleaching upon cation intercalation. We studied the effect of the electrochemical insertion of small cations (H+, Li+ or Na+) in WO3 thin films by means of `in situ' transmittance and reflectance
Enrico Masetti; D. Dini; Franco Decker
National Aeronautics and Space Administration MAVEN Orbit Insertion
National Aeronautics and Space Administration NASAfacts MAVEN Orbit Insertion Mars Atmosphere. MAVEN: Exploring Mars' climate history #12;National Aeronautics and Space Administration Goddard Space and ionosphere of Mars, characterizing their current states and determining the rates of loss of gas to space
Waliser, Duane E.
PiggyBac Transgene Insertion Site GIZA Strain
Technology Transfer Automated Retrieval System (TEKTRAN)
Genetically modified New World screwworms were analyzed by inverse PCR and several sequences of genomic DNA flanking the transposon insertion sites were obtained. One transgenic strain, designated GIZA, exhibited the properties of a lethal genetic mutation. Classic genetic crosses of these insects r...
Noname manuscript No. (will be inserted by the editor)
marketing strategy is the e-mail service Hotmail: Every mail sent via a Hotmail account contained some,000 for advertise- ment, Hotmail grew to 12 million users in 18 months (Jurvetson, 2000). The name viral marketingNoname manuscript No. (will be inserted by the editor) On Tractable Cases of Target Set Selection
Wichmann, Felix
31. Photocopy of map. Insert from Master Plan Notes Prepared ...
Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey
31. Photocopy of map. Insert from Master Plan Notes Prepared by Environmental Planning and Design, Pittsburgh, Pa. Original at Missouri Botanical Garden. 1972 MASTER PLAN 32. 'THE TREE,' SCULPTURE BY ALEXANDER CALDER, 1966 - Missouri Botanical Garden, 2345 Tower Grove Avenue, Saint Louis, Independent City, MO
30. Photocopy of map. Insert, p. 138, Master Plan Report, ...
Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey
30. Photocopy of map. Insert, p. 138, Master Plan Report, 1960, Vol. II. Layton, Layton and Rohrbach, General Planning and Resource Consultants. 1960 MASTER PLAN, SHOWING PROPOSED LOCATION OF LINNAEAN HOUSE - Missouri Botanical Garden, 2345 Tower Grove Avenue, Saint Louis, Independent City, MO
RNA editing by cytidine insertion in mitochondria of Physarum polycephalum
A COROLLARY of the central dogma of molecular biology is that genetic information passes from DNA to RNA by the continuous synthesis of RNA on a DNA template. The demonstration of RNA editing1 (the specific insertion, deletion or substitution of residues in RNA to create an RNA with a sequence different from its own template) raised the possibility that in
R. Mahendran; M. R. Spottswood; D. L. Miller
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Pendubot: Combining of Energy and Intuitive. This double-link pendulum, usually called pendubot, is an un- deractuated system. The double-link pendulum system, Pendubot, Swing up, Unstable equilibrium, Stabilization, Satu- rated control. 1 Introduction
Paris-Sud XI, Universit� de
Otological Findings Ten Years after Myringotomy with Tympanostomy Tube Insertion
Introduction: To study the long-term complications of tympanostomy tube insertion in young children 10 years after surgery. Materials and Methods: In September 2011, the medical records of all patients who had undergone myringotomy with tympanostomy tube insertion between February 2000 and March 2001 at the two general hospitals of Isfahan University of Medical Sciences were studied. Of the 98 patients who fulfilled the inclusion criteria, 82 patients agreed to participate and were enrolled in the study. The complications of the operation were evaluated in these patients. Results: Of the 164 ears that were operated on, myringosclerosis was found in 17.1%, atrophy of the tympanic membrane in 1.2%, permanent perforation of the tympanic membrane in 0.6% and tympanic membrane atelectasis in 0.6%. None of the patients developed cholesteatoma as a complication of tympanostomy tube insertion. Conclusion: Considering the low risk of serious complications after 10 years, tympanostomy tube insertion is a safe and effective treatment option in the treatment of otitis media with effusion. PMID:24303407
Barati, Behrouz; Hashemi, Seyed Mostafa; Goljanian Tabrizi, Ali
Noname manuscript No. (will be inserted by the editor)
Noname manuscript No. (will be inserted by the editor) Web-GIS tool for the management of rural land markets. Application to the Land Bank of Galicia (NW Spain) Juan Porta · Jorge Parapar · Paula of rural markets in the last few years have become one of the main concerns of rural administrations
Touriño, Juan
Exp Fluids manuscript No. (will be inserted by the editor)
Exp Fluids manuscript No. (will be inserted by the editor) Three-dimensional Schlieren Measurements Abstract The `synthetic schlieren' method has been used to measure the amplitude of internal waves of a Gaussian or a Gaussian multiplied in amplitude by a sine function. Keywords Synthetic schlieren
Sutherland, Bruce
Insertion and Partition of Sodium Taurocholate into Egg Phosphatidylcholine Vesicles
Purpose. To get a continuous description of the insertion and partition processes of sodium taurocholate (TC) into the lipid bilayers of vesicles that can serve as a model for understanding the mechanism of destabilization by the bile salts of liposomes used as drug carriers for oral administration.
Karine Andrieux; Laura Forte; Sylviane Lesieur; Mait� Paternostre; Michel Ollivon; C�cile Grabielle-Madelmont
African origin of human-specific polymorphic Alu insertions.
Alu elements are a family of interspersed repeats that have mobilized throughout primate genomes by retroposition from a few "master" genes. Among the 500,000 Alu elements in the human genome are members of the human-specific subfamily that are not fixed in the human species; that is, not all chromosomes carry an Alu element at a particular locus. Four such polymorphic human-specific Alu insertions were analyzed by a rapid, PCR-based assay that uses primers that flank the insertion point to determine genotypes based on the presence or absence of the Alu element. These four polymorphic Alu insertions were shown to be absent from the genomes of a number of nonhuman primates, consistent with their arising as human genetic polymorphisms sometime after the human/African ape divergence. Analysis of 664 unrelated individuals from 16 population groups from around the world revealed substantial levels of variation within population groups and significant genetic differentiation among groups. No significant associations were found among the four loci, consistent with their location on different chromosomes. A maximum-likelihood tree of population relationships showed four major groupings consisting of Africa, Europe, Asia/Americas, and Australia/New Guinea, which is concordant with similar trees based on other loci. A particularly useful feature of the polymorphic Alu insertions is that the ancestral state is known to be the absence of the Alu element, and the presence of the Alu element at a particular chromosomal site reflects a single, unique event in human evolution. A hypothetical ancestral group can then be included in the tree analysis, with the frequency of each insertion set to zero. The ancestral group connected to the maximum-likelihood tree within the African branch, which suggests an African origin of these polymorphic Alu insertions. These data are concordant with other diverse data sets, which lends further support to the recent African origin hypothesis for modern humans. Polymorphic Alu insertions represent a source of genetic variation for studying human population structure and evolution. Images PMID:7991620
Batzer, M A; Stoneking, M; Alegria-Hartman, M; Bazan, H; Kass, D H; Shaikh, T H; Novick, G E; Ioannou, P A; Scheer, W D; Herrera, R J
A Transposable Element Insertion Confers Xenobiotic Resistance in Drosophila
The increase in availability of whole genome sequences makes it possible to search for evidence of adaptation at an unprecedented scale. Despite recent progress, our understanding of the adaptive process is still very limited due to the difficulties in linking adaptive mutations to their phenotypic effects. In this study, we integrated different levels of biological information to pinpoint the ecologically relevant fitness effects and the underlying molecular and biochemical mechanisms of a putatively adaptive TE insertion in Drosophila melanogaster: the pogo transposon FBti0019627. We showed that other than being incorporated into Kmn1 transcript, FBti0019627 insertion also affects the polyadenylation signal choice of CG11699 gene. Consequently, only the short 3?UTR transcript of CG11699 gene is produced and the expression level of this gene is higher in flies with the insertion. Our results indicated that increased CG11699 expression leads to xenobiotic stress resistance through increased ALDH-III activity: flies with FBti0019627 insertion showed increased survival rate in response to benzaldehyde, a natural xenobiotic, and to carbofuran, a synthetic insecticide. Although differences in survival rate between flies with and without the insertion were not always significant, when they were, they were consistent with FBti0019627 mediating resistance to xenobiotics. Taken together, our results provide a plausible explanation for the increase in frequency of FBti0019627 in natural populations of D. melanogaster and add to the limited number of examples in which a natural genetic mutation has been linked to its ecologically relevant phenotype. Furthermore, the widespread distribution of TEs across the tree of life and conservation of stress response pathways across organisms make our results relevant not only for Drosophila, but for other organisms as well. PMID:25122208
Mateo, Lidia; Ullastres, Anna; Gonz�lez, Josefa
Reversible insertion of carbenes into ruthenium-silicon bonds.
The five-coordinate carbene complexes [Ru{?P,P,Si-Si(Me)(C6H4-2-PiPr2)2}Cl(?CHR)] (2, R = Ph; 3, R = SiMe3), analogues of the Grubbs catalyst, were prepared from the dimer [Ru(?-Cl){?P,P,Si-Si(Me)(C6H4-2-PiPr2)2}]2 (1) and the corresponding diazoalkane N2CHR. The particular structural features that result from the presence of a strongly trans directing silyl group at the pincer ligand of these complexes are discussed on the basis of NMR information and the crystal structure of the vinylidene analogue [Ru{?P,P,Si-Si(Me)(C6H4-2-PiPr2)2}Cl(?C?CHPh)] (4), which was also obtained from 1 and phenylacetylene. The reactions of 3 with reagents such as P(OMe)3, CO, NCMe, and K(acac) illustrate that the first response of these carbene complexes to an increase of the coordination number around ruthenium is the insertion of the carbene ligand into the Ru-Si bond. These reactions also indicate that the insertion process is reversible and allows typical transformations of carbene ligands such as C-H functionalizations via carbene insertion (in the acac ligand) or the formation of ketene from CO. In addition, the reactions of 3 with terminal alkynes such as phenylacetylene or 3,3-dimethyl-1-butyne show that the inserted carbenes can also undergo reactions typical of metal-bound alkyls such as alkyne insertion and C-H reductive elimination. PMID:24289787
Bernal, Mar�a J; Torres, Olga; Mart�n, Marta; Sola, Eduardo
A transposable element insertion confers xenobiotic resistance in Drosophila.
The increase in availability of whole genome sequences makes it possible to search for evidence of adaptation at an unprecedented scale. Despite recent progress, our understanding of the adaptive process is still very limited due to the difficulties in linking adaptive mutations to their phenotypic effects. In this study, we integrated different levels of biological information to pinpoint the ecologically relevant fitness effects and the underlying molecular and biochemical mechanisms of a putatively adaptive TE insertion in Drosophila melanogaster: the pogo transposon FBti0019627. We showed that other than being incorporated into Kmn1 transcript, FBti0019627 insertion also affects the polyadenylation signal choice of CG11699 gene. Consequently, only the short 3'UTR transcript of CG11699 gene is produced and the expression level of this gene is higher in flies with the insertion. Our results indicated that increased CG11699 expression leads to xenobiotic stress resistance through increased ALDH-III activity: flies with FBti0019627 insertion showed increased survival rate in response to benzaldehyde, a natural xenobiotic, and to carbofuran, a synthetic insecticide. Although differences in survival rate between flies with and without the insertion were not always significant, when they were, they were consistent with FBti0019627 mediating resistance to xenobiotics. Taken together, our results provide a plausible explanation for the increase in frequency of FBti0019627 in natural populations of D. melanogaster and add to the limited number of examples in which a natural genetic mutation has been linked to its ecologically relevant phenotype. Furthermore, the widespread distribution of TEs across the tree of life and conservation of stress response pathways across organisms make our results relevant not only for Drosophila, but for other organisms as well. PMID:25122208
Mateo, Lidia; Ullastres, Anna; Gonz�lez, Josefa
... Poison Help Line The toll-free Poison Help line, 1-800-222-1222 , which connects you to your ... and Services Administration (HRSA) , funds the Poison Help line ( 1-800-222-1222 ), which connects you to your ...
Characterization and event specific-detection by quantitative real-time PCR of T25 maize insert.
T25 is one of the 4 maize transformation events from which commercial lines have so far been authorized in Europe. It was created by polyethylene glycol-mediated transformation using a construct bearing one copy of the synthetic pat gene associated with both promoter and terminator of the 35S ribosomal gene from cauliflower mosaic virus. In this article, we report the sequencing of the whole T25 insert and the characterization of its integration site by using a genome walking strategy. Our results confirmed that one intact copy of the initial construct had been integrated in the plant genome. They also revealed, at the 5' junction of the insert, the presence of a second truncated 35S promoter, probably resulting from rearrangements which may have occurred before or during integration of the plasmid DNA. The analysis of the junction fragments showed that the integration site of the insert presented high homologies with the Huck retrotransposon family. By using one primer annealing in the maize genome and the other in the 5' end of the integrated DNA, we developed a reliable event-specific detection system for T25 maize. To provide means to comply with the European regulation, a real-time PCR test was designed for specific quantitation of T25 event by using Taqman chemistry. PMID:15859082
Collonnier, C�cile; Schattner, Alexandra; Berthier, Georges; Boyer, Francine; Cou�-Philippe, G�raldine; Diolez, Annick; Duplan, Marie-No�lle; Fernandez, Sophie; Kebdani, Na�ma; Kobilinsky, Andr�; Romaniuk, Marcel; de Beuckeleer, Marc; de Loose, Marc; Windels, Pieter; Bertheau, Yves
The Zebrafish Insertion Collection (ZInC): a web based, searchable collection of zebrafish mutations generated by DNA insertion.
ZInC (Zebrafish Insertional Collection, http://research.nhgri.nih.gov/ZInC/) is a web-searchable interface of insertional mutants in zebrafish. Over the last two decades, the zebrafish has become a popular model organism for studying vertebrate development as well as for modeling human diseases. To facilitate such studies, we are generating a genome-wide knockout resource that targets every zebrafish protein-coding gene. All mutant fish are freely available to the scientific community through the Zebrafish International Resource Center (ZIRC). To assist researchers in finding mutant and insertion information, we developed a comprehensive database with a web front-end, the ZInC. It can be queried using multiple types of input such as ZFIN (Zebrafish Information Network) IDs, UniGene accession numbers and gene symbols from zebrafish, human and mouse. In the future, ZInC may include data from other insertional mutation projects as well. ZInC cross-references all integration data with the ZFIN (http://zfin.org/). PMID:23180778
Varshney, Gaurav K; Huang, Haigen; Zhang, Suiyuan; Lu, Jing; Gildea, Derek E; Yang, Zhongan; Wolfsberg, Tyra G; Lin, Shuo; Burgess, Shawn M
Hydrogen insertion reaction with restricted diffusion. Part 1. Potential step�EIS theory and review for the direct insertion mechanism
Electrochemical impedance spectroscopy (EIS) theory is presented for the direct hydrogen insertion reaction with restricted diffusion. The limiting shapes of the Nyquist impedance plots are given and the changes in the impedance diagrams due to the double-layer capacitance are presented. The responses to small potential steps are also considered. All the limiting expressions for the current transients are given and
J. S. Chen; J.-P. Diard; R. Durand; C. Montella
Genetic determinants of feline leukemia virus-induced lymphoid tumors: patterns of proviral insertion and gene rearrangement.
The genetic basis of feline leukemia virus (FeLV)-induced lymphoma was investigated in a series of 63 lymphoid tumors and tumor cell lines of presumptive T-cell origin. These were examined for virus-induced rearrangements of the c-myc, flvi-2 (bmi-1), fit-1, and pim-1 loci, for T-cell receptor (TCR) gene rearrangements, and for the presence of env recombinant FeLV (FeLV-B). The myc locus was most frequently affected in naturally occurring lymphomas (32%; n = 38) either by transduction (21%) or by proviral insertion (11%). Proviral insertions were also common at flvi-2 (24%). The two other loci were occupied in a smaller number of the natur