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Chromosomal translocations are a common phenomenon in Arabidopsis thaliana T-DNA insertion lines
SUMMARY Ordered collections of Arabidopsis thaliana lines containing mapped T-DNA insertions have become an important resource for plant scientists performing genetic studies. Previous reports have indicated that T-DNA insertion lines can have chromosomal translocations associated with the T-DNA insertion site, but the prevalence of these rearrangements has not been well documented. To determine the frequency with which translocations are present in a widely-used collection of T-DNA insertion lines, we analyzed 64 independent lines from the Salk T-DNA mutant collection. Chromosomal translocations were detected in 12 of the 64 lines surveyed (19%). Two assays were used to screen the T-DNA lines for translocations: pollen viability and genome-wide genetic mapping. Although the measurement of pollen viability is an indirect screen for the presence of a translocation, all 11 of the T-DNA lines showing an abnormal pollen phenotype were found to contain a translocation when analyzed using genetic mapping. A normal pollen phenotype does not, however, guarantee the absence of a translocation. We observed one T-DNA line with normal pollen that nevertheless had a translocation based on genetic mapping results. One additional phenomenon that we observed through our genetic mapping experiments was that the T-DNA junctions on the 5?- and 3?-sides of a targeted gene can genetically separate from each other in some cases. Two of the lines in our survey displayed this ĹT-DNA borders separateĺ phenomenon. Experimental procedures for efficiently screening T-DNA lines for the presence of chromosomal abnormalities are presented and discussed.
Clark, Katie A.; Krysan, Patrick J.
T-DNA insertional mutagenesis for activation tagging in rice.
We have developed a new T-DNA vector, pGA2715, which can be used for promoter trapping and activation tagging of rice (Oryza sativa) genes. The binary vector contains the promoterless beta-glucuronidase (GUS) reporter gene next to the right border. In addition, the multimerized transcriptional enhancers from the cauliflower mosaic virus 35S promoter are located next to the left border. A total of 13,450 T-DNA insertional lines have been generated using pGA2715. Histochemical GUS assays have revealed that the GUS-staining frequency from those lines is about twice as high as that from lines transformed with the binary vector pGA2707, which lacks the enhancer element. This result suggests that the enhancer sequence present in the T-DNA improves the GUS-tagging efficiency. Reverse transcriptase-PCR analysis of a subset of randomly selected pGA2715 lines shows that expression of the genes immediately adjacent to the inserted enhancer is increased significantly. Therefore, the large population of T-DNA-tagged lines transformed with pGA2715 could be used to screen for promoter activity using the gus reporter, as well as for creating gain-of-function mutants. PMID:12481047
Jeong, Dong-Hoon; An, Suyoung; Kang, Hong-Gyu; Moon, Sunok; Han, Jong-Jin; Park, Sunhee; Lee, Hyun Sook; An, Kyungsook; An, Gynheung
T-DNA Insertional Mutagenesis for Activation Tagging in Rice1
We have developed a new T-DNA vector, pGA2715, which can be used for promoter trapping and activation tagging of rice (Oryza sativa) genes. The binary vector contains the promoterless ?-glucuronidase (GUS) reporter gene next to the right border. In addition, the multimerized transcriptional enhancers from the cauliflower mosaic virus 35S promoter are located next to the left border. A total of 13,450 T-DNA insertional lines have been generated using pGA2715. Histochemical GUS assays have revealed that the GUS-staining frequency from those lines is about twice as high as that from lines transformed with the binary vector pGA2707, which lacks the enhancer element. This result suggests that the enhancer sequence present in the T-DNA improves the GUS-tagging efficiency. Reverse transcriptase-PCR analysis of a subset of randomly selected pGA2715 lines shows that expression of the genes immediately adjacent to the inserted enhancer is increased significantly. Therefore, the large population of T-DNA-tagged lines transformed with pGA2715 could be used to screen for promoter activity using the gus reporter, as well as for creating gain-of-function mutants.
Jeong, Dong-Hoon; An, Suyoung; Kang, Hong-Gyu; Moon, Sunok; Han, Jong-Jin; Park, Sunhee; Lee, Hyun Sook; An, Kyungsook; An, Gynheung
Molecular analysis of T-DNA insertion mutants identified putative regulatory elements in the AtTERT gene
Analysis of plants bearing a T-DNA insertion is a potent tool of modern molecular biology, providing valuable information about the function and involvement of genes in metabolic pathways. A collection of 12 Arabidopsis thaliana lines with T-DNA insertions in the gene coding for the catalytic subunit of telomerase (AtTERT) and in adjacent regions was screened for telomerase activity [telomere repeat amplification protocol (TRAP) assay], telomere length (terminal restriction fragments), and AtTERT transcription (quantitative reverse transcription-PCR). Lines with the insertion located upstream of the start codon displayed unchanged telomere stability and telomerase activity, defining a putative minimal AtTERT promoter and the presence of a regulatory element linked to increased transcription in the line SALK_048471. Lines bearing a T-DNA insertion inside the protein-coding region showed telomere shortening and lack of telomerase activity. Transcription in most of these lines was unchanged upstream of the T-DNA insertion, while it was notably decreased downstream. The expression profile varied markedly in mutant lines harbouring insertions at the 5' end of AtTERT which showed increased transcription and abolished tissue specificity. Moreover, the line FLAG_385G01 (T-DNA insertion inside intron 1) revealed the presence of a highly abundant downstream transcript with normal splicing but without active telomerase. The role of regulatory elements found along the AtTERT gene is discussed in respect to natural telomerase expression and putative intron-mediated enhancement.
Fojtova, Miloslava; Peska, Vratislav; Dobsakova, Zuzana; Mozgova, Iva; Fajkus, Jiri; Sykorova, Eva
Spectrum of T-DNA integrations for insertional mutagenesis of Histoplasma capsulatum
Agrobacterium-mediated transformation is being increasingly used for insertional mutagenesis of fungi. To better evaluate its effectiveness as a mutagen for the fungal pathogen Histoplasma capsulatum, we analyzed a collection of randomly selected T-DNA insertion mutants. Testing of different T-DNA element vectors engineered for transformation of fungi showed that pBHt2 provides the highest transformation efficiency and the lowest rate of vector backbone carryover. Sixty-eight individual T-DNA integrations were characterized by recovery of T-DNA ends and flanking genomic sequences. The right border end of the T-DNA is largely preserved whereas the left border end is frequently truncated. Analysis of T-DNA insertion sites confirms the lack of any integration hotspots in the Histoplasma genome. Relative to genes, T-DNA integrations show significant bias towards promoter regions at the expense of coding sequences. With consideration for potential promoter interruption and the demonstrated efficacy of intronic insertions, 61% of mapped T-DNA insertions should impair gene expression or function. Mapping of T-DNA flanking sequences demonstrates 67% of T-DNA integrations are integrations at a single chromosomal site and 31% of T-DNA integrations are associated with large-scale chromosomal rearrangements. This characterization of T-DNA insertions in mutants selected without regard to phenotype supports application of Agrobacterium-mediated transformation as an insertional mutagen for genome-based screens and functional discovery of genes in Histoplasma.
Kemski, Megan M.; Stevens, Bryan; Rappleye, Chad A.
Studies on rice seed quality through analysis of a large-scale T-DNA insertion population
A rice (Oryza sativa) T-DNA insertion population, which included more than 63 000 independent transgenic lines and 8 840 identified flanking sequence tags (FSTs) that were mapped onto the rice genome, was developed to systemically study the rice seed quality control. Genome-wide analysis of the FST distribution showed that T-DNA insertions were positively correlated with expressed genes, but negatively with
Fang-Fang Fu; Rui Ye; Shu-Ping Xu; Hong-Wei Xue
Analysis of T-DNA insertion site distribution patterns in Arabidopsis thaliana reveals special features of genes without insertions
Large collections of sequence-indexed T-DNA insertion mutants are invaluable resources for plant functional genomics. Flanking sequence tag (FST) data from these collections indicated that T-DNA insertions are not randomly distributed in the Arabidopsis thaliana genome and that there are still a fairly high number of annotated genes without T-DNA insertions. We have analyzed FST data from the FLAGdb, GABI-Kat, and
Yong Li; Mario G. Rosso; Bekir ▄lker; Bernd Weisshaar
High throughput T-DNA insertion mutagenesis in rice: a first step towards in silico reverse genetics.
A library of 29,482 T-DNA enhancer trap lines has been generated in rice cv. Nipponbare. The regions flanking the T-DNA left border from the first 12,707 primary transformants were systematically isolated by adapter anchor PCR and sequenced. A survey of the 7480 genomic sequences larger than 30 bp (average length 250 bp), representing 56.4% of the total readable sequences and matching the rice bacterial artificial chromosome/phage artificial chromosome (BAC/PAC) sequences assembled in pseudomolecules allowed the assigning of 6645 (88.8%) T-DNA insertion sites to at least one position in the rice genome of cv. Nipponbare. T-DNA insertions appear to be rather randomly distributed over the 12 rice chromosomes, with a slightly higher insertion frequency in chromosomes 1, 2, 3 and 6. The distribution of 723 independent T-DNA insertions along the chromosome 1 pseudomolecule did not differ significantly from that of the predicted coding sequences in exhibiting a lower insertion density around the centromere region and a higher density in the subtelomeric regions where the gene density is higher. Further establishment of density graphs of T-DNA inserts along the recently released 12 rice pseudomolecules confirmed this non-uniform chromosome distribution. T-DNA appeared less prone to hot spots and cold spots of integration when compared with those revealed by a concurrent assignment of the Tos17 retrotransposon flanking sequences deposited in the National Center for Biotechnology Information (NCBI). T-DNA inserts rarely integrated into repetitive sequences. Based on the predicted gene annotation of chromosome 1, preferential insertion within the first 250 bp from the putative ATG start codon has been observed. Using 4 kb of sequences surrounding the insertion points, 62% of the sequences showed significant similarity to gene encoding known proteins (E-value < 1.00 e(-05)). To illustrate the in silico reverse genetic approach, identification of 83 T-DNA insertions within genes coding for transcription factors (TF) is presented. Based both on the estimated number of members of several large TF gene families (e.g. Myb, WRKY, HD-ZIP, Zinc-finger) and on the frequency of insertions in chromosome 1 predicted genes, we could extrapolate that 7-10% of the rice gene complement is already tagged by T-DNA insertion in the 6116 independent transformant population. This large resource is of high significance while assisting studies unravelling gene function in rice and cereals, notably through in silico reverse genetics. PMID:15255873
Sallaud, Christophe; Gay, CÚline; Larmande, Pierre; BŔs, Martine; Piffanelli, Pietro; PiÚgu, Benoit; Droc, GaÚtan; Regad, Farid; Bourgeois, Emmanuelle; Meynard, Donaldo; PÚrin, Christophe; Sabau, Xavier; GhesquiŔre, Alain; Glaszmann, Jean Christophe; Delseny, Michel; Guiderdoni, Emmanuel
T-DNA insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus Laccaria bicolor
Summary Ectomycorrhiza is a mutualistic symbiosis formed between fine roots of trees and the mycelium of soil fungi. This symbiosis plays a key role in forest ecosystems for the mineral nutrition of trees and the biology of the fungal communities associated. The characterization of genes involved in developmental and metabolic processes is important to understand the complex interactions that control the ectomycorrhizal symbiosis. Agrobacterium?mediated gene transfer (AMT) in fungi is currently opening a new era for fungal research. As whole genome sequences of several fungi are being released studies about T?DNA integration patterns are needed in order to understand the integration mechanisms involved and to evaluate the AMT as an insertional mutagenesis tool for different fungal species. The first genome sequence of a mycorrhizal fungus, the basidiomycete Laccaria bicolor, became public in July 2006. Release of Laccaria genome sequence and the availability of AMT makes this fungus an excellent model for functional genomic studies in ectomycorrhizal research. No data on the integration pattern in Laccaria genome were available, thus we optimized a plasmid rescue approach for this fungus. To this end the transformation vector (pHg/pBSk) was constructed allowing the rescue of the T?DNA right border (RB)ľgenomic DNA junctions in Escherichia coli. Fifty?one Agrobacterium?transformed fungal strains, picked up at random from a larger collection of T?DNA tagged strains (about 500), were analysed. Sixty?nine per cent were successfully rescued for the RB of which 87% were resolved for genomic integration sequences. Our results demonstrate that the plasmid rescue approach can be used for resolving T?DNA integration sites in Laccaria. The RB was well conserved during transformation of this fungus and the integration analysis showed no clear sequence homology between different genomic sites. Neither obvious sequence similarities were found between these sites and the T?DNA borders indicating non?homologous integration of the transgenes. Majority (75%) of the integrations were located in predicted genes. Agrobacterium?mediated gene transfer is a powerful tool that can be used for functional gene studies in Laccaria and will be helpful along with plasmid rescue in searching for relevant fungal genes involved in the symbiotic process.
Kemppainen, Minna; Duplessis, Sebastien; Martin, Francis; Pardo, Alejandro G.
Analyses of single-copy Arabidopsis T-DNA-transformed lines show that the presence of vector backbone sequences, short inverted repeats and DNA methylation is not sufficient or necessary for the induction of transgene silencing
In genetically transformed plants, transgene silen- cing has been correlated with multiple and complex insertions of foreign DNA, e.g. T-DNA and vector backbone sequences. Occasionally, single-copy transgenes also suffer transgene silencing. We have compared integration patterns and T-DNA\\/plant DNA junctions in a collection of 37 single-copy T-DNA-transformed Arabidopsis lines, of which 13 displayed silencing. Vector sequences were found integrated in
Trine J. Meza; Biljana Stangeland; Inderjit S. Mercy; Magne Skarn; Dag A. Nymoen; Anita Berg; Melinka A. Butenko; Anne-Mari Hakelien; Camilla Haslekas; Leonardo A. Meza-Zepeda; Reidunn B. Aalen
A candidate gene OsAPC6 of anaphase-promoting complex of rice identified through T-DNA insertion.
A dwarf mutant (Oryza sativa anaphase-promoting complex 6 (OsAPC6)) of rice cultivar Basmati 370 with 50% reduced plant height as compared to the wild type was isolated by Agrobacterium tumefaciens-mediated transformation using Hm(R) Ds cassette. This mutant was found to be insensitive to exogenous gibberellic acid (GA(3)) application. Homozygous mutant plants showed incomplete penetrance and variable expressivity for plant height and pleiotropic effects including gibberellic acid insensitivity, reduced seed size, panicle length, and female fertility. Single copy insertion of T-DNA and its association with OsAPC6 was confirmed by Southern hybridization, germination on hygromycin, and 3:1 segregation of HPT gene in F(2) from OsAPC6 x Basmati 370 cross. The T-DNA flanking region sequenced through thermal asymmetric interlaced polymerase chain reaction showed a single hit on chromosome 3 of japonica rice cultivar Nipponbare in the second exonic region of a gene which encodes for sixth subunit of anaphase-promoting complex/cyclosome. The candidate gene of 8.6-kb length encodes a 728-amino acid protein containing a conserved tetratricopeptide repeat (TPR) domain and has only a paralog, isopenicillin N-synthase family protein on the same chromosome without the TPR domain. There was no expression of the gene in the mutant while in Basmati 370, it was equal in both roots and shoots. The knockout mutant OsAPC6 interferes with the gibberellic acid signaling pathway leading to reduced height and cell size probably through ubiquitin-mediated proteolysis. Further functional validation of the gene through RNAi is in progress. PMID:20091079
Kumar, Mankesh; Basha, P Osman; Puri, Anju; Rajpurohit, Deepak; Randhawa, Gursharn Singh; Sharma, Tilak Raj; Dhaliwal, Harcharan Singh
Effect of T-DNA insertions on mRNA transcript copy numbers upstream and downstream of the insertion site in Arabidopsis thaliana explored by surface enhanced Raman spectroscopy.
We report the effect of a T-DNA insertion on the expression level of mRNA transcripts of the TWISTED DWARF 1 (TWD1) gene upstream and downstream of the T-DNA insertion site in Arabidopsis. A novel approach based on surface-enhanced Raman spectroscopy (SERS) was developed to detect and quantify the effect of a T-DNA insertion on mRNA transcript accumulation at 5'- and 3'-ends of the TWD1 gene. A T-DNA insertion mutant in the TWD1 gene (twd1-2) was chosen to test the sensitivity and the feasibility of the approach. The null mutant of the FK506-like immunophilin protein TWD1 in Arabidopsis shows severe dwarfism and strong disoriented growth of plant organs. A spontaneous arising suppressor allele of twd1-2 called twd-sup displayed an intermediate phenotype between wild type and the knockout phenotype of twd1-2. Both twd1 mutant alleles have identical DNA sequences at the TWD1 locus including the T-DNA insertion in the fourth intron of the TWD1 gene but they show clear variability in the mutant phenotype. We present here the development and application of SERS-based mRNA detection and quantification using the expression of the TWD1 gene in wild type and both mutant alleles. The hallmarks of our SERS approach are a robust and fast assay to detect up to 0.10áfm of target molecules including the ability to omit in vitro transcription and amplification steps after RNA isolation. Instead we perform direct quantification of RNA molecules. This enables us to detect and quantify rare RNA molecules at high levels of precision and sensitivity. PMID:24460907
Kadam, Ulhas; Moeller, Claudia A; Irudayaraj, Joseph; Schulz, Burkhard
Analysis of T-DNA/Host-Plant DNA Junction Sequences in Single-Copy Transgenic Barley Lines
Sequencing across the junction between an integrated transfer DNA (T-DNA) and a host plant genome provides two important pieces of information. The junctions themselves provide information regarding the proportion of T-DNA which has integrated into the host plant genome, whilst the transgene flanking sequences can be used to study the local genetic environment of the integrated transgene. In addition, this information is important in the safety assessment of GM crops and essential for GM traceability. In this study, a detailed analysis was carried out on the right-border T-DNA junction sequences of single-copy independent transgenic barley lines. T-DNA truncations at the right-border were found to be relatively common and affected 33.3% of the lines. In addition, 14.3% of lines had rearranged construct sequence after the right border break-point. An in depth analysis of the host-plant flanking sequences revealed that a significant proportion of the T-DNAs integrated into or close to known repetitive elements. However, this integration into repetitive DNA did not have a negative effect on transgene expression.
Bartlett, Joanne G.; Smedley, Mark A.; Harwood, Wendy A.
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.
Larmande, Pierre; Gay, Celine; Lorieux, Mathias; Perin, Christophe; Bouniol, Matthieu; Droc, Gaetan; Sallaud, Christophe; Perez, Pascual; Barnola, Isabelle; Biderre-Petit, Corinne; Martin, Jerome; Morel, Jean Benoit; Johnson, Alexander A. T.; Bourgis, Fabienne; Ghesquiere, Alain; Ruiz, Manuel; Courtois, Brigitte; Guiderdoni, Emmanuel
An Arabidopsis T-DNA insertion mutant for galactokinase (AtGALK, At3g06580) hyperaccumulates free galactose and is insensitive to exogenous galactose.
Galactokinase (GALK, EC 220.127.116.11) is a cytosolic enzyme with a wide occurrence across the taxonomic kingdoms. It catalyzes the phosphorylation of ?-d-galactose (Gal) to ?-d-Gal-1-P. The cytotoxicity of free (unphosphorylated) Gal is well documented in plants and causes marked defects. An Arabidopsis GALK (AtGALK, At3g06580) was previously identified, cloned and functionally characterized in Escherichia coli and was suggested to occur as a single copy gene in Arabidopsis. We identified an AtGALK T-DNA insertion mutant (atgalk) that (i) is AtGALK transcript deficient; (ii) displays no GALK activity in vegetative tissues; and (iii) accumulates Gal up to 6.8 mg g(-1) FW in vegetative tissues, in contrast to wild-type plants. By constitutively overexpressing the AtGALK cDNA, atgalk was functionally rescued. Three independent transformed lines showed restored AtGALK transcripts and GALK activity and had low leaf Gal concentrations comparable with those observed in wild-type plants. Surprisingly, in vitro grown atgalk plants were largely insensitive to the exogenous application of up to 100 mM free Gal, while wild-type plants exhibited sensitivity to low Gal concentrations (10 mM). Furthermore, atgalk seedlings retained the capacity for uptake of exogenously supplied Gal (100 mM), accumulating up to 57 mg g(-1) FW in leaves. Leaves from soil-grown atgalk plants that exhibited no growth or morphological defects were used to demonstrate that the accumulating Gal occurred exclusively in the vacuoles of mesophyll protoplasts. Collectively, these findings suggest a novel Gal detoxification pathway that targets free Gal to the vacuole and is active in the atgalk mutant background. PMID:22437845
Egert, AurÚlie; Peters, Shaun; Guyot, Christelle; Stieger, Bruno; Keller, Felix
Selection of T-DNA-Tagged Male and Female Gametophytic Mutants by Segregation Distortion in Arabidopsis
As a strategy for the identification of T-DNA-tagged gametophytic mutants, we have used T-DNA inser- tional mutagenesis based on screening for distorted segregation ratios by antibiotic selection. Screening of z1000 transgenic Arabidopsis families led to the isolation of eight lines showing reproducible segregation ratios of z1:1, suggesting that these lines are putative gametophytic mutants caused by T-DNA insertion at a
Ross Howden; Soon Ki Park; James M. Moore; James Orme; Ueli Grossniklaus; David Twell
Major chromosomal rearrangements induced by T-DNA transformation in Arabidopsis.
We show that major chromosomal rearrangements can occur upon T-DNA transformation of Arabidopsis thaliana. In the ACL4 line, two T-DNA insertion loci were found; one is a tandem T-DNA insert in a head-to-head orientation, and the other is a truncated insert with only the left part of the T-region. The four flanking DNA regions were isolated and located on the Arabidopsis chromosomes; for both inserts, one side of the T-DNA maps to chromosome 2, whereas the other side maps to chromosome 3. Both chromosome 3 flanking regions map to the same location, despite a 1.4-kb deletion at this point, whereas chromosome 2 flanking regions are located 40 cM apart on the bottom arm of chromosome 2. These results strongly suggest a reciprocal translocation between chromosomes 2 and 3, with the breakpoints located at the T-DNA insertion sites. The interchanged fragments roughly correspond to the 20-cM distal ends of both chromosomes. Moreover, a large inversion, spanning 40 cM on the genetic map, occurs on the bottom arm of chromosome 2. This was confirmed by genetic analyses that demonstrated a strong reduction of recombination in the inverted region. Models for T-DNA integration and the consequences for T-DNA tagging are discussed in light of these results.
Nacry, P; Camilleri, C; Courtial, B; Caboche, M; Bouchez, D
In-depth molecular and phenotypic characterization in a rice insertion line library facilitates gene identification through reverse and forward genetics approaches.
We report here the molecular and phenotypic features of a library of 31,562 insertion lines generated in the model japonica cultivar Nipponbare of rice (Oryza sativa L.), called Oryza Tag Line (OTL). Sixteen thousand eight hundred and fourteen T-DNA and 12,410 Tos17 discrete insertion sites have been characterized in these lines. We estimate that 8686 predicted gene intervals--i.e. one-fourth to one-fifth of the estimated rice nontransposable element gene complement--are interrupted by sequence-indexed T-DNA (6563 genes) and/or Tos17 (2755 genes) inserts. Six hundred and forty-three genes are interrupted by both T-DNA and Tos17 inserts. High quality of the sequence indexation of the T2 seed samples was ascertained by several approaches. Field evaluation under agronomic conditions of 27,832 OTL has revealed that 18.2% exhibit at least one morphophysiological alteration in the T1 progeny plants. Screening 10,000 lines for altered response to inoculation by the fungal pathogen Magnaporthe oryzae allowed to observe 71 lines (0.7%) developing spontaneous lesions simulating disease mutants and 43 lines (0.4%) exhibiting an enhanced disease resistance or susceptibility. We show here that at least 3.5% (four of 114) of these alterations are tagged by the mutagens. The presence of allelic series of sequence-indexed mutations in a gene among OTL that exhibit a convergent phenotype clearly increases the chance of establishing a linkage between alterations and inserts. This convergence approach is illustrated by the identification of the rice ortholog of AtPHO2, the disruption of which causes a lesion-mimic phenotype owing to an over-accumulation of phosphate, in nine lines bearing allelic insertions. PMID:22369597
Lorieux, Mathias; Blein, MÚlisande; Lozano, Jaime; Bouniol, Mathieu; Droc, GaÚtan; DiÚvart, Anne; PÚrin, Christophe; Mieulet, Delphine; Lanau, NadŔge; BŔs, Martine; RouviŔre, Claire; Gay, CÚline; Piffanelli, Pietro; Larmande, Pierre; Michel, Corinne; Barnola, Isabelle; Biderre-Petit, Corinne; Sallaud, Christophe; Perez, Pascual; Bourgis, Fabienne; GhesquiŔre, Alain; Gantet, Pascal; Tohme, Joe; Morel, Jean Benoit; Guiderdoni, Emmanuel
Mapped Ds/T-DNA launch pads for functional genomics in barley.
A system for targeted gene tagging and local saturation mutagenesis based on maize transposable elements (Ac/Ds) was developed in barley (Hordeum vulgare L.). We generated large numbers of transgenic barley lines carrying a single copy of the non-autonomous maize Ds element at defined positions in the genome. Independent Ds lines were either generated by activating Ds elements in existing single-copy lines after crossing with AcTPase-expressing plants or by Agrobacterium-mediated transformation. Genomic DNA flanking Ds and T-DNA insertion sites from over 200 independent lines was isolated and sequenced, and was used for a sequence based mapping strategy in a barley reference population. More than 100 independent Ds insertion sites were mapped and can be used as launch pads for future targeted tagging of genes in the vicinity of the insertion sites. Sequence analysis of Ds and T-DNA flanking regions revealed a sevenfold preference of both mutagens for insertion into non-redundant, gene-containing regions of the barley genome. However, whilst transposed Ds elements preferentially inserted adjacent to regions with a high number of predicted and experimentally validated matrix attachment regions (nuclear MARs), this was not the case for T-DNA integration sites. These findings and an observed high transposition frequency from mapped launch pads demonstrate the future potential of gene tagging for functional genomics and gene discovery in barley. PMID:16889649
Zhao, Tiehan; Palotta, Margaret; Langridge, Peter; Prasad, Manoj; Graner, Andreas; Schulze-Lefert, Paul; Koprek, Thomas
Forward genetics screening of Medicago truncatula Tnt1 insertion lines.
A large population of Medicago truncatula insertion lines has been generated using the Tnt1 retrotransposon. More than 21,000 insertion lines have been generated, representing more than 500,000 insertion events. This mutant population is being used by the legume research community to screen for various different mutants using a forward genetics approach. Some of the phenotypes that have been screened using this population include developmentally abnormal phenotypes in leaves, stem, flowers, and roots. In addition to these, mutants with defects in symbiosis with Rhizobium and mycorrhiza, mutants with altered nonhost resistance against Asian Soybean Rust and switch grass rust pathogens, mutants with altered lignin content, mutants with altered cell wall structure, etc. have been identified. Here, we describe the high throughput methodology that is being used to identify these M. truncatula mutants. PMID:23996311
Yarce, Juan Carlos Serrani; Lee, Hee-Kyung; Tadege, Million; Ratet, Pascal; Mysore, Kirankumar S
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.
Identification of pathogenesis-associated genes by T-DNA-mediated insertional mutagenesis in Botrytis cinerea: a type 2A phosphoprotein phosphatase and an SPT3 transcription factor have significant impact on virulence.
Agrobacterium tumefaciens-mediated transformation (ATMT) was used to generate an insertional mutant library of the gray mold fungus Botrytis cinerea. From a total of 2,367 transformants, 68 mutants showing significant reduction in virulence on tomato and bean plants were analyzed in detail. As reported for other fungal ATMT libraries, integrations were mostly single copy, occurred preferentially in noncoding (regulatory) regions, and were frequently accompanied by small deletions of the target sequences and loss of parts of the border sequence. Two T-DNA integration events that were found to be linked to virulence were characterized in more detail: a catalytic subunit of a PP2A serine/threonine protein phosphatase (BcPP2Ac) and the SPT3 subunit of a Spt-Ada-Gcn5-acetyltransferase (SAGA-like) transcriptional regulator complex. Gene replacement and silencing approaches revealed that both Bcpp2Ac and SPT3 are crucial for virulence, growth, and differentiation as well as for resistance to H(2)O(2) in B. cinerea. PMID:22112214
Giesbert, S; Schumacher, J; Kupas, V; Espino, J; SegmŘller, N; Haeuser-Hahn, I; Schreier, P H; Tudzynski, P
Risk factors for pneumothorax during percutaneous hickman line insertion in patients with solid and haematological tumours
The purpose of this study was to identify risk factors for pneumothorax during percutaneous subclavian Hickman line insertion in patients with haematological (HT) or solid tumours (ST). One hundred and twelve patients (55 HT, 57 ST) had 132 subclavian Hickman lines inserted under fluoroscopic control. Lines were inserted on the left on 116 occasions and the right in 16. Thirty-five
K. J. Harrington; H. S. Pandha; J. S. Hollyer; S. A. Kelly; A. R. Bateman; C. Lewanski; P. Morris; J. E. Jackson
Germinal Transpositions of the Maize Element Dissociation from T-DNA Loci in Tomato
We have analyzed the pattern of germinal transpositions of artificial Dissociation (Ds) transposons in tomato. T-DNA constructs carrying Ds were transformed into tomato, and the elements were trans-activated by crossing to lines transformed with a stabilized Activator (sAc) that expressed the transposase gene. The sAc T-DNA carried a GUS gene to monitor its segregation. The Ds elements were inserted in a marker gene so that excision from the T-DNA could be monitored. The Ds elements also carried a genetic marker that was intended to be used for reinsertion selection of the elements after excision. Unfortunately, this gene was irreversibly inactivated on crossing to sAc. Germinal excision frequencies of Ds averaged 15-40%, but there was large variation between and within plants. Southern hybridization analysis of stable transposed Ds elements indicated that although unique transpositions predominate, early transposition events can lead to large clonal sectors in the germline of developing plants and to sibling offspring carrying the same transposition event. Multiple germinal transpositions from three different loci were examined for uniqueness, and 15 different transpositions were identified from each of three T-DNA loci that carried a single independent Ds. These were mapped relative to the donor T-DNA loci, and for each locus 70-80% of the transposed elements were closely linked to the donor site.
Carroll, B. J.; Klimyuk, V. I.; Thomas, C. M.; Bishop, G. J.; Harrison, K.; Scofield, S. R.; Jones, JDG.
Technical note: development of a tool to insert abomasal infusion lines into dairy cows.
A tool was developed to aid in ruminal insertion of abomasal infusion lines into dairy cows. The tool consisted of 2 pieces cut from polyvinyl chloride pipe. The first piece of pipe, the insertion tool, contained a groove that held the flexible plastic flange that is on the end of the infusion line. The insertion tool containing the flange was inserted into the ruminal cannula, through the sulcus omasi, and into the abomasum. The second piece of pipe, the delivery tool, was threaded through the insertion tool, and it was used to dislodge the flange from the insertion tool and into the abomasum. PMID:16960071
Gressley, T F; Reynal, S M; Colmenero, J J Olmos; Broderick, G A; Armentano, L E
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.
Blood Cultures at Central Line Insertion in the Intensive Care Unit: Comparison with Peripheral Venipuncture?
Blood cultures are a key diagnostic test for intensive care unit (ICU) patients; however, contaminants complicate interpretations and lead to unnecessary antibiotic administration and costs. Indications for blood cultures and central venous catheter (CVC) insertions often overlap for ICU patients. Obtaining blood cultures under the strict sterile precautions utilized for CVC insertion might be expected to decrease culture contamination. This retrospective study compared the results of blood cultures taken at CVC insertion, at arterial line insertion, and from peripheral venipuncture in order to validate the advantage of CVC insertion cultures. Cultures from indwelling lines were excluded. Results of 14,589 blood cultures, including 2,736 (19%) CVC, 1,513 (10%) arterial line, and 10,340 (71%) peripheral cultures taken over 5.5 years in two ICUs (general and medical) were analyzed. CVC cultures were contaminated more frequently than arterial line or peripheral cultures (225/2,736 [8%] CVC, 48/1,513 [3%] arterial line, and 378/10,340 (4%) peripheral cultures [P < 0.001 for CVC versus peripheral and CVC versus arterial line cultures]). True pathogens were found more frequently in CVC insertion cultures (334/2,736 [12%] CVC, 155/1,513 [10%] arterial line, and 795/10,340 [8%] peripheral cultures [P < 0.001 for CVC versus peripheral cultures; P = 0.055 for CVC versus arterial line cultures; P < 0.001 for peripheral versus arterial line cultures]). Contamination and true-positive rates were similar for culture sets from the two ICUs for each given culture source. Despite superior sterile precautions, cultures taken at the time of central line insertion had a higher contamination rate than did either peripheral or arterial line blood cultures. This may be related to the increased manipulations required for CVC insertion.
Stohl, Sheldon; Benenson, Shmuel; Sviri, Sigal; Avidan, Alexander; Block, Colin; Sprung, Charles L.; Levin, Phillip D.
Central line insertion bundle: experiences and challenges in an adult ICU.
Background: Central venous catheters are used frequently in the intensive care unit (ICU). However, there is an associated morbidity, mortality and cost derived from their infectious and mechanical complications. The Scottish Patient Safety Programme (SPSP) has developed a multi faceted care bundle, with the aim of reducing catheter-related blood stream infections. Aim: This paper aims to identify and describe the experience and challenges in implementing the SPSP central line insertion bundle in one adult ICU, in a large inner city teaching hospital. Interventions:'Plan-Do-Study-Act' cycles, checklists for insertion and a standardized trolley were adopted to implement the central line insertion bundle in clinical practice. Conclusion/Implications: Improving the reliability of the central line insertion bundle has reduced infections. Key steps in the process were setting clear aims and ensuring staff understand the change process and measurement of results. This is fundamental to the success of any quality improvement process. PMID:22497916
McPeake, Joanne; Cantwell, Shelia; Booth, Malcolm G; Daniel, Malcolm
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
Genomic rearrangements by LINE-1 insertion-mediated deletion in the human and chimpanzee lineages.
Long INterspersed Elements (LINE-1s or L1s) are abundant non-LTR retrotransposons in mammalian genomes that are capable of insertional mutagenesis. They have been associated with target site deletions upon insertion in cell culture studies of retrotransposition. Here, we report 50 deletion events in the human and chimpanzee genomes directly linked to the insertion of L1 elements, resulting in the loss of approximately 18 kb of sequence from the human genome and approximately 15 kb from the chimpanzee genome. Our data suggest that during the primate radiation, L1 insertions may have deleted up to 7.5 Mb of target genomic sequences. While the results of our in vivo analysis differ from those of previous cell culture assays of L1 insertion-mediated deletions in terms of the size and rate of sequence deletion, evolutionary factors can reconcile the differences. We report a pattern of genomic deletion sizes similar to those created during the retrotransposition of Alu elements. Our study provides support for the existence of different mechanisms for small and large L1-mediated deletions, and we present a model for the correlation of L1 element size and the corresponding deletion size. In addition, we show that internal rearrangements can modify L1 structure during retrotransposition events associated with large deletions. PMID:16034026
Han, Kyudong; Sen, Shurjo K; Wang, Jianxin; Callinan, Pauline A; Lee, Jungnam; Cordaux, Richard; Liang, Ping; Batzer, Mark A
Genomic rearrangements by LINE-1 insertion-mediated deletion in the human and chimpanzee lineages
Long INterspersed Elements (LINE-1s or L1s) are abundant non-LTR retrotransposons in mammalian genomes that are capable of insertional mutagenesis. They have been associated with target site deletions upon insertion in cell culture studies of retrotransposition. Here, we report 50 deletion events in the human and chimpanzee genomes directly linked to the insertion of L1 elements, resulting in the loss of ?18 kb of sequence from the human genome and ?15 kb from the chimpanzee genome. Our data suggest that during the primate radiation, L1 insertions may have deleted up to 7.5 Mb of target genomic sequences. While the results of our in vivo analysis differ from those of previous cell culture assays of L1 insertion-mediated deletions in terms of the size and rate of sequence deletion, evolutionary factors can reconcile the differences. We report a pattern of genomic deletion sizes similar to those created during the retrotransposition of Alu elements. Our study provides support for the existence of different mechanisms for small and large L1-mediated deletions, and we present a model for the correlation of L1 element size and the corresponding deletion size. In addition, we show that internal rearrangements can modify L1 structure during retrotransposition events associated with large deletions.
Han, Kyudong; Sen, Shurjo K.; Wang, Jianxin; Callinan, Pauline A.; Lee, Jungnam; Cordaux, Richard; Liang, Ping; Batzer, Mark A.
Formation of Complex Extrachromosomal T-DNA Structures in Agrobacterium tumefaciens-Infected Plants1[C][W][OA
Agrobacterium tumefaciens is a unique plant pathogenic bacterium renowned for its ability to transform plants. The integration of transferred DNA (T-DNA) and the formation of complex insertions in the genome of transgenic plants during A. tumefaciens-mediated transformation are still poorly understood. Here, we show that complex extrachromosomal T-DNA structures form in A. tumefaciens-infected plants immediately after infection. Furthermore, these extrachromosomal complex DNA molecules can circularize in planta. We recovered circular T-DNA molecules (T-circles) using a novel plasmid-rescue method. Sequencing analysis of the T-circles revealed patterns similar to the insertion patterns commonly found in transgenic plants. The patterns include illegitimate DNA end joining, T-DNA truncations, T-DNA repeats, binary vector sequences, and other unknown ôfillerö sequences. Our data suggest that prior to T-DNA integration, a transferred single-stranded T-DNA is converted into a double-stranded form. We propose that termini of linear double-stranded T-DNAs are recognized and repaired by the plantĺs DNA double-strand break-repair machinery. This can lead to circularization, integration, or the formation of extrachromosomal complex T-DNA structures that subsequently may integrate.
Singer, Kamy; Shiboleth, Yoel M.; Li, Jianming; Tzfira, Tzvi
Trichome Development in Arabidopsis thaliana. I. T-DNA Tagging of the GLABROUS1 Gene
Progeny from a transformed Arabidopsis plant (produced by the Agrobacterium-mediated seed transformation procedure) were found to be segregating for an altered trichome phenotype. lhe mutant plants have normal leaf trichomes but completely lack trichomes usually found on the stem. The mutation is tightly linked to a T-DNA insert. Complementation analysis with genetically characterized trichome mutants revealed that the new mutation
M. David Marks; Kenneth A. Feldmann
Analysis of T-DNA alleles of flavonoid biosynthesis genes in Arabidopsis ecotype Columbia
Background The flavonoid pathway is a long-standing and important tool for plant genetics, biochemistry, and molecular biology. Numerous flavonoid mutants have been identified in Arabidopsis over the past several decades in a variety of ecotypes. Here we present an analysis of Arabidopsis lines of ecotype Columbia carrying T-DNA insertions in genes encoding enzymes of the central flavonoid pathway. We also provide a comprehensive summary of various mutant alleles for these structural genes that have been described in the literature to date in a wide variety of ecotypes. Findings The confirmed knockout lines present easily-scorable phenotypes due to altered pigmentation of the seed coat (or testa). Knockouts for seven alleles for six flavonoid biosynthetic genes were confirmed by PCR and characterized by UPLC for altered flavonol content. Conclusion Seven mutant lines for six genes of the central flavonoid pathway were characterized in ecotype, Columbia. These lines represent a useful resource for integrating biochemical and physiological studies with genomic, transcriptomic, and proteomic data, much of which has been, and continues to be, generated in the Columbia background.
Superconductive tapped delay lines for low-insertion-loss wideband analog signal-processing filters
Transversal filters have been realized as miniature superconductive tapped electromagnetic delay lines. The low loss of superconductors permits the use of a compact stripline structure which offers useful delay at microwave frequencies. Taps are realized as backward-wave couplers or a impedance discontinuities. Chirp filters are formed by cascading such couplers of increasing length. Pulse expansion and compression have been demonstrated over a 2.6-GHz bandwidth. By weighting the taps, side-lobe levels have been reduced to 25 dB below the peak output. A coupled-mode analysis has been employed for predicting the response of this class of filters and has achieved very good agreement with experiment. The analysis also predicts that, if coupling were increased to reduce insertion loss below about 10 dB, significant phase distortion results with a commensurate degradation of side-lobe levels.
Withers, R.S.; Wright, P.V.
Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana.
A recessive pale mutation, designated as cs, was identified by transferred-DNA (T-DNA)-mediated insertional mutagenesis in Arabidopsis thaliana. The pale mutation, cosegregating with the hygromycin resistance marker of the T-DNA, was mapped to the position of the ch-42 (chlorata) locus on chromosome 4. Lack of genetic complementation between cs and ch-42 mutants indicated allelism. Plant boundaries of the T-DNA insert rescued from the pale mutant were used as probes for the isolation of genomic and full-length cDNA clones of the wild-type cs gene. Transformation of the pale mutant with T-DNA vectors carrying these clones resulted in a normal green phenotype, thus demonstrating positive complementation of the T-DNA induced mutation. DNA sequence comparison of the cs mutant and its wild-type allele revealed that the T-DNA insertion occurred 11 bp upstream of the stop codon. A fusion protein, seven amino acids longer than its wild-type counterpart of Mr 46,251, is therefore synthesized in the pale mutant. Transcript analysis during dark-light transition, in vitro protein transport assay, and the absence of DNA sequence homology between cs and known genes indicates that the light regulated expression of the cs gene results in the synthesis of a novel chloroplast protein. Images Fig.1 Fig.5 Fig.2 Fig.6
Koncz, C; Mayerhofer, R; Koncz-Kalman, Z; Nawrath, C; Reiss, B; Redei, G P; Schell, J
Identification and characterization of novel polymorphic LINE-1 insertions through comparison of two human genome sequence assemblies.
Mobile elements represent a relatively new class of markers for the study of human evolution. Long interspersed elements (LINEs) belong to a group of retrotransposons comprising approximately 21% of the human genome. Young LINE-1 (L1) elements that have integrated recently into the human genome can be polymorphic for insertion presence/absence in different human populations at particular chromosomal locations. To identify putative novel L1 insertion polymorphisms, we computationally compared two draft assemblies of the whole human genome (Public and Celera Human Genome assemblies). We identified a total of 148 potential polymorphic L1 insertion loci, among which 73 were candidates for novel polymorphic loci. Based on additional analyses we selected 34 loci for further experimental studies. PCR-based assays and DNA sequence analysis were performed for these 34 loci in 80 unrelated individuals from four diverse human populations: African-American, Asian, Caucasian, and South American. All but two of the selected loci were confirmed as polymorphic in our human population panel. Approximately 47% of the analyzed loci integrated into other repetitive elements, most commonly older L1s. One of the insertions was accompanied by a BC200 sequence. Collectively, these mobile elements represent a valuable source of genomic polymorphism for the study of human population genetics. Our results also suggest that the exhaustive identification of L1 insertion polymorphisms is far from complete, and new whole genome sequences are valuable sources for finding novel retrotransposon insertion polymorphisms. PMID:17034961
Konkel, Miriam K; Wang, Jianxin; Liang, Ping; Batzer, Mark A
Aberrant pre-mRNA maturation is caused by LINE insertions into introns of the white gene of Drosophila melanogaster.
Insertional mutagenesis screens have provided thousands of mutant alleles for analysing genes of varied functions in Drosophila melanogaster. We here document mechanisms of insertional mutagenesis by a LINE element, the I factor, by determining the molecular structure of RNAs produced from two alleles of the white gene of D.melanogaster, wIR1 and wIR6. These alleles result from insertion of the I factor into introns of the gene. We show that sequences present within the element direct aberrant splicing and termination events. When the I factor is inserted within the white first intron it may lead to the use of a cryptic 3' splice site which does not contain the dinucleotide AG. This splicing gives rise to a chimeric messenger RNA whose synthesis is controlled differently in tissues where the mutated gene is expressed. When the I factor is inserted within the white last intron it induces synthesis of truncated mRNAs. These results provide, for the first time, mechanisms for I factor insertional mutagenesis. They are discussed in the more general context of RNA processing in Drosophila and the evolution of eukaryotic gene introns. Images
Lajoinie, O; Drake, M E; Dastugue, B; Vaury, C
Proteins encoded by Agrobacterium tumefaciens Ti plasmid DNA (T-DNA) in crown gall tumors
In order to detect proteins that may be produced in crown gall tumors as a result of expression of incorporated Agrobacterium tumefaciens Ti plasmid DNA (T-DNA), we have isolated mRNA complementary to T-DNA and translated this in a protein-synthesizing system derived from wheat germ. mRNA prepared from cultured E1 tumor from Nicotiana tabacum hybridized with HindIII fragment 1 sequences of T-DNA immobilized on cellulose nitrate filters. Two proteins of 30,000 and 16,500 Mr were produced when this selected RNA was released and translated. Other tumor lines from N. tabacum were investigated, and a protein of slightly less than 30,000 Mr was encoded by HindIII fragment 1 sequences of 15955/01 tumor. No products were observed for 15955/1 tumor line, which differs from E1/B6-806 and 15955/01 in that it does not produce octopine. mRNA species of each of the tumor lines hybridized to Bst I fragment 8 sequences of T-DNA and produced a common protein of 15,000 Mr. Because this protein is derived from the region of the T-DNA that is conserved in octopine- and nopaline-type crown gall tumors, it may play a role in oncogenicity. Images
McPherson, Joan C.; Nester, Eugene W.; Gordon, Milton P.
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 H(2)O(2). 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 H(2)O(2). 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
Symbiotic mutants deficient in nodule establishment identified after T-DNA transformation of Lotus japonicus
Nitrogen-fixing root nodules develop on legumes as a result of an interaction between host plants and soil bacteria collectively\\u000a referred to as rhizobia. The organogenic process resulting in nodule development is triggered by the bacterial microsymbiont,\\u000a but genetically controlled by the host plant genome. Using T-DNA insertion as a tool to identify novel plant genes that regulate\\u000a nodule ontogeny, we
L. Schauser; K. Handberg; N. Sandal; J. Stiller; T. ThykjŠr; E. Pajuelo; A. Nielsen; J. Stougaard
Characterization and isolation of a T-DNA tagged banana promoter active during in vitro culture and low temperature stress
Background Next-generation transgenic plants will require a more precise regulation of transgene expression, preferably under the control of native promoters. A genome-wide T-DNA tagging strategy was therefore performed for the identification and characterization of novel banana promoters. Embryogenic cell suspensions of a plantain-type banana were transformed with a promoterless, codon-optimized luciferase (luc+) gene and low temperature-responsive luciferase activation was monitored in real time. Results Around 16,000 transgenic cell colonies were screened for baseline luciferase activity at room temperature 2 months after transformation. After discarding positive colonies, cultures were re-screened in real-time at 26░C followed by a gradual decrease to 8░C. The baseline activation frequency was 0.98%, while the frequency of low temperature-responsive luciferase activity was 0.61% in the same population of cell cultures. Transgenic colonies with luciferase activity responsive to low temperature were regenerated to plantlets and luciferase expression patterns monitored during different regeneration stages. Twenty four banana DNA sequences flanking the right T-DNA borders in seven independent lines were cloned via PCR walking. RT-PCR analysis in one line containing five inserts allowed the identification of the sequence that had activated luciferase expression under low temperature stress in a developmentally regulated manner. This activating sequence was fused to the uidA reporter gene and back-transformed into a commercial dessert banana cultivar, in which its original expression pattern was confirmed. Conclusion This promoter tagging and real-time screening platform proved valuable for the identification of novel promoters and genes in banana and for monitoring expression patterns throughout in vitro development and low temperature treatment. Combination of PCR walking techniques was efficient for the isolation of candidate promoters even in a multicopy T-DNA line. Qualitative and quantitative GUS expression analyses of one tagged promoter in a commercial cultivar demonstrated a reproducible promoter activity pattern during in vitro culture. Thus, this promoter could be used during in vitro selection and generation of commercial transgenic plants.
Santos, Efren; Remy, Serge; Thiry, Els; Windelinckx, Saskia; Swennen, Rony; Sagi, Laszlo
Retroviral Elements and Their Hosts: Insertional Mutagenesis in the Mouse Germ Line
The inbred mouse is an invaluable model for human biology and disease. Nevertheless, when considering genetic mechanisms of variation and disease, it is important to appreciate the significant differences in the spectra of spontaneous mutations that distinguish these species. While insertions of transposable elements are responsible for only ~0.1% of de novo mutations in humans, the figure is 100-fold higher
Irina A Maksakova; Mark T Romanish; Liane Gagnier; Catherine A Dunn; Louie N. van de Lagemaat; Dixie L Mager
Incidence of Genome Structure, DNA Asymmetry, and Cell Physiology on T-DNA Integration in Chromosomes of the Phytopathogenic Fungus Leptosphaeria maculans
The ever-increasing generation of sequence data is accompanied by unsatisfactory functional annotation, and complex genomes, such as those of plants and filamentous fungi, show a large number of genes with no predicted or known function. For functional annotation of unknown or hypothetical genes, the production of collections of mutants using Agrobacterium tumefaciensľmediated transformation (ATMT) associated with genotyping and phenotyping has gained wide acceptance. ATMT is also widely used to identify pathogenicity determinants in pathogenic fungi. A systematic analysis of T-DNA borders was performed in an ATMT-mutagenized collection of the phytopathogenic fungus Leptosphaeria maculans to evaluate the features of T-DNA integration in its particular transposable element-rich compartmentalized genome. A total of 318 T-DNA tags were recovered and analyzed for biases in chromosome and genic compartments, existence of CG/AT skews at the insertion site, and occurrence of microhomologies between the T-DNA left border (LB) and the target sequence. Functional annotation of targeted genes was done using the Gene Ontology annotation. The T-DNA integration mainly targeted gene-rich, transcriptionally active regions, and it favored biological processes consistent with the physiological status of a germinating spore. T-DNA integration was strongly biased toward regulatory regions, and mainly promoters. Consistent with the T-DNA intranuclear-targeting model, the density of T-DNA insertion correlated with CG skew near the transcription initiation site. The existence of microhomologies between promoter sequences and the T-DNA LB flanking sequence was also consistent with T-DNA integration to host DNA mediated by homologous recombination based on the microhomology-mediated end-joining pathway.
Bourras, Salim; Meyer, Michel; Grandaubert, Jonathan; Lapalu, Nicolas; Fudal, Isabelle; Linglin, Juliette; Ollivier, Benedicte; Blaise, Francoise; Balesdent, Marie-Helene; Rouxel, Thierry
Routine chest radiographs after central line insertion: Mandatory postprocedural evaluation or unnecessary waste of resources?
Purpose: To study the cost and impact on patient management of the routine performance of chest radiographs in patients undergoing\\u000a imaged-guided central venous catheter insertion.\\u000a \\u000a \\u000a Methods: Six hundred and twenty-one catheters placed in 489 patients over a 42-month period formed the study group. Catheters were\\u000a placed in the right internal jugular vein (425), left internal jugular vein (133), and subclavian
Brian Lucey; Jose C. Varghese; Philip Haslam; Michael J. Lee
Analysis of the D? spectral line shape on the carbon limiter insertion experiments in Heliotron J
In order to investigate the velocity distribution function of deuterium atoms, which gives information about the recycling process, in the Heliotron J edge plasma region, spectral line shapes of D? emitted in front of a carbon limiter have been measured with a high-resolution spectrometer. Measured spectral profiles have an asymmetric structure and are decomposed into the broad and narrow Gaussian
H. Kawazome; K. Takahashi; S. Tsuboi; H. Arimoto; T. Mizuuchi; N. Nishino; H. Okada; K. Nagasaki; S. Kobayashi; S. Yamamoto; Y. Suzuki; K. Ohashi; S. Nakazawa; M. Kaneko; H. Shidara; Y. Fukagawa; S. Nishio; M. Yamada; H. Yamazaki; K. Kondo; F. Sano
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.
Gibaud, Anne; Vogt, Nicolas; Brison, Olivier; Debatisse, Michelle; Malfoy, Bernard
Automatic computation of wing-fuselage intersection lines and fillet inserts with fixed-area constraint
NASA Technical Reports Server (NTRS)
Procedures for automatic computation of wing-fuselage juncture geometry are described. These procedures begin with a geometry in wave-drag format. First, an intersection line is computed by extrapolating the wing to the fuselage. Then two types of filleting procedures are described, both of which utilize a combination of analytical and numerical techniques appropriate for automatic calculation. An analytical technique for estimating the added volume due to the fillet is derived, and an iterative procedure for revising the fuselage to compensate for this additional volume is given. Sample results are included in graphical form.
Barger, Raymond L.; Adams, Mary S.
Controlled insertional mutagenesis using a LINE-1 (ORFeus) gene-trap mouse model
A codon-optimized mouse LINE-1 element, ORFeus, exhibits dramatically higher retrotransposition frequencies compared with its native long interspersed element 1 counterpart. To establish a retrotransposon-mediated mouse model with regulatable and potent mutagenic capabilities, we generated a tetracycline (tet)-regulated ORFeus element harboring a gene-trap cassette. Here, we show that mice expressing tet-ORFeus broadly exhibit robust retrotransposition in somatic tissues when treated with doxycycline. Consistent with a significant mutagenic burden, we observed a reduced number of double transgenic animals when treated with high-level doxycycline during embryogenesis. Transgene induction in skin resulted in a white spotting phenotype due to somatic ORFeus-mediated mutations that likely disrupt melanocyte development. The data suggest a high level of transposition in melanocyte precursors and consequent mutation of genes important for melanoblast proliferation, differentiation, or migration. These findings reveal the utility of a retrotransposon-based mutagenesis system as an alternative to existing DNA transposon systems. Moreover, breeding these mice to different tet-transactivator/reversible tet-transactivator lines supports broad functionality of tet-ORFeus because of the potential for dose-dependent, tissue-specific, and temporal-specific mutagenesis.
O'Donnell, Kathryn A.; An, Wenfeng; Schrum, Christina T.; Wheelan, Sarah J.; Boeke, Jef D.
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.
Analysis of T-DNA- Xa21 loci and bacterial blight resistance effects of the transgene Xa21 in transgenic rice
The genetic loci and phenotypic effects of the transgene Xa21, a bacterial blight (BB) resistance gene cloned from rice, were investigated in transgenic rice produced through an Agrobacterium-mediated transformation system. The flanking sequences of integrated T-DNAs were isolated from Xa21 transgenic rice lines using thermal asymmetric interlaced PCR. Based on the analysis of 24 T-DNA- Xa21 flanking sequences, T-DNA loci
Wenxue Zhai; Caiyan Chen; Xuefeng Zhu; Xuewei Chen; Dechun Zhang; Xiaobing Li; Lihuang Zhu
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-MartÝnez, Ramfis; Hamdy, Osama; Marante, Daniel; Marulanda, MarÝa InÚs; Marchetti, Albert; Hegazi, Refaat A; Mechanick, Jeffrey I
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.
Nieto-Martinez, Ramfis; Hamdy, Osama; Marante, Daniel; Ines Marulanda, Maria; Marchetti, Albert; Hegazi, Refaat A.; Mechanick, Jeffrey I.
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
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
Analysis of crosstalk of coupled transmission lines by inserting additional traces grounded with vias on printed circuit boards
In analog and digital electronic systems, crosstalk between traces on the printed circuit boards (PCB) can degrade the performance of equipment operations. To reduce crosstalk between two parallel traces of which one is called aggressor trace and the other one is called victim trace, two additional traces grounded by vias are inserted and the aggressor trace is in between the
Shulan Li; Yuan'an Liu; Zhanhai Song; Hefei Hu
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
C Garion; A Poncet; F Seyvet; M Sitko; B Skoczen; J P Tock
US Patent & Trademark Office Database
A shoe insert for adjusting the fit of a shoe on a person's foot. The insert is formed from resilient material and is secured within the heel area of a shoe. The insert is elongated and has a narrow central portion for engaging a person's heel between the heel bone and the achilles tendon, and a pair of enlarged lateral portions for engaging the sides of the heel. The insert also has a tapered thickness which decreases from a thick upper part toward a thin lower part of the insert.
The angiotensin converting enzyme insertion/deletion polymorphism alters the response of muscle energy supply lines to exercise.
The presence of a silencing sequence (the I-allele) in the gene for the upstream regulator of blood flow, angiotensin I-converting enzyme (ACE), is associated with superior endurance performance and its trainability. We tested in a retrospective study with 36 Caucasian men of Swiss descent whether carriers of the ACE I-allele demonstrate a modified adaptive response of energy supply lines in knee extensor muscle, and aerobic fitness, to endurance training based on 6 weeks of supervised bicycle exercise or 6 months of self-regulated running (p value Vaughan, David; Huber-Abel, Felicitas A; Graber, Franziska; Hoppeler, Hans; FlŘck, Martin 2013-07-01
Vaughan, David; Huber-Abel, Felicitas A; Graber, Franziska; Hoppeler, Hans; FlŘck, Martin
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.
An intronic LINE-1 element insertion in the dystrophin gene aborts dystrophin expression and results in Duchenne-like muscular dystrophy in the corgi breed.
Duchenne muscular dystrophy (DMD) is a dystrophin-deficient lethal muscle disease. To date, the catastrophic muscle wasting phenotype has only been seen in dystrophin-deficient humans and dogs. Although Duchenne-like symptoms have been observed in more than a dozen dog breeds, the mutation is often not known and research colonies are rarely established. Here, we report an independent canine DMD model originally derived from the Pembroke Welsh corgi breed. The affected dogs presented clinical signs of muscular dystrophy. Immunostaining revealed the absence of dystrophin and upregulation of utrophin. Histopathologic examination showed variable fiber size, central nucleation, calcification, fibrosis, neutrophil and macrophage infiltration and cardiac focal vacuolar degeneration. Carrier dogs also displayed mild myopathy. The mutation was identified as a long interspersed repetitive element-1 (LINE-1) insertion in intron 13, which introduced a new exon containing an in-frame stop codon. Similar mutations have been seen in human patients. A colony was generated by crossing carrier females with normal males. Affected puppies had a normal birth weight but they experienced a striking growth delay in the first 5 days. In summary, the new corgi DMD model offers an excellent opportunity to study DMD pathogenesis and to develop novel therapies. PMID:20714321
Smith, Bruce F; Yue, Yongping; Woods, Philip R; Kornegay, Joe N; Shin, Jin-Hong; Williams, Regina R; Duan, Dongsheng
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.
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
Insertion element IS1296 in Mycoplasma mycoides subsp. mycoides small colony identifies a European clonal line distinct from African and Australian strains.
Strains of Mycoplasma mycoides subsp. mycoides small colony (SC) type, the agent of contagious bovine pleuropneumonia (CBPP), were analysed with respect to the polymorphism of distribution of a newly discovered insertion element, IS1296, on the chromosome. Analysis of 64 strains isolated from Europe, Africa and Australia, including four vaccine strains and the type strain PG1, revealed ten different IS patterns, forming two main clusters. The European strains originated from outbreaks of CBPP in different countries, and from various other sources such as semen and preputial washings from cattle, lungs from goats and buffalo, and milk from sheep. They showed identical IS1296 patterns, except one strain which had an additional IS1296 element, but the pattern belonged to the same cluster. This shows that the strains from Europe form a clonal lineage. The strains originating from different geographical parts of the African continent and from Australia showed four closely related IS1296 patterns which belong to a separate cluster. This indicates that strains from Africa and Australia form a clonal lineage different from that of the European strains, suggesting that the sporadic cases of CBPP that have re-emerged in Europe almost 15 years after the last declared endemic case in 1967 arose from an established reservoir within Europe rather than being the result of repeated importation from Africa and Australia. While most strains from Africa and Australia had the same IS1296 pattern, all vaccine strains could be distinguished by an individual pattern. The type strain PG1 also had a particular IS1296 pattern which belongs to the cluster of the strains from Africa and Australia. The molecular definition of clonality of M. mycoides subsp. mycoides SC strains with IS1296 represents a rapid and reproducible method for subtyping and differentiation of vaccine strains. It permits at the present time the definition of two main clonal lines, one including the strains from the European continent and a second with strains from Africa and Australia. PMID:8574413
Cheng, X; Nicolet, J; Poumarat, F; Regalla, J; Thiaucourt, F; Frey, J
In vivo formation of double-stranded T-DNA molecules by T-strand priming.
During plant genetic transformation, Agrobacterium transfers a single-stranded DNA (T-strand) into the host cell. Increasing evidence suggests that double-stranded (ds) T-DNA, converted from T-strands, are potent substrates for integration. Nevertheless, the molecular mechanism governing T-strand conversion to dsT-DNA is unknown. Integrated T-DNA molecules typically exhibit deletions at their 3' end as compared with their 5' end. We hypothesize that this may result from asymmetric polymerization of T-DNA's ends. Here we show that ?-glucuronidase (GUS) expression from sense T-strands is more efficient than from antisense T-strands, supporting asymmetric conversion. Co-transfection with two partially complementary, truncated GUS-encoding T-strands results in GUS expression, which suggests functional hybridization of the T-strands via complementary annealing and supports the notion that T-strands can anneal with primers. Indeed, red fluorescent protein (RFP) expression from mutated T-strand can be restored by delivery of synthetic DNA and RNA oligonucleotides with partial wild-type RFP sequence, implying the involvement of plant DNA repair machinery. PMID:23963047
Liang, Zhuobin; Tzfira, Tzvi
Random T-DNA mutagenesis identifies a Cu/Zn superoxide dismutase gene as a virulence factor of Sclerotinia sclerotiorum.
Agrobacterium-mediated transformation (AMT) was used to identify potential virulence factors in Sclerotinia sclerotiorum. Screening AMT transformants identified two mutants showing significantly reduced virulence. The mutants showed growth rate, sclerotial formation, and oxalate production similar to that of the wild type. The mutation was due to a single T-DNA insertion at 212 bp downstream of the Cu/Zn superoxide dismutase (SOD) gene (SsSOD1, SS1G_00699). Expression levels of SsSOD1 were significantly increased under oxidative stresses or during plant infection in the wild-type strain but could not be detected in the mutant. SsSOD1 functionally complemented the Cu/Zn SOD gene in a ?sod1 Saccharomyces cerevisiae mutant. The SOD mutant had increased sensitivity to heavy metal toxicity and oxidative stress in culture and reduced ability to detoxify superoxide in infected leaves. The mutant also had reduced expression levels of other known pathogenicity genes such as endo-polygalacturanases sspg1 and sspg3. The functions of SsSOD1 were further confirmed by SsSOD1-deletion mutation. Like the AMT insertion mutant, the SsSOD1-deletion mutant exhibited normal growth rate, sclerotial formation, oxalate production, increased sensitivity to metal and oxidative stress, and reduced virulence. These results suggest that SsSOD1, while not being required for saprophytic growth and completion of the life cycle, plays critical roles in detoxification of reactive oxygen species during host-pathogen interactions and is an important virulence factor of Sclerotinia sclerotiorum. PMID:23252459
Xu, Liangsheng; Chen, Weidong
Dihalocarbene Insertion Experiment
ERIC Educational Resources Information Center
Describes the insertion reaction using the insertion of carbenes into carbon-hydrogen bonds as an example. Outlines an experiment that will illustrate dihalocarbene insertions into diisopropyl ether. (GS)
Goh, S. H.
National Technical Information Service (NTIS)
This patent application discusses removable inserts of Raney nickel catalysts adapted for insertion in a tube wall reactor. The inserts are massive to provide a heat sink, to prevent overheating, to provide physical strength and to present sufficient mate...
S. A. O'Hare J. E. Mauser C. E. Armentrout
Chest drainage tube insertion; Insertion of tube into chest; Tube thoracostomy ... When your chest tube is inserted, you will lie on your side or sit partly upright, with one arm over your head. The ...
The Agrobacterium tumefaciens virulence D2 protein is responsible for precise integration of T-DNA into the plant genome.
The VirD2 protein of Agrobacterium tumefaciens was shown to pilot T-DNA during its transfer to the plant cell nucleus. We analyze here its participation in the integration of T-DNA by using a virD2 mutant. This mutation reduces the efficiency of T-DNA transfer, but the efficiency of integration of T-DNA per se is unaffected. Southern and sequence analyses of integration events obtained with the mutated VirD2 protein revealed an aberrant pattern of integration. These results indicate that the wild-type VirD2 protein participates in ligation of the 5'-end of the T-strand to plant DNA and that this ligation step is not rate limiting for T-DNA integration. Images
Tinland, B; Schoumacher, F; Gloeckler, V; Bravo-Angel, A M; Hohn, B
Genetic analysis of the virD operon of Agrobacterium tumefaciens: a search for functions involved in transport of T-DNA into the plant cell nucleus and in T-DNA integration.
The transferred DNA (T-DNA) is transported from Agrobacterium tumefaciens to the nucleus and is stably integrated into the genome of many plant species. It has been proposed that the VirD2 protein, tightly attached to the T-DNA, pilots the T-DNA into the plant cell nucleus and that it is involved in integration. Using agroinfection and beta-glucuronidase expression as two different very sensitive transient assays for T-DNA transfer, together with assays for stable integration, we have shown that the C-terminal half of the VirD2 protein and the VirD3 protein are not involved in T-DNA integration. However, the bipartite nuclear localization signal, which is located within the C terminus of the VirD2 protein and which has previously been shown to be able to target a foreign protein into the plant cell nucleus, was shown to be required for efficient T-DNA transfer. virD4 mutants were shown by agroinfection to be completely inactive in T-DNA transfer. Images
Koukolikova-Nicola, Z; Raineri, D; Stephens, K; Ramos, C; Tinland, B; Nester, E W; Hohn, B
T-DNA analysis of plants regenerated from hairy root tumors
Plants regenerated from hairy root tumors induced on Nicotiana glauca and Nicotiana tabacum by Agrobacterium rhizogenes strain A4 were examined for the presence of T-DNA. Regenerated N. tabacum plants contained intact copies of both TL-DNA and TR-DNA. However, plants regenerated from N. glauca tumors did not contain the TR-DNA region corresponding to the tms (auxin synthesis) genes. Some of the
Brian H. Taylor; Richard M. Amasino; Frank F. White; Eugene W. Nester; Milton P. Gordon
Laparoscopic-assisted uterovaginal anastomosis for uterine cervix atresia with vaginal aplasia using a silicone stent lined with acellular porcine small intestinal submucosa graft inserted using a 16F Foley catheter.
Herein is reported a novel technique for cervical reconstruction of congenital cervicovaginal atresia. The patient was a 16-year-old girl with congenital atresia of the cervix and vagina, didelphic uterus, and right hematosalpinx. At laparoscopic-assisted creation of a neocervix, a silicone stent was inserted using a 16F Foley catheter and lined with an acellular porcine small intestinal submucosa graft under ultrasound guidance. At 3-month clinical follow-up after placement of the stent, the patient had regular menstrual flow. The neocervix was completely mucosalized on the inner surface at 4 months after surgery. There were no complications related to the silicone stent or the cervical stent. Cervical reconstruction using a vaginal mucosa-lined silicone stent is accessible and effective, and provides an alternative option to preserve reproductive potential in patients with cervicovaginal atresia. PMID:23680519
Zhou, Qiongjie; Chen, Xiaojun; Luo, Xuezhen; Ding, Jinxin; Zhang, Guofu; Ren, Yunyun; Liu, Zhi; Hua, Keqin
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.
Samsonov, Andrey; Zenser, Nathan; Zhang, Fan; Zhang, Hongyi; Fetter, John; Malkov, Dmitry
Transgene-specific and event-specific molecular markers for characterization of transgenic papaya lines resistant to Papaya ringspot virus.
The commercially valuable transgenic papaya lines carrying the coat protein (CP) gene of Papaya ringspot virus (PRSV) and conferring virus resistance have been developed in Hawaii and Taiwan in the past decade. Prompt and sensitive protocols for transgene-specific and event-specific detections are essential for traceability of these lines to fulfill regulatory requirement in EU and some Asian countries. Here, based on polymerase chain reaction (PCR) approaches, we demonstrated different detection protocols for characterization of PRSV CP-transgenic papaya lines. Transgene-specific products were amplified using different specific primer pairs targeting the sequences of the promoter, the terminator, the selection marker, and the transgene, and the region across the promoter and transgene. Moreover, after cloning and sequencing the DNA fragments amplified by adaptor ligation-PCR, the junctions between plant genomic DNA and the T-DNA insert were elucidated. The event-specific method targeting the flanking sequences and the transgene was developed for identification of a specific transgenic line. The PCR patterns using primers designed from the left or the right flanking DNA sequence of the transgene insert in three selected transgenic papaya lines were specific and reproducible. Our results also verified that PRSV CP transgene is integrated into transgenic papaya genome in different loci. The copy number of inserted T-DNA was further confirmed by real-time PCR. The event-specific molecular markers developed in this investigation are crucial for regulatory requirement in some countries and intellectual protection. Also, these markers are helpful for prompt screening of a homozygote-transgenic progeny in the breeding program. PMID:19526355
Fan, Ming-Jen; Chen, Shu; Kung, Yi-Jung; Cheng, Ying-Huey; Bau, Huey-Jiunn; Su, Tien-Tsai; Yeh, Shyi-Dong
High Efficiency Transgene Segregation in Co-Transformed Maize Plants using an Agrobacterium Tumefaciens 2 T-DNA Binary System
For regulatory issues and research purposes it would be desirable to have the ability to segregate transgenes in co-transformed maize. We have developed a highly efficient system to segregate transgenes in maize that was co-transformed using an Agrobacterium tumefaciens 2 T-DNA binary system. Three vector treatments were compared in this study; (1) a 2 T-DNA vector, where the selectable marker
Michael Miller; Laura Tagliani; Ning Wang; Benjamin Berka; Dennis Bidney; Zuo-Yu Zhao
T-DNA tagged knockout mutation of rice OsGSK1 , an orthologue of Arabidopsis BIN2 , with enhanced tolerance to various abiotic stresses
T-DNA-tagged rice plants were screened under cold- or salt-stress conditions to determine the genes involved in the molecular\\u000a mechanism for their abiotic-stress response. Line 0-165-65 was identified as a salt-responsive line. The gene responsible\\u000a for this GUS-positive phenotype was revealed by inverse PCR as OsGSK1 (O\\u000a ryza \\u000a s\\u000a ativa\\u000a g\\u000a lycogen \\u000a s\\u000a ynthase \\u000a k\\u000a inase3-like gene \\u000a 1), a member of
Serry Koh; Sang-Choon Lee; Min-Kyung Kim; Jun Ho Koh; Sichul Lee; Gynheung An; Sunghwa Choe; Seong-Ryong Kim
Peripherally inserted central venous catheters: frequency of complications in premature newborn depends on the insertion site.
Objective:The use of peripherally inserted central venous catheters (PICC lines) has reduced the mortality and morbidity of premature newborns. The usual sites of insertion are the veins in the upper arms but other locations are being used as well.Study Design:To examine whether using the axillary vein as a site of insertion of a PICC line affects the frequency of complications. Our study has a clinical trial design. A total of 62 neonates that had a PICC line inserted were recruited and randomly divided equally in two groups: in Group A (mean birth weight, standard deviation (s.d.)=1353 (142)?g), the PICC line was inserted through the axillary vein, and in Group B (birth weight=1308 (112)?g), the PICC line was inserted in other sites further from the axillary vein. The outcomes measured were the total PICC line-related complications, the reason for removing the catheter, the number of total attempts until successful insertion and the mean duration of stay of the catheter. The likelihood of having an adverse outcome was assessed with Mantel-Haenszel odds ratio (OR).Results:Premature neonates with axillary PICC lines were 12 times less likely to have line-related complications (inflammation, blockage, edema, infection) as compared with any other site of insertion (OR= 95%, confidence interval (CI)=0.10 (0.01 to 0.8)) and they were seven times more likely to have the PICC line removed because they achieved full enteral nutrition as compared with the other causes (OR 95%, confidence interval (CI)=10.35 (4.88 to 21.96)). There was no statistical difference between the two groups in the number of attempts until successful PICC line insertion (P=0.667) and the mean duration of stay of the PICC line (P=0.97).Conclusion:The use of the axillary vein as a site of insertion of a PICC line was correlated with significantly less complications in premature newborns as opposed to the other sites of insertion. PMID:24625519
Panagiotounakou, P; Antonogeorgos, G; Gounari, E; Papadakis, S; Labadaridis, J; Gounaris, A K
Central Vascular Catheter Insertion in the NICU: A Lean Approach.
Purpose: To apply Lean principles to the process of inserting central lines into neonates in the NICU.Design: The authors used standard interviews and live observations to obtain concrete data on the current process of central line insertions. Recommendations for improvement were then suggested based on Lean principles.Sample: NICU care providers.Main Outcome Variable: Non-value-added time (waste), provider confidence, and variation in placing central lines.Results: There was large variation in how providers inserted central lines, and providers were least confident with placing peripherally inserted central catheter lines (55 percent confident). Live observations showed that 53 percent of the current process consisted of waste, with the line insertion and radiography phases of the process as the most wasteful. Lean principles can be applied to a neonatal clinical setting and can be an effective methodology for NICU care providers to improve the way that we care for our patients. PMID:24985110
Liu, Xinran; Suresh, Gautham
[Sequence analysis of Bt insertion flanking fragments in transgenic Bt cotton].
The copies of outside gene and DNA structure of integration locus are important in high expression and avoidance of gene silence. Many research results showed that outside genes were inserted into the plant genome with recombinant type, and the integration was related to border T-DNA sequence in the course of Agrobaterium mediated transformation. SAR structure was also found in the integration location of transformants with direct DNA transformation method. The pollen-tube pathway transformation, one of direct DNA transformation method, was very successfully used in Bt transgenic cotton in China. But until now there has been no report about Bt gene integration. The aim of this research was to investigate flanking DNA structure in order to explain the mechanism of direct transformation method in the future. The structure of flanking DNA fragments of Bt integration in four different transgenic Bt cotton varieties including Simian-3, 161 resistant breeding line, 161 sensitive breeding line and Guokangmian-1 was analyzed with TAIL-PCR and nested PCR. The flanking DNA sequences of different self-crossed progenies from one plant are the same. In contrary, their DNA sequences are diverse for different breeding lines. Upstream flanking fragments in some transgenic cotton contained short transformation plasmid sequences, downstream flanking fragments in all the transgenic cotton varieties were composed of high percentage AT. The percentage of AT in Simian-3 transgenic Bt variety was as high as 92%. All the flanking fragments were of multi-copies and similar to repetitive sequences. No recognition sites of TOP enzyme were found in these fragments. PMID:12200866
Zuo, Kai-Jing; Zhang, Xiao-Long; Nie, Yi-Chun; Liu, Jin-Lan; Sun, Ji-Zhong
Disassembly of synthetic Agrobacterium T-DNA-protein complexes via the host SCFVBF ubiquitin-ligase complex pathway
One the most intriguing, yet least studied, aspects of the bacteriumľhost plant interaction is the role of the host ubiquitin/proteasome system (UPS) in the infection process. Increasing evidence indicates that pathogenic bacteria subvert the host UPS to facilitate infection. Although both mammalian and plant bacterial pathogens are known to use the host UPS, the first prokaryotic F-box protein, an essential component of UPS, was identified in Agrobacterium. During its infection, which culminates in genetic modification of the host cell, Agrobacterium transfers its T-DNAŚas a complex (T-complex) with the bacterial VirE2 and host VIP1 proteinsŚinto the host cell nucleus. There the T-DNA is uncoated from its protein components before undergoing integration into the host genome. It has been suggested that the host UPS mediates this uncoating process, but there is no evidence indicating that this activity can unmask the T-DNA molecule. Here we provide support for the idea that the plant UPS uncoats synthetic T-complexes via the Skp1/Cullin/F-box protein VBF pathway and exposes the T-DNA molecule to external enzymatic activity.
Zaltsman, Adi; Lacroix, Benoit; Gafni, Yedidya; Citovsky, Vitaly
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
Direct trocar insertion versus veress needle insertion in laparoscopic cholecystectomy
BACKGROUND: Direct insertion of the trocar is an alternative method to Veress needle insertion for the creation of pneumoperitoneum. The safety of direct disposable shielded trocar insertion for the creation of pneumoperitoneum was assessed by comparing with Veress needle insertion during laparoscopic cholecystectomy (LC).METHODS:One thousand five hundred patients undergoing LC with pneumoperitoneum were included in this study. In 470 patients
Mehmet Ali Yerdel; Kaan Karayalcin; Ayhan Koyuncu; Baris Akin; Cuneyt Koksoy; Ahmet G Turkcapar; Nezih Erverdi; Iskender Alašayir; Cihan Bumin; Nusret Aras
Extensive changes in DNA methylation patterns accompany activation of a silent T-DNA ipt gene in Agrobacterium tumefaciens-transformed plant cells.
We crossed a male-sterile, Agrobacterium-transformed Nicotiana tabacum plant that contains a silent, hypermethylated T-DNA ipt oncogene with a normal tobacco plant and found that the methylated state of the ipt gene was stably inherited through meiosis in the offspring. However, when tissues of these plants were placed in cell culture, the ipt gene was spontaneously reactivated in a very small fraction of the cells; if 5-azacytidine was added to the culture medium, ipt gene reactivation occurred at high frequency. We analyzed the pattern of DNA methylation in a region spanning the ipt gene in a line that does not express the ipt gene, in five derivatives of this line that reexpressed the ipt gene either spontaneously or after 5-azacytidine treatment, and in tissues of a sibling of this line that reexpressed ipt spontaneously. We found that the ipt locus was highly methylated in the unexpressed state but that spontaneous or 5-azacytidine-induced reexpression always resulted in extensive demethylation of a region including 5' upstream, coding, and 3' downstream regions of the ipt gene. The role of DNA methylation in gene regulation in this system is discussed. Images
John, M C; Amasino, R M
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.
Double-stranded cleavage of T-DNA and generation of single-stranded T-DNA molecules in Escherichia coli by a virD-encoded border-specific endonuclease from Agrobacterium tumefaciens.
The virD locus of Agrobacterium tumefaciens Ti plasmid pTiA6 was sequenced. Computer analysis of the sequence indicated five possible open reading frames (ORFs) within this locus. Two additional ORFs were identified distal to this locus. However, only two polypeptides of apparent molecular masses 16 and 56 kilodaltons, the products of ORFs 1 and 2, were detected in Escherichia coli, both in vivo and in an in vitro coupled transcription-translation system. The virD locus was cloned in expression vector pKK223.3 under control of a tac promoter and introduced into an E. coli strain harboring mini-Ti plasmid pAL1050. When induced with isopropyl-beta-D-thiogalactopyranoside, the virD gene products exhibited double-stranded T-DNA border-specific endonuclease activity. Deletion analysis demonstrated that this activity is encoded within the 5'-proximal 1.7-kilobase-pair portion of the virD locus that carries ORF 1 and most of ORF 2. Neither ORF 1 nor ORF 2 independently showed endonuclease activity; complementation studies indicated that the products of ORFs 1 and 2 together have this activity. The expression of this 1.7-kilobase-pair region of the virD locus caused double-stranded cleavage of the T-DNA at or near the borders and generated single-stranded T-DNA molecules with approximately equal frequencies in E. coli. Images
Jayaswal, R K; Veluthambi, K; Gelvin, S B; Slightom, J L
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
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
Trudy F. C. Mackay; Richard F. Lyman; Michael S. Jackson
T-DNA trapping of a cryptic promoter identifies an ortholog of highly conserved SNZ growth arrest response genes in
A T-DNA tagged Arabidopsis locus, A37, identified by a promoter-trap aph(3%)II reporter gene fusion expressed in calli and roots, encodes an ortholog of evolutionarily conserved SNZ growth arrest response proteins. Gene A37 is located on chromosome 3-35, lacks introns, and shares considerable sequence identity with HEVER1 from rubber tree, SLEXORFA-1 from Stellaria longipes, SNZ1 from yeast, and SNZ-homologs from bacteria
Csaba Mathea; Eva Horvath; Jeff Schell; Csaba Koncz
Nucleotide sequences of the Pseudomonas savastanoi indoleacetic acid genes show homology with Agrobacterium tumefaciens T-DNA
We report the nucleotide sequences of iaaM and iaaH, the genetic determinants for, respectively, tryptophan 2-monooxygenase and indoleacetamide hydrolase, the enzymes that catalyze the conversion of L-tryptophan to indoleacetic acid in the tumor-forming bacterium Pseudomonas syringae pv. savastanoi. The sequence analysis indicates that the iaaM locus contains an open reading frame encoding 557 amino acids that would comprise a protein with a molecular weight of 61,783; the iaaH locus contains an open reading frame of 455 amino acids that would comprise a protein with a molecular weight of 48,515. Significant amino acid sequence homology was found between the predicted sequence of the tryptophan monooxygenase of P. savastanoi and the deduced product of the T-DNA tms-1 gene of the octopine-type plasmid pTiA6NC from Agrobacterium tumefaciens. Strong homology was found in the 25 amino acid sequence in the putative FAD-binding region of tryptophan monooxygenase. Homology was also found in the amino acid sequences representing the central regions of the putative products of iaaH and tms-2 T-DNA. The results suggest a strong similarity in the pathways for indoleacetic acid synthesis encoded by genes in P. savastanoi and in A. tumefaciens T-DNA. Images
Yamada, Tetsuji; Palm, Curtis J.; Brooks, Bob; Kosuge, Tsune
A radiometric method for measuring the insertion loss of radome materials
NASA Technical Reports Server (NTRS)
Radiometer system measures effective noise temperature directed towards sky, with and without radome over antenna horn. Data is then translated into computer format. With additional transmission line insertion loss data from other measurements, computer calculates insertion loss of radome material.
Seidel, B. L.; Stelzried, C. T.
Implanon«: Insertion and Removal
\\u000a The etonogestrel implant, Implanon« is a sterile 4-cm Î 2-mm single rod that is implanted subdermally and can provide up to\\u000a 3 years of continuous effective contraception. Each rod contains 68 mg of the progestin etonogestrel encased in an ethylene\\u000a vinylacetate copolymer skin [1]. It is supplied in a sterile, preloaded disposable applicator. The applicator greatly facilitates\\u000a proper insertion in
Jeffrey P. Levine
This patent describes an improved floating spacer grid in combination with a nuclear fuel assembly of the type having parallel fuel rods and at least one guide tube maintained in a laterally spaced array by axially spaced spacer grids. The fuel rods and guide tube extend through the spacer grids. The guide tube has a uniform circular outer surface, the spacer grids are movably mounted to the guide tube to permit axial displacement of the spacer grids relative the guide tube. Saddles form from and within the spacer grids at top and bottom edges of the grids, a spacer sleeve positioned concentrically about the guide tube intermediate the adjacent ones of the axially-spaced spacer grids for limiting axial displacement of the spacer grids, and the spacer sleeve has opposite ends closely spaced from the edges of the adjacent spacer grids to define a displacement space. The improvement comprises in combination; tubular inserts, each insert mounted to the guide tube within each of the spacer grids. Each insert includes a tubular portion positioned by the saddles to circumscribe the guide tube within the spacer grid and a circular flange portion at one end of the tubular portion. The flange portion is positioned in the displacement space and abutted to the edge of the spacer grid on the saddles, intermediate the spacer grid and spacer sleeve, and the circular flange portion has substantially the same outer diameter as the spacer sleeve thus precluding direct contact of the spacer sleeve with the spacer grid. The inserts carry all axial through-grid loads and prevent a horizontal vector force component applied against the grids.
King, R.A.; Larson, J.G.; Walton, L.A.
Cryptococcus neoformans Virulence Gene Discovery through Insertional Mutagenesis
Insertional mutagenesis was applied to Cryptococcus neoformans to identify genes associated with virulence attributes. Using biolistic transformation, we generated 4,300 nourseothricin (NAT)-resistant strains, of which 590 exhibited stable resistance. We focused on mutants with defects in established virulence factors and identified two with reduced growth at 37░C, four with reduced production of the antioxidant pigment melanin, and two with an increased sensitivity to nitric oxide (NO). The NAT insertion and mutant phenotypes were genetically linked in five of eight mutants, and the DNA flanking the insertions was characterized. For the strains with altered growth at 37░C and altered melanin production, mutations were in previously uncharacterized genes, while the two NO-sensitive strains bore insertions in the flavohemoglobin gene FHB1, whose product counters NO stress. Because of the frequent instability of nourseothricin resistance associated with biolistic transformation, Agrobacterium-mediated transformation was tested. This transkingdom DNA delivery approach produced 100% stable nourseothricin-resistant transformants, and three melanin-defective strains were identified from 576 transformants, of which 2 were linked to NAT in segregation analysis. One of these mutants contained a T-DNA insertion in the promoter of the LAC1 (laccase) gene, which encodes a key enzyme required for melanin production, while the second contained an insertion in the promoter of the CLC1 gene, encoding a voltage-gated chloride channel. Clc1 and its homologs are required for ion homeostasis, and in their absence Cu+ transport into the secretory pathway is compromised, depriving laccase and other Cu+-dependent proteins of their essential cofactor. The NAT resistance cassette was optimized for cryptococcal codon usage and GC content and was then used to disrupt a mitogen-activated protein kinase gene, a predicted gene, and two putative chloride channel genes to analyze their contributions to fungal physiology. Our findings demonstrate that both insertional mutagenesis methods can be applied to gene identification, but Agrobacterium-mediated transformation is more efficient and generates exclusively stable insertion mutations.
Idnurm, Alexander; Reedy, Jennifer L.; Nussbaum, Jesse C.; Heitman, Joseph
Ear tube insertion - series (image)
Ear tube insertion is recommended for: fluid in the middle ear (present for more than 12 weeks) recurrent ear ... the accumulated fluid is suctioned out. A small tube is inserted through the incised eardrum in order ...
Agrobacterium rhizogenes pRi8196 T-DNA: Mapping and DNA Sequence of Functions Involved in Mannopine Synthesis and Hairy Root Differentiation
This paper presents the map and DNA sequence analysis of pRi8196 transferred DNA (T-DNA) genes encoding root-inducing and mannopine synthesis functions. A canonical 24-base-pair border repeat as well as two \\
Genevieve Hansen; Mireille Larribe; Daniele Vaubert; Jacques Tempe; Brenda J. Biermann; Alice L. Montoya; Mary-Dell Chilton; Jean Brevet
Generation of selectable marker-free sheath blight resistant transgenic rice plants by efficient co-transformation of a cointegrate vector T-DNA and a binary vector T-DNA in one Agrobacterium tumefaciens strain
Co-transformation of Oryza sativa L. var. Pusa Basmati1 was done using an Agrobacterium\\u000a tumefaciens strain harbouring a single-copy cointegrate vector and a multi-copy binary vector in the same cell. The T-DNA of the cointegrate\\u000a vector pGV2260::pSSJ1 carried the hygromycin phosphotransferase (hph) and ?-glucuronidase (gus) genes. The binary vector pCam-chi11, without a plant selectable marker gene, harboured the rice chitinase (chi11)
Rajasekaran Sripriya; Vengoji Raghupathy; Karuppannan Veluthambi
Virulence genes, borders, and overdrive generate single-stranded T-DNA molecules from the A6 Ti plasmid of Agrobacterium tumefaciens.
Agrobacterium tumefaciens transfers the T-DNA portion of its Ti plasmid to the nuclear genome of plant cells. Upon cocultivation of A. tumefaciens A348 with regenerating tobacco leaf protoplasts, six distinct single-stranded T-DNA molecules (T strands) were generated in addition to double-stranded T-DNA border cleavages which we have previously reported (K. Veluthambi, R.K. Jayaswal, and S.B. Gelvin, Proc. Natl. Acad. Sci. USA 84:1881-1885, 1987). The T region of an octopine-type Ti plasmid has four border repeats delimiting three T-DNA regions defined as T left (TL), T center (TC), and T right (TR). The six T strands generated upon induction corresponded to the TL, TC, TR, TL + TC, TC + TR, and TL + TC + TR regions, suggesting that the initiation and termination of T-strand synthesis can occur at each of the four borders. Most TL + TC + TR T-strand molecules corresponded to the top T-DNA strand, whereas the other five T strands corresponded to the bottom T-DNA strand. Generation of T strands required the virA, virG, and virD operons. Extra copies of vir genes, harbored on cosmids within derivatives of A. tumefaciens A348, enhanced production of T strands. The presence of right and left border repeats in their native orientation is important for the generation of full-length T strands. When a right border repeat was placed in the opposite orientation, single-stranded T-DNA molecules that corresponded to the top strand were generated. Deletion of overdrive, a sequence that flanks right border repeats and functions as a T-DNA transmission enhancer, reduced the level of T-strand generation. Induction of A. tumefaciens cells by regenerating tobacco protoplasts increased the copy number of the Ti plasmid relative to the bacterial chromosome. Images
Veluthambi, K; Ream, W; Gelvin, S B
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.
Michielse, Caroline B; van Wijk, Ringo; Reijnen, Linda; Cornelissen, Ben JC; Rep, Martijn
Analysis of characters coded for by T-DNA using hybridization between tumorous and normal plant cells.
In somatic hybrids between tumourous Nicotiana tabacum (B6S3) and normal mesophyll Atropa belladonna cells, the following traits, directly or indirectly connected with T-DNA gene expression and tumourous growth, were analysed: lysopine dehydrogenase activity (LpDH), shoot suppression, root suppression, ability to grow on media with D-lactose as a sole carbon source and resistance to 2-aminoethylcysteine, 5-bromodeoxyuridine and 5-methyltryptophan. Dominant (semidominant) expression was observed for all but one trait studied, e.g. shoot suppression which behaved as a recessive character. PMID:24248297
Kanevsky, I F; Gleba, Y Y
Agrobacterium -mediated transformation system for large-scale producion of transgenic chinese cabbage ( Brassica rapa L. ssp. pekinensis ) plants for insertional mutagenesis
In order to better utilize insertional mutagenesis and functional genomics in Chinese cabbage, we have developed an improved\\u000a transformation system that more efficiently produces a large number of transgenic plants. Hypocotyl explants were inoculated\\u000a withAgrobacterium tumefaciens LBA4404. This strain harbors tagging vector pRCV2, which contains a hygromycin-resistance gene, an ampicillin resistance\\u000a gene, and a bacterial replication origin within the T-DNA.
Mi-Kyung Lee; Hyoung-Seok Kim; Jung-Sun Kim; Sung-Hoon Kim; Young-Doo Park
Experimental Attempts for Deep Insertion in Ultrasonically Forced Insertion Process
NASA Astrophysics Data System (ADS)
In this paper, we describe two attempts of obtaining deep insertion in an ultrasonically forced insertion (USFI) process. One was to correct the inclination of an inserted rod by passively generated bending vibrations. The inclination causes a partial plastic deformation, which decreases the holding power of processing materials. Two types of horn with grooves for excitation of bending vibrations were examined. The other was to make differences in vibration velocity and the phase of a rod and a metal plate by damping the vibration of a metal plate by using a rubber sheet. As results, the attempts proposed in this study were confirmed to be effective to obtain a deep insertion.
Ono, Satoshi; Aoyagi, Manabu; Tamura, Hideki; Takano, Takehiro
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
Needle insertion modeling and simulation
AbstractŚA methodology for estimating the force distribution that occurs along a needle shaft during insertion is described. An experimental system for measuring planar tissue phantom defor- mation during needle insertions has been developed and is pre- sented. A two-dimensional linear elastostatic material model, dis- cretised using the finite element method, is used to derive con- tact force information that is
Simon Peter DiMaio; Septimiu E. Salcudean
Insertion device vacuum system designs
Synchrotron light source insertion device vacuum systems now in operation and systems proposed for the future are reviewed. An overview of insertion devices is given and four generic vacuum chamber designs, transition section design and pumping considerations are discussed. Examples of vacuum chamber systems are presented.
Hoyer, E.
Agrobacterium proteins VirD2 and VirE2 mediate precise integration of synthetic T-DNA complexes in mammalian cells
Agrobacterium tumefaciens-mediated plant transformation, a unique example of interkingdom gene transfer, has been widely adopted for the generation of transgenic plants. In vitro synthesized transferred DNA (T-DNA) complexes comprising single-stranded DNA and Agrobacterium virulence proteins VirD2 and VirE2, essential for plant transformation, were used to stably transfect HeLa cells. Both proteins positively influenced efficiency and precision of transgene integration by increasing overall transformation rates and by promoting full-length single-copy integration events. These findings demonstrate that the virulence proteins are sufficient for the integration of a T-DNA into a eukaryotic genome in the absence of other bacterial or plant factors. Synthetic T-DNA complexes are therefore unique protein:DNA delivery vectors with potential applications in the field of mammalian transgenesis.
Pelczar, Pawel; Kalck, Veronique; Gomez, Divina; Hohn, Barbara
A selectively terminable transgenic rice line expressing human lactoferrin.
Human lactoferrin (hLF) is a multifunctional milk protein which could be utilized for promoting human health. Transgenic rice has been used as a bioreactor for mass production of recombinant hLF. However, one major concern over such transgenic rice is the risk of its unintended spreading into environment and into our food supplies. Here we report the development of selectively terminable transgenic rice expressing human lactoferrin in seeds. These transgenic rice plants could be selectively terminated by bentazon, a common herbicide used for rice weed control. The hLF expression cassette was constructed into a T-DNA containing the RNA interference cassette suppressing the expression of the rice gene CYP81A6 which detoxifies herbicide bentazon, and the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) cassette which confers to glyphosate tolerance. A transgenic line, named as G281, was identified for its high sensitivity to bentazon, high tolerance to glyphosate, and high expression of hLF. Southern analysis suggested G281 is a single copy insertion event. Field tests demonstrated that G281 plants can be completely killed by a single spray of bentazon at 1000 mg/L, which is safe to regular rice and represents only half of the dose recommended by manufacturer for rice field weed control. Therefore, any G281 contaminations in regular rice could be selectively terminated to make sure it will not enter food or feed supplies. PMID:20433928
Lin, Chaoyang; Nie, Peng; Lu, Wei; Zhang, Qing; Li, Jing; Shen, Zhicheng
Tool Removes Coil-Spring Thread Inserts
NASA Technical Reports Server (NTRS)
Tool removes coil-spring thread inserts from threaded holes. Threads into hole, pries insert loose, grips insert, then pulls insert to thread it out of hole. Effects essentially reverse of insertion process to ease removal and avoid further damage to threaded inner surface of hole.
Collins, Gerald J., Jr.; Swenson, Gary J.; Mcclellan, J. Scott
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).
Insertion Devices, Future Developments, Limitations.
National Technical Information Service (NTIS)
The first generation of insertion devices having reached maturity (or almost), it is appropriate to survey directions along which new developments are expected. These include: Beam cooling with undulators, elementary particle physics with wigglers and und...
P. L. Csonka
Gas turbine vane cooling air insert
This patent describes a gas turbine. It comprises turbine vanes, each of the vanes supplied with cooling air and having: an airfoil portion forming a first cavity having an insert disposed therein for directing the flow of the cooling air, the insert having first and second insert ends; a shroud portion from which the airfoil portion extends, the insert attached to the shroud portion at the first insert end; an insert extension extending through a portion of the insert and extending beyond the first insert end, the insert extension and the insert forming an annular gap therebetween separating the insert from the insert extension; a plate covering at least a portion of the shroud, the plate having a first hole formed therein through which the insert extension extends; and at least a first seal extending between the insert extension and the insert, and sealing the annular gap therebetween. This patent also describes a method of making a gas turbine. It comprises welding a first tubular insert adjacent its first end to a vane outer shroud; partially inserting a second tubular insert into the first tubular member and attaching the second tubular insert thereto; placing a plate having a hole formed therein on the outer shroud so that the hole surrounds the second tubular insert; and attaching the second tubular insert to the plate by placing a first seal between the first and second tubular inserts and attaching the first seal to each of the first and second tubular inserts, and placing a second seal between the second tubular insert and the plate and welding the second seal to the second tubular insert and the plate.
North, W.E.; Hultgren, K.G.; Dishman, C.D.; Van Heusden, G.S.
Molecular analysis of high-copy insertion sites in maize
High-copy transposon mutagenesis is an effective tool for creating gene disruptions in maize. In order to molecularly define transposon-induced disruptions on a genome-wide scale, we optimized TAIL-PCR to amplify genomic DNA flanking maize Robertsonĺs Mutator insertions. Sample sequencing from 43 Mutator stocks and the W22 inbred line identified 676 non-redundant insertions, and only a small fraction of the flanking sequences showed significant similarity to maize repetitive sequences. We further designed and tested 79 arbitrary primers to identify 12 primers that amplify all Mutator insertions within a DNA sample at 3.1-fold redundancy. Importantly, the products are of sufficient size to use as substrates or probes for hybridization-based identification of gene disruptions. Our adaptation simplifies previously published TAIL-PCR protocols and should be transferable to other high-copy insertional mutagens.
Settles, A. Mark; Latshaw, Susan; McCarty, Donald R.
Agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in medicinal fungus Cordyceps militaris.
Cordyceps militaris is an insect-born fungus with various biological and pharmacological activities. The mutant library of C. militaris was constructed by improved Agrobacterium tumefaciens-mediated transformation (ATMT), for the ultimate identification of genes involved in isolate degeneration during fruiting body production. Successful transformation of C.ámilitaris JM4 by A. tumefaciens AGL-1 carrying vector pATMT1 was performed, with efficiency in the range of 30-600 transformants per 1Î10(5) conidia. Acetosyringone (AS) supplement in C. militaris ATMT was not necessary during either precultivation or cocultivation. The transformation procedure was optimised based on the ratios between donor A. tumefaciens and recipient conidia, and pH value of cocultivation media. The integration of the hyg gene into C. militaris genome was determined by PCR and Southern blot analysis, suggesting that 67-88% resulting transformants in cultivation conditions with or without AS were inserted by T-DNA and 55-80% were single-copy. Special mutants with altered phenotypes and growth potentials were characterised. The efficient TAIL-PCR approach was established for identifying T-DNA flanking sequences from C. militaris mutants. The successful construction of the mutant library indicated the usefulness of this approach for functional genetic analysis in this important fungus. PMID:21354533
Zheng, Zhuangli; Huang, Chuanhua; Cao, Li; Xie, Cuihong; Han, Richou
Genetic and physiological characterization of the arlequin insertional mutant reveals a key regulator of reproductive development in tomato.
The genetic and phenotypic characterization of a new tomato (Solanum lycopersicum) insertional mutant, Arlequin (Alq) is reported. Alq mutant plants were affected in reproductive development and their sepals were homeotically converted into fleshy fruit-like organs. Molecular analysis demonstrated that a single copy of T-DNA was present in the mutant genome while genetic analysis confirmed that the mutant phenotype co-segregated with the T-DNA insertion and was inherited as a monogenic semi-dominant trait. The histological and scanning electron microscope analyses revealed cell identity changes in both external and internal tissues of Alq sepals. Flowers developed by Alq homozygous plants showed a severe mutant phenotype, since after fruit set, not only did the sepals become succulent but they also followed a ripening pattern similar to that of normal fruits. From a metabolic viewpoint, Alq sepals also behaved like a fruit, as they acquired the properties of a sink that acted alternatively and independently to the fruit. In fact, expression of regulatory genes controlling tomato fruit ripening was detected in Alq sepals at similar levels to those observed in mature fruits. Furthermore, the Alq mutation inhibited the development of the abscission zone in tomato flowers indicating that the JOINTLESS gene is regulated by ALQ. Results from the genetic and developmental characterization of the Alq tomato mutant suggest that the ALQ gene participates in the regulatory pathway controlling fruit ripening of tomato. PMID:20081209
Pineda, Benito; GimÚnez-Caminero, Estela; GarcÝa-Sogo, Bego˝a; Antˇn, MarÝa Teresa; AtarÚs, Alejandro; Capel, Juan; Lozano, Rafael; Angosto, Trinidad; Moreno, Vicente
A DNA insertional mutation results in microphthalmia in transgenic mice
Transgenic mice were produced by microinjection of a humanA?-globin gene construct containing site 2 of the locus control region and theA?-globin gene with its 3? enhancer sequence. One transgenic mouse line 95?HS2?en91) displayed an altered phenotype when the insertion event of this transgenic line was homozygous. These animals lack the normal pigmentation seen in their hemizygous and non-transgenic littermates, thus
Joan M. Krakowsky; Raymond E. Boissy; Jon C. Neumann; Jerry B. Lingrel
Summary of Experimental Insertions Workshop.
National Technical Information Service (NTIS)
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 ...
J. Sandweiss M. Month
Needle Insertion Modelling and Simulation
A methodology for estimating the force dis- tribution that occurs along a needle shaft during inser- tion is described. To validate the approach, an exper- imental system for measuring planar tissue deforma- tion during needle insertions has been developed and is presented. The planar motion of a soft tissue phan- tom is measured during needle penetration and used in conjunction
Simon P. Dimaio; S. E. Salcudean
T-DNA trapping of a cryptic promoter identifies an ortholog of highly conserved SNZ growth arrest response genes in Arabidopsis
A T-DNA tagged Arabidopsis locus, A37, identified by a promoter-trap aph(3?)II reporter gene fusion expressed in calli and roots, encodes an ortholog of evolutionarily conserved SNZ growth arrest response proteins. Gene A37 is located on chromosome 3ľ35, lacks introns, and shares considerable sequence identity with HEVER1 from rubber tree, SLEXORFA-1 from Stellaria longipes, SNZ1 from yeast, and SNZ-homologs from bacteria
Lßszlˇ ÍkrÚsz; Csaba MßthÚ; ╔va Horvßth; Jeff Schell; Csaba Koncz; Lßszlˇ Szabados
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
A coumaroyl-ester-3-hydroxylase Insertion Mutant Reveals the Existence of Nonredundant meta-Hydroxylation Pathways and Essential Roles for Phenolic Precursors in Cell Expansion and Plant Growth1[W][OA
Cytochromes P450 monooxygenases from the CYP98 family catalyze the meta-hydroxylation step in the phenylpropanoid biosynthetic pathway. The ref8 Arabidopsis (Arabidopsis thaliana) mutant, with a point mutation in the CYP98A3 gene, was previously described to show developmental defects, changes in lignin composition, and lack of soluble sinapoyl esters. We isolated a T-DNA insertion mutant in CYP98A3 and show that this mutation leads to a more drastic inhibition of plant development and inhibition of cell growth. Similar to the ref8 mutant, the insertion mutant has reduced lignin content, with stem lignin essentially made of p-hydroxyphenyl units and trace amounts of guaiacyl and syringyl units. However, its roots display an ectopic lignification and a substantial proportion of guaiacyl and syringyl units, suggesting the occurrence of an alternative CYP98A3-independent meta-hydroxylation mechanism active mainly in the roots. Relative to the control, mutant plantlets produce very low amounts of sinapoyl esters, but accumulate flavonol glycosides. Reduced cell growth seems correlated with alterations in the abundance of cell wall polysaccharides, in particular decrease in crystalline cellulose, and profound modifications in gene expression and homeostasis reminiscent of a stress response. CYP98A3 thus constitutes a critical bottleneck in the phenylpropanoid pathway and in the synthesis of compounds controlling plant development. CYP98A3 cosuppressed lines show a gradation of developmental defects and changes in lignin content (40% reduction) and structure (prominent frequency of p-hydroxyphenyl units), but content in foliar sinapoyl esters is similar to the control. The purple coloration of their leaves is correlated to the accumulation of sinapoylated anthocyanins.
Abdulrazzak, Nawroz; Pollet, Brigitte; Ehlting, Jurgen; Larsen, Kim; Asnaghi, Carole; Ronseau, Sebastien; Proux, Caroline; Erhardt, Mathieu; Seltzer, Virginie; Renou, Jean-Pierre; Ullmann, Pascaline; Pauly, Markus; Lapierre, Catherine; Werck-Reichhart, Daniele
Agrobacterium tumefaciens VirC2 enhances T-DNA transfer and virulence through its C-terminal ribbon-helix-helix DNA-binding fold.
Agrobacterium tumefaciens VirC2 stimulates processing of single-stranded T-DNA that is translocated into plants to induce tumor formation, but how VirC2 functions is unclear. Here, we report the 1.7-A X-ray crystal structure of its trypsin-resistant C-terminal domain, VirC2(82-202), which reveals a form of the ribbon-helix-helix (RHH) DNA-binding fold contained within a single polypeptide chain. DNA-binding assays and mutagenesis indicate that VirC2 uses this RHH fold to bind double-stranded DNA but not single-stranded DNA. Mutations that severely affect VirC2 DNA binding are highly deleterious for both T-DNA transfer into yeast and the virulence of A. tumefaciens in different plants including Nicotiana glauca and Kalanchoe daigremontiana. These data suggest that VirC2 enhances T-DNA transfer and virulence through DNA binding with its RHH fold. The RHH fold of VirC2 is the first crystal structure representing a group of predicted RHH proteins that facilitate endonucleolytic processing of DNA for horizontal gene transfer. PMID:19482939
Lu, Jun; den Dulk-Ras, Amke; Hooykaas, Paul J J; Glover, J N Mark
1046. Insertional Leukemogenesis after Retroviral Gene Transfer into Murine Hematopoietic Stem Cells
Replication defective retroviruses deliver transgenes into hematopoietic stem cells (HSC) with high efficacy. The first successful clinical study revealed an unexpectedly high frequency of insertional leukemogenesis, not predicted by extensive preclinical experimental studies. In cell lines, the frequency of insertional oncogenic events has been estimated at approximately 10?7 per transduced cell. To examine the in vivo incidence, we monitored mice
Martijn H. Brugman; Monique M. A. Verstegen; Trudi P. Visser; Erna Frńnzel-Luiten; Manfred Schmidt; Albertus W. Wognum; Christof vonKalle; Gerard Wagemaker
Rapid tooling of die DMLS inserts for shoot-squeeze moulding (DISA) system
The rapid tooling (RT) technology applied to die-plates for moulding in DISAmaticę cast line was investigated. Die inserts made by RT direct techniques such as: direct metal laser sintering (DMLS); stereolithography (SL); laminated object manufacturing (LOM); and selective laser sintering of polymer powder (SLS-P), are compared. Mechanical properties and accuracy are critical on choosing the rapid tooling die inserts as
J. C. Ferreira
New insert layer structure OLEDs
NASA Astrophysics Data System (ADS)
An insert layer structure organic electroluminescent device(OLED) based on a new luminescent material (Zn(salen)) is fabricated. The configuration of the device is ITO/CuPc/NPD/Zn(salen)/Liq/LiF/Al/CuPc/NPD/Zn(salen)/Liq/LiF/Al. Effective insert electrode layers comprising LiF(1 nm)/Al(5 nm) are used as a single semitransparent mirror, and bilayer cathode LiF(1 nm)/Al(100 nm) is used as a reflecting mirror. The two mirrors form a Fabry-Perot microcavity and two emissive units. The maximum brightness and luminous efficiency reach 674 cd/m2 and 2.652 cd/A, respectively, which are 2.1 and 3.7 times higher than the conventional device, respectively. The superior brightness and luminous efficiency over conventional single-unit devices are attributed to microcavity effect.
Gao, Zhi-Xiang; Hao, Yu-Ying; Lei, Jun-Feng; Ma, Cheng; Xu, Bing-She
Ultrasound guided spine needle insertion
NASA Astrophysics Data System (ADS)
An ultrasound (US) guided, CT augmented, spine needle insertion navigational system is introduced. The system consists of an electromagnetic (EM) sensor, an US machine, and a preoperative CT volume of the patient anatomy. Three-dimensional (3D) US volume is reconstructed intraoperatively from a set of two-dimensional (2D) freehand US slices, and is coregistered with the preoperative CT. This allows the preoperative CT volume to be used in the intraoperative clinical coordinate. The spatial relationship between the patient anatomy, surgical tools, and the US transducer are tracked using the EM sensor, and are displayed with respect to the CT volume. The pose of the US transducer is used to interpolate the CT volume, providing the physician with a 2D "x-ray vision" to guide the needle insertion. Many of the system software components are GPU-accelerated, allowing real-time performance of the guidance system in a clinical setting.
Chen, Elvis C. S.; Mousavi, Parvin; Gill, Sean; Fichtinger, Gabor; Abolmaesumi, Purang
Insert type milling cutter evaluation
The purpose of this test was to obtain data on the major manufacturers of Insert Type Milling Cutters. Each manufacturers' representative was contacted and informed of the pending test. They were told of the type of material to be machined and the parameters of the operation. The manufacturers were asked to supply us with a cutter and inserts; they recommended to meet our needs for machining 304 stainless steel as specified in the Machining Data Handbook. Only three manufacturers responded; Carboloy, Kennametal, and Valenite., and cutters from each were tested. Each cutter was exposed to the exact same conditions, varying only where the cutter parameters dictated, to assure equality. The test results show that the Valenite cutter and inserts would require the least amount of maintenance, the greatest amount of wear life, and the best finishes. This cutter also required the least amount of horsepower in each cut. This is also advantageous from a power consumption outlook. The Valenite used a maximum of 8.01 Hp, which is 5968.48 W of power, where the Kennametal cutter used a maximum of 17.443 Hp, which is 13,012.48 W of power consumed. This is almost two times the energy consumption.
Gonzales, J.F.
PAIR: polymorphic Alu insertion recognition
Background Alu polymorphisms are some of the most common polymorphisms in the genome, yet few methods have been developed for their detection. Methods We present algorithms to discover Alu polymorphisms using paired-end high throughput sequencing data from multiple individuals. We consider the problem of identifying sites containing polymorphic Alu insertions. Results We give efficient and practical algorithms that detect polymorphic Alus, both those that are inserted with respect to the reference genome and those that are deleted. The algorithms have a linear time complexity and can be run on a standard desktop machine in a very short amount of time on top of the output of tools standard for sequencing analysis. Conclusions In our simulated dataset we are able to locate 98.1% of Alus inserted with respect to the reference and 97.7% of Alus deleted, our simulations also show an excellent correlations between the deletions detected in parents and children. We further run our algorithms on publicly available data from the 1000 genomes project and find several thousand Alu polymorphisms in each individual.
Insert sleeve prevents tube soldering contamination
NASA Technical Reports Server (NTRS)
Teflon sleeve insert prevents contamination of internal tube surfaces by solder compound during soldering operations that connect and seal the tube ends. The sleeve insert is pressed into the mating tube ends with a slight interference fit.
Stein, J.
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.
Duangpan, Saowapa; Zhang, Wenli; Wu, Yufang; Jansky, Shelley H.; Jiang, Jiming
Large-scale mapping of transposable element insertion sites using digital encoding of sample identity.
Determining the genomic locations of transposable elements is a common experimental goal. When mapping large collections of transposon insertions, individualized amplification and sequencing is both time consuming and costly. We describe an approach in which large numbers of insertion lines can be simultaneously mapped in a single DNA sequencing reaction by using digital error-correcting codes to encode line identity in a unique set of barcoded pools. PMID:24374352
Gohl, Daryl M; Freifeld, Limor; Silies, Marion; Hwa, Jennifer J; Horowitz, Mark; Clandinin, Thomas R
BayGenomics: a resource of insertional mutations in mouse embryonic stem cells
The BayGenomics gene-trap resource (http:\\/\\/ baygenomics.ucsf.edu) provides researchers with access to thousands of mouse embryonic stem (ES) cell lines harboring characterized insertional mutations in both known and novel genes. Each cell line contains an insertional mutation in a specific gene. The identity of the gene that has been interrupted can be determined from a DNA sequence tag. Approximately 75% of
Doug Stryke; Michiko Kawamoto; Conrad C. Huang; Susan J. Johns; Leslie A. King; Courtney A. Harper; Elaine C. Meng; Roy E. Lee; Alice Yee; Larry L'italien; Pao-tien Chuang; Stephen G. Young; William C. Skarnes; Patricia C. Babbitt; Thomas E. Ferrin
Networking senescence-regulating pathways by using Arabidopsis enhancer trap lines.
The last phase of leaf development, generally referred to as leaf senescence, is an integral part of plant development that involves massive programmed cell death. Due to a sharp decline of photosynthetic capacity in a leaf, senescence limits crop yield and forest plant biomass production. However, the biochemical components and regulatory mechanisms underlying leaf senescence are poorly characterized. Although several approaches such as differential cDNA screening, differential display, and cDNA subtraction have been employed to isolate senescence-associated genes (SAGs), only a limited number of SAGs have been identified, and information regarding the regulation of these genes is fragmentary. Here we report on the utilization of enhancer trap approach toward the identification and analysis of SAGs. We have developed a sensitive large-scale screening method and have screened 1,300 Arabidopsis enhancer trap lines and have identified 147 lines in which the reporter gene GUS (beta-glucuronidase) is expressed in senescing leaves but not in non-senescing ones. We have systematically analyzed the regulation of beta-glucuronidase expression in 125 lines (genetically, each contains single T-DNA insertion) by six senescence-promoting factors, namely abscisic acid, ethylene, jasmonic acid, brassinosteroid, darkness, and dehydration. This analysis not only reveals the complexity of the regulatory circuitry but also allows us to postulate the existence of a network of senescence-promoting pathways. We have also cloned three SAGs from randomly selected enhancer trap lines, demonstrating that reporter expression pattern reflects the expression pattern of the endogenous gene. PMID:11402199
He, Y; Tang, W; Swain, J D; Green, A L; Jack, T P; Gan, S
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.
Automatic insertion of BIST hardware using VHDL
We present a system which automatically inserts BIST hardware to a circuit described in VHDL. An appropriate VHDL modeling style for automatic insertion of BIST hardware is investigated. Use of BILBO is primarily pursued in the system. Algorithmic and rule-based approaches are used in the insertion of BILBO. Test scheduling and control signal distribution are also performed by the system.
Kwanghyun Kim; Joseph G. Tront; Dong Sam Ha
Recent advances in insertion devices
Demand for more and better insertion devices (IDs) at new third-generation synchrotron radiation facilities has led to significant advances in ID technology at different laboratories around the world. In this overview of this progress, focus is on those results that apply to IDs in general rather than one specific ID or laboratory. The advances fall into two general categories: those that reduce the net effect that the ID has on the particle beam, and those that enhance the quality of the emitted light spectrum. The need for these advances, factors that are most important inaachieving them, and the current state of the art are discussed.
Gluskin, E.; Moog, E.R.
Exchange of peripherally inserted central catheters is associated with an increased risk for bloodstream infection.
It is not uncommon that the peripherally inserted central catheter (PICC) needs to be replaced either due to blockage or migration to a peripheral position. In such circumstances, there are two methods of PICC placement: new-site insertion and exchange by using the old PICC as a guide wire. Our objective was to investigate risk of infection associated with the exchange method. In this retrospective study, data on all PICC insertions in the neonatal intensive care unit in 2004 to 2008 were obtained. In the population who needed removal of existing PICC and insertion of a new one, we compared central line-associated bloodstream infection (CLABSI) within 1 week of insertion between the two insertion methods. Of 1148 PICC insertions reviewed, 164 (103 new-site and 61 exchange insertions) were performed after removal of a blocked/malpositioned PICC and therefore comprised the study population. The rate of CLABSI was higher in the exchange method (9.8% versus 1%, P < 0.007). After adjusting for the confounders, the odds for CLABSI within 7 days of PICC insertion was higher with the exchange method (odds ratio 25.2, 95% confidence interval: 2.17 to 292.98; P = 0.01). In infants, insertion of PICCs using the exchange method carries an increased risk of bloodstream infection. PMID:21082537
McCoy, Michael; Bedwell, Susan; Noori, Shahab
Abnormal endosperm development causes female sterility in rice insertional mutant OsAPC6.
A T-DNA insertional mutant OsAPC6 of rice, with gibberellic acid insensitivity and reduced height, had up to 45% reduced seed set. The insertion occurred on chromosome 3 of rice in the gene encoding one of the subunits of anaphase promoting complex/Cyclosome APC6. The primary mother cells of the mutant plants had normal meiosis, male gametophyte development and pollen viability. Confocal laser scanning microscopic (CLSM) studies of megagametophyte development showed abnormal mitotic divisions with reduced number or total absence of polar nuclei in about 30-35% megagametophytes of OsAPC6 mutant leading to failure of endosperm and hence embryo and seed development. Abnormal female gametophyte development, high sterility and segregation of tall and gibberellic acid sensitive plants without selectable marker Hpt in the selfed progeny of OsAPC6 mutant plants indicate that the mutant could be maintained in heterozygous condition. The abnormal mitotic divisions during megagametogenesis could be attributed to the inactivation of the APC6/CDC16 of anaphase promoting complex of rice responsible for cell cycle progression during megagametogenesis. Functional validation of the candidate gene through transcriptome profiling and RNAi is in progress. PMID:22195590
Awasthi, Anjali; Paul, Priyanka; Kumar, Satish; Verma, Shailendra Kumar; Prasad, R; Dhaliwal, H S
Retrieval analysis of posterior stabilized polyethylene tibial inserts and its clinical relevance.
This was a retrieval analysis of 83 PS inserts to assess the effect of limb alignment, implant position and joint line position on the pattern of wear in posterior stabilized (PS) tibial inserts. The total damage score was significantly higher in knees with postoperative varus alignment more than 3░ (P = 0.03). The total damage score to the post was significantly more in knees with joint line elevation more than 5 mm (9.7 ▒ 3.9, compared to 6.5 ▒ 3.7 in knees with less joint line elevation) (P = 0.05). Limb malalignment and joint line elevation resulted in more damage in PS inserts. An external rotation subluxation damage pattern was found in joint line elevation. PMID:23809707
Pang, Hee-Nee; Jamieson, Paul; Teeter, Matthew G; McCalden, Richard W; Naudie, Douglas D R; MacDonald, Steven J
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: email@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)
Management of irritant contact dermatitis and peripherally inserted central catheters.
Cutaneous skin changes are common in patients undergoing treatment for cancer. However, changes in central line care, maintenance practices, and chemotherapy protocols in the early 2000s may have led to the development of a common problem of irritant contact dermatitis (ICD) at peripherally inserted central catheter (PICC) insertion sites. Repeated exposure to chlorhexidine gluconate topical antiseptic solution, used in the general dressing care and maintenance with PICCs, may be the leading contributor to the development of ICD at the insertion site. A number of additional factors theoretically contribute to the development of ICD at the PICC insertion site in patients receiving chemotherapy. In this article, ICD will be defined, incidence and potential risk factors will be identified, and diagnostic framework will be explored; in addition, pathophysiology, onset, presentation, evaluation, and differential diagnosis of ICD at PICC sites will be analyzed. Finally, a synopsis of three different treatment approaches from healthcare facilities in Canada as well as implications for nursing practice and research will be presented. PMID:22459537
Kutzscher, Lia
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)
The distribution of transgene insertion sites in barley determined by physical and genetic mapping.
The exact site of transgene insertion into a plant host genome is one feature of the genetic transformation process that cannot, at present, be controlled and is often poorly understood. The site of transgene insertion may have implications for transgene stability and for potential unintended effects of the transgene on plant metabolism. To increase our understanding of transgene insertion sites in barley, a detailed analysis of transgene integration in independently derived transgenic barley lines was carried out. Fluorescence in situ hybridization (FISH) was used to physically map 23 transgene integration sites from 19 independent barley lines. Genetic mapping further confirmed the location of the transgenes in 11 of these lines. Transgene integration sites were present only on five of the seven barley chromosomes. The pattern of transgene integration appeared to be nonrandom and there was evidence of clustering of independent transgene insertion events within the barley genome. In addition, barley genomic regions flanking the transgene insertion site were isolated for seven independent lines. The data from the transgene flanking regions indicated that transgene insertions were preferentially located in gene-rich areas of the genome. These results are discussed in relation to the structure of the barley genome.
Salvo-Garrido, Haroldo; Travella, Silvia; Bilham, Lorelei J; Harwood, Wendy A; Snape, John W
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
Gas fired fireplace insert with heat extractor
A combination glass door fireplace closure and heat exchanger which can be inserted into the cavity of a gas fired fireplace to permit more efficient and economical operation is disclosed. The insert includes a plurality of substantially parallel c-shaped exchanger tubes. The lower and upper ends of the tube are coupled to a glass door fireplace closure which is adapted
Reliability of inserts in sandwich composite panels
Inserts are commonly used to transfer loads to sandwich composite structures. Local stress concentrations due to inserts are known to cause structural failure, and experimental pull-out tests show that the failure load can vary by 20% between batches of sandwich panels. Clearly, uncertainty in the mechanical properties of the constituent materials needs to be addressed in the design and optimization
Bryan Smith; Biswajit Banerjee
Design Loads for Inserts Embedded in Concrete.
National Technical Information Service (NTIS)
Detailed test procedures are presented for a research program on cast-in-place inserts embedded in reinforced concrete. Three types of inserts, two of malleable iron and one of ductile steel, capable of receiving a 3/4 inch threaded rod were tested. Other...
T. W. Reichard E. F. Carpenter E. V. Leyendecker
Insertion Sequence Diversity in Archaea
Insertion sequences (ISs) can constitute an important component of prokaryotic (bacterial and archaeal) genomes. Over 1,500 individual ISs are included at present in the ISfinder database (www-is.biotoul.fr), and these represent only a small portion of those in the available prokaryotic genome sequences and those that are being discovered in ongoing sequencing projects. In spite of this diversity, the transposition mechanisms of only a few of these ubiquitous mobile genetic elements are known, and these are all restricted to those present in bacteria. This review presents an overview of ISs within the archaeal kingdom. We first provide a general historical summary of the known properties and behaviors of archaeal ISs. We then consider how transposition might be regulated in some cases by small antisense RNAs and by termination codon readthrough. This is followed by an extensive analysis of the IS content in the sequenced archaeal genomes present in the public databases as of June 2006, which provides an overview of their distribution among the major archaeal classes and species. We show that the diversity of archaeal ISs is very great and comparable to that of bacteria. We compare archaeal ISs to known bacterial ISs and find that most are clearly members of families first described for bacteria. Several cases of lateral gene transfer between bacteria and archaea are clearly documented, notably for methanogenic archaea. However, several archaeal ISs do not have bacterial equivalents but can be grouped into Archaea-specific groups or families. In addition to ISs, we identify and list nonautonomous IS-derived elements, such as miniature inverted-repeat transposable elements. Finally, we present a possible scenario for the evolutionary history of ISs in the Archaea.
Filee, J.; Siguier, P.; Chandler, M.
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.
Mitochondrial DNA transfer to the nucleus generates extensive insertion site variation in maize.
Mitochondrial DNA (mtDNA) insertions into nuclear chromosomes have been documented in a number of eukaryotes. We used fluorescence in situ hybridization (FISH) to examine the variation of mtDNA insertions in maize. Twenty overlapping cosmids, representing the 570-kb maize mitochondrial genome, were individually labeled and hybridized to root tip metaphase chromosomes from the B73 inbred line. A minimum of 15 mtDNA insertion sites on nine chromosomes were detectable using this method. One site near the centromere on chromosome arm 9L was identified by a majority of the cosmids. To examine variation in nuclear mitochondrial DNA sequences (NUMTs), a mixture of labeled cosmids was applied to chromosome spreads of ten diverse inbred lines: A188, A632, B37, B73, BMS, KYS, Mo17, Oh43, W22, and W23. The number of detectable NUMTs varied dramatically among the lines. None of the tested inbred lines other than B73 showed the strong hybridization signal on 9L, suggesting that there is a recent mtDNA insertion at this site in B73. Different sources of B73 and W23 were examined for NUMT variation within inbred lines. Differences were detectable, suggesting either that mtDNA is being incorporated or lost from the maize nuclear genome continuously. The results indicate that mtDNA insertions represent a major source of nuclear chromosomal variation. PMID:18202357
Lough, Ashley N; Roark, Leah M; Kato, Akio; Ream, Thomas S; Lamb, Jonathan C; Birchler, James A; Newton, Kathleen J
Mitochondrial DNA Transfer to the Nucleus Generates Extensive Insertion Site Variation in Maize
Mitochondrial DNA (mtDNA) insertions into nuclear chromosomes have been documented in a number of eukaryotes. We used fluorescence in situ hybridization (FISH) to examine the variation of mtDNA insertions in maize. Twenty overlapping cosmids, representing the 570-kb maize mitochondrial genome, were individually labeled and hybridized to root tip metaphase chromosomes from the B73 inbred line. A minimum of 15 mtDNA insertion sites on nine chromosomes were detectable using this method. One site near the centromere on chromosome arm 9L was identified by a majority of the cosmids. To examine variation in nuclear mitochondrial DNA sequences (NUMTs), a mixture of labeled cosmids was applied to chromosome spreads of ten diverse inbred lines: A188, A632, B37, B73, BMS, KYS, Mo17, Oh43, W22, and W23. The number of detectable NUMTs varied dramatically among the lines. None of the tested inbred lines other than B73 showed the strong hybridization signal on 9L, suggesting that there is a recent mtDNA insertion at this site in B73. Different sources of B73 and W23 were examined for NUMT variation within inbred lines. Differences were detectable, suggesting either that mtDNA is being incorporated or lost from the maize nuclear genome continuously. The results indicate that mtDNA insertions represent a major source of nuclear chromosomal variation.
Lough, Ashley N.; Roark, Leah M.; Kato, Akio; Ream, Thomas S.; Lamb, Jonathan C.; Birchler, James A.; Newton, Kathleen J.
High-throughput semiquantitative analysis of insertional mutations in heterogeneous tumors
Retroviral and transposon-based insertional mutagenesis (IM) screens are widely used for cancer gene discovery in mice. Exploiting the full potential of IM screens requires methods for high-throughput sequencing and mapping of transposon and retroviral insertion sites. Current protocols are based on ligation-mediated PCR amplification of junction fragments from restriction endonuclease-digested genomic DNA, resulting in amplification biases due to uneven genomic distribution of restriction enzyme recognition sites. Consequently, sequence coverage cannot be used to assess the clonality of individual insertions. We have developed a novel method, called shear-splink, for the semiquantitative high-throughput analysis of insertional mutations. Shear-splink employs random fragmentation of genomic DNA, which reduces unwanted amplification biases. Additionally, shear-splink enables us to assess clonality of individual insertions by determining the number of unique ligation points (LPs) between the adapter and genomic DNA. This parameter serves as a semiquantitative measure of the relative clonality of individual insertions within heterogeneous tumors. Mixing experiments with clonal cell lines derived from mouse mammary tumor virus (MMTV)-induced tumors showed that shear-splink enables the semiquantitative assessment of the clonality of MMTV insertions. Further, shear-splink analysis of 16 MMTV- and 127 Sleeping Beauty (SB)ľinduced tumors showed enrichment for cancer-relevant insertions by exclusion of irrelevant background insertions marked by single LPs, thereby facilitating the discovery of candidate cancer genes. To fully exploit the use of the shear-splink method, we set up the Insertional Mutagenesis Database (iMDB), offering a publicly available web-based application to analyze both retroviral- and transposon-based insertional mutagenesis data.
Koudijs, Marco J.; Klijn, Christiaan; van der Weyden, Louise; Kool, Jaap; ten Hoeve, Jelle; Sie, Daoud; Prasetyanti, Pramudita R.; Schut, Eva; Kas, Sjors; Whipp, Theodore; Cuppen, Edwin; Wessels, Lodewyk; Adams, David J.; Jonkers, Jos
Mechanism of alamethicin insertion into lipid bilayers.
Alamethicin adsorbs on the membrane surface at low peptide concentrations. However, above a critical peptide-to-lipid ratio (P/L), a fraction of the peptide molecules insert in the membrane. This critical ratio is lipid dependent. For diphytanoyl phosphatidylcholine it is about 1/40. At even higher concentrations P/L > or = 1/15, all of the alamethicin inserts into the membrane and forms well-defined pores as detected by neutron in-plane scattering. A previous x-ray diffraction measurement showed that alamethicin adsorbed on the surface has the effect of thinning the bilayer in proportion to the peptide concentration. A theoretical study showed that the energy cost of membrane thinning can indeed lead to peptide insertion. This paper extends the previous studies to the high-concentration region P/L > 1/40. X-ray diffraction shows that the bilayer thickness increases with the peptide concentration for P/L > 1/23 as the insertion approaches 100%. The thickness change with the percentage of insertion is consistent with the assumption that the hydrocarbon region of the bilayer matches the hydrophobic region of the inserted peptide. The elastic energy of a lipid bilayer including both adsorption and insertion of peptide is discussed. The Gibbs free energy is calculated as a function of P/L and the percentage of insertion phi in a simplified one-dimensional model. The model exhibits an insertion phase transition in qualitative agreement with the data. We conclude that the membrane deformation energy is the major driving force for the alamethicin insertion transition. Images FIGURE 1
He, K; Ludtke, S J; Heller, W T; Huang, H W
Multipacting Analysis of a Quarter Wave Choke Joint Used for Insertion of a Demountable Cathode into a SRF Photoinjector.
National Technical Information Service (NTIS)
The multipacting phenomena in accelerating structures and coaxial lines are well documented and methods of mitigating or suppressing it are understood. The multipacting that occurs in a quarter wave choke joint designed to mount a cathode insertion stalk ...
A. Burrill I. Ben-Zvi J. Rathke M. Cole P. Kneisel R. Manus R. Rimmer
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)
Effects of Single P-Element Insertions on Olfactory Behavior in Drosophila Melanogaster
Single P-element (P[lArB]) insertional mutagenesis of an isogenic strain was used to identify autosomal loci affecting odor-guided behavior of Drosophila melanogaster. The avoidance response to benzaldehyde of 379 homozygous P[lArB] element-containing insert lines was evaluated quantitatively. Fourteen smell impaired (smi) lines were identified in which P[lArB] element insertion caused different degrees of hyposmia in one or both sexes. The smi loci map to different cytological locations and probably are novel olfactory genes. Enhancer trap analysis of the smi lines indicates that expression of at least 10 smi genes is controlled by olfactory tissue-specific promoter/enhancer elements.
Anholt, RRH.; Lyman, R. F.; Mackay, TFC.
Performance Analysis of 'Star' Pack Cushion Inserts.
National Technical Information Service (NTIS)
The objectives of the study were two-fold: (1) Modify 'Star' pack cushion inserts to provide more uniform cushioning protection and most cost effective pack; and (2) determine adequacy of previously published 'Star' pack performance data for representing ...
R. V. Brown
Compiler-Assisted Static Checkpoint Insertion.
National Technical Information Service (NTIS)
This paper describes a compiler-assisted approach for static checkpoint insertion. Instead of fixing the checkpoint location before program execution, a compiler enhanced polling mechanism is utilized to maintain both the desired checkpoint intervals and ...
J. Long W. K. Fuchs J. A. Abraham
Compiler-Assisted Static Checkpoint Insertion.
National Technical Information Service (NTIS)
This paper describes a compiler-assisted approach for static checkpoint insertion. Instead of fixing the checkpoint location before program execution, a compiler enhanced polling mechanism is utilized to maintain both the desired checkpoint intervals and ...
J. Long J. A. Abraham W. K. Fuchs
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)
Transosseous fixation in insertional Achilles tendonitis.
The surgical approach of severe insertional Achilles tendonitis involves debridement, repair, and reattachment. Previously described techniques for bone-tendon reattachment involved the use of suture anchors in the sparsely trabeculated calcaneus. The following technique tip is a reintroduction of bicortical transosseous fixation previously used in rotator cuff repairs. Bone tunnels are a viable option in the treatment of insertional Achilles tendonitis; however, additional research is necessary to validate its efficacy in this application. PMID:22841157
Sundararajan, Priya Ponnapula
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
Sensing membrane stresses by protein insertions.
Protein domains shallowly inserting into the membrane matrix are ubiquitous in peripheral membrane proteins involved in various processes of intracellular membrane shaping and remodeling. It has been suggested that these domains sense membrane curvature through their preferable binding to strongly curved membranes, the binding mechanism being mediated by lipid packing defects. Here we make an alternative statement that shallow protein insertions are universal sensors of the intra-membrane stresses existing in the region of the insertion embedding rather than sensors of the curvature per se. We substantiate this proposal computationally by considering different independent ways of the membrane stress generation among which some include changes of the membrane curvature whereas others do not alter the membrane shape. Our computations show that the membrane-binding coefficient of shallow protein insertions is determined by the resultant stress independently of the way this stress has been produced. By contrast, consideration of the correlation between the insertion binding and the membrane curvature demonstrates that the binding coefficient either increases or decreases with curvature depending on the factors leading to the curvature generation. To validate our computational model, we treat quantitatively the experimental results on membrane binding by ALPS1 and ALPS2 motifs of ArfGAP1. PMID:24722359
Campelo, Felix; Kozlov, Michael M
Cancer gene discovery: exploiting insertional mutagenesis
Insertional mutagenesis has been utilized as a functional forward genetics screen for the identification of novel genes involved in the pathogenesis of human cancers. Different insertional mutagens have been successfully used to reveal new cancer genes. For example, retroviruses (RVs) are integrating viruses with the capacity to induce the deregulation of genes in the neighborhood of the insertion site. RVs have been employed for more than 30 years to identify cancer genes in the hematopoietic system and mammary gland. Similarly, another tool that has revolutionized cancer gene discovery is the cut-and-paste transposons. These DNA elements have been engineered to contain strong promoters and stop cassettes that may function to perturb gene expression upon integration proximal to genes. In addition, complex mouse models characterized by tissue-restricted activity of transposons have been developed to identify oncogenes and tumor suppressor genes that control the development of a wide range of solid tumor types, extending beyond those tissues accessible using RV-based approaches. Most recently, lentiviral vectors (LVs) have appeared on the scene for use in cancer gene screens. LVs are replication defective integrating vectors that have the advantage of being able to infect non-dividing cells, in a wide range of cell types and tissues. In this review, we describe the various insertional mutagens focusing on their advantages/limitations and we discuss the new and promising tools that will improve the insertional mutagenesis screens of the future.
Ranzani, Marco; Annunziato, Stefano; Adams, David J.; Montini, Eugenio
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)
Sensing Membrane Stresses by Protein Insertions
Protein domains shallowly inserting into the membrane matrix are ubiquitous in peripheral membrane proteins involved in various processes of intracellular membrane shaping and remodeling. It has been suggested that these domains sense membrane curvature through their preferable binding to strongly curved membranes, the binding mechanism being mediated by lipid packing defects. Here we make an alternative statement that shallow protein insertions are universal sensors of the intra-membrane stresses existing in the region of the insertion embedding rather than sensors of the curvature per se. We substantiate this proposal computationally by considering different independent ways of the membrane stress generation among which some include changes of the membrane curvature whereas others do not alter the membrane shape. Our computations show that the membrane-binding coefficient of shallow protein insertions is determined by the resultant stress independently of the way this stress has been produced. By contrast, consideration of the correlation between the insertion binding and the membrane curvature demonstrates that the binding coefficient either increases or decreases with curvature depending on the factors leading to the curvature generation. To validate our computational model, we treat quantitatively the experimental results on membrane binding by ALPS1 and ALPS2 motifs of ArfGAP1.
Campelo, Felix; Kozlov, Michael M.
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, StÚphane; Trap-Gentil, Marie-VÚronique; HÚbrard, Claire; Weyens, Guy; Delaunay, Alain; Barnes, Steve; Lefebvre, Marc; Joseph, Claude
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. 3 figures.
Carr, R.
IUD insertions following cesarean deliveries examined.
A paper presented at the 111th annual meeting of the American Public Health Association in November 1983 reported on the performance of IUDs inserted immediately following cesarean deliveries as well as those inserted in interval women with previous cesarean sections. Physicians at Xuan Wu Hospital in Beijing, China manually inserted IUDs through the incisions of 52 cesarean-sectioned women immediately after delivery of the placenta. In 147 women the IUDs were inserted vaginally immediately after normal vaginal delivery. Delta T and Delta Loop IUDs were inserted in equal proportions in both groups of women. These IUDs are Family Health International's (FHI's) modifications of the standard Lippes Loop D and Copper T 220C. Biodegradable No. 2 chromic sutures are added to the upper crossarm to enhance IUD retention when inserted in the puerperium, thereby reducing the risk of expulsion as uterine involution occurs. All of the 199 study participants were currently married, admitted for delivery of their 1st birth, and between 25-29 years. All but 1 woman in the cesarean group but only 57% of the vaginal delivery group had their IUDs inserted by obstetricians/gynecologists. The follow-up rate at 6 months postinsertion exceeded 95% for both groups. Expulsion rates at 1, 3, and 6 months postinsertion were significantly lower and thus continuation rates significantly higher for the cesarean group than for the vaginal group. The difference in rates did not change substantially even when the 2 groups were compared considering only those women whose IUD was inserted by the obstetricians/gynecologists. There were no serious adverse effects with immediate postcesarean insertions. The 2 groups showed no statistically significant differences in pregnancy rates or in bleeding, pain, or other causes of removal. Another study from China and one from Mexico support this finding, but neither has provided a comparison group. 2 data sets were used to examine the effects of interval insertions with women with previous cesareans. 288 of the women who received locally designed Szontagh IUDs in a Hungarian Center had had 1 or more previous cesareans. Each was matched by age, parity, and date of insertion with 2 controls who each had at least 1 birth but not a cesarean. The cumulative expulsion rates were higher among the cesarean-sectioned women than among the nonsectioned women at 1, 3, 6, and 12 months postinsertion. The other data set used was a pooled file of over 32,000 IUD users from which 318 interval women reported 1 or more previous cesarean deliveries. 299 of these women, inserted with either the Lippes Loop, Copper T, or Multiload IUDs, were matched with two, and the remaining 19 with one, controls from the same data set by center, IUD type, and the other matching criteria of the Hungarian study. There were no significant differences in expulsion rates between the 2 groups. PMID:12265941
Rack Insertion End Effector (RIEE) automation
NASA Technical Reports Server (NTRS)
NASA is developing a mechanism to manipulate and insert Racks into the Space Station Logistic modules. The mechanism consists of the following: a base with three motorized degrees of freedom, a 3 section motorized boom that goes from 15 to 44 feet in length, and a Rack Insertion End Effector (RIEE) with 5 hand wheels for precise alignment. The robotics section was tasked with the automation of the RIEE unit. In this report, for the automation of the RIEE unit, application of the Perceptics Vision System was conceptually developed to determine the position and orientation of the RIEE relative to the logistic module, and a MathCad program is written to display the needed displacements for precise alignment and final insertion of the Rack. The uniqueness of this report is that the whole report is in fact a MathCad program including text, derivations, and executable equations with example inputs and outputs.
Malladi, Narasimha
Deformation of femoral nails with intramedullary insertion.
Current methods of distal interlocking of intramedullary femoral nails are dependent on image intensification. However, radiation exposure to the patient, the operating room staff, and the surgeon remains a concern. Proximally mounted, radiation-free aiming systems for distal interlocking of femoral nails have reportedly failed because of nail deformation with insertion. To better understand this deformation, a three-dimensional magnetic motion tracking system was used to determine the position of the distal interlocking hole following nail insertion. The amount and direction of deformation of commercially available small-diameter implants (unslotted 9-mm nails inserted without reaming) and large-diameter implants (slotted 13-mm nails inserted with reaming) from a single manufacturer were analyzed. Measurements of deformation (three translations and three angles), based on the center of the distal transverse locking hole, were performed on 10 paired intact human cadaveric femora before and after insertion. The technique produced the following results for the small and large-diameter nails, respectively: lateral translations of 18.1+/-10.0 mm (mean+/-SD, range: 47.8 mm) and 21.5+/-7.9 mm (range: 26.4 mm), dorsal translations of -3.1+/-4.3 mm (range: 15.2 mm) and 0.4+/-9.8 mm (range: 30.1 mm), and rotation about the longitudinal axes of -0.1+/-0.2 degrees (range: 0.7 degrees) and 10.0+/-3.1 degrees (range: 7.8 degrees). This technique is useful for measuring insertion-related femoral nail deformation. The data for the nails tested suggest that a simple aiming arm, mounted on the proximal end of the femoral nail alone, will not sufficiently provide accurate distal aiming. PMID:9820280
Krettek, C; Mannss, J; Miclau, T; Schandelmaier, P; Linnemann, I; Tscherne, H
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 ôpresenceö of a retroelement at an orthologous genomic position in two species indicates their common ancestry in contrast to its ôabsenceö in more distant species. To resolve the placental origin controversy we extracted ?2 million potentially phylogenetically informative, retroposon-containing loci from representatives of the major placental mammalian lineages and found highly significant evidence challenging all current single hypotheses of their basal origin. The Exafroplacentalia hypothesis (Afrotheria as the sister group to all remaining placentals) is significantly supported by five retroposon insertions, the Epitheria hypothesis (Xenarthra as the sister group to all remaining placentals) by nine insertion patterns, and the Atlantogenata hypothesis (a monophyletic clade comprising Xenarthra and Afrotheria as the sister group to Boreotheria comprising all remaining placentals) by eight insertion patterns. These findings provide significant support for a ôsoftö 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.
Churakov, Gennady; Kriegs, Jan Ole; Baertsch, Robert; Zemann, Anja; Brosius, Jurgen; Schmitz, Jurgen
PBmice: an integrated database system of piggyBac (PB) insertional mutations and their characterizations in mice
DNA transposon piggyBac (PB) is a newly established mutagen for large-scale mutagenesis in mice. We have designed and implemented an integrated database system called PBmice (PB Mutagenesis Information CEnter) for storing, retrieving and displaying the information derived from PB insertions (INSERTs) in the mouse genome. This system is centered on INSERTs with information including their genomic locations and flanking genomic sequences, the expression levels of the hit genes, and the expression patterns of the trapped genes if a trapping vector was used. It also archives mouse phenotyping data linked to INSERTs, and allows users to conduct quick and advanced searches for genotypic and phenotypic information relevant to a particular or a set of INSERT(s). Sequence-based information can be cross-referenced with other genomic databases such as Ensembl, BLAST and GBrowse tools used in PBmice offer enhanced search and display for additional information relevant to INSERTs. The total number and genomic distribution of PB INSERTs, as well as the availability of each PB insertional LINE can also be viewed with user-friendly interfaces. PBmice is freely available at http://www.idmshanghai.cn/PBmice or http://www.scbit.org/PBmice/.
Sun, Ling V.; Jin, Ke; Liu, Yiming; Yang, Wenwei; Xie, Xing; Ye, Lin; Wang, Li; Zhu, Lin; Ding, Sheng; Su, Yi; Zhou, Jie; Han, Min; Zhuang, Yuan; Xu, Tian; Wu, Xiaohui; Gu, Ning; Zhong, Yang
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
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. (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
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.
The BDGP gene disruption project: single transposon insertions associated with 40% 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 >30,000 fly strains were localized and analyzed relative to predicted Drosophila gene structures. Approximately 6300 lines that maximize genomic coverage were selected to be sent to the Bloomington Stock Center for public distribution, bringing the size of the BDGP gene disruption collection to 7140 lines. It now includes individual lines predicted to disrupt 5362 of the 13,666 currently annotated Drosophila genes (39%). Other lines contain an insertion at least 2 kb from others in the collection and likely mutate additional incompletely annotated or uncharacterized genes and chromosomal regulatory elements. The remaining strains contain insertions likely to disrupt alternative gene promoters or to allow gene misexpression. The expanded BDGP gene disruption collection provides a public resource that will facilitate the application of Drosophila genetics to diverse biological problems. Finally, the project reveals new insight into how transposons interact with a eukaryotic genome and helps define optimal strategies for using insertional mutagenesis as a genomic tool.
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
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
Hugo J. Bellen; Robert W. Levis; Guochun Liao; Yuchun He; Joseph W. Carlson; Garson Tsang; Martha Evans-Holm; P. Robin Hiesinger; Karen L. Schulze; Gerald M. Rubin; Roger A. Hoskins; Allan C. Spradling
Non-Threaded Structural Insert for Component Attachment.
National Technical Information Service (NTIS)
The present invention provides a non-threaded insert operable to transfer or couple a mechanical load to a structure. This non-threaded insert includes an upper insert having a noncircular cross section and a lower insert. The coupled upper and lower inse...
M. J. DeLaune
Analyse d'inserts pour les structures sandwich composites
Analysis of insert in sandwich composite structures. While designing sandwich structures, a major issue is how to introduce loads. This is usually done by several kinds of inserts (resin, metallic, etc.). Five kinds of inserts for sandwich structure panels are dealt with in this study. An experimental study is performed in order to describe the breaking points of inserts while
F. Noirot; J.-F. Ferrero; J.-J. Barrau; B. Castanie; M. Sudre
Analysis of human immunodeficiency virus type 1 promoter insertion in vivo.
Our study reports the occurrence of HIV-1 insertional activation events in vivo. Using a previously described PCR assay, small aliquots of broncho-alveolar lavage (BAL) cells obtained from AIDS patients were analyzed. Nine percent (5/54) of the aliquots contained proviral-host sequence transcripts indicating HIV-1 promoter insertion, whereas multiply spliced HIV-1 mRNAs were found in 28% (15/54) of the aliquots. In four of five events, insertions affect distinct cellular transcription units expressed in a T-cell line. To establish a ratio between provirus integration and promoter insertion events, an in vitro infection study was performed and transcripts containing HIV-1 and K-ras or CD4 gene sequences, respectively, were monitored. Given the randomness of retrovirus integration, 170 sense-oriented HIV integrations into these gene loci were predicted to occur. Three distinct promoter insertion events were observed, indicating that 1.8% of integrated proviruses transcribed adjacent genes. Based on this result and a mean of 257 proviral copies per 10(6) BAL cells, we would expect to observe 25 promoter insertion events in our in vivo study. That only five events were found may be due to the lower transcriptional activity of HIV-1 in vivo than that in cell cultures. PMID:11831720
Raineri, I; SolŔr, M; Senn, H P
Coldmass for lhc dipole insertion magnets..
National Technical Information Service (NTIS)
Brookhaven National Laboratory (BNL) is building a number of magnets for the insertion regions of the Large Hadron Collider (LHC). This paper presents the magnetic design and the expected field quality in 2-in-1 dipole magnets. A unique feature of this co...
A. Gupta Alforque Anerella Kelly Plate Rufer
Dynamic analysis of vibratory insertion process
Purpose ľ The paper aims to investigate theoretically and experimentally the process of compliantly supported peg insertion into a bush for high-speed assembly, when vibrations are provided to the bush in the axial direction, and to analyse the influence of the parameters of the dynamic system and excitation on the assembly process. Design\\/methodology\\/approach ľ The mathematical model of parts vibratory
Sigitas Kilikevicius; Bronius Baksys
Method for Making a Resilient Storage Insert.
National Technical Information Service (NTIS)
The patent application relates to a method for making plastic items, such as resilient storage inserts, and is particularly suitable for making short production runs of plastic items. A relatively inexpensive male die of wood or metal is first made to pro...
C. K. Shaffer J. B. Weiss
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)
Apparatus for inserting flexible pipe into wells
An apparatus and method are described for inserting soft flexible pipe or hole into oil and gas wells under pressure. A capstan-type drive mechanism in combination with a pair of stuffing boxes and a reel guide a soft flexible pipe into the well. The capstan pulls the pipe from a storage reel into a sealed pressure chamber of the capstan
Anatomy and dimensions of rotator cuff insertions
The purpose of this study was to devise and implement an accurate and reproducible method of measuring the area and dimensions of the rotator cuff tendon insertions and their distance from the articular surface. Twenty fresh-frozen cadaveric upper-extremity specimens were divided into 2 groups of 10. In group 1 the specimens were dissected, leaving only the most distal rotator cuff
Jeffrey R Dugas; Deirdre A Campbell; Russell F Warren; Bruce H Robie; Peter J Millett
Thermal Performance of the XRS Helium Insert
NASA Technical Reports Server (NTRS)
The X-Ray Spectrometer (XRS) is an instrument on the Japanese Astro-E satellite, scheduled for launch early in the year 2000. The XRS Helium Insert comprises a superfluid helium cryostat, an Adiabatic Demagnetization Refrigerator (ADR), and the XRS calorimeters with their cold electronics. The calorimeters are capable of detecting X-rays over the energy range 0.1 to 10 keV with a resolution of 12 eV. The Helium Insert completed its performance and verification testing at Goddard in January 1999. It was shipped to Japan, where it has been integrated with the neon dewar built by Sumitomo Heavy Industries. The Helium Insert was given a challenging lifetime requirement of 2.0 years with a goal of 2.5 years. Based on the results of the thermal performance tests, the predicted on-orbit lifetime is 2.6 years with a margin of 30%. This is the result of both higher efficiency in the ADR cycle and the low temperature top-off, more than compensating for an increase in the parasitic heat load. This paper presents a summary of the key design features and the results of the thermal testing of the XRS Helium Insert.
Breon, Susan R.; DiPirro, Michael J.; Tuttle, James G.; Shirron, Peter J.; Warner, Brent A.; Boyle, Robert F.; Canavan, Edgar R.
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.
Pressure-Localizing Inserts for Bagging Laminations
NASA Technical Reports Server (NTRS)
Devices compress composite laminates to conform to tight inside corners of molds. Proposed use of pressure-localizing insert allows composite laminates to be compacted into tight corners by conventional pressure or vacuum-bagging techniques. Because of manufacturing technique, larger selection of part shapes becomes amenable to lamination.
Schmaling, Dave; Carter, Donald
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.
Consolidation and disposal of PWR fuel inserts
Design and licensing of the Surry Power Station Independent Spent Fuel Storage Installation was initiated in 1982 by Virginia Power as part of a comprehensive strategy to increase spent fuel storage capacity at the Station. Designed to use large, metal dry storage casks, the Surry Installation will accommodate 84 such casks with a total storage capacity of 811 MTU of spent pressurized water reactor fuel assemblies. Virginia Power provided three storage casks for testing at the Idaho National Engineerinq Laboratory's Test Area North and the testing results have been published by the Electric Power Research Institute. Sixty-nine spent fuel assemblies were transported in truck casks from the Surry Power Station to Test Area North for testing in the three casks. Because of restrictions imposed by the cask testing equipment at Test Area North, the irradiated insert components stored in these fuel assemblies at Surry were removed prior to transport of the fuel assemblies. Retaining these insert components proved to be a problem because of a shortage of spent fuel assemblies in the spent fuel storage pool that did not already contain insert components. In 1987 Virginia Power contracted with Chem-Nuclear Systems, Inc. to process and dispose of 136 irradiated insert components consisting of 125 burnable poison rod assemblies, 10 thimble plugging devices and 1 part-length rod cluster control assembly. This work was completed in August and September 1987, culminating in the disposal at the Barnwell, SC low-level radioactive waste facility of two CNS 3-55 liners containing the consolidated insert components.
Wakeman, B.H. (Virginia Electric and Power Co., Glen Allen, VA (United States))
Sex-specific aspects of endogenous retroviral insertion and deletion
Background We wish to understand how sex and recombination affect endogenous retroviral insertion and deletion. While theory suggests that the risk of ectopic recombination will limit the accumulation of repetitive DNA in areas of high meiotic recombination, the experimental evidence so far has been inconsistent. Under the assumption of neutrality, we examine the genomes of eighteen species of animal in order to compute the ratio of solo-LTRs that derive from insertions occurring down the male germ line as opposed to the female one (male bias). We also extend the simple idea of comparing autosome to allosome in order to predict the ratio of full-length proviruses we would expect to see under conditions of recombination linked deletion or otherwise. Results Using our model, we predict the ratio of allosomal to autosomal full-length proviruses to lie between 32 and 23 under increasing male bias in mammals and between 1 and 2 under increasing male bias in birds. In contrast to our expectations, we find that a pattern of male bias is not universal across species and that there is a frequent overabundance of full-length proviruses on the allosome beyond the ratios predicted by our model. Conclusions We use our data as a whole to argue that full-length proviruses should be treated as deleterious mutations or as effectively neutral mutations whose persistence in a full-length state is linked to the rate of meiotic recombination and whose origin is not universally male biased. These conclusions suggest that retroviral insertions on the allosome may be more prolific and that it might be possible to identify mechanisms of replication that are enhanced in the female sex.
Mapping of transposable element Dissociation inserts in Brassica oleracea following plant regeneration from streptomycin selection of callus.
To investigate the potential of heterologous transposons as a gene-tagging system in broccoli (Brassica oleracea var. italica), we have introduced a Dissociation ( Ds)-based two-element transposon system. Ds has been cloned into a 35S-SPT excision-marker system, with transposition being driven by an independent 35S-transposase gene construct. In three successive selfed generations of plants, there was no evidence of germinal-excision events. In a previous study, we overcame this apparent inability to produce B. oleracea plants with germinal excisions by performing a novel tissue-culture technique to select for fully green shoots from seed with somatic excision events. The results showed a very high efficiency of regeneration of fully green plants (up to 65%), and molecular analysis showed that the plants contained the equivalent of a germinal-excision event. In this study, we followed the previous work by using inverse and nested PCR to generate probes of flanking genomic DNA adjacent to independently reinserted Ds elements, and these were hybridised to DNA from a double-haploid mapping population of B. oleracea. Seventeen Ds insertions and the original Ds T-DNA site have been localised, and these are spread over six (out of nine) linkage groups. Distribution of inserts show that 15 were found on a different linkage group to the original 'launch' site, and of these 11 were found to be clustered on two separate groups. Previous studies in other plant species have found that germinal excision of Ds predominantly moves to sites linked close to the donor site. However, this study shows a potential to produce plants with Ds insertion scattered over many unlinked sites. PMID:15014879
McKenzie, Neil; Dale, Philip J
DNA representation of variegating heterochromatic P-element inserts in diploid and polytene tissues of Drosophila melanogaster
Position-effect variegation (PEV) is the mosaic expression of a euchromatic gene brought into juxtaposition with heterochromatin. Fourteen different transformedDrosophila melanogaster lines with variegating P-element inserts were used to examine the DNA levels of these transgenes. Insert sites include pericentric, telomeric and fourth chromosome regions. Southern blot analyses showed that the heterochromatichsp26 transgenes are underrepresented 1.3- to 33-fold in polytene tissue
Lori L. Wallrath; Vamsi P. Guntur; Lisa E. Rosman; Sarah C. R. Elgin
National Synchrotron Light Source-II (NSLS-II) will be a medium energy storage ring of 3GeV electron beam energy with sub-nm.rad horizontal emittance and top-off capability at 500mA. Damping wigglers will be used not only to reduce the beam emittance but also used as broadband sources for users. Cryo-Permanent Magnet Undulators (CPMUs) are considered for hard X-ray linear device, and permanent magnet based elliptically polarized undulators (EPUs) for variable polarization devices for soft X-ray. 6T superconducting wiggler with minimal fan angle will be installed in the second phase as well as quasi-periodic EPU for VUV and possibly high-temperature superconducting undulator. R&D plans have been established to pursue the performance enhancement of the baseline devices and to design new types of insertion devices. A new insertion device development laboratory will also be established.
A small, insertable oven for boronization
A small insertable oven for benchmarking the boronizing characteristics of solid compounds, such as decaborane and carborane, has been developed for the Phaedrus-T tokamak. Assembly and installation of the oven are relatively easy as the oven design utilizes a Langmuir probe drive assembly, which is standard equipment on most tokamaks and allows the oven to be inserted into the tokamak without requiring a vent. Films deposited by heating carborane into the vapor state with the oven are found to be spatially nonuniform in both thickness and in the ratio of boron to carbon as compared to films deposited with trimethylboron, a gaseous compound. Overall plasma performance is not found to be greatly affected by whether decaborane, carborane or trimethylboron is used for boronization in Phaedrus-T. {copyright} {ital 1996 American Institute of Physics.} {lt}ii;010512{gt}
Brouchous, D.A.; Diebold, D.A.; Doczy, M.L. [Department of Nuclear Engineering and Engineering Physics, University of Wisconsin, 1500 Johnson Drive, 337 ERB, Madison, Wisconsin 53706-1687 (United States)] [Department of Nuclear Engineering and Engineering Physics, University of Wisconsin, 1500 Johnson Drive, 337 ERB, Madison, Wisconsin 53706-1687 (United States)
Insertional Achilles tendinosis: pathogenesis and treatment.
Insertional Achilles tendinopathy can be a painful debilitating condition that should initially be treated non-operatively. If pain becomes chronic and debilitating, despite appropriate conservative treatment, debridement of the diseased portion of the Achilles tendon and removal of the impinging calcaneal prominence and transfer of the flexor hallucis longus through a single incision can be a reliable pain relieving procedure with relatively high patient satisfaction. PMID:19857838
Den Hartog, Bryan D
Assessment of the ultrasonic dental scaler insert
The ultrasonic dental scaler is an invaluable tool in the fight against dental calculus and periodontal disease. The purpose of this study was to assess the natural frequency of the straight internal ultrasonic scaler insert 30KÖ FSI«-SLIÖ-10S (Dentsply International, York, USA) and to measure the displacement observed at the tip when operated under different generator settings. Two approaches were used:
S. C Trenter; A. D Walmsley; G Landini; J. M Shippen
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.
Coldmass for LHC Dipole Insertion Magnets
Brookhaven National Laboratory (BNL) is building a number of magnets for the insertion regions of the Large Hadron Collider (LHC). This paper presents the magnetic design and the expected field quality in 2-in-1 dipole magnets. A unique feature of this cold mass design is the use of an oblate-shaped yoke. This concept permits a variety of BNL-built magnets to have
R. Gupta; R. Alforque; M. Anerella; E. Kelly; S. Plate; C. Rufer; P. Wanderer; E. Willen; K. C. Wu
Magnetic measurements on insertion devices at NSLS
Magnetic measurements on three insertion devices for use in the NSLS x-ray ring are described. Hall probes were used for field mapping, and rotating long coils were used to determine integrated multipole coefficients. Results are presented for an iron pole-REC hybrid undulator, a vanadium permendur pole-REC wiggler and a 5 Tesla superconducting wiggler. 6 refs., 8 figs., 2 tabs.
G. Decker; J. Galayda; M. Kitamura; L. Solomon
Rack Insertion End Effector (RIEE) guidance
NASA Astrophysics Data System (ADS)
NASA-KSC has developed a mechanism to handle and insert Racks into the Space Station Logistic Modules. This mechanism consists of a Base with 3 motorized degrees of freedom, a 3 section motorized Boom that goes from 15 to 44 feet in length, and a Rack Insertion End Effector (RIEE) with 5 hand wheels for precise alignment. During the 1993 NASA-ASEE Summer Faculty Fellowship Program at KSC, I designed an Active Vision (Camera) Arrangement and developed an algorithm to determine (1) the displacements required by the Room for its initial positioning and (2) the rotations required at the five hand-wheels of the RIEE, for the insertion of the Rack, using the centroids fo the Camera Images of the Location Targets in the Logistic Module. Presently, during the summer of '94, I completed the preliminary design of an easily portable measuring instrument using encoders to obtain the 3-Dimensional Coordinates of Location Targets in the Logistics Module relative to the RIEE mechanism frame. The algorithm developed in '93 can use the output of this instrument also. Simplification of the '93 work and suggestions for the future work are discussed.
Malladi, Narasimha S.
Rack Insertion End Effector (RIEE) guidance
NASA Technical Reports Server (NTRS)
NASA-KSC has developed a mechanism to handle and insert Racks into the Space Station Logistic Modules. This mechanism consists of a Base with 3 motorized degrees of freedom, a 3 section motorized Boom that goes from 15 to 44 feet in length, and a Rack Insertion End Effector (RIEE) with 5 hand wheels for precise alignment. During the 1993 NASA-ASEE Summer Faculty Fellowship Program at KSC, I designed an Active Vision (Camera) Arrangement and developed an algorithm to determine (1) the displacements required by the Room for its initial positioning and (2) the rotations required at the five hand-wheels of the RIEE, for the insertion of the Rack, using the centroids fo the Camera Images of the Location Targets in the Logistic Module. Presently, during the summer of '94, I completed the preliminary design of an easily portable measuring instrument using encoders to obtain the 3-Dimensional Coordinates of Location Targets in the Logistics Module relative to the RIEE mechanism frame. The algorithm developed in '93 can use the output of this instrument also. Simplification of the '93 work and suggestions for the future work are discussed.
Malladi, Narasimha S.
A Miniaturized Multilayered Marchand Balun Using Coupled Artificial Transmission Lines
A miniaturized Marchand balun is investigated in this letter. The Marchand balun is realized on a six-layered printed circuit board by integration of two coupled artificial transmission line sections. An additional artificial line is inserted in-between the coupled lines for compensating the asymmetry. With the help of the even odd mode analysis, the electrical characteristics of the coupled lines are
Tzyh-Ghuang Ma; Yu-Ting Cheng
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))
Software-implemented fault insertion: An FTMP example
NASA Technical Reports Server (NTRS)
This report presents a model for fault insertion through software; describes its implementation on a fault-tolerant computer, FTMP; presents a summary of fault detection, identification, and reconfiguration data collected with software-implemented fault insertion; and compares the results to hardware fault insertion data. Experimental results show detection time to be a function of time of insertion and system workload. For the fault detection time, there is no correlation between software-inserted faults and hardware-inserted faults; this is because hardware-inserted faults must manifest as errors before detection, whereas software-inserted faults immediately exercise the error detection mechanisms. In summary, the software-implemented fault insertion is able to be used as an evaluation technique for the fault-handling capabilities of a system in fault detection, identification and recovery. Although the software-inserted faults do not map directly to hardware-inserted faults, experiments show software-implemented fault insertion is capable of emulating hardware fault insertion, with greater ease and automation.
Czeck, Edward W.; Siewiorek, Daniel P.; Segall, Zary Z.
Detection of transgenes in three genetically modified rice lines by fluorescence in situ hybridization
Fluorescence in situ hybridization (FISH) using T-DNA probes was applied to localize transgenes onto specific chromosomes\\u000a and confirm the steady integration of transferred genes in three genetically modified (GM) rice lines, LS28 (event LS30-32-20-1),\\u000a Cry1Ac1 (event C7-1-9-1) and LS28ÎCry1Ac1 (event L\\/C1-1-3-1), which are a rice leaf blast-resistant single trait GM line,\\u000a a leaf folder-resistant single trait GM line, and a
Hye Mi Park; Eun Jin Jeon; Nomar Espinosa Waminal; Kong Sik Shin; Soon Jong Kweon; Beom-Seok Park; Seok Cheol Suh; Hyun Hee Kim
Numerical Simulation of Milling Using Inserts with Structural Gradients.
National Technical Information Service (NTIS)
Temperatures and stresses in milling inserts with structural gradients were calculated with finite element analysis. Experimentally determined cutting forces, chip thickness and tool/chip contact length were used as input data to the simulations. Inserts ...
A. Thuvander H. Thoors
Operation of synchrotron light sources with multiple insertion devices.
National Technical Information Service (NTIS)
The stability requirements of the next generation of synchrotron radiation facilities have been achieved on insertion device beamlines of existing rings. However, one insertion device (ID) affects the stored beam and hence the performance of all the other...
J. Galayda A. M. Fauchet
Aggregation of prion protein with insertion mutations is proportional to the number of inserts
Mutation in the prion gene, PRNP, accounts for approx. 10ľ15% of human prion diseases. However, little is known about the mechanisms by which a mutant prion protein (PrP) causes disease. We compared the biochemical properties of a wild-type human prion protein, rPrPC (recombinant wild-type PrP), which has five octapeptide-repeats, with two recombinant human prion proteins with insertion mutations, one with three more octapeptide repeats, rPrP8OR, and the other with five more octapeptide repeats, rPrP10OR. We found that the insertion mutant proteins are more prone to aggregate, and the degree and kinetics of aggregation are proportional to the number of inserts. The octapeptide-repeat and ?-helix 1 regions are important in aggregate formation, because aggregation is inhibited with monoclonal antibodies that are specific for epitopes in these regions. We also showed that a small amount of mutant protein could enhance the formation of mixed aggregates that are composed of mutant protein and wild-type rPrPC. Accordingly, rPrP10OR is also more efficient in promoting the aggregation of rPrPC than rPrP8OR. These findings provide a biochemical explanation for the clinical observations that the severity of the disease in patients with insertion mutations is proportional to the number of inserts, and thus have implications for the pathogenesis of inherited human prion disease.
Yu, Shuiliang; Yin, Shaoman; Li, Chaoyang; Wong, Poki; Chang, Binggong; Xiao, Fan; Kang, Shin-Chung; Yan, Huimin; Xiao, Gengfu; Tien, Po; Sy, Man-Sun
Inserts ľ megafillers in composite restorations: a literature review
Modern insert systems have been available on the dental market since the late 1980s. In general, two major systems can be\\u000a distinguished: those with and those without preparation instruments in combination with matching standardized inserts. It\\u000a is claimed that one of the advantages of insert systems is the better marginal adaptation of insert\\/composite restorations\\u000a compared to mere composite restorations: the
M. Federlin; B. Thonemann; G. Schmalz
Transposon Insertion Finder (TIF): a novel program for detection of de novo transpositions of transposable elements
Background Transposition event detection of transposable element (TE) in the genome using short reads from the next-generation sequence (NGS) was difficult, because the nucleotide sequence of TE itself is repetitive, making it difficult to identify locations of its insertions by alignment programs for NGS. We have developed a program with a new algorithm to detect the transpositions from NGS data. Results In the process of tool development, we used next-generation sequence (NGS) data of derivative lines (ttm2 and ttm5) of japonica rice cv. Nipponbare, regenerated through cell culture. The new program, called a transposon insertion finder (TIF), was applied to detect the de novo transpositions of Tos17 in the regenerated lines. TIF searched 300 million reads of a line within 20ámin, identifying 4 and 12 de novo transposition in ttm2 and ttm5 lines, respectively. All of the transpositions were confirmed by PCR/electrophoresis and sequencing. Using the program, we also detected new transposon insertions of P-element from NGS data of Drosophila melanogaster. Conclusion TIF operates to find the transposition of any elements provided that target site duplications (TSDs) are generated by their transpositions.
Low-Weight Inserts for Aluminum Honeycomb Panels
NASA Technical Reports Server (NTRS)
Fiber/epoxy composites used in place of solid aluminum. New proposal suggests to make inserts out of such lightweight, high strength materials as fiber/epoxy composites or high density aluminum honeycomb. Composite insert is same size and shape as aluminum insert it replaces.
Bumgarner, G. S.; Reed, M. W.
Development of the hybrid insert for composite sandwich satellite structures
Composite sandwich structures which are widely employed in satellite structures require many inserts for assembly. In this study, a new lightweight insert for sandwich structures was developed by reinforcing the web of insert with high strength carbon composite to increase the loading capability with reduced mass. Finite element analysis was performed to numerically predict the load capability of the new
Jun Woo Lim; Dai Gil Lee
Insertion loss of sound waves through composite acoustic window materials
This study evaluated the insertion loss of sound waves through composite acoustic window materials for sonar dome applications, and examined the influence of dome shape on the sonar performance. The insertion loss of sound waves through acoustic window materials was experimentally measured as a function of frequency at normal incidence. The insertion loss was also theoretically estimated with three- and
Ji-Hye Lee; Byoung-Nam Kim; Ku-Kyun Shin; Suk Wang Yoon
Chronic insertional Achilles tendinopathy: surgical outcomes
Summary Background and objective: insertional Achilles tendinopathy is a common condition among athletes and joggers. One fifth of the injuries involves the insertion of the tendon. The etiology is either due to mechanical overuse related to sports activity, or a systemic inflammatory disease. The clinical appearance includes pain and movement restriction. The primary treatment is conservative. The surgery referred to in this study (Calcaneal Osteotomy) is performed by decompression of the posterior margin of the calcaneus. If the tendon is degenerative, debridement is needed. There is controversy on the surgical outcome and the surgical approach. A retrospective analytic observational study. Methods: 20 patients who were diagnosed with IAT (21 feet) and were operated on between the years 2000ľ2007 by calcaneal osteotomy. Main outcome measures: the primary measure of success was diminished pain. It was demonstrated in the AOFAS score and in the VAS scale of pain. Results: the average grade in the AOFAS questionnaire improved by 20 points, and the average grading of pain in the VAS scale was decreased by 4.21. The median satisfaction rate was 8, the average was 5.81. 62% of the patients would repeat the surgery/recommend it. We found a significant relationship (p=0.022) between patients who avoided sports activity while suffering from insertional Achilles tendinopathy and the satisfaction rate from the surgery. Conclusions: using the calcaneal osteotomy technique as a surgical treatment for IAT is successful, and improves measures of pain and function. Low compliance with avoiding sports activity while suffering from an IAT might lead to a need for surgery.
Oshri, Yael; Palmanovich, Ezequiel; Brin, Yaron Shagra; Karpf, Ronen; Massarwe, Sabri; Kish, Benny; Nyska, Meir
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.
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
Compiler-assisted static checkpoint insertion
NASA Technical Reports Server (NTRS)
This paper describes a compiler-assisted approach for static checkpoint insertion. Instead of fixing the checkpoint location before program execution, a compiler enhanced polling mechanism is utilized to maintain both the desired checkpoint intervals and reproducible checkpoint 1ocations. The technique has been implemented in a GNU CC compiler for Sun 3 and Sun 4 (Sparc) processors. Experiments demonstrate that the approach provides for stable checkpoint intervals and reproducible checkpoint placements with performance overhead comparable to a previously presented compiler assisted dynamic scheme (CATCH) utilizing the system clock.
Long, Junsheng; Fuchs, W. K.; Abraham, Jacob A.
Cassini magnetometer observations during Saturn orbit insertion.
Cassini's successful orbit insertion has provided the first examination of Saturn's magnetosphere in 23 years, revealing a dynamic plasma and magnetic environment on short and long time scales. There has been no noticeable change in the internal magnetic field, either in its strength or its near-alignment with the rotation axis. However, the external magnetic field is different compared with past spacecraft observations. The current sheet within the magnetosphere is thinner and more extended, and we observed small diamagnetic cavities and ion cyclotron waves of types that were not reported before. PMID:15731444
Dougherty, M K; Achilleos, N; Andre, N; Arridge, C S; Balogh, A; Bertucci, C; Burton, M E; Cowley, S W H; Erdos, G; Giampieri, G; Glassmeier, K-H; Khurana, K K; Leisner, J; Neubauer, F M; Russell, C T; Smith, E J; Southwood, D J; Tsurutani, B T
Acute Pancreatitis Complicating Intragastric Balloon Insertion
We present a case of acute pancreatitis that happened as a complication of intragastric balloon insertion used for weight reduction in a young obese lady. Although many complications can happen with this type of procedure, this presentation is very rare and may not have been reported previously. The patient responded well to removal of the balloon since her pain resolved completely and her serum amylase normalized. She did not have any obvious cause of pancreatitis other than a significant radiologically proven compression of the pancreas by the balloon.
Mohammed, AbdulRahman Elsheikh; Benmousa, Ali
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)
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)
Use of the Endopath Optiview after Failed Conventional Cannula Insertion
Cannula insertion is considered to fail after three unsuccessful attempts to enter the abdominal cavity with standard cannulas. A prospective clinical trial was conducted to evaluate the usefulness of Endopath Optiview after failed insertion of conventional cannulas. Cannula insertion was unsuccessful in 4 of 267 women undergoing elective laparoscopic sterilization. In all four women the Endopath Optiview was introduced successfully into the abdominal cavity. In two women omental adhesions had to be transversed. The Endopath Optiview is valuable for recognizing and evaluating the path of failed cannula insertions in the abdominal wall. It is crucial to verify intraabdominal adhesions and safely separate them under direct visualization during the insertion. PMID:9074141
Deep insertion - round window approach by using SRA electrode.
Hearing preservation surgery requires specially a traumatic technique. Having some preoperative anatomical data of the size of patient's cochlea surgeon can design his or her insertion depth. In the study we have evaluated a relation between hearing preservation rate and angular insertion depth estimated intraoperatively and postoperatively having measured insertion angle from radiological assessment and calculations given by Escude. There has not been no statistically significant difference between insertion depth angle, either estimated intraoperatively and measured and calculated post-operatively, and hearing preservation rate in the group. This analysis confirms a traumaticy of insertion in hearing preservation surgery. PMID:24869441
Skar?y?ski, H; Matusiak, M; Furmanek, M; Skarzy?ski, P H
Model of a Hollow Cathode Insert Plasma
NASA Technical Reports Server (NTRS)
A 2-D axisymmetric fluid model of the plasma in the insert region of a hollow cathode is presented. The level of sophistication included in the model is motivated in part by the need to determine quantitatively plasma fluxes to the emitter surface. The ultimate goal is to assess whether plasma effects can degrade the life of impregnated inserts beyond those documented throughout the 30-50 year history of vacuum cathode technologies. Results from simulations of a 1.2-cm diameter cathode operating at a discharge current of 25 A, and a gas flow rate of 5 sccm, suggest that approximately 10 A of electron current, and 3.5 A of ion current return to the emitter surface. The total emitted electron current computed by the model is about 35 A. Comparisons with plasma measurements suggest that anomalous heating of the plasma due to two-stream instabilities is possible near the orifice region. Solution to the heavy species energy equation, with classical transport and no viscous effects, predicts heavy species temperatures as high as 2640 K.
Mikellides, Ioannis G.; Katz, Ira; Goebel, Dan M.; Polk, James E.
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.
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)
Mammalian NUMT insertion is non-random
It is well known that remnants of partial or whole copies of mitochondrial DNA, known as Nuclear MiTochondrial sequences (NUMTs), are found in nuclear genomes. Since whole genome sequences have become available, many bioinformatics studies have identified putative NUMTs and from those attempted to infer the factors involved in NUMT creation. These studies conclude that NUMTs represent randomly chosen regions of the mitochondrial genome. There is less consensus regarding the nuclear insertion sites of NUMTs Ś previous studies have discussed the possible role of retrotransposons, but some recent ones have reported no correlation or even anti-correlation between NUMT sites and retrotransposons. These studies have generally defined NUMT sites using BLAST with default parameters. We analyze a redefined set of human NUMTs, computed with a carefully considered protocol. We discover that the inferred insertion points of NUMTs have a strong tendency to have high-predicted DNA curvature, occur in experimentally defined open chromatin regions and often occur immediately adjacent to A?+?T oligomers. We also show clear evidence that their flanking regions are indeed rich in retrotransposons. Finally we show that parts of the mitochondrial genome D-loop are under-represented as a source of NUMTs in primate evolution.
Tsuji, Junko; Frith, Martin C.; Tomii, Kentaro; Horton, Paul
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
Migration of tendinous insertions. I. Cause and mechanism.
The cause and mechanisms of the migration of tendons and ligaments were studied in young rabbits. Three techniques were used: (1) Marking of insertions, the neighbouring periosteum and the diaphysis with metallic markers. (2) Marking of insertion sites by tetracycline as an indicator of osteogenesis. (3) Histological examination. The insertions used in the study were of three different characters: (1) Insertions subject to muscular traction (patellar ligament, quadratus femoris muscle, tibialis anterior muscle). (2) The distal insertions of the medial collateral ligament of the knee, stretched by the activity of the proximal epiphyseal cartilage of the tibia. (3) The proximal and distal insertions of the anterior annular ligament of the tibia, inserted solely in bone and periosteum. The cause of migration is the growth of periosteum dragging the insertions during its stretching, caused itself by the activity of the epiphyseal plates. The local mechanism governing migration while ensuring a continuous connexion with the bone is not the same in all sites. It depends upon the character of the bony surface at the insertion and of the function of the insertion zone, which can be osteogenic, resorptive or both. A plexus of precollagenous fibres is present at all resorptive insertion sites, and at some of the osteogenic sites. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8
Dorfl, J
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.
Transcutaneous insertion of the Chitwood« clamp in case of minimally invasive cardiac surgery. Personal experience
Summary We describe a simple trick consisting of an insertion of a trans-thoracic Chitwood« clamp (Scanlan International, Inc, St Paul, MN, USA) through a small skin incision (<1cm), via the second intercostal space along the anterior axillary line, in case of minimally invasive mitral surgery, through right thoracotomy. This trick has been used in more than 100 patients and no complication or discomfort have been reported so far.
Transcutaneous insertion of the Chitwood« clamp in case of minimally invasive cardiac surgery. Personal experience.
We describe a simple trick consisting of an insertion of a trans-thoracic Chitwood« clamp (Scanlan International, Inc, St Paul, MN, USA) through a small skin incision (<1cm), via the second intercostal space along the anterior axillary line, in case of minimally invasive mitral surgery, through right thoracotomy. This trick has been used in more than 100 patients and no complication or discomfort have been reported so far. PMID:24629816
Sansone, F; Ceresa, F; PatanŔ, F
Human LINE retrotransposons generate processed pseudogenes
Long interspersed elements (LINEs) are endogenous mobile genetic elements1-4 that have dispersed and accumulated in the genomes of higher eukaryotes via germline transposition, with up to 100,000 copies in mammalian genomes. In humans, LINEs are the major source of insertional mutagenesis, being involved in both germinal and somatic mutant phenotypes4. Here we show that the human LINE retrotransposons, which transpose
CÚcile Esnault; JoŰl Maestre; Thierry Heidmann
Non-insertional tendinopathy of the subscapularis
Purpose: (1) Describe a previously unreported finding involving the intra-articular portion of the subscapularis, the Conrad lesion. (2) Describe a novel classification system for the spectrum of non-insertional tendinopathy of the subscapularis. (3) Report the outcomes of surgical treatment of this spectrum of pathology. Materials and Methods: Outcomes of 34 patients (23 males and 11 females, mean age 60.5 ▒ 7.5) with non-insertional tendinopathy of the subscapularis treated arthroscopically were retrospectively reviewed. All patients had anterior shoulder pain with no weakness during belly-press testing and no subscapularis footprint involvement on magnetic resonance imaging. All patients were managed with subscapularis tendon debridement and side-to-side repair along with treatment of concomitant pathology. Results: Seven patients had a Type I lesion (so-called Conrad lesion) ľ a nodule on the leading edge of the subscapularis. Eighteen patients had a Type II lesion ľ a visible split tear with degeneration in the upper Ż of the intra-articular tendon. Nine patients had a Type III lesion ľ more extensive splitting in the tendon with advanced tendon degeneration. At a mean follow-up of 24 months, 97% of patients were completely satisfied. Significant improvements were seen in forward elevation (152 ▒ 12░ to 172 ▒ 5░, P < 0.001) and visual analog scale pain scores (5.9 ▒ 1.7-0.6 ▒ 1.0, P < 0.001). Internal rotation strength and external rotation motion at the side were maintained. ASES scores averaged 95.4 ▒ 7.4, disabilities of arm, shoulder and hand scores averaged 6.19 ▒ 9.8, Western Ontario Rotator Cuff scores averaged 91.7 ▒ 9.3 and the average University of California at Los Angeles score was 33.1 ▒ 2.4. Conclusions: We present a previously unreported finding of the subscapularis, the Conrad lesion, along with a novel classification system for non-insertional tendinopathy of the subscapularis. Arthroscopic treatment of this spectrum of tendinopathy along with concomitant shoulder pathology eliminated pain and improved patient outcomes without detrimental effects. Level of Evidence: IV, Retrospective Case Series.
Dierckman, Brian D.; Shah, Nirav R.; Larose, Connor R.; Gerbrandt, Stacey; Getelman, Mark H.
Intermetallic insertion anodes for lithium batteries.
Binary intermetallic compounds containing lithium, or lithium alloys, such as Li{sub x}Al, Li{sub x}Si and Li{sub x}Sn have been investigated in detail in the past as negative electrode materials for rechargeable lithium batteries. It is generally acknowledged that the major limitation of these systems is the large volumetric expansion that occurs when lithium reacts with the host metal. Such large increases in volume limit the practical use of lithium-tin electrodes in electrochemical cells. It is generally recognized that metal oxide electrodes, MO{sub y}, in lithium-ion cells operate during charge and discharge by means of a reversible lithium insertion/extraction process, and that the cells offer excellent cycling behavior when the crystallographic changes to the unit cell parameters and unit cell volume of the Li{sub x}MO{sub y} electrode are kept to a minimum. An excellent example of such an electrode is the spinel Li{sub 4}Ti{sub 5}O{sub 12}, which maintains its cubic symmetry without any significant change to the lattice parameter (and hence unit cell volume) during lithium insertion to the rock-salt composition Li{sub 7}Ti{sub 5}O{sub 12}. This spinel electrode is an example of a ternary Li{sub x}MO{sub y} system in which a binary MO{sub y} framework provides a stable host structure for lithium. With this approach, the authors have turned their attention to exploring ternary intermetallic systems Li{sub x}MM{prime} in the hope of finding a system that is not subject to the high volumetric expansion that typifies many binary systems. In this paper, the authors present recent data of their investigations of lithium-copper-tin and lithium-indium-antimonide electrodes in lithium cells. The data show that lithium can be inserted reversibly into selected intermetallic compounds with relatively small expansion of the lithiated intermetallic structures.
Thackeray, M. M.; Vaughey, J.; Johnson, C. S.; Kepler, K. D.
This paper describes plans to upgrade insertion devices (IDs) at the National Synchrotron Light Source (NSLS), Brookhaven National Laboratory, U.S.A. The aging wiggler (W120) at X25 is being replaced by a 1 m long in-vacuum mini-gap undulator (MGU-X25) optimized for a dedicated macromolecular crystallography program. A new, 1/3 m long, undulator (MGU or SCU-X9), will be installed between a pair of RF cavities at X9, and will serve a new beamline dedicated for small angle x-ray scattering (SAXS). Both IDs will have provision for cryocooling the NdFeB hybrid arrays to 150K to raise the field and K-value and to obtain better spectral coverage. Design issues of the devices and other considerations, especially magnetic measurement at low temperature, will be discussed.
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.
Petri, Michelina; Flores, Noe; Rogge, Ryan A.; Cascarina, Sean M.; Ross, Eric D.
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.
Detecting Statistically Significant Common Insertion Sites in Retroviral Insertional Mutagenesis Screens
Retroviral insertional mutagenesis screens, which identify genes involved in tumor development in mice, have yielded a substantial number of retroviral integration sites, and this number is expected to grow substantially due to the introduction of high-throughput screening techniques. The data of various retroviral insertional mutagenesis screens are compiled in the publicly available Retroviral Tagged Cancer Gene Database (RTCGD). Integrally analyzing these screens for the presence of common insertion sites (CISs, i.e., regions in the genome that have been hit by viral insertions in multiple independent tumors significantly more than expected by chance) requires an approach that corrects for the increased probability of finding false CISs as the amount of available data increases. Moreover, significance estimates of CISs should be established taking into account both the noise, arising from the random nature of the insertion process, as well as the bias, stemming from preferential insertion sites present in the genome and the data retrieval methodology. We introduce a framework, the kernel convolution (KC) framework, to find CISs in a noisy and biased environment using a predefined significance level while controlling the family-wise error (FWE) (the probability of detecting false CISs). Where previous methods use one, two, or three predetermined fixed scales, our method is capable of operating at any biologically relevant scale. This creates the possibility to analyze the CISs in a scale space by varying the width of the CISs, providing new insights in the behavior of CISs across multiple scales. Our method also features the possibility of including models for background bias. Using simulated data, we evaluate the KC framework using three kernel functions, the Gaussian, triangular, and rectangular kernel function. We applied the Gaussian KC to the data from the combined set of screens in the RTCGD and found that 53% of the CISs do not reach the significance threshold in this combined setting. Still, with the FWE under control, application of our method resulted in the discovery of eight novel CISs, which each have a probability less than 5% of being false detections.
de Ridder, Jeroen; Uren, Anthony; Kool, Jaap; Reinders, Marcel; Wessels, Lodewyk
49 CFR Appendix E to Part 227 - Use of Insert Earphones for Audiometric Testing
Code of Federal Regulations, 2011 CFR
...EXPOSURE Pt. 227, App. E Appendix E to Part 227ŚUse of Insert...insertion depth (fit). C. Insert earphones shall...with insert earphones. C. Transitioning Employees...process of developing an STS (e.g., demonstrate a 7...
49 CFR Appendix E to Part 227 - Use of Insert Earphones for Audiometric Testing
Code of Federal Regulations, 2012 CFR
...EXPOSURE Pt. 227, App. E Appendix E to Part 227ŚUse of Insert...insertion depth (fit). C. Insert earphones shall...with insert earphones. C. Transitioning Employees...process of developing an STS (e.g., demonstrate a 7...
Ocular inserts -- Advancement in therapy of eye diseases
The ocular insert represents a significant advancement in the therapy of eye disease. Ocular inserts are defined as sterile, thin, multilayered, drug-impregnated, solid or semisolid consistency devices placed into the cul-de-sac or conjuctival sac, whose size and shape are especially designed for ophthalmic application. They are composed of a polymeric support that may or may not contain a drug. The drug can later be incorporated as dispersion or a solution in the polymeric support. They offer several advantages as increased ocular residence and sustained release of medication into the eye. The insert includes a body portion sized to position within a lachrymal canaliculus of the eyelid. The inserts are classified according to their solubility as insoluble, soluble, or bioerodible inserts. The release of drug from the insert depends upon the diffusion, osmosis, and bioerosion of the drug, and this article is an attempt to present a brief about this newer drug delivery system.
Kumari, Anita; Sharma, Pramod K.; Garg, Vipin K.; Garg, Garima
Insertional mutagenesis of Drosophila heterochromatin with single P elements.
Insertional mutagenesis with transposable P elements has greatly facilitated the identification and analysis of genes located throughout the 70% of the Drosophila melanogaster genome classified as euchromatin. In contrast, genetically marked P elements have only rarely been shown to transpose into heterochromatin. By carrying out single P element insertional mutagenesis under conditions where position-effect variegation was suppressed, we efficiently generated strains containing insertions at diverse sites within centromeric and Y-chromosome heterochromatin. The tendency of P elements to transpose locally was shown to operate within heterochromatin, and it further enhanced the recovery of heterochromatic insertions. Three of the insertions disrupted vital genes known to be present at low density in heterochromatin. Strains containing single P element insertions will greatly facilitate the structural and functional analysis of this poorly understood genomic component. PMID:8170943
Zhang, P; Spradling, A C
ôProactiveö Management of Percutaneously Inserted Central Catheters Results in Decreased Incidence of Infection in the ELBW Population
OBJECTIVE: Extremely low birth weight (ELBW) infants often acquire catheter-related infections (CRIs) when a percutaneously inserted central catheter (PICC) is used for parenteral nutrition or drug administration. Our objective was to compare the incidence of CRIs after we established a ôPICC Maintenance Teamö for the proactive management Ś compared to expectant management Ś of these lines.STUDY DESIGN: We did a
Sergio G Golombek; Annie J Rohan; Boriana Parvez; Anne L Salice; Edmund F LaGamma
Rule insertion and rule extraction from evolving fuzzy neural networks: algorithms and applications for building adaptive, intelligent expert systems
Discusses the concept of intelligent expert systems and suggests tools for building an adaptable, in an online or in an off-line mode, rule base during the system operation in a changing environment. It applies evolving fuzzy neural networks (EFuNNs) as associative memories for the purpose of dynamic storing and modifying a rule base. Algorithms for rule extraction and rule insertion
Nikola Kasabov; Brendon Woodford
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
Needle Insertion Modelling for the Interactive Simulation of Percutaneous Procedures
A novel interactive virtual needle insertion simulation is presented. The insertion model simulates three-degree-of-freedom\\u000a needle motion, physically-based needle forces, linear elastostatic tissue deformation and needle flexibility for the planning\\u000a and training of percutaneous therapies and procedures. To validate the approach, an experimental system for measuring planar\\u000a tissue deformation during needle insertions has been developed and is presented. A real-time simulation
Simon P. Dimaio; S. E. Salcudean
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
A method for timing constrained redundant via insertion
NASA Astrophysics Data System (ADS)
Redundant via (RV) insertion is a useful mechanism to enhance via reliability. However, when extra vias are inserted into the design, the circuit timing might be changed. Therefore, how to insert RV under the timing constraints is the main challenge. In this paper, we introduce a new model to compute the distance between a RV and the corresponding single via, put forward a new RV type, which is called the long length via (LLV), and then present an improved RV insertion method considering the timing constraints. This computing model can certify that the timing, which is obtained after inserting a RV, is not greater than the original timing. Meanwhile, the new RV type LLV can increase the possibility of RV insertion; this method provides a global perspective for the RV insertion. Considering the timing constraints, the total redundant via insertion rate is 85.38% in the MIS-based method, while our proposed method can obtain a high insertion rate 88.79% for the tested circuits.
Junping, Wang; Dan, Xu; Yongbang, Su
Does Feeding Tube Insertion and its Timing Improve Survival?
Background/Objectives The objective of this study was to examine survival with and without a percutaneous endoscopic gastrostomy (PEG) feeding tube using rigorous methods to account for selection bias. A second objective was to examine whether the timing of feeding tube insertion affected survival. Design Prospective cohort study Setting All U.S. Nursing Homes Participants 36,492 nursing home residents with advanced cognitive impairment from dementia and new problems eating studied between 1999ľ2007. Measurements Survival following the development of the need for eating assistance and feeding tube insertion. Results Of the 36,492 nursing home (NH) residents (88.4% white, mean age 84.9, 87.4% with one feeding tube risk factor), 1,957 (5.4%) had a feeding tube inserted within 1 year of developing eating problems. After multivariate analysis correcting for selection bias with propensity score weights, no difference was found in the survival of the 2 groups (AHR 1.03, 95% CI 0.94ľ1.13). Among residents who were tube-fed, the timing of PEG tube insertion relative to the onset of eating problems was not associated with improved survival post feeding tube insertion (AHR 1.01, 95% CI 0.86ľ1.20) comparing those persons with a PEG inserted within a month of developing an eating problem compared to later (4 months) insertion. Conclusion This national study confirms that neither insertion of PEG feeding tubes nor the timing of the insertion improve survival.
Teno, Joan M.; Gozalo, Pedro L.; Mitchell, Susan L.; Kuo, Sylvia; Rhodes, Ramona L.; Bynum, Julie P.W.; Mor, Vincent
In vivo oxidation in retrieved highly crosslinked tibial inserts.
The current study determined (I) the environment where oxidation in a series of retrieved, HXL UHMWPE tibial inserts occurred (in vivo or postexplant); and (II) the effect of fabrication variables (irradiation source, irradiation dose) and thermal processing after irradiation (annealing or remelting) on oxidation resistance. Hypotheses examined are (1) HXL UHMWPE tibial inserts have potential to oxidize in vivo, and (2) annealed HXL UHMWPE oxidizes at a higher rate in vivo than remelted HXL UHMWPE. Highly crosslinked UHMWPE tibial inserts (87), received by an IRB-approved retrieval laboratory from 20 surgeons at 10 institutions across the U.S., were analyzed from 2005 to 2011. Fourier transform infrared spectroscopy was used to determine oxidation and trans-vinylene index for each retrieved insert. Measured oxidation that was maximum subsurface was found in 56% of all HXL tibial inserts. This maximum oxidation correlated significantly with in vivo time, trans-vinylene index, and thermal processing after irradiation. Articular oxidation rate correlated with crosslinking irradiation dose and thermal processing after irradiation. Retrieved below-melt annealed tibial inserts had significantly higher articular oxidation rates than remelted tibial inserts (p < 0.001). Articular oxidation rates correlated positively with cross-linking irradiation dose and postirradiation thermal processing. Edge oxidation correlated with postirradiation thermal processing. Oxidation of HXL UHMWPE may have clinical implications for tibial inserts, since loss of UHMWPE toughness resulting from oxidation has led to fatigue damage in gamma-sterilized tibial inserts. PMID:22997097
Currier, B H; Van Citters, D W; Currier, J H; Carlson, E M; Tibbo, M E; Collier, J P
Development of collimator insert for linac based stereotactic irradiation.
The aim of this study is to develop collimator inserts of various sizes which are either not commercially available or are expensive to import. The dosimetry parameters such as tissue maximum ratio (TMR), off-axis ratio (OAR) and output factor of the developed collimator insert are compared with that of the commercial collimator insert (Radionics). In order to check the suitability of the collimator insert developed locally for clinical use and to standardize the method of development, a collimator insert of 15 mm identical to the one supplied by Radionics is developed with low-melting alloy (Cerrobend). Moreover for the clinical use of the developed collimator insert, certain acceptance tests are performed which include a collimator concentricity test, beam size check and radiation leakage test. The dose verification is carried out with a thermoluminescent dosimeter (7LiF rods) and an FBX chemical dosimeter in a human-head-shaped Perspex phantom filled with water. The variation between the calculated and measured dose is found to be within +2.4% for 7LiF rods and -2.0% for the FBX chemical dosimeter thus ensuring the suitability of the developed collimator insert for clinical use. This has encouraged us to standardize the method adapted to develop the collimator insert and to develop collimator inserts of different field sizes. PMID:10533938
Singh, I R; Brindha, S; Ravindran, B P; John, S; Rajshekhar, V; Rangad, F V; Roul, R K
NOTE: Development of collimator insert for linac based stereotactic irradiation
NASA Astrophysics Data System (ADS)
The aim of this study is to develop collimator inserts of various sizes which are either not commercially available or are expensive to import. The dosimetry parameters such as tissue maximum ratio (TMR), off-axis ratio (OAR) and output factor of the developed collimator insert are compared with that of the commercial collimator insert (Radionics). In order to check the suitability of the collimator insert developed locally for clinical use and to standardize the method of development, a collimator insert of 15 mm identical to the one supplied by Radionics is developed with low-melting alloy (Cerrobend). Moreover for the clinical use of the developed collimator insert, certain acceptance tests are performed which include a collimator concentricity test, beam size check and radiation leakage test. The dose verification is carried out with a thermoluminescent dosimeter (7LiF rods) and an FBX chemical dosimeter in a human-head-shaped Perspex phantom filled with water. The variation between the calculated and measured dose is found to be within +2.4% for 7LiF rods and -2.0% for the FBX chemical dosimeter thus ensuring the suitability of the developed collimator insert for clinical use. This has encouraged us to standardize the method adapted to develop the collimator insert and to develop collimator inserts of different field sizes.
Rabi Raja Singh, I.; Brindha, S.; Ravindran, B. Paul; John, Subhashini; Rajshekhar, V.; Rangad, Faith V.; Roul, R. K.
Changes in the chromosomal insertion pattern of the copia element during the process of making chromosomes homozygous in Drosophila melanogaster.
In situ hybridization on polytene chromosomes of Drosophila melanogaster was used to compare the insertion patterns of copia and mdg1 transposable elements on chromosome 2 in male gametes sampled by two different methods: (i) by crossing the males tested with females from a highly inbred line with known copia and mdg1 insertion profiles; (ii) by crossing the same males with females from a marked strain, and analysing the resulting homozygous chromosomes. Crossing of the males with the inbred line led to homogeneous insertion profiles for both the copia and mdg1 elements in larvae, thus giving an accurate estimation of the patterns in the two gamete classes of each male. Crossing with the marked strain led, however, to heterogeneity in insertion patterns of the copia transposable element, while no significant polymorphism was observed for mdg1. The use of balancer chromosomes is thus not an adequate way of inferring transposable element insertion patterns of Drosophila males, at least for the copia element. This technique could, however, be powerful for investigating the control of movements of this element. PMID:7862091
Garcia Guerreiro, M P; BiÚmont, C
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.
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.
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
Antibody binding loop insertions as diversity elements
In the use of non-antibody proteins as affinity reagents, diversity has generally been derived from oligonucleotide-encoded random amino acids. Although specific binders of high-affinity have been selected from such libraries, random oligonucleotides often encode stop codons and amino acid combinations that affect protein folding. Recently it has been shown that specific antibody binding loops grafted into heterologous proteins can confer the specific antibody binding activity to the created chimeric protein. In this paper, we examine the use of such antibody binding loops as diversity elements. We first show that we are able to graft a lysozyme-binding antibody loop into green fluorescent protein (GFP), creating a fluorescent protein with lysozyme-binding activity. Subsequently we have developed a PCR method to harvest random binding loops from antibodies and insert them at predefined sites in any protein, using GFP as an example. The majority of such GFP chimeras remain fluorescent, indicating that binding loops do not disrupt folding. This method can be adapted to the creation of other nucleic acid libraries where diversity is flanked by regions of relative sequence conservation, and its availability sets the stage for the use of antibody loop libraries as diversity elements for selection experiments.
Kiss, Csaba; Fisher, Hugh; Pesavento, Emanuele; Dai, Minghua; Valero, Rosa; Ovecka, Milan; Nolan, Rhiannon; Phipps, M. Lisa; Velappan, Nileena; Chasteen, Leslie; Martinez, Jennifer S.; Waldo, Geoffrey S.; Pavlik, Peter; Bradbury, Andrew R.M.
Small insertions are more deleterious than small deletions in human genomes.
Although lines of evidence suggest that small insertions and deletions differ in their mechanisms of formation, there remains the debate on whether natural selection acts differently on the two indel types. Currently available personal genomes and the 1000 Genomes Project permit population level and genome scale comparison of the selection regimes on the two indel types. We first developed a statistical model to evaluate the indel frequency biases of the 1000 Genomes Project phase 1 data. We then identified four independent lines of evidence demonstrating that human small (1-4ábp) insertions are on average more deleterious than deletions. This genome-wide selection pattern is not affected by methodology, demography, and regional differences including indel density, introns versus exons, repeats versus nonrepeats, recombination rates, and the timing of DNA replication. This selection pattern has a profound effect on indel frequency spectra, deletional bias, and local single-nucleotide mutation rates. Finally, we observed that small insertions appear to be more actively implicated in shaping fast-evolving genomic sequences (or nonconserved regions). PMID:24000181
Huang, Shengfeng; Li, Jie; Xu, Anlong; Huang, Guangrui; You, Leiming
The ROSA26 LacZ-neo(R) insertion confers resistance to mammary tumors in Apc(Min/+) mice.
B6.129S7-Gtrosa26 (ROSA26) mice carry a LacZ-neo(R) insertion on Chromosome (Chr) 6, made by promoter trapping with AB1 129 ES cells. Female C57BL/6J Apc(Min/+) (B6 Min/+) mice are very susceptible to the induction of mammary tumors after treatment with ethylnitrosourea (ENU). However, ENU-treated B6 mice carrying both Apc(Min) and ROSA26 are resistant to mammary tumor formation. Thus, ROSA26 mice carry a modifier of Min-induced mammary tumor susceptibility. We have previously mapped the modifier to a 4-cM interval of 129-derived DNA that also contains the ROSA26 insertion. Here we report additional evidence for the effect of the ROSA26 insertion on mammary tumor formation. To test the hypothesis that the resistance was due to a linked modifier locus, we utilized two approaches. We have derived and tested two lines of mice that are congenic for 129-derived DNA within the minimal modifier interval and show that they are as susceptible to mammary tumors as are B6 mice. Additionally, we analyzed a backcross population segregating for the insertion and show that mice carrying the insertion are more resistant to mammary tumor development than are mice not carrying the insertion. Thus, the resistance is not due to a 129-derived modifier allele, but must be due to the ROSA26 insertion. In addition, the effect of the ROSA26 insertion can be detected in a backcross population segregating for other mammary modifiers. PMID:11471054
Kohlhepp, R L; Hegge, L F; Moser, A R
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.
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
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.
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.
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
Metal inserts in structural composite materials manufactured by RTM
Load transfer on a structural composite part can be carried out by means of metal inserts. Apart from their detachable features, their introduction in a composite material is not without consequence. This paper examines the impact of the inserts in a structural composite material obtained by the Resin Transfer Moulding process. Numerical flow simulations, using RTMFLOT software developed by the
B. Ferret; M. Anduze; C. Nardari
Characterizations of Recursively Enumerable Languages by Means of Insertion Grammars
An insertion grammar is based on pure rules of the form uv ? uxv (the string x is inserted in the context (u, v)). A strict subfamily of the context-sensitive family is obtained, incomparable with the family of linear languages. We prove here that each recursively enumerable language can be written as the weak coding of the image by an
Carlos MartÝn-vide; Gheorghe Paun; Arto Salomaa
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2010 CFR
...2009-04-01 false Patient package inserts...Requirements for Specific New Drugs or Devices ┬ž 310.515 Patient package inserts...injectables in multiple-dose vials, a sufficient...injectables, with each dose administered to the patient. This...
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2010 CFR
...2010-04-01 false Patient package inserts...Requirements for Specific New Drugs or Devices ┬ž 310.515 Patient package inserts...injectables in multiple-dose vials, a sufficient...injectables, with each dose administered to the patient. This...
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 passive mechanism for insertion of convex pegs
The author expands on some recent results for more generalized insertions in robotics to develop and analyze a passive mechanism capable of inserting arbitrary convex shaped parts. He implemented the strategies of H. Inoue (1979) and S. Simunovic (1979) on a system comprised of commercially available components (a PUMA 560 robot and an Astek six-axis force-torque sensor). It was not
David R. Strip
Simultaneous routing and buffer insertion with restrictions on buffer locations
During the routing of global interconnects, macro blocks formuseful routing regions which allow wires to go through but forbidbuffers to be inserted. They give restrictions on bufferlocations. In this paper, we take these buffer location restrictionsinto consideration and solve the simultaneous mazerouting and buffer insertion problem. Given a block placementdefining buffer location restrictions and a pair of pins(a source and
Hai Zhou; D. F. Wong; I-Min Liu; Adnan Aziz
Insertional Mutagenesis of Drosophila Heterochromatin with Single P Elements
Insertional mutagenesis with transposable P elements has greatly facilitated the identification and analysis of genes located throughout the 70% of the Drosophila melanogaster genome classified as euchromatin. In contrast, genetically marked P elements have only rarely been shown to transpose into heterochromatin. By carrying out single P element insertional mutagenesis under conditions where position-effect variegation was suppressed, we efficiently generated
Ping Zhang; Allan C. Spradling
A Metallic Insert and Locking Hole Location System
This paper describes a preliminary study that was performed to look at the possible use of a low power microwave transmission system to determine the location of metallic orthopaedic inserts placed inside the human body for the purpose of repairing fractures or breaks in bones and also to determine the location of locking holes contained within the insert. For this
C. P. Hancock
Interactive simulation of surgical needle insertion and steering
We present algorithms for simulating and visualizing the insertion and steering of needles through deformable tissues for surgical training and planning. Needle insertion is an essential component of many clinical procedures such as biopsies, injections, neurosurgery, and brachytherapy cancer treatment. The success of these procedures depends on accurate guidance of the needle tip to a clinical target while avoiding vital
Nuttapong Chentanez; Ron Alterovitz; Daniel Ritchie; Lita Cho; Kris K. Hauser; Ken Goldberg; Jonathan R. Shewchuk; James F. O'Brien
Downstream insertion of the adenovirus tripartite leader sequence enhances expression in universal eukaryotic vectors.
A series of universal eukaryotic gene expression vectors was constructed. All vectors contain a viral promoter and enhancer, a polylinker for insertion of the gene of interest and a polyadenylation sequence. To enhance translation, we inserted the tripartite leader sequence of an adenovirus downstream of the promoter. Using the chloramphenicol acetyl transferase (cat) gene as a marker, we show that the strength of various promoters/enhancers in different cell lines differed by two orders of magnitude. The presence of the tripartite leader increased the efficiency of gene expression up to 18-fold. The level of increase is promoter specific and is most likely influenced by additional sequences flanking the tripartite leader sequence. PMID:8267981
Sheay, W; Nelson, S; Martinez, I; Chu, T H; Bhatia, S; Dornburg, R
... for adults A A A Beau's lines (crosswise nail plate grooves) are due to severe illness or ... Beau's lines are horizontal (transverse) depressions in the nail plate that run parallel to the shape of ...
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
Method for improving the durability of ion insertion materials
The invention provides a method of protecting an ion insertion material from the degradative effects of a liquid or gel-type electrolyte material by disposing a protective, solid ion conducting, electrically insulating, layer between the ion insertion layer and the liquid or gel-type electrolyte material. The invention further provides liquid or gel-type electrochemical cells having improved durability having a pair of electrodes, a pair of ion insertion layers sandwiched between the pair of electrodes, a pair of solid ion conducting layers sandwiched between the ion insertion layers, and a liquid or gel-type electrolyte material disposed between the solid ion conducting layers, where the solid ion conducting layer minimizes or prevents degradation of the faces of the ion insertion materials facing the liquid or gel-type electrolyte material. Electrochemical cells of this invention having increased durability include secondary lithium batteries and electrochromic devices.
Lee, Se-Hee (Lakewood, CO) [Lakewood, CO; Tracy, C. Edwin (Golden, CO) [Golden, CO; Cheong, Hyeonsik M. (Seoul, KR) [Seoul, KR
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)
Smart structures for shock wave attenuation using ER inserts
NASA Astrophysics Data System (ADS)
This Paper demonstrates the possibility of shock wave attenuation propagating through a smart structure that incorporates ER insert. The wave transmission of ER inserted beam is theoretically derived using Mead & Markus model and the theoretical results are compared with the finite element analysis results. To experimentally verify the shock wave attenuation, ER insert in an aluminum plate is made and two piezoceramic disks are used as transmitter and receiver of the wave. The transmitter sends a sine pulse signal such that a component of shock wave travels through the plate structure and the receiver gets the transmitted wave signal. Wave propagation of the ER insert can be adjusted by changing the applied electric field on the ER insert. Details of the experiment are addressed and the possibility of shock wave attenuation is experimentally verified. This kind of smart structure can be used for warship and submarine hull structures to protect fragile and important equipment.
Kim, Jaehwan; Kim, Jung-Yup; Choi, Seung-Bok; Kim, Kyung-Su
ERIC Educational Resources Information Center
One tool for enhancing students' work with data in the science classroom is the measure line. As a coteacher and curriculum developer for The Inquiry Project, the author has seen how measure lines--a number line in which the numbers refer to units of measure--help students not only represent data but also analyze it in ways that generateů
Crissman, Sally
Tunable Nonlinear Transmission Lines with switched varactors
Nonlinear Transmission Lines (NLTLs) have been used in many applications for waveform shaping and impedance engineering. To minimize insertion loss and jitter, adaptive impedance is desirable. Here we propose to use switched varactors on NLTL to achieve impedance-tuning. We present an analytical model to examine the impact of switch parasitics on the performance of the NLTL. We find that for
Fan Yu; K. G. Lyon; E. C. Kan
Efficient screening of transgenic plant lines for ecological research.
Plants stably transformed to manipulate the expression of genes mediating ecological performance have profoundly altered research in plant ecology. Agrobacterium-mediated transformation remains the most effective method of creating plants harbouring a limited number of transgene integrations of low complexity. For ecological/physiological research, the following requirements must be met: (i) the regenerated plants should have the same ploidy level as the corresponding wild-type plant and (ii) contain a single transgene copy in a homozygous state; (iii) the T-DNA must be completely inserted without vector backbone sequence and all its elements functional; and (iv) the integration should not change the phenotype of the plant by interrupting chromosomal genes or by mutations occurring during the regeneration procedure. The screening process to obtain transformed plants that meet the above criteria is costly and time-consuming, and an optimized screening procedure is presented. We developed a flow chart that optimizes the screening process to efficiently select transformed plants for ecological research. It consists of segregational analyses, which select transgenic T? and T? generation plants with single T-DNA copies that are homozygous. Indispensable molecular genetic tests (flow cytometry, diagnostic PCRs and Southern blotting) are performed at the earliest and most effective times in the screening process. qPCR to quantify changes in transcript accumulation to confirm gene silencing or overexpression is the last step in the selection process. Because we routinely transform the wild tobacco, Nicotiana attenuata, with constructs that silence or ectopically overexpress ecologically relevant genes, the proposed protocol is supported by examples from this system. PMID:21518300
Gase, Klaus; Weinhold, Arne; Bozorov, Tohir; Schuck, Stefan; Baldwin, Ian T
Modulation of the transmembrane helix insertion pathway by polar cargo
NASA Astrophysics Data System (ADS)
In an earlier study, we found a series of kinetic steps in the pH-triggered insertion of the pHLIP (pH (Low) Insertion Peptide). In the present work we observe that the polarity of the inserting end, including its cargo, modulates the number of intermediates, and that insertion can be described as a two state process for a simple case. Each investigated pHLIP variant preserve the pH-dependent properties of surface binding to membrane at neutral pH and insertion at low pH to form a transmembrane helix. However, there are thermodynamic and kinetic properties that are determined by the degree of cargo polarity. The presence of a polar cargo at the peptide's inserting end leads to the appearance of two additional intermediate states on the insertion pathway of the pHLIP-2E peptide, which itself (when no cargo is attached) shows an all-or-none transition from the partially unstructured membrane-surface to the transmembrane state described well by the two-state model at 800 ms timescale. We discuss the utility of our observations for the design of new delivery agents for the direct translocation of polar therapeutic and diagnostic cargo molecules across cellular membranes. The work is supported by NIH grants CA133890 to OAA, DME, YRK.
Wijesinghe, Dayanjali; Karabadzhak, Alexander; Markin, Vladislav; Engelman, Donald; Andreev, Oleg; Reshetnyak, Yana
Force of cochlear implant electrode insertion performed by a robotic insertion tool: comparison of traditional versus Advance Off-Stylet techniques
Objective Robotic cochlear implant electrode array insertion offers substantial potential advantages, namely repeatability and minimization of insertion forces leading to decreased intracochlear trauma. Using such a robotic insertion tool, we sought to analyze force profiles during deployment of stylet-containing electrode arrays using either traditional insertion, in which the stylet is withdrawn after complete insertion of the electrode, or Advance Off-Stylet (AOS) insertion, in which the stylet is withdrawn simultaneous with electrode array insertion. Study design Prospective. Setting Tertiary referral center. Intervention(s) A robotic cochlear implant insertion tool coupled with a force sensing carriage was used to perform electrode array insertions into an anatomically-correct, 3-dimensional scala tympani model during either straight insertion (n=4) or AOS insertion (n=4). Main outcome measure(s) Both insertion techniques begin with a 7mm straight insertion during which forces were similar averaging approximately 0.006N. For insertion from 7ľ17mm, traditional insertion forces averaged 0.046 ▒ 0.027N with a peak of 0.093N and AOS insertion forces averaged 0.008 ▒ 0.006N with a peak of 0.034N. Beyond 9.74 mm, the difference between traditional and AOS insertion forces were highly significant. Conclusions With the use of a robotic insertion tool, which minimizes operator variability and maximizes repeatability, we have shown that cochlear implant electrode insertion via AOS is associated with lower average and maximum insertion forces, compared to traditional insertion. These findings support the use of AOS over traditional, straight insertion.
Webster, Robert J.; Dietrich, Mary S.; Labadie, Robert F.
Prevention of Transformer Saturation at a Black Start by Resistor Insertion of 500kV CB
NASA Astrophysics Data System (ADS)
A generating plant can not energize longer transmission lines at once after blackout. The generating plant energize shorter transmission lines firstly, then circuit breaker turns on to connect other transmission lines through the transformer. Transient over voltage oscillation may occur in the transmission lines caused by the resonance of the transmission line. It is reported that the peak voltage exceeds more than 2p.u. when the non-sinusoidal wave voltage caused by saturation in the transformer core excites the resonated transmission line. This paper describes the effect for preventing saturation of transformer by turning on contactors with appropriate resistors before main contactors of the 500kV circuit breaker. The first part presents the mechanism of preventing saturation in the transformer core by pre-resistor insertion of 500kV circuit breaker. The second part presents the expected effect in the actual power system simulated by analytical model using a transients simulation program, EMTP.
Nagashima, Hiroshi; Kosugi, Narifumi; Shimojo, Toshikazu; Furukawa, Nobuhiko; Kusunoki, Kiyoshi; Sugeta, Takayuki
Industrial stator vane with sequential impingement cooling inserts
A turbine stator vane for an industrial engine, the vane having two impingement cooling inserts that produce a series of impingement cooling from the pressure side to the suction side of the vane walls. Each insert includes a spar with a row of alternating impingement cooling channels and return air channels extending in a radial direction. Impingement cooling plates cover the two sides of the insert and having rows of impingement cooling holes aligned with the impingement cooling channels and return air openings aligned with the return air channel.
Jones, Russell B; Fedock, John A; Goebel, Gloria E; Krueger, Judson J; Rawlings, Christopher K; Memmen, Robert L
Visually Guided Z Technique to Insert the Endopath Optiview
Visually guided Z insertion of the Endopath Optiview was evaluated in 200 consecutive women undergoing elective laparoscopic sterilization in an ambulatory setting. The technique was employed to establish the primary intraabdominal surgical port. All cases were completed successfully. There were no failed insertions and no multiple attempts to enter the abdominal cavity. Direct visualization while inserting the Endopath Optiview is important to localize the tip of the advancing instrument at any given moment. It lets the surgeon choose the shortest route in the abdominal wall and change the angle of the instrument when appropriate. PMID:9074142
Preferential insertion of lactose permease in phospholipid domains: AFM observations
We report the insertion of a transmembrane protein, lactose permease (LacY) from Escherichia coli (E. coli), in supported lipid bilayers (SLBs) of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), in biomimetic molar proportions. We provide evidence of the preferential insertion of LacY in the fluid domains. Analysis of the self-assembled protein arrangements showed that LacY: (i) is inserted as a monomer within
Laura Picas; Adrißn Carretero-Genevrier; M. Teresa Montero; J. L. Vßzquez-Ibar; Bastien Seantier; Pierre-Emmanuel Milhiet; Jordi Hernßndez-Borrell
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.
Insertional mutagenesis of Listeria monocytogenes with a novel Tn917 derivative that allows direct cloning of DNA flanking transposon insertions.
To carry out efficient insertional mutagenesis in Listeria monocytogenes and to facilitate the characterization of disrupted genes, two novel derivatives of Tn917 were constructed, Tn917-LTV1 and Tn917-LTV3. The derivatives (i) transpose at a significantly elevated frequency, (ii) generate transcriptional lacZ fusions when inserted into a chromosomal gene in the appropriate orientation, and (iii) allow the rapid cloning in Escherichia coli of chromosomal DNA flanking transposon insertions. The rapid cloning of DNA flanking insertions is possible because the transposon derivatives carry ColE1 replication functions, a cluster of polylinker cloning sites, and antibiotic resistance genes selectable in E. coli (bla in the case of Tn917-LTV1; neo and ble in the case of Tn917-LTV3). The enhanced transposition frequency of Tn917-LTV1 and Tn917-LTV3 (about 100-fold in Bacillus subtilis) is believed to be due to the fortuitous placement of vector-derived promoters upstream from the Tn917 transposase gene. In L. monocytogenes, Tn917-LTV3 transposed at a frequency of 8 x 10(-4) when introduced on a pE194Ts-derived vector and generated at least eight different auxotrophic mutations. Two nonhemolytic insertion mutants of L. monocytogenes were isolated, and DNA flanking the transposon insertions was cloned directly into E. coli, making use of the ColE1 rep functions and neo gene carried by Tn917-LTV3. Both insertions were shown to be within hlyA, the L. monocytogenes hemolysin structural gene. Although Tn917-LTV1 and Tn917-LTV3 were constructed specifically for genetic analysis of L. monocytogenes, their enhanced transposition frequency and convenience for cloning of DNA adjacent to sites of insertions make them the transposon derivatives of choice for insertional mutagenesis in any gram-positive bacteria that support replication of pE194Ts.
Camilli, A; Portnoy, A; Youngman, P
Post insertion catheter care in peritoneal dialysis centers across Europe: results of the Post Insertion Project of the Research Board.
The EDTNA/ERCA survey of Post Insertion Catheter Care in Peritoneal Dialysis (PICC) was a project organised through the Collaborative Research Programme (CRP) of the EDTNA/ERCA. In this survey, data were collected from 54 participating centres in 20 countries. From this survey it became clear that there is no standardised approach to immediate post-catheter insertion treatment protocols. If we want to reduce technique failure of PD related to catheter failure, a first step will be to investigate the different policies used in Europe in order to evaluate the outcome results derived from different policies in post insertion catheter care. PMID:15163035
Castro, M J; Vijt, D; Endall, G; Elseviers, M; Lindley, E
Insertion of nitriles into complexes of tantalum pentachloride with amines
Data from the study of the insertion of aceto-, propio-, and benzonitrile into complexes of TaCl/sub 5/ with diethyl- and triethylamine in CCl/sub 4/ solutions have been discussed. It has been established that the nature of the ligands, rather than the nitrile being inserted into the complex, has a significant influence on the composition of the final products. In contrast to the case of acetonitrile, the insertion of propionitrile and benzonitrile into a complex is observed already at room temperature. On the basis of spectroscopic data it has been postulated that the carbon tetrachloride employed as a solvent accelerates the insertion of nitriles and promotes their deprotonation in the presence of an aprotic amine.
Glushkova, M.A.; Chumaevskii, N.A.; Khmelevskaya, L.V.; Ershova, M.M.; Buslaev, Yu.A.
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
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.
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2012 CFR
...distributing additional patient labeling pieces to the dispensor. (3) Patient package inserts for...dispensed in acute-care hospitals or long-term care facilities will be considered...section if provided to the patient before...
21 CFR 310.515 - Patient package inserts for estrogens.
Code of Federal Regulations, 2011 CFR
...distributing additional patient labeling pieces to the dispensor. (3) Patient package inserts for...dispensed in acute-care hospitals or long-term care facilities will be considered...section if provided to the patient before...
Spontaneous Radiation Emission from Short, High Field Strength Insertion Devices
Since the earliest papers on undulaters were published, it has been known how to calculate the spontaneous emission spectrum from ''short'' undulaters when the magnetic field strength parameter is small compared to unity, or in ''single'' frequency sinusoidal undulaters where the magnetic field strength parameter is comparable to or larger than unity, but where the magnetic field amplitude is constant throughout the undulater. Fewer general results have been obtained in the case where the insertion device is both short, i.e., the magnetic field strength parameter changes appreciably throughout the insertion device, and the magnetic field strength is high enough that ponderomotive effects, radiation retardation, and harmonic generation are important physical phenomena. In this paper a general method is presented for calculating the radiation spectrum for short, high-field insertion devices. It is used to calculate the emission from some insertion device designs of recent interest.
Geoffrey Krafft
Shoe inserts and orthotics for sport and physical activities.
The purposes of this paper were to discuss the perceived benefits of inserts and orthotics for sport activities and to propose a new concept for inserts and orthotics. There is evidence that inserts or orthotics reduce or prevent movement-related injuries. However, there is limited knowledge about the specific functioning an orthotic or insert provides. The same orthotic or insert is often proposed for different problems. Changes in skeletal movement due to inserts or orthotics seem to be small and not systematic. Based on the results of a study using bone pins, one may question the idea that a major function of orthotics or inserts consists in aligning the skeleton. Impact cushioning with shoe inserts or orthotics is typically below 10%. Such small reductions might not be important for injury reduction. It has been suggested that changes in material properties might produce adjustments in the muscular response of the locomotor system. The foot has various sensors to detect input signals with subject specific thresholds. Subjects with similar sensitivity threshold levels seem to respond in their movement pattern in a similar way. Comfort is an important variable. From a biomechanical point of view, comfort may be related to fit, additional stabilizing muscle work, fatigue, and damping of soft tissue vibrations. Based on the presented evidence, the concept of minimizing muscle work is proposed when using orthotics or inserts. A force signal acts as an input variable on the shoe. The shoe sole acts as a first filter, the insert or orthotic as a second filter, the plantar surface of the foot as a third filter for the force input signal. The filtered information is transferred to the central nervous system that provides a subject specific dynamic response. The subject performs the movement for the task at hand. For a given movement task, the skeleton has a preferred path. If an intervention supports/counteracts the preferred movement path, muscle activity can/must be reduced/increased. Based on this concept, an optimal insert or orthotic would reduce muscle activity, feel comfortable, and should increase performance. PMID:10416543
Nigg, B M; Nurse, M A; Stefanyshyn, D J
Identification of insertion hot spots for non-LTR retrotransposons: computational and biochemical application to Entamoeba histolytica
The genome of the human pathogen Entamoeba histolytica contains non-long terminal repeat (LTR) retrotransposons, the EhLINEs and EhSINEs, which lack targeted insertion. We investigated the importance of local DNA structure, and sequence preference of the element-encoded endonuclease (EN) in selecting target sites for retrotransposon insertion. Pre-insertion loci were tested computationally to detect unique features based on DNA structure, thermodynamic considerations and protein interaction measures. Target sites could readily be distinguished from other genomic sites based on these criteria. The contribution of the EhLINE1-encoded EN in target site selection was investigated biochemically. The sequence-specificity of the EN was tested in vitro with a variety of mutated substrates. It was possible to assign a consensus sequence, 5?-GCATT-3?, which was efficiently nicked between A-T and T-T. The upstream G residue enhanced EN activity, possibly serving to limit retrotransposition in the A+T-rich E.histolytica genome. Mutated substrates with poor EN activity showed structural differences compared with normal substrates. Analysis of retrotransposon insertion sites from a variety of organisms showed that, in general, regions of favorable DNA structure were recognized for retrotransposition. A combination of favorable DNA structure and preferred EN nicking sequence in the vicinity of this structure may determine the genomic hotspots for retrotransposition.
Mandal, Prabhat K.; Rawal, Kamal; Ramaswamy, Ram; Bhattacharya, Alok; Bhattacharya, Sudha
Kinematic Analysis of Six Different Insertion Sites for FDS Opponensplasty
Six variations of opponensplasty were biomechanically studied to determine if there is a significant difference in the amount\\u000a of opposition produced by altering the insertion site of the tendon transfer while keeping the tendon source and pulley location\\u000a constant. Five cadaveric arms were used to simulate six described insertion sites for opponensplasty tendon transfers. The\\u000a arms were mounted through the
Martin C. Skie; Thomas Parent; Kenneth Mudge; Qiang Dai
Pressure drop analysis in the CS insert coil
The Central Solenoid Model Coil (CSMC) and the CS Insert Coil (CSIC) were tested during the spring and summer of 2000 at JAERI Naka, Japan, within the framework of the ITER large projects. The CSIC is a single-layer one-in-hand solenoid inserted in the bore of the CSMC. It uses a Nb3Sn dual-channel cable-in-conduit conductor (CICC), about 140 m long, cooled
R. Zanino; C. Y. Gung; K. Hamada; L. Savoldi
The Structure of Fully H-Inserted ?-Manganese Dioxide Compounds
Four materials forming a structural series of ?-manganese dioxides were identified by an increasing proportion of micro-twinning (17ľ100%) and a nearly constant amount of de Wolff disorder (Pr? 0.41) as determined from their XRD patterns. Chemical H-insertion was performed in a nonaqueous environment and the structure of the fully H-inserted compounds investigated by XRD. In contrast to the starting materials
L. A. H. MacLean; F. L. Tye
Evaluation of antitubercular drug insertion into preformed dipalmitoylphosphatidylcholine monolayers.
Insertion profiles of antitubercular drugs isoniazid (INH), rifampicin (RFM) and ethambutol (ETH) into dipalmitoylphosphatidylcholine (DPPC) membrane models were evaluated by Langmuir monolayer technique. Maximum drug insertion into DPPC monolayer was observed with rifampicin with a surface pressure increase (Deltapi(max)) in the range of 21-33 mN/m depending upon rifampicin concentration. Isoniazid had minimal insertion resulting in a lower Deltapi(max) of about 2-3 mN/m, suggestive of minimal interactions between INH and DPPC. Ethambutol surface pressure increment on insertion resulted in an intermediate rise in the Deltapi(max) (6-10 mN/m). Antitubercular drug combination in the ratio of 2 mM:0.7 mM:4.5 mM for INH:RFM:ETH, attained Deltapi(max) between 25 and 33 mN/m. Insertion profiles similar to rifampicin were exhibited by the antitubercular drug mixture suggestive of predominant rifampicin insertion into the DPPC monolayer. The extent of drug insertion into the DPPC monolayer is suggestive of the drug penetration potential into biological membranes in vivo. Higher RFM Deltapi(max) is suggestive of excellent cell membrane penetration, which explains broad reach of the drug to all the organs including the cerebrospinal fluid while lower Deltapi(max) of INH suggests poor membrane penetration restricting the entry of the drug in different biological membranes. DPPC membrane destabilization was observed at higher antitubercular drug concentrations indicated by the negative slopes of the surface pressure-time curves. This may correlate with the dose related toxic effects observed in tuberculosis affected patients. Drug insertion studies offer a potential tool in understanding the pharmacotoxicological behavior of the various pharmacological agents. PMID:18082382
Chimote, G; Banerjee, R
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
Metal-Matrix Composite Parts With Metal Inserts
NASA Technical Reports Server (NTRS)
Developmental fabrication process produces metal-matrix composite (MMC) parts with integral metal inserts. With inserts, MMC parts readily joined to similar parts by use of brazing, welding, or mechanical fasteners. Process conceived to make strong, lightweight components of structures erected in outer space. Also useful on Earth, in such automotive parts as rocker arms, cylinder liners, and pistons. Potential industrial applications include parts subjected to high stresses at high temperatures, as in power-generation, mining, and oil-drilling equipment.
Majkowski, T.; Kashalikar, U.
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.
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
Cervical Lidocaine for IUD Insertional Pain: a Randomized Controlled Trial
Objective Anticipated pain with intrauterine device (IUD) insertion may be a barrier to widespread use. Our objective was to evaluate the efficacy of intracervical 2% lidocaine gel for pain relief with IUD insertion. Study Design We performed a double-blind, randomized controlled trial of women undergoing IUD insertion. Participants were randomly assigned to 2% lidocaine or placebo gel. Study gel (3ccs) wase placed 3 minutes prior to IUD insertion. Pain scores were measured at various time points using a 10-point visual analog scale. Results Of the 200 participants randomized, 199 completed the study. Pain scores among lidocaine and placebo arms were similar at tenaculum placement (lidocaine and placebo; median 4, range 0ľ10 p=0.15) as well as with insertion (lidocaine: median 5 range 1ľ10, placebo: median 6 range 0ľ10 p=0.16). These results did not differ by parity. Conclusions Topical or intracervical 2% lidocaine gel prior to IUD insertion does not decrease pain scores.
McNicholas, Colleen P.; Madden, Tessa; Zhao, Qiuhong; Secura, Gina; Allsworth, Jenifer E.; Peipert, Jeffrey F.
Detecting motifs and patterns at mobile genetic element insertion site
Mobile genetic elements (MGEs) occupy major proportion of eukaryotic genomes and are present in significant numbers in prokaryote genomes also. Here we report a new method which extracts a motif at the site of insertion of MGE using tools such as DNA SCANNER. The flanking region of the insertion site is extracted and is analyzed in DNA Scanner for physiochemical properties like protein-interaction measures, energy profiles as well as structural parameters. In case significant signals are observed, the most frequently occurring n-mer (5
Dev, Bhuvan Bhaskar; Malik, Aman; Rawal, Kamal
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.
Alterovitz, Ron; Goldberg, Kenneth Y.; Pouliot, Jean; Hsu, I-Chow
21 CFR 872.3900 - Posterior artificial tooth with a metal insert.
Code of Federal Regulations, 2010 CFR
...2012-04-01 false Posterior artificial tooth with a metal insert. 872.3900 ...┬ž 872.3900 Posterior artificial tooth with a metal insert. (a) Identification. A posterior artificial tooth with a metal insert is a...
Altered imino diacid synthesis and transcription in crown gall tumors with transposon Tn5 insertions in the 3' end of the octopine synthase gene.
Octopine synthase encoded by the T-DNA (transferred DNA) locus ocs synthesizes N2-(D-1-carboxyethyl)-L-amino acids in octopine-type crown gall tumors. So far, derivatives of only basic amino acids have been isolated. We have detected a glutamine derivative and called it heliopine. Tumors induced by several Ti plasmids with transposon Tn5 insertions in the 3' end of ocs still synthesized small quantities of N2-(1-carboxyethyl)-arginine and N2-(1-carboxyethyl)-glutamine. In addition, N2-(1,3-dicarboxypropyl)-asparagine, which is absent in wild-type octopine tumors, was detected in these tumors. These three imino diacids (octopine, heliopine, and asparaginopine, respectively, or their isomers) were undetectable in tumors induced by Ti plasmids harboring deletions of the ocs gene. Poly(A)+ RNAs which hybridize to the ocs sequence can also be detected in the ocs::Tn5 tumors; these RNAs, however, were heterogeneous in size and shorter in length than the normal ocs mRNA. These results indicate that mutant ocs products synthesize imino diacids in these ocs::Tn5 tumors. PMID:2478521
Chang, C C; Jayaswal, R K; Chen, C M; Gelvin, S B
Positioning long lines: contrast versus plain radiography
AIMŚTo assess the value of contrast versus plain radiography in determining radio-opaque long line tip position in neonates.?METHODSŚIn a prospective study, plain radiography was performed after insertion of radio-opaque long lines. If the line tip was not visible on the plain film, a second film with contrast was obtained in an attempt to visualise the tip.?RESULTSŚSixty eight lines were inserted during the study period, 62 of which were included in the study. In 31, a second radiographic examination with contrast was necessary to determine position of the tip. In 29 of these, the line tip was clearly visualised with contrast. On two occasions, the line tip could not be seen because the contrast had filled the vein and obscured the tip from view. Eight of the lines that required a second radiograph with contrast were repositioned.?CONCLUSIONŚIntravenous contrast should be routinely used in the assessment of long line position in the neonate.??
Reece, A; Ubhi, T; Craig, A; Newell, S
Evolutionary Dynamics of Large Numts in the Human Genome: Rarity of Independent Insertions and Abundance of Post-Insertion Duplications
ááAbstract\\u000a \\u000a We determined the phylogenetic positions of 82 large nuclear pseudogenes of mitochondrial origin (numts) within the human genome. For each numt, two possibilities pertaining to its origin were considered: (1) independent insertion from the mitochondria into the nucleus,\\u000a or (2) genomic duplication subsequent to the insertion. A significant increase in the rate of numt accumulation is seen after the
Einat Hazkani-Covo; Rotem Sorek; Dan Graur
P-Element Insertion Alleles of Essential Genes on the Third Chromosome of Drosophila Melanogaster: Correlation of Physical and Cytogenetic Maps in Chromosomal Region 86e-87f
We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover ~25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty-one of these 38 complementation groups have at least one allele that has a P-element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven ``supercontigs'' that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the ?? element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms.
Deak, P.; Omar, M. M.; Saunders, RDC.; Pal, M.; Komonyi, O.; Szidonya, J.; Maroy, P.; Zhang, Y.; Ashburner, M.; Benos, P.; Savakis, C.; Siden-Kiamos, I.; Louis, C.; Bolshakov, V. N.; Kafatos, F. C.; Madueno, E.; Modolell, J.; Glover, D. M.
The heterozygous abp1/ABP1 insertional mutant has defects in functions requiring polar auxin transport and in regulation of early auxin-regulated genes.
AUXIN-BINDING PROTEIN 1 (ABP1) is not easily accessible for molecular studies because the homozygous T-DNA insertion mutant is embryo-lethal. We found that the heterozygous abp1/ABP1 insertion mutant has defects in auxin physiology-related responses: higher root slanting angles, longer hypocotyls, agravitropic roots and hypocotyls, aphototropic hypocotyls, and decreased apical dominance. Heterozygous plants flowered earlier than wild-type plants under short-day conditions. The length of the main root, the lateral root density and the hypocotyl length were little altered in the mutant in response to auxin. Compared to wild-type plants, transcription of early auxin-regulated genes (IAA2, IAA11, IAA13, IAA14, IAA19, IAA20, SAUR9, SAUR15, SAUR23, GH3.5 and ABP1) was less strongly up-regulated in the mutant by 0.1, 1 and 10 ?m IAA. Surprisingly, ABP1 was itself an early auxin-up-regulated gene. IAA uptake into the mutant seedlings during auxin treatments was indistinguishable from wild-type. Basipetal auxin transport in young roots was slower in the mutant, indicating a PIN2/EIR1 defect, while acropetal transport was indistinguishable from wild-type. In the eir1 background, three of the early auxin-regulated genes tested (IAA2, IAA13 and ABP1) were more strongly induced by 1 ?m IAA in comparison to wild-type, but eight of them were less up-regulated in comparison to wild-type. Similar but not identical disturbances in regulation of early auxin-regulated genes indicate tight functional linkage of ABP1 and auxin transport regulation. We hypothesize that ABP1 is involved in the regulation of polar auxin transport, and thus affects local auxin concentration and early auxin gene regulation. In turn, ABP1 itself is under the transcriptional control of auxin. PMID:21223392
Effendi, Yunus; Rietz, Steffen; Fischer, Urs; Scherer, GŘnther F E
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
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.
NSDL National Science Digital Library
This activity gives students practice drawing straight lines with a ruler and looking for and categorizing shapes, for example, by the number of sides in polygons. The Teachers' Notes page includes suggestions for implementation, discussion questions and ideas for extension.
Line segment matching based on the depth face of building
NASA Astrophysics Data System (ADS)
The paper presents an algorithm to find homonymous line segments from images based on the depth face of building. TIN is formed from the object points calculated by homonymous points in stereopair, and contour lines are inserted from TIN with a given interval. With image and contour lines, the different depth faces of building can be partitioned approximately. Homonymous line segments are found in different depth faces by matching along epipolar line, the method is tested feasible. It solves the problem of line matching in the changed area of building depth face simultaneously.
Liu, Yawen
The Rice Miniature Inverted Repeat Transposable Element mPing Is an Effective Insertional Mutagen in Soybean1[C][W][OA
Insertional mutagenesis of legume genomes such as soybean (Glycine max) should aid in identifying genes responsible for key traits such as nitrogen fixation and seed quality. The relatively low throughput of soybean transformation necessitates the use of a transposon-tagging strategy where a single transformation event will produce many mutations over a number of generations. However, existing transposon-tagging tools being used in legumes are of limited utility because of restricted transposition (Ac/Ds: soybean) or the requirement for tissue culture activation (Tnt1: Medicago truncatula). A recently discovered transposable element from rice (Oryza sativa), mPing, and the genes required for its mobilization, were transferred to soybean to determine if it will be an improvement over the other available transposon-tagging tools. Stable transformation events in soybean were tested for mPing transposition. Analysis of mPing excision at early and late embryo developmental stages revealed increased excision during late development in most transgenic lines, suggesting that transposition is developmentally regulated. Transgenic lines that produced heritable mPing insertions were identified, with the plants from the highest activity line producing at least one new insertion per generation. Analysis of the mPing insertion sites in the soybean genome revealed that features displayed in rice were retained including transposition to unlinked sites and a preference for insertion within 2.5 kb of a gene. Taken together these findings indicate that mPing has the characteristics necessary for an effective transposon-tagging resource.
Hancock, C. Nathan; Zhang, Feng; Floyd, Kristen; Richardson, Aaron O.; LaFayette, Peter; Tucker, Donna; Wessler, Susan R.; Parrott, Wayne A.
Retrotransposon-Induced Heterochromatin Spreading in the Mouse Revealed by Insertional Polymorphisms
The ôarms raceö relationship between transposable elements (TEs) and their host has promoted a series of epigenetic silencing mechanisms directed against TEs. Retrotransposons, a class of TEs, are often located in repressed regions and are thought to induce heterochromatin formation and spreading. However, direct evidence for TEľinduced local heterochromatin in mammals is surprisingly scarce. To examine this phenomenon, we chose two mouse embryonic stem (ES) cell lines that possess insertionally polymorphic retrotransposons (IAP, ETn/MusD, and LINE elements) at specific loci in one cell line but not the other. Employing ChIP-seq data for these cell lines, we show that IAP elements robustly induce H3K9me3 and H4K20me3 marks in flanking genomic DNA. In contrast, such heterochromatin is not induced by LINE copies and only by a minority of polymorphic ETn/MusD copies. DNA methylation is independent of the presence of IAP copies, since it is present in flanking regions of both full and empty sites. Finally, such spreading into genes appears to be rare, since the transcriptional start sites of very few genes are less than one Kb from an IAP. However, the B3galtl gene is subject to transcriptional silencing via IAP-induced heterochromatin. Hence, although rare, IAP-induced local heterochromatin spreading into nearby genes may influence expression and, in turn, host fitness.
Rebollo, Rita; Karimi, Mohammad M.; Bilenky, Misha; Gagnier, Liane; Miceli-Royer, Katharine; Zhang, Ying; Goyal, Preeti; Keane, Thomas M.; Jones, Steven; Hirst, Martin; Lorincz, Matthew C.; Mager, Dixie L.
Lithium insertion in carbonaceous materials containing silicon
NASA Astrophysics Data System (ADS)
Three different series of silicon-containing carbonaceous materials were synthesized for use as anodes in lithium ion cells. Disordered (or pregraphitic) carbons containing nanodispersed silicon were prepared by the chemical vapour deposition (CVD) of various chlorosilanes (SiClsb4, (CHsb3)sb2Clsb2Si, and (CHsb3)sb3ClSi) with benzene in two different apparatuses. Silicon oxycarbide glasses were synthesized by the pyrolysis of over 50 silicon-containing polymers at various temperatures, although the principal materials in the study were prepared at 1000sp░C. Finally, materials which we believe to be similar to disordered carbons containing nanodispersed silicon were prepared by the pyrolysis of various blends of pitches with polysilanes. Powder X-ray diffraction was used to learn about the structure of all the materials made. Thermal gravimetric analysis was used to determine the silicon content in the CVD materials and, when coupled to a residual gas analyzer, to study the decomposition process of the polymers. Near edge X-ray absorption spectroscopy measurements of the silicon L- and K-edges of CVD materials and the silicon K-edges of silicon oxycarbides were used to learn about local chemical environments of the silicon atoms. Lithium metal electrochemical test cells of the silicon-containing CVD materials showed larger capacities (up to 500 mAh/g) than pure carbons prepared in the same way (ś300 mAh/g). The additional capacity was observed to be centered near 0.4 V on charge, the average voltage observed for the removal of lithium from a silicon-lithium alloy. Chemical analysis showed that the stoichiometries of materials made by polymer pyrolysis were distributed over a well-defined region in the Si-O-C Gibbs phase diagram. An interesting series of materials is found near the line in the Si-O-C Gibbs triangle connecting carbon to SiOsb{1.3}. Lithium metal electrochemical test cells made using all the silicon oxycarbides synthesized showed that a stoichiometry of about Sisb{.25}Csb{.45}Osb{.30} gave the maximum reversible capacity (about 900 mAh/g). However, materials near this stoichiometry exhibit large irreversible capacities (>350 mAh/g) and significant hysteresis (the voltage difference between charge and discharge) in the voltage profile (ś0.8 V). In an attempt to reduce the oxygen content in one of the silicon oxycarbide glasses, a sample was washed in a dilute solution of hydrofluoric acid (HF) for times ranging from 2 minutes to 24 hours. The material lost, at most, 40 percent of its initial mass, although there was only a small change in its stoichiometry. In addition to the techniques mentioned above, small angle X-ray scattering and BET surface area measurements were used to study the microscopic pore network that was created by the HF washing. Lithium metal electrochemical test cells made using the product of pyrolysing pitch-polysilane blends showed that the capacity increased with silicon content from 340 mAh/g for pure carbon to a maximum of 600 mAh/g for samples with about 15 atomic % silicon (Sisb{.14}Osb{.09}Csb{.77}). The capacity then decreased to near zero as the composition approached SiC. These materials contain oxygen which is correlated to irreversible capacity loss. (Abstract shortened by UMI.)
Wilson, Alfred Macdonald
Timing of femoral prosthesis insertion during cemented arthroplasty: cement curing and static mechanical strength in an in vivo model
Background Modern cementing techniques aim to fix the implanted femoral prosthesis in the medullary cavity to minimize long-term complications such as aseptic loosening. The cure stage of bone cement into which the femoral component is being inserted is an important variable that is decided at the time of surgery. Late-cure cement is more viscous than early-cure cement and requires greater force on the part of the surgeon to insert the femoral prosthesis. We compared 2 cementing techniques, femoral component insertion into early-cure cement and insertion into late-cure cement, using an in vivo model to identify if cement cure stage affects the strength of the boneľcement interface. Methods We performed bilateral hemiarthroplasties using only the femoral component in vivo on paired porcine femora. The femora were harvested and cross-sectioned in preparation for strength testing. We measured bond strength by peak load required to push the femoral prosthesis and surrounding cement mantle free of the cancellous bone. Results All radiographs showed good cement interdigitation with no evidence of radiolucent lines at the boneľcement interface. We could not differentiate the early-cure and late-cure groups on postoperative radiographs. The mean failure load for the late-cure arthroplasties was 908 N (standard deviation [SD] 420 N), whereas the mean failure load for the conjugate early-cure arthroplasties was 503 N (SD 342 N). Conclusion Femoral component insertion into late-cure cement required significantly higher loads for push-out than femoral component insertion into early-cure cement.
Hunt, Stephen; Stone, Craig; Seal, Shane
Extra-anatomical complications of antegrade double-J insertion
Introduction: Insertion of a double-J (JJ) stent is a common procedure often carried out in the retrograde route by the urologists and the antegrade route by the radiologists. Reported complications include stent migration, encrustation, and fracture. Extra-anatomic placement of an antegrade JJ stent is a rare but infrequently recognized complication. Materials and Methods: We performed a retrospective audit of 165 antegrade JJ stent insertions performed over three consecutive years by a single interventional radiologist. All renal units were hydronephrotic at the time of nephrostomy. All procedures were performed under local anaesthetic with antibiotic prophylaxis. Results: Antegrade stent insertion was carried out simultaneously at the time of nephrostomy in 55 of the 165 cases (33%). The remainder were inserted at a mean of 2 weeks following decompression. In five (3%) patients, who had delayed antegrade stenting following nephrostomy, the procedure was complicated by silent ureteric perforation and an extra-anatomic placement of the stent. These complications had delayed manifestations, which included two retroperitoneal abscesses, a pelvic urinoma, a case each of ureterorectal fistula, and ureterovaginal fistula. Risk factors for ureteric perforation include previous pelvic malignancy, pelvic surgery, pelvic radiation, and a history of ureteric manipulation. Conclusion: Antegrade ureteric JJ stenting is a procedure not without complications. Extra-anatomic placement of the antegrade stent is a hitherto the infrequently reported complication but needs a high index of suspicion to be diagnosed. Risk factors for ureteric perforation at the time of stent insertion have to be considered to prevent this potential complication.
Rao, A. R.; Alleemudder, A.; Mukerji, G.; Mishra, V.; Motiwala, H.; Charig, M.; Karim, O. M. A.
Insertion of Alzheimer's A?40 Peptide into Lipid Monolayers
The amyloid beta (A?) peptide is the major component found in the amyloid deposits in the brains of Alzheimer's disease patients. In vitro studies have demonstrated that the aggregation of A? can take place at three orders of magnitude lower concentrations in the presence of phospholipid molecules compared to bulk peptide studies, suggesting that membrane lipids may mediate A? toxicity. To understand the interaction of A? with lipid membranes, we have examined A?40 with anionic dipalmitoylphosphatidylglycerol (DPPG), zwitterionic dipalmitoylphosphatidylcholine (DPPC), and cationic dipalmitoyltrimethylammonium propane (DPTAP) monolayers under different subphase conditions. We have used a constant surface pressure insertion assay to assess the degree of peptide insertion into the lipids. Simultaneously, we monitored the surface morphology of the monolayers with fluorescence microscopy. We have also performed dual-probe fluorescence measurements where both the peptide and lipid are tagged with chromophores. Isotherm measurements show that A? inserts into both DPTAP and DPPG monolayers under physiologically relevant conditions. Insertion into DPPC occurs at lipid densities below that found in a bilayer. The level of insertion is inversely proportional to the lipid packing density. Our results indicate that lipids need not be anionic to interact with A?. Electrostatic effects involved in A?40-lipid interaction are discussed.
Ege, Canay; Lee, Ka Yee C.
GAPDH regulates cellular heme insertion into inducible nitric oxide synthase
Heme proteins play essential roles in biology, but little is known about heme transport inside mammalian cells or how heme is inserted into soluble proteins. We recently found that nitric oxide (NO) blocks cells from inserting heme into several proteins, including cytochrome P450s, hemoglobin, NO synthases, and catalase. This finding led us to explore the basis for NO inhibition and to identify cytosolic proteins that may be involved, using inducible NO synthase (iNOS) as a model target. Surprisingly, we found that GAPDH plays a key role. GAPDH was associated with iNOS in cells. Pure GAPDH bound tightly to heme or to iNOS in an NO-sensitive manner. GAPDH knockdown inhibited heme insertion into iNOS and a GAPDH mutant with defective heme binding acted as a dominant negative inhibitor of iNOS heme insertion. Exposing cells to NO either from a chemical donor or by iNOS induction caused GAPDH to become S-nitrosylated at Cys152. Expressing a GAPDH C152S mutant in cells or providing a drug to selectively block GAPDH S-nitrosylation both made heme insertion into iNOS resistant to the NO inhibition. We propose that GAPDH delivers heme to iNOS through a process that is regulated by its S-nitrosylation. Our findings may uncover a fundamental step in intracellular heme trafficking, and reveal a mechanism whereby NO can govern the process.
Chakravarti, Ritu; Aulak, Kulwant S.; Fox, Paul L.; Stuehr, Dennis J.
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.
Costantini, Maria; Bernardi, Giorgio
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.
Krempaskř, 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)
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 [open quotes]master[close quotes] 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. Four such polymorphic human-specific Alu insertions were analyzed by a rapid, PCR-based assay. 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. 45 refs., 2 figs., 1 tab.
Batzer, M.A.; Alegria-Hartman, M. (Lawrence Livermore National Lab., CA (United States)); Stoneking, M. (Pennsylvania State Univ., University Park, PA (United States)); Bazan, H.; Kass, D.H.; Shaikh, T.H.; Scheer, W.D. (Louisiana State Univ. Medical Center, New Orleans, LA (United States)); Novick, G.E.; Herrera, R.J. (Florida International Univ., Miami, FL (United States)); Ioannou, P.A. (Cyprus Institute of Neurology and Genetics, Nicosia (Cyprus)) (and others)
A technique for simultaneous needle insertion in prostate seed implantation.
The purpose was to develop a fast needle insertion system to shorten the implantation time and to restrain prostate swelling during the implantation, thus reducing the seed setup error. The basic idea is to insert all the needles simultaneously using ultrasound guidance. The developed system consists of two similar templates that are connected. All the needles are set and locked to a moveable rear template according to the dose plan. The needle pack is then pushed into position, the lock released and seeds implanted needle by needle. A test and training phantom was also built. PMID:18263939
Heikkilń, Vesa-Pekka; Suorsa, Niina
Integral fill yarn insertion and beatup method using inflatable membrane
NASA Technical Reports Server (NTRS)
An apparatus and method for integral fill yarn insertion and beatup are disclosed. A modified rapier contains a channel for holding fill yarn. The channel is covered with a flexible and inflatable boot, and an inflating apparatus for this boot is also attached. Fill yarn is inserted into the channel, and the rapier is extended into a shed formed by warp yarn. Next, the rapier is pushed into the fell of the fabric, and the flexible and inflatable cover inflated, which both pushes the yarn into the fell of the fabric and performs beatup. The rapier is withdrawn and the shed closed to complete one step of the weaving process.
Farley, Gary L. (inventor)
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)
Synthesis and characterization of conducting polymer inserted carbon nanotubes
NASA Astrophysics Data System (ADS)
The carbon nanotubes filled with the photo-conducting polymer poly(N-vinyl carbazole) and the conducting polymer polypyrrole were prepared by polymerizing the monomers inside the nanotubes using the supercritical carbon dioxide. The endohedral nanotubes were characterized by HRTEM and ^1H NMR, which confirmed that the inserted material was indeed the conducting polymer [1]. I-V characteristics of the polymer inserted carbon nanotubes are presented. [1] Johannes Steinmetz, Soyoung Kwon, Hyun-Jung Lee, Edy Abou-Hamad, Robert Almairac, Christophe Goze-Bac, Hwayong Kim, Yung-Woo Park,, Chem. Phys. Lett., 431, 139 (2006)
Choi, A. Jeong; Nam, Young Woo; Park, Yung Woo
Viral insertion in Evi12 causes expression of aberrant Grp94 mRNAs containing the viral gag myristylation motif
Ecotropic Virus Integration site 12 (Evi12) is a common virus insertion site (cVIS) in retrovirally induced murine models of leukemia and lymphoma, suggesting an important role for this locus in these hematopoietic disorders. Evi12 is located near the promoter of the ER chaperone protein and Hsp90 family member Grp94. Here we show that viral insertion in Evi12 results in the expression of aberrant Grp94 transcripts in Cas-Br-MuLV as well as in AKXD induced hematopoietic tumors, demonstrating that Grp94 is a common viral target gene. While most transcripts encode for truncated forms of Grp94, transcripts containing viral gag sequences were detected in the leukemia cell line NFS107. Interestingly, these fusion transcripts encode for myristylated viral-Grp94 fusion proteins that localize to the plasma membrane. Combined with recent evidence that myristylated forms of Hsp90 transform cells, our data suggest that myristylation of target genes may be an important mechanism in retrovirally mediated oncogenesis. Since retroviral insertion in Evi12 also affects the expression of a recently identified novel gene Grp94 neighboring nucleotidase (Gnn), located at the other side of Evi12, it appears that proviral insertion can lead to deregulation of two genes present in the same locus.
Akker, Eric van den; Aarts, Lambertus H.J. [Department of Hematology, Erasmus University Medical Center, Dr. Molewaterplein 50, 3015 GE Rotterdam (Netherlands); Delwel, Ruud [Department of Hematology, Erasmus University Medical Center, Dr. Molewaterplein 50, 3015 GE Rotterdam (Netherlands)], E-mail: email@example.com
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.
Spradling, A C; Stern, D; Beaton, A; Rhem, E J; Laverty, T; Mozden, N; Misra, S; Rubin, G M
Method of installing a metallic threaded insert in a composite\\/rubber panel
A method of installing a metallic threaded insert in a composite\\/rubber panel consists of threading a first insert section into a preformed hole in a fiberglass composite panel and bonding a rubber panel over the fiberglass composite panel. The rubber panel includes an opening which is positioned over the first insert section. A second insert section is threaded onto an
Stanley Izdebski
NSDL National Science Digital Library
This lesson from Illuminations asks students to measure the diameter and circumference of various circular objects, plot the measurements on a graph, and relate the slope of the line to ├┬, the ratio of circumference to diameter. A student activity sheet is included. The material is appropriate for grades 9-12 and should require 1 class period to complete.
Trapping Cardiac Recessive Mutants via Expression-based Insertional Mutagenesis Screening
Rationale Mutagenesis screening is a powerful genetic tool for probing biological mechanisms underlying vertebrate development and human diseases. However, the increased colony management efforts in vertebrates impose a significant challenge for identifying genes affecting a particular organ such as the heart, especially those exhibiting adult phenotypes upon depletion. Objective We aim to develop a facile approach that streamlines colony management efforts via enriching cardiac mutants, which enables us to screen for adult phenotypes. Methods and Results The transparency of the zebrafish embryos enabled us to score 67 stable transgenic lines generated from an insertional mutagenesis screen using a transposon-based protein trapping vector. Fifteen lines with cardiac monomeric red fluorescent protein (mRFP) reporter expression were identified. We defined the molecular nature for 10 lines and bred them to homozygosity, which led to the identification of one embryonic lethal, one larval lethal, and one adult recessive mutant exhibiting cardiac hypertrophy at one year of age. Further characterization of these mutants uncovered an essential function of methionine adenosyltransferase II, alpha a (mat2aa) in cardiogenesis, an essential function of mitochondrial ribosomal protein S18B (mrps18b) in cardiac mitochondrial homeostasis, as well as a function of DnaJ (Hsp40) homolog, subfamily B, member 6b (dnajb6b) in adult cardiac hypertrophy. Conclusions We demonstrate that transposon-based gene trapping is an efficient approach for identifying both embryonic and adult recessive mutants with cardiac expression. The generation of a Zebrafish Insertional Cardiac (ZIC) mutant collection shall facilitate the annotation of a vertebrate cardiac genome, as well as enable heart-based adult screens.
Ding, Yonghe; Liu, Weibin; Deng, Yun; Jomok, Beninio; Yang, Jingchun; Huang, Wei; Clark, Karl J.; Zhong, Tao P.; Lin, Xueying; Ekker, Stephen C.; Xu, Xiaolei
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
Measurement of prostate rotation during insertion of needles for brachytherapy
Background and purposeThe purpose of this study is to investigate whether prostate rotation due to needle insertion for prostate brachytherapy is predictable and if so, to quantify this rotation, and to see whether locking needles reduce the magnitude of prostate rotation.
Vera Lagerburg; Marinus A. Moerland; Jan J. W. Lagendijk; Jan J. Battermann
Meningococcal septic shock after IUD insertion, a case presentation.
Neisseria meningitidis is a normal commensal of human mucous membranes that is no longer considered to be restricted to the nasopharynx. Due to the practice of oral sex, the mucous membranes of the cervix, urethra or anus have become a potential infection site for this bacterium. Inserting an intrauterine device (IUD), can alter the protective barrier of the endocervical mucosa, allowing for bacterial infection and systemic spread. We present a case report of a 40-year-old woman who presented with abdominal pain, spotting and fever after inserting an IUD and developed a fulminant septic shock. Blood cultures and cultures from ascites showed the presence of Neisseria meningitidis group Y. From our knowledge, there are a few cases presented in the literature of toxic shock syndrome after IUD insertion, caused by Staphylococcus aureus or Streptococcus group A, but this is the first case of meningococcal sepsis after IUD insertion described. So, even though IUDs rarely cause significant infection, physicians should consider this device as a possible source in reproductive-age women with the clinical features of sepsis. PMID:24502071
Romosan, Gina; Blidisel, A; Grigoras, D; Houtsios, A; Ionac, M
Modeling of external ear acoustics for insert headphone usage
Although acoustics of the external ear has been studied extensively for auralization and hearing aids, the acoustic behavior with insert headphones is not as well known. Our research focused on the eects of outer ear physical dimensions, particularly to sound pressure at the eardrum. The main parameter was the length of the canal, but eardrum's damping of resonances was also
Marko Hiipakka; Miikka Tikander; Matti Karjalainen
Hydrogen insertion compounds of UO/sub 3/
Hydrogen-insertion compounds, H /sub x/ UO/sub 3/, of two polymorphs of UO/sub 3/ have been prepared. Unit-cell dimensions have been determined by Guinier powder X-ray analysis and are compared with those of the parent oxide. Infrared spectra of these phases are also presented and discussed.
Dickens, P.G.; Hawke, S.V.; Weller, M.T.
Computerized Planning of Prostate Cryosurgery Using Variable Cryoprobe Insertion Depth
The current study presents a computerized planning scheme for prostate cryosurgery using a variable insertion-depth strategy. This study is a part of an ongoing effort to develop computerized tools for cryosurgery. Based on typical clinical practices, previous automated planning schemes have required that all cryoprobes be aligned at a single insertion depth. The current study investigates the benefit of removing this constraint, in comparison with results based on uniform insertion-depth planning as well as the so-called ôpullback procedureö. Planning is based on the so-called ôbubble-packing methodö, and its quality is evaluated with bioheat transfer simulations. This study is based on five 3D prostate models, reconstructed from ultrasound imaging, and cryoprobe active length in the range of 15 mm to 35 mm. The variable insertion depth technique is found to consistently provide superior results when compared to the other placement methods. Furthermore, it is shown that both the optimal active length and the optimal number of cryoprobes vary among prostate models, based on the size and shape of the target region. Due to its low computational cost, the new scheme can be used to determine the optimal cryoprobe layout for a given prostate model in real time.
Rossi, Michael R.; Tanaka, Daigo; Shimada, Kenji; Rabin, Yoed
Astronaut Rich Clifford inserts tape into payload high data recorder
NASA Technical Reports Server (NTRS)
On the Space Shuttle Endeavour's aft flight deck, Astronaut Michael R. (Rich) Clifford, mission specialist, inserts a tape in the payload high rate recorder. Three of these state-of-the-art recorders captured four times the amount of data that could be radioed to the ground.
Test Results for LHC Insertion Region Dipole Magnets.
National Technical Information Service (NTIS)
The Superconducting Magnet Division at Brookhaven National Laboratory (BNL) has made 20 insertion region dipoles for the Large Hadron Collider (LHC) at CERN. These 9.45 m-long, 8 cm aperture magnets have the same coil design as the arc dipoles now operati...
J. Muratore A. Jain M. Anerella J. Cozzolino G. Ganetis A. Ghosh R. Gupta
Magnesium oxide inserts for the LECO Carbon Analyzer
LECO carbon analysis of plutonium metal and plutonium oxide at the Rocky Flats Plant generates several hundred kilograms of high silica residues each year. The plutonium in these residues is difficult and expensive to recover using production dissolution processes. A magnesium oxide (MgO) insert has been developed that significantly lowers the plutonium recovery costs without adversely affecting accuracy of the carbon analysis.
Bagaasen, L.M.; Jensen, C.M.
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
In search of polymorphic Alu insertions with restricted geographic distributions
Summary Alu elements are transposable elements that have reached over one million copies in the human genome. Some Alu elements inserted in the genome so recently that they are still polymorphic for insertion presence or absence in human populations. Recently, there has been an increasing interest in using Alu variation for studies of human population genetic structure and inference of individual geographic origin. Currently, this requires a high number of Alu loci. Here, we used a linker-mediated PCR method to preferentially identify low frequency Alu elements in various human DNA samples with different geographic origins. The candidate Alu loci were subsequently genotyped in 18 worldwide human populations (~370 individuals), resulting in the identification of two new Alu insertions restricted to populations of African ancestry. Our results suggest that it may ultimately become possible to correctly infer the geographic affiliation of unknown samples with high levels of confidence without having to genotype as many as 100 Alu loci. This is desirable if Alu insertion polymorphisms are to be used for human evolution studies or forensic applications.
Cordaux, Richard; Srikanta, Deepa; Lee, Jungnam; Stoneking, Mark; Batzer, Mark A.
In search of polymorphic Alu insertions with restricted geographic distributions.
Alu elements are transposable elements that have reached over one million copies in the human genome. Some Alu elements inserted in the genome so recently that they are still polymorphic for insertion presence or absence in human populations. Recently, there has been an increasing interest in using Alu variation for studies of human population genetic structure and inference of individual geographic origin. Currently, this requires a high number of Alu loci. Here, we used a linker-mediated polymerase chain reaction method to preferentially identify low-frequency Alu elements in various human DNA samples with different geographic origins. The candidate Alu loci were subsequently genotyped in 18 worldwide human populations (approximately 370 individuals), resulting in the identification of two new Alu insertions restricted to populations of African ancestry. Our results suggest that it may ultimately become possible to correctly infer the geographic affiliation of unknown samples with high levels of confidence without having to genotype as many as 100 Alu loci. This is desirable if Alu insertion polymorphisms are to be used for human evolution studies or forensic applications. PMID:17498924
Cordaux, Richard; Srikanta, Deepa; Lee, Jungnam; Stoneking, Mark; Batzer, Mark A
Indentation and needle insertion properties of the human eye.
PurposeCharacterization of the biomechanical properties of the human eye has a number of potential utilities. One novel purpose is to provide the basis for development of suitable tissue-mimicking material. The purpose of this study was to determine the indentation and needle insertion characteristics on human eye globes and tissue strips.MethodsAn indenter assessed the elastic response of human eye globes and tissue strips under increasing compressive loads. Needle insertion determined the force (N) needed to penetrate various areas of the eye wall.ResultsThe results demonstrated that globes underwent slightly greater indentation at the midline than at the central cornea, and corneal strips indented twofold more than scleral strips, although neither difference was significant (P=0.400 and P=0.100, respectively). Significant differences were observed among various areas of needle insertion (P<0.001). Needle insertion through the anterior sclera (adjacent to the limbus) and posterior sclera (adjacent to the optic nerve) required the greatest amount of force (0.954 and 1.005?N, respectively). The force required to penetrate the central cornea (0.518?N) was significantly lower than all other areas except the midline sclera (0.700?N)ConclusionThese data form the basis for further research into the development of a tissue-mimicking human eye construct with potential utility as a model for use in ophthalmology research and surgical teaching. PMID:24810571
Matthews, A; Hutnik, C; Hill, K; Newson, T; Chan, T; Campbell, G
Axial-power-shaping rod-insertion test
On 23 to 25 June 1982 the eight TMI Unit 2 Axial Power Shaping Rods (APSRs) underwent insertion testing to determine the mechanicsl motion of the drive and rod systems. The purpose of the test was twofold: to gain insight into the extent of core and upper plenum damage from knowing the ability to move some or all of the
K. Parlee; J. Weissburg; W. Austin; G. Carter
Axial-Power-Shaping Rod-Insertion Test.
National Technical Information Service (NTIS)
On 23 to 25 June 1982 the eight TMI Unit 2 Axial Power Shaping Rods (APSRs) underwent insertion testing to determine the mechanicsl motion of the drive and rod systems. The purpose of the test was twofold: to gain insight into the extent of core and upper...
K. Parlee J. Weissburg W. Austin G. Carter
Astronaut Scott Carpenter inserted into Aurora 7 spacecraft
NASA Technical Reports Server (NTRS)
Astronaut M. Scott Carpenter, pilot of the Mercury-Atlas 7 space flight, is inserted into Aurora 7 spacecraft during the prelaunch countdown. Carpenter is assisted into the spacecraft by Astronaut John Glenn and Gunter Vendt, McDonnell Douglas pad capsule test conducter.
The Insertion of Halocarbenes into the Mercury-Halogen Bond.
National Technical Information Service (NTIS)
The previously postulated possibility of inserting CX2 into a mercury-halogen bond was demonstrated experimentally. An example of such a process is: CHCl3 + photon (yields) .CHCl2 + .Cl (yields) HCl + :CCl2; :CCl2 + HgCl (yields) CCl3HgCl.
M. E. Gordon K. V. Darragh D. Seyferth
Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey
33. INTERIOR VIEW OF FEEDER PREPARING TO INSERT THE FEEDER ROD INTO ONE OF THE TWO LARGER NAIL OR SPIKE CUTTING MACHINES WHICH PRODUCE THE LARGER SIZE NAILS; NOTE THE OTHER SPIKE CUTTING MACHINE IN THE BACKGROUND - LaBelle Iron Works, Thirtieth & Wood Streets, Wheeling, Ohio County, WV
21 CFR 529.1940 - Progesterone intravaginal inserts.
Code of Federal Regulations, 2013 CFR
...should be disposed after a single use. Administration of vaginal inserts for periods greater than 7 days may result in reduced fertility. Dinoprost solution provided by No. 000009 in ┬ž 510.600(c) of this chapter. (2) Ewes Ô(i) Amount ....
Embedded Micromirror Inserts for Optical Printed Circuit Boards
We present the use of an embedded 45deg micromirror, which is patterned in polymer photoresist using deep proton writing. The micromirror is metallized and inserted in a laser ablated cavity in the optical layer and in a next step covered with cladding material. Surface roughness measurements confirm the excellent quality of the mirror facet. Loss measurements have been carried out
Nina Hendrickx; J▄rgen Van Erps; Erwin Bosman; Christof Debaes; Hugo Thienpont; Peter Van Daele
Developmental Outcomes after Early or Delayed Insertion of Tympanostomy Tubes
background To prevent later developmental impairments, myringotomy with the insertion of tympa- nostomy tubes has often been undertaken in young children who have persistent otitis media with effusion. We previously reported that prompt as compared with delayed in- sertion of tympanostomy tubes in children with persistent effusion who were younger than three years of age did not result in improved
Jack L. Paradise; Thomas F. Campbell; Christine A. Dollaghan; Heidi M. Feldman; Beverly S. Bernard; D. Kathleen Colborn; Howard E. Rockette; Janine E. Janosky; Dayna L. Pitcairn; Marcia Kurs-Lasky; Diane L. Sabo; Clyde G. Smith
Hearing Aid Evaluation: Predicting Speech Gain from Insertion Gain.
ERIC Educational Resources Information Center
This study, involving 11 older males with mild or moderate sensorineural loss, tested whether speech gain that results from a hearing aid fitting is predictable on the basis of unaided performance intensity curve, unaided narrow-band sound field thresholds, hearing aid insertion gain as a function of frequency, ambient noise, and internal hearingů
Dillon, Harvey
Analysis of HIV-2 Vpx by modeling and insertional mutagenesis
Vpx facilitates HIV-2 nuclear localization by a poorly understood mechanism. We have compared Vpx to an NMR structure HIV-1 Vpr in a central helical domain and probed regions of Vpx by insertional mutagenesis. A predicted loop between helices two and three appears to be unique, overlapping with a known novel nuclear localization signal. Overall, Vpx was found to be surprisingly flexible, tolerating a series of large insertions. We found that insertion within the polyproline-containing C-terminus destabilizes nuclear localization, whereas mutating a second helix in the central domain disrupts viral packaging. Other insertional mutants in the predicted loop and in a linker region between the central domain and the C-terminus may be useful as sites of intramolecular tags as they could be packaged adequately and retained preintegration complex associated integration activity in a serum starvation assay. An unexpected result was found within a previously defined nuclear localization motif near aa 71. This mutant retained robust nuclear localization in a GFP fusion assay and was competent for preintegration complex associated nuclear import. In summary, we have modeled helical content in Vpx and assessed potential sites of intramolecular tags which may prove useful for protein-protein interactions studies.
Mahnke, Lisa A. [Department of Medicine, Divisions of Oncology and Infections Diseases, Washington University School of Medicine, PO Box 8069, 660 South Euclid Avenue, Saint Louis, MO 63110 (United States)]. E-mail: firstname.lastname@example.com; Belshan, Michael [Department of Medicine, Divisions of Oncology and Infections Diseases, Washington University School of Medicine, PO Box 8069, 660 South Euclid Avenue, Saint Louis, MO 63110 (United States); Ratner, Lee [Department of Medicine, Divisions of Oncology and Infections Diseases, Washington University School of Medicine, PO Box 8069, 660 South Euclid Avenue, Saint Louis, MO 63110 (United States)]. E-mail: email@example.com
Characteristics of joining inserts for composite sandwich panels
Composite sandwich constructions are widely employed in various light weight structures, because composite sandwich panels have high specific stiffness and high specific bending strength compared to solid panels. Since sandwich panels are basically unsuited to carry localized loads, the sandwich structure should provide joining inserts to transfer the localized loads to other structures.In this work, the load transfer characteristics of
Byoung Jung Kim; Dai Gil Lee
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.
Bone registration method for robot assisted surgery: pedicle screw insertion
SUMMARY A registration method that identifies bone geometry with respect to a robotic manipulator arm is presented. Although the method is generally applicable to many orthopaedic internal fixation, it was only demonstrated for the insertion of pedicle screws in vertebral bodies for spine fixation. The method relies upon obtaining an impression of the vertebral bodies. Computed Tomography scans of both
K Abdel-Malek; D P McGowan; V K Goel; D Kowalski; A Hager
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
Preparation of Boron-Substituted Carboranes by Boron Insertion Reactions.
National Technical Information Service (NTIS)
The boron insertion reaction of Hawthorne was employed in preparing 3-fluoro-, 3-bromo- and 3-diphenylamino-o-carborane. This reaction was further extended to the m-carborane system to produce 2-fluoro- and 2-diphenylamino-m-carborane. 3-Fluoro-o-carboran...
J. S. Roscoe S. Kongpricha S. Papetti
Mobilizing diversity: transposable element insertions in genetic variation and disease
Transposable elements (TEs) comprise a large fraction of mammalian genomes. A number of these elements are actively jumping in our genomes today. As a consequence, these insertions provide a source of genetic variation and, in rare cases, these events cause mutations that lead to disease. Yet, the extent to which these elements impact their host genomes is not completely understood.
Kathryn A O'Donnell; Kathleen H Burns
Low-cost waveguide filters with printed-circuit inserts
An easy to manufacture waveguide filter is presented. This type of filter is based on printed-circuit-board inserts with resonator strips. The electromagnetic properties of the strips interacting with the electromagnetic field inside the waveguide are described by equivalent circuit elements. Filter synthesis and computer optimization is applied to design band-stop filters. Finally simulated results are compared with measurements
H. Schmiedel; M. Muller; A. Anufriev
hiTRAN« Wire Matrix Inserts in Fouling Applications
Fouling characteristics are dictated largely by the properties of the thermal and hydrodynamic boundary layers. As a result, fouling mitigation strategies must take into account the conditions in this region. hiTRAN wire matrix tube inserts are a useful tool in altering the conditions near the tube wall, especially in the laminar and transition flow regions. This review article considers particle
J. M. Ritchie; P. Droegemueller; M. J. H. Simmons
Insertional Mutagenesis of the Drosophila Genome with Single P Elements
A versatile genetic method for identifying and cloning Drosophila melanogaster genes affecting any recognizable phenotype is described. Strains are constructed in which the insertion of a single P transposable element has caused a new mutation, greatly simplifying the genetic and molecular analysis of the affected gene. Mutagenesis is initiated by crossing two strains, each of which contains a specially designed
Lynn Cooley; Richard Kelley; Allan Spradling
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
Placental and cord insertion pathologies: screening, diagnosis, and management.
Placenta previa, low-lying placenta, and placenta accreta are aberrations in the normal development of the placenta. Diagnosis in the prenatal period is essential because each of these pathologies can have a profound impact on the management of labor, birth, and the third stage. In recent years, there has been an increase in the occurrence of these placental abnormalities, with the increase in the cesarean rate considered to be a main cause of this phenomenon. Comprehensive risk assessment, combined with recent advances in ultrasonography, can provide earlier detection of impaired placental implantation. Umbilical cord insertion pathologies are also of concern. Velamentous cord insertion is a defect in the insertion site of the umbilical cord resulting from the atrophy of portions of the developing placenta. In this condition, the blood vessels of the umbilical cord are not protected by Wharton's jelly, resulting in a potential for increased risk of breakage when the amniotic membranes rupture. Vasa previa is a velamentous insertion of the umbilical cord in which the blood vessels are present over the cervical os. If these blood vessels rupture during labor, it can have catastrophic effects on the fetus. Prenatal diagnosis of this condition can allow the certified nurse-midwife/certified midwife (CNM/CM) to plan for the safe birth of the newborn and avoid fetal hemorrhage. This article provides a review of risk factors, diagnosis pathophysiology, and management options for these conditions, thus enabling the CNM/CM to provide safe, effective care and management. PMID:24751147
Wiedaseck, Susan; Monchek, Ruth
Insertion site specificity of the transposon Tn3.
The Tn3-deletion method [Davies and Hutchison, Nucleic Acids Res. 19, 5731-5738, (1991)] was used to sequence a 9.4 kb DNA fragment. Transpositional 'warm' spots were not a limiting factor but a 935 bp 'cold' spot was completed using a synthetic oligonucleotide primer. Two hundred and twenty three miniTn3 insertion sites from three sequencing projects were aligned and a 19 bp asymmetric consensus site was identified. There is no absolute sequence requirement at any position in this consensus, so insertion occurs promiscuously (approximately 37% of sites are potential targets). In our sequencing projects, multiply targeted sites always closely matched the consensus, although not all close matches were targeted frequently. The 935 bp cold spot showed no unusual features when analysed with the consensus sequence. The consensus can be used to accurately predict likely insertion sites in a new sequence. Synthetic oligonucleotides based on the consensus and a known hot spot for Tn3 were mutagenised. These sequences were not hot spots in our vectors, suggesting that the primary sequence alone is not sufficient to create an insertional hot spot. We conclude that some other factor, such as DNA secondary structure, also plays an important role in target site selection for the transposon Tn3. PMID:7885847
Davies, C J; Hutchison, C A
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
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
Fluid flow in a tube with an elastic membrane insertion
The unsteady flow of a viscous incompressible fluid in a circular tube with an elastic insertion is studied numerically. The deformation of the elastic membrane is obtained by the theory of nite elasticity whose equations are solved simultaneously with the fluid equations in the axisymmetric approximation. The elastic wall expands outwards due to the positive transmural pressure and represents an
G IANNI P EDRIZZETTI; Ingegneria Civile
Insert earphone modeling and measurement by IEC-60711 coupler.
In this study, an analytical model based on the equivalent circuit method is developed to simulate the frequency response of an insert earphone. This earphone incorporates a miniature loudspeaker commonly used in computer, communication, consumer, and car electronics. Through the laser triangulation method, electroacoustic parameters of a miniature loudspeaker are obtained. Several earphone design configurations are analyzed in accordance with the open and closed states of front leakage hole, vent, and back leakage hole. To validate the analysis, an insert earphone that is attached to IEC-60711 coupler and a specially designed fixture tube is experimentally measured for frequency response using electroacoustic equipment in the air. Simulation and experimental results show good agreement over the complete audible frequency range. Analysis indicates that states of front leakage hole, vent, and back leakage hole of an insert earphone have significant effects on frequency response. The front leakage hole affects the low frequency response, whereas the vent affects the fundamental resonance. Detailed analysis has been provided to further improve the design of insert earphones. PMID:21342831
Huang, Chen-Hung; Pawar, S; Hong, Zih-Jyun; Huang, Jin
Use of dimorphic Alu insertions in human DNA fingerprinting.
National Technical Information Service (NTIS)
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 pop...
G. E. Novick T. Gonzalez J. Garrison C. C. Novick R. J. Herrera
Vitamer Levels, Stress Response, Enzyme Activity, and Gene Regulation of Arabidopsis Lines Mutant in the Pyridoxine/Pyridoxamine 5?-Phosphate Oxidase (PDX3) and the Pyridoxal Kinase (SOS4) Genes Involved in the Vitamin B6 Salvage Pathway1[W][OA
PDX3 and SALT OVERLY SENSITIVE4 (SOS4), encoding pyridoxine/pyridoxamine 5?-phosphate oxidase and pyridoxal kinase, respectively, are the only known genes involved in the salvage pathway of pyridoxal 5?-phosphate in plants. In this study, we determined the phenotype, stress responses, vitamer levels, and regulation of the vitamin B6 pathway genes in Arabidopsis (Arabidopsis thaliana) plants mutant in PDX3 and SOS4. sos4 mutant plants showed a distinct phenotype characterized by chlorosis and reduced plant size, as well as hypersensitivity to sucrose in addition to the previously noted NaCl sensitivity. This mutant had higher levels of pyridoxine, pyridoxamine, and pyridoxal 5?-phosphate than the wild type, reflected in an increase in total vitamin B6 observed through HPLC analysis and yeast bioassay. The sos4 mutant showed increased activity of PDX3 as well as of the B6 de novo pathway enzyme PDX1, correlating with increased total B6 levels. Two independent lines with T-DNA insertions in the promoter region of PDX3 (pdx3-1 and pdx3-2) had decreased PDX3 activity. Both also had decreased activity of PDX1, which correlated with lower levels of total vitamin B6 observed using the yeast bioassay; however, no differences were noted in levels of individual vitamers by HPLC analysis. Both pdx3 mutants showed growth reduction in vitro and in vivo as well as an inability to increase growth under high light conditions. Increased expression of salvage and some of the de novo pathway genes was observed in both the pdx3 and sos4 mutants. In all mutants, increased expression was more dramatic for the salvage pathway genes.
Gonzalez, Eugenia; Danehower, David; Daub, Margaret E.
Long acting betaxolol ocular inserts based on polymer composite.
Poor bioavailability and therapeutic response of conventional therapy due to many pre-corneal constraints necessitate the development of novel controlled and sustained ocular drug delivery to become a standard one in modern pharmaceutical era. This investigation aimed to study the drug release kinetics of betaxolol hydrochloride from a hydrophobic matrix system of PMMA cast with incorporating different proportions of polyethylene oxide (PEO) and evaluate its ability to improve ocular bioavailability and duration of action for the drug. Matrix type ocular inserts were prepared by the film casting technique and characterized in vitro by drug release studies using a flow through apparatus that simulated the eye conditions. All the formulations were subjected to physicochemical evaluation. Rabbit model with steroid induced glaucoma was used to establish in vivo efficacy of inserts. Polymer composition and concentration significantly affected the drug release based on change in diffusional path length and formation of gelaneous pores by polymer erosion. Formulations released the drug by non-fickian diffusion including anomalous transport (0.5
Gevariya, Hitesh B; Patel, Jayvadan K
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 ônanosilosö 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 1ľ25, SP-B 9ľ25, SP-B 11ľ25, and SP-B 1ľ25Nflex (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 1ľ25Nflex) 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 1ľ25 (as a monomer or dimer) and synthetic lung lavage lipids quickly improved oxygenation and dynamic compliance, whereas SP-B 11ľ25 surfactants showed oxygenation and dynamic compliance values similar to that of lipids alone, demonstrating a positive correlation between formation of stable, but reversible, nanosilos and in vivo efficacy.
Frey, Shelli L.; Pocivavsek, Luka; Waring, Alan J.; Walther, Frans J.; Hernandez-Juviel, Jose M.; Ruchala, Piotr
Recent insertion of a 52-kb mitochondrial DNA segment in the wheat lineage.
The assembly of a 1.3-Mb size region of the wheat genome has provided the opportunity to study a recent nuclear mitochondrial DNA insertion (NUMT). In the present study, we have studied two bacterial artificial chromosomes (BACs) and characterized a 52-kb NUMT segment from the tetraploid and hexaploid wheat BAC libraries. The conserved orthologous NUMT regions from tetraploid and hexaploid wheat Langdon and Chinese Spring shared identical gene haplotypes even though mutations (insertions, deletions, and substitutions) had occurred. The 52-kb NUMT was present in hexaploid variety Chinese Spring, but absent in variety Hope, by sequence comparison of their corresponding region. Amplifying the NUMT junctions using a set of the wheat materials including diploid, tetraploid, and hexaploid lines showed that none of the diploid wheat carried the region and only some tetraploid and hexaploid wheat were positive for the NUMT. Age estimation of the NUMT displayed the mean ages of Langdon NUMT and Chinese Spring NUMT to be 378,000 and 416,000áyears ago, respectively. Reverse transcription PCR and sequencing of the nad7 gene showed 28 C ? U RNA editing sites and four partial editing sites, as expected for mitochondrial DNA expression. Specific SNPs discriminated between cDNA from the nucleus and the mitochondria and suggested that the nuclear copy was not expressed. The mitochondrial DNA studied was inserted into the genome quite recently within the wheat lineage and gave rise to the non-coding nuclear nad7 gene. The NUMT segment could be lost and acquired frequently during the wheat evolution. PMID:21761280
Zhang, Juncheng; Jia, Jizeng; Breen, James; Kong, Xiuying
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.
Suttiprapa, Sutas; Kines, Kristine J.; Tort, Jose F.; Mann, Victoria H.; Turner, Daniel J.; Berriman, Matthew; Brindley, Paul J.
NSDL National Science Digital Library
All About Circuits is a website that ├ó┬┬provides a series of online textbooks covering electricity and electronics.├ó┬┬ Written by Tony R. Kuphaldt, the textbooks available here are wonderful resources for students, teachers, and anyone who is interested in learning more about electronics. This specific section, Transmission Lines, is the fourteenth chapter in Volume II ├ó┬┬ Alternating Current. A few of the topics covered in this chapter include: Circuits and the speed of light; Finite-length transmission lines; and Waveguides. Diagrams and detailed descriptions of concepts are included throughout the chapter to provide users with a comprehensive lesson. Visitors to the site are also encouraged to discuss concepts and topics using the All About Circuits discussion forums (registration with the site is required to post materials).
Kuphaldt, Tony R.
NSDL National Science Digital Library
Children can use this interactive applet to practice counting and ordering numbers up to 24 in groups of 10 at a time. An empty clothesline appears along with 9 number cards and a mystery card. Students drag the numbers onto the line in the correct sequence and then determine the mystery number. After the first round, 1 - 10, users may select from three ranges, starting at 1, 7, or 15.
Analysis of Primary Stability of Dental Implants Inserted in Different Substrates Using the Pullout Test and Insertion Torque
The aim of the study was to evaluate mechanical behavior of implants inserted in three substrates, by measuring the pullout strength and the relative stiffness. 32 implants (Master Porous-Conexao, cylindrical, external hexagon, and surface treatment) were divided into 4 groups (n = 8): pig rib bone, polyurethane Synbone, polyurethane Nacional 40 PCF, and pinus wood. Implants were installed with the exact distance of 5?mm of another implant. The insertion torque (NĚcm) was quantified using the digital Kratos torque meter and the pullout test (N) was performed by an axial traction force toward the long axis of the implant (2?min/mm) through mount implant devices attached to a piece adapted to a load cell of 200?Kg of a universal testing machine (Emic DL10000). Data of insertion torque and maximum pullout force were submitted to one-way ANOVA and Bonferroni tests (? = 0.05). Polyurethane Nacional 40 PCF and pinus wood showed the highest values of insertion torque and pullout force, with significant statistical difference (P < 0.05) with other groups. The analysis showed stiffness materials with the highest values for primary stability.
Oliscovicz, Nathalia Ferraz; Shimano, Antonio Carlos; Marcantonio Junior, Elcio; Lepri, Cesar Penazzo; dos Reis, Andrea Candido
This article presents the results of an experimental investigation of heat transfer augmentation by porous media in a natural gas-fired radiant tube burner. The results show that significant heat transfer augmentation is possible with the use of porous ceramic inserts in both premixed and nonpremixed gas-fired radiant tube burners. Furthermore, this work has shown that geometry variations in the porous
T. E. Peard; J. E. Peters; M. Q. Brewster; R. O. Buckius
Chromosomal integration of phage lambda by means of a DNA insertion element.
Phage lambdacam112, which contains the chloramphenicol resistance transposon Tn9 and has a deletion of attP and the int gene, will lysogenize Escherichia coli K-12. Prophage integration occurs at different chromosomal sites, including lacY and malB, but not at attB. All lambdacam112 prophages are excised from the chromosome after induction but with various efficiencies for different locations. Heteroduplex analysis of lambdaplacZ transducing phages isolated from a lacY::lambdacam112 prophage reveals an insertion sequence 1 (IS1) element at the joint of viral and chromosomal DNA. Two lines of evidence indicate that lambdacam112 encodes an excision activity that recognizes the IS1 element: (i) prophage derepression increases the frequency of excision from lacY to yield lac+ revertants, and (ii) lambdacam112 infection increases reversion of a galT::IS1 mutation about 50-fold. Our results indicate that the IS1 termini of TN9 can replace attP as a site for lambda insertion in the bacterial chromosome and that excision events are catalyzed by an IS1-encoded protein under lambda repressor and N gene control.
MacHattie, L A; Shapiro, J A
Sequencing human-gibbon breakpoints of synteny reveals mosaic new insertions at rearrangement sites
The gibbon genome exhibits extensive karyotypic diversity with an increased rate of chromosomal rearrangements during evolution. In an effort to understand the mechanistic origin and implications of these rearrangement events, we sequenced 24 synteny breakpoint regions in the white-cheeked gibbon (Nomascus leucogenys, NLE) in the form of high-quality BAC insert sequences (4.2 Mbp). While there is a significant deficit of breakpoints in genes, we identified seven human gene structures involved in signaling pathways (DEPDC4, GNG10), phospholipid metabolism (ENPP5, PLSCR2), ?-oxidation (ECH1), cellular structure and transport (HEATR4), and transcription (ZNF461), that have been disrupted in the NLE gibbon lineage. Notably, only three of these genes show the expected evolutionary signatures of pseudogenization. Sequence analysis of the breakpoints suggested both nonclassical nonhomologous end-joining (NHEJ) and replication-based mechanisms of rearrangement. A substantial number (11/24) of humanľNLE gibbon breakpoints showed new insertions of gibbon-specific repeats and mosaic structures formed from disparate sequences including segmental duplications, LINE, SINE, and LTR elements. Analysis of these sites provides a model for a replication-dependent repair mechanism for double-strand breaks (DSBs) at rearrangement sites and insights into the structure and formation of primate segmental duplications at sites of genomic rearrangements during evolution.
Girirajan, Santhosh; Chen, Lin; Graves, Tina; Marques-Bonet, Tomas; Ventura, Mario; Fronick, Catrina; Fulton, Lucinda; Rocchi, Mariano; Fulton, Robert S.; Wilson, Richard K.; Mardis, Elaine R.; Eichler, Evan E.
Analysis of cell poration by femtosecond laser for particle insertion by optical manipulation
NASA Astrophysics Data System (ADS)
The introduction and subsequent expression of external DNA inside single living mammalian cell (transfection) can be achieved by photoporation with femtosecond laser. After photoporation, external DNA can be introduced by trapping and successive insertion of DNA coated nanoparticle in the cell using optical tweezers. To maximize the transfection efficiency, one of the major aspects is that the photoporated cell should not be damaged and cell membrane should heal itself immediately or after sometime while the cells are healed in the CO2 incubator. Furthermore, the size of hole created as a result of photoporation should be more than the size of DNA coated nanoparticle to be inserted inside the cell. In this paper, an analysis has been done on single cell of important breast cancer cell lines named MCF-7 and MDAMB231. Size of holes created in cell membrane after photoporation has been measured and the required optimum energy with sustained cell life were determined. Using this analysis, most favorable conditions for maximum transfection efficiency can be determined.
Muhammad, Waleed; Kim, Jung-Dae; Lee, Yong-Gu
The effect of insertion depth on the theoretical SAR patterns of 915 MHz dipole antenna arrays for hyperthermia.
Interstitial microwave antenna array hyperthermia (IMAAH) is presently used clinically in the treatment of cancer. This paper presents the theoretical specific absorption rate (SAR) patterns of 915 MHz microwave antenna arrays for varying insertion depths. The antennas were oriented in a 2 x 2 cm square array. Insertion depth, defined as distance from skin to antenna tip, ranged from 5.9 to 17.6 cm. Two different antenna configurations were considered. In the first the antenna had a distal section a quarter-wavelength long (resonant case), while the second had a distal section approximately 13% longer than a quarter-wavelength (non-resonant case). SAR patterns were calculated from theoretical expressions, and displayed as lines of constant SAR normalized to the maximum SAR value in the array. The results show that regions of concentrated power deposition or 'hotspots' occurred in the centre of the array and moved in a complex but predictable fashion as insertion depth was varied. For insertion depths shorter than a resonant half-wavelength, there occurred one hotspot distal to the antenna junctions. As insertion depth was increased beyond a resonant half-wavelength, the hotspot moved proximal to the antenna junctions and eventually split in two. For depths very much longer than a resonant half-wavelength a hotspot centred about the antenna junction dominated the SAR pattern. For the resonant case the maximum SAR was often along the central axis of the array, while for the non-resonant case the maximum SAR was at the antennas with a local maximum on the central axis. PMID:2592787
James, B J; Strohbehn, J W; Mechling, J A; Trembly, B S
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
Cassini observations of Saturn's inner plasmasphere: Saturn orbit insertion results
NASA Astrophysics Data System (ADS)
We present new and definitive results of Cassini plasma spectrometer (CAPS) data acquired during passage through Saturn's inner plasmasphere by the Cassini spacecraft during the approach phase of the Saturn orbit insertion period. This analysis extends the original analysis of Sittler et al. [2005. Preliminary results on Saturn's inner plasmasphere as observed by Cassini: comparison with Voyager. Geophys. Res. Lett. 32, L14S07, doi:10.1029/2005GL022653] to Lś10 along with also providing a more comprehensive study of the interrelationship of the various fluid parameters. Coincidence data are sub-divided into protons and water group ions. Our revised analysis uses an improved convergence algorithm which provides a more definitive and independent estimate of the spacecraft potential ?SC for which we enforce the protons and water group ions to co-move with each other. This has allowed us to include spacecraft charging corrections to our fluid parameter estimations and allow accurate estimations of fluctuations in the fluid parameters for future correlative studies. In the appendix we describe the ion moments algorithm, and minor corrections introduced by not weighting the moments with sin ? term in Sittler et al. [2005] (Correction offset by revisions to instruments geometric factor). Estimates of the spacecraft potential and revised proton densities are presented. Our total ion densities are in close agreement with the electron densities reported by Moncuquet et al. [2005. Quasi-thermal noise spectroscopy in the inner magnetosphere of Saturn with Cassini/RPWS: electron temperatures and density. Geophys. Res. Lett. 32, L20S02, doi:10.1029/2005GL022508] who used upper hybrid resonance (UHR) emission lines observed by the radio and plasma wave science (RPWS) instrument. We show a positive correlation between proton temperature and water group ion temperature. The proton and thermal electron temperatures track each with both having a positive radial gradient. These results are consistent with pickup ion energization via Saturn's rotational electric field. We see evidence for an anti-correlation between radial flow velocity VR and azimuthal velocity V ?, which is consistent with the magnetosphere tending to conserve angular momentum. Evidence for MHD waves is also present. We show clear evidence for outward transport of the plasma via flux tube interchange motions with the radial velocity of the flow showing positive radial gradient with Vś0.12(L/4)5.5km/s functional dependence for 4< L<10 (i.e., if we assume to be diffusive transport then DśD0L11 for fixed stochastic time step ?t). Previous models with centrifugal transport have used DśD0L3 dependence. The radial transport seems to begin at Enceladus' L shell, Lś4, where we also see a minimum in the W+ ion temperature Tś35eV. For the first time, we are measuring the actual flux tube interchange motions in the magnetosphere and how it varies with radial distance. These observations can be used as a constraint with regard to future transport models for Saturn's magnetosphere. Finally, we evaluate the thermodynamic properties of the plasma, which are all consistent with the pickup process being the dominant energy source for the plasma.
Sittler, E. C.; Thomsen, M.; Johnson, R. E.; Hartle, R. E.; Burger, M.; Chornay, D.; Shappirio, M. D.; Simpson, D.; Smith, H. T.; Coates, A. J.; Rymer, A. M.; McComas, D. J.; Young, D. T.; Reisenfeld, D.; Dougherty, M.; Andre, N.
Influence of inserting a thin fullerene layer on pentacene organic thin-film transistor
NASA Astrophysics Data System (ADS)
The performance of organic thin-film transistors (TFTs) with a pentacene/fullerene(C60)/pentacene (PCP) sandwich structure is presented. Using a 3.5 nm-thick C60 layer inserted between the pentacene films, the obtained hole mobility is improved by more than six times. By applying atomic force microscopy, x-ray diffraction, Raman spectrum, and transmission line method analysis, one can reasonably infer that the smoother surface of the pentacene film covered with thin C60 layer delays the phase transformation of the upper pentacene film, resulting in stronger intermolecular coupling and the reduction of channel resistance of the PCP TFTs from 3.03 to 1.72 M?, and, therefore, improving the device performance.
Li, Yu-Chang; Lin, Yu-Ju; Wei, Chia-Yu; Chou, Dei-Wei; Tsao, Chun-Ho; Wang, Yeong-Her
CUDA accelerated simulation of needle insertions in deformable tissue
NASA Astrophysics Data System (ADS)
This paper presents a stiff needle-deformable tissue interaction model. The model uses a mesh-less discretization of continuum; avoiding thus the expensive remeshing required by the finite element models. The proposed model can accommodate both linear and nonlinear material characteristics. The needle-deformable tissue interaction is modeled through fundamental boundaries. The forces applied by the needle on the tissue are divided in tangent forces and constraint forces. The constraint forces are adaptively computed such that the material is properly constrained by the needle. The implementation is accelerated using NVidia CUDA. We present detailed analysis of the execution timing in both serial and parallel case. The proposed needle insertion model was integrated in a custom software that loads DICOM images, generate the deformable model, and can simulate different insertion strategies.
Patriciu, Alexandru
Large-scale, saturating insertional mutagenesis of the mouse genome
We describe the construction of a large-scale, orderly assembly of mutant ES cells, generated with retroviral insertions and having mutational coverage in >90% of mouse genes. We also describe a method for isolating ES cell clones with mutations in specific genes of interest from this library. This approach, which combines saturating random mutagenesis with targeted selection of mutations in the genes of interest, was successfully applied to the gene families of G protein-coupled receptors (GPCRs) and nuclear receptors. Mutant mouse strains in 60 different GPCRs were generated. Applicability of the technique for the GPCR genes, which on average represent fairly small targets for insertional mutagenesis, indicates the general utility of our approach for the rest of the genome. The method also allows for increased scale and automation for the large-scale production of mutant mice, which could substantially expedite the functional characterization of the mouse genome.
Gragerov, Alexander; Horie, Kyoji; Pavlova, Maria; Madisen, Linda; Zeng, Hongkui; Gragerova, Galina; Rhode, Alex; Dolka, Io; Roth, Patricia; Ebbert, Amanda; Moe, Stephanie; Navas, Christopher; Finn, Eric; Bergmann, John; Vassilatis, Demetrios K.; Pavlakis, George N.; Gaitanaris, George A.
Mediastinal abscess and esophageal stricture following voice prosthesis insertion.
The use of a tracheo-esophageal voice prosthesis is a well-established procedure to restore the voice in total laryngectomees. The insertion of the prosthesis is not a risk-free procedure, various complications having been reported especially in irradiated patients. Here described is a case of an esophageal rupture after secondary tracheo-esophageal puncture with mediastinal abscess in a patient previously treated with pharyngo-laryngectomy and subsequent radiotherapy for a left pyriform sinus carcinoma, which required immediate surgical drainage through a left cervical approach. Few weeks after surgical drainage an esophageal stricture at the site of the rupture developed, which was only temporarily resolved after the insertion of biodegradable esophageal stents, followed by re-stenosis once the reabsorption of the stent took place. PMID:24210965
Bozzo, Corrado; Meloni, Francesco; Trignano, Mario; Profili, Stefano
Intentional ingestion and insertion of foreign objects: a forensic perspective.
Intentional ingestion and insertion of foreign objects is a topic that has generated mounting interest among medical professionals over the past two decades. When featured in the literature, it has been typically discussed in medical subdisciplines, such as emergency medicine, surgery, gastroenterology, and urology. However, in-depth exploration of this multidimensional phenomenon in the field of psychiatry has thus far been limited. This article presents illustrative clinical vignettes from forensic practice of deliberate ingestion/insertion of objects and then examines specific aspects of this behavior that are critical to achieving a better understanding of it. The clinical, legal, and ethics-related implications surrounding this conduct are also explored. By taking a comprehensive approach, the aim is to foster a greater appreciation of this syndrome by clinicians and ultimately to arrive at improved practice guidelines surrounding these cases, including a more informed therapeutic plan and an enhanced management approach. PMID:22396349
Klein, Carolina A
Amyloid protein unfolding and insertion kinetics on neuronal membrane mimics
NASA Astrophysics Data System (ADS)
Atomistic details of beta-amyloid (A? ) 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 A? in binary lipid mixtures with and without cholesterol that mimic the cholesterol-depleted and cholesterol-enriched lipid nanodomains of neurons. The protein conformational transition kinetics was evaluated from the secondary structure profile versus simulation time plot. The extent of membrane disruption was examined by the calculated order parameters of lipid acyl chains and cholesterol fused rings as well as the density profiles of water and lipid headgroups at defined regions across the lipid bilayer from our simulations. Our results revealed that both the cholesterol content and the length of the protein affect the protein-insertion and membrane stability in our model lipid bilayer systems.
Qiu, Liming; Buie, Creighton; Vaughn, Mark; Cheng, Kwan
Supporting device and apparatus for inserting the device
US Patent & Trademark Office Database
In connection with herniotomia or laparoscopy a device (12) for supporting the abdominal wall may be passed into the abdominal cavity through a percutaneously arranged guard tube (23) that is preferably inserted in the abdominal cavity through the hernial canal. The supporting device (12) includes a barrier material (13) which may for instance be a synthetic net. The supporting device has a stiffener (14) allowing such a controlled deformation of the barrier material that the supporting device may pass through the guard tube (23), following which the stiffener is capable of unfolding the barrier material into a substantially plane condition. After insertion and unfolding the supporting device (12) may be tightened against the peritoneum (26) by means of a thread (19) that may be fastened on the surface of the patient's skin.
Out of nowhere: thought insertion, ownership and context-integration.
We argue that thought insertion primarily involves a disruption of the sense of ownership for thoughts and that the lack of a sense of agency is but a consequence of this disruption. We defend the hypothesis that this disruption of the sense of ownership stems from a failure in the online integration of the contextual information related to a thought, in particular contextual information concerning the different causal factors that may be implicated in their production. Loss of unity of consciousness, manifested by incoherent subjective experiences is a general phenomenal characteristic of schizophrenia. This loss of coherence has been hypothesized to reflect a generalized deficit of contextual information integration not conveyed by, but related to, a target event. This deficit is manifested across many cognitive domains. We argue that it is also manifested in the process of thinking itself, resulting in causally decontextualized thoughts that are experienced as inserted thoughts. PMID:23262256
Martin, Jean-Remy; Pacherie, Elisabeth
Objective evaluation of insert material for diabetic and athletic footwear.
Five of the most commonly used materials for shoe inserts (soft Plastazote, medium Pelite, PPT, Spenco, and Sorbothane) were objectively evaluated in the laboratory to characterize their behavior in the following three specific functions that correspond to clinical use: (1) the effect on the materials of repeated compression. (2) the effect of a combination of repetitive shear and compression. (3) the force-distribution (force-attenuation) properties of these materials, both when new and after repeated compression. The last function represents a model for relief of pressure beneath plantar bony prominences, a topic of special concern for the insensitive foot. All materials were effective in reducing transmitted force over the simulated bony prominence with a rank order of effectiveness. Other factors considered were: amount and rate of permanent deformation offset by considerations of enhanced moldability when comparing the neoprene and urethane materials with the polyethylene foams. The ideal insert represents a combination of material to achieve both durability and moldability. PMID:3229697
Brodsky, J W; Kourosh, S; Stills, M; Mooney, V
[Neonatology nurses' knowledge about Peripherally Inserted Central Venous Catheter].
The Peripherally Inserted Central Catheter (PICC) has been used as a safe venous access for infants at risk. The aim of this study was to describe the knowledge and practice of nurses from the five public Neonatal Intensive Care Units, of Recife-PE, Brazil, about the use of the PICC. The sample was comprised by 52 nurses; data were collected from January to February/2010. It was found that 64,8% of nurses did not have license for insertion of the PICC. Only two units routinely used the PICC. About the indication of the access, the accuracy was above 70%. In unit B only 8,3% of nurses reported adequate initial location of the catheter tip. It was concluded that is necessary greater incentives to train nurses to use the PICC. PMID:22751707
Belo, Marcela Patricia Macŕdo; Silva, Roberta Albuquerque Mello de Castro; Nogueira, Isis Larissa Maia; Mizoguti, Daniele Pereira; Ventura, Claudiane Maria Urbano
Deep Insertion Probe Measurements of the Hall Dynamo on MST
NASA Astrophysics Data System (ADS)
Fluctuation-induced forces and stresses have been shown to be of importance in the self-organization processes of the RFP. These forces have been extensively studied in the edge of MST, (ra>0.8), and have shown that the MHD and Hall dynamo terms, (
Triana, J. C.; Almagri, A. F.; Sarff, J. S.; Sauppe, J. P.; Sovinec, C. R.
Intravenous insertion site protection: moisture accumulation in intravenous site protectors.
Stabilizing the intravenous catheter after insertion is a significant part of intravenous therapy. Dislodgments of the cannula from its optimal position in the vein can lead to complications such as phlebitis, thrombophlebitis, infiltration, and infection. Intravenous site protector shields are designed to protect the catheter from impact and tissue trauma at the insertion site. Nurses have requested ventilation in these shields to avoid moisture build up that may increase the risk of infections. To address this issue, experimental laboratory testing was performed to determine if moisture accumulation as evidenced by increased weight of the shield and visible evidence of condensation occurred. No moisture condensation problems with the ventilated intravenous site protectors were found. PMID:8852177
Lee, W E; Vallino, L M
Random tag insertions by Transposon Integration mediated Mutagenesis (TIM).
Transposon Integration mediated Mutagenesis (TIM) is a broadly applicable tool for protein engineering. This method combines random integration of modified bacteriophage Mu transposons with their subsequent defined excision employing type IIS restriction endonuclease AarI. TIM enables deletion or insertion of an arbitrary number of bases at random positions, insertion of functional sequence tags at random positions, replacing randomly selected triplets by a specific codon (e.g. scanning) and site-saturation mutagenesis. As a proof of concept a transposon named GeneOpenerAarIKan was designed and employed to introduce 6xHis tags randomly into the esterase EstC from Burkholderia gladioli. A TIM library was screened with colony based assays for clones with an integrated 6xHis tag and for clones exhibiting esterase activity. The employed strategy enables the isolation of randomly tagged active enzymes in single mutagenesis experiments. PMID:18627780
Hoeller, Brigitte M; Reiter, Birgit; Abad, Sandra; Graze, Ina; Glieder, Anton
Neurosteroids promote phosphorylation and membrane insertion of extrasynaptic GABAA receptors.
Neurosteroids are synthesized within the brain and act as endogenous anxiolytic, anticonvulsant, hypnotic, and sedative agents, actions that are principally mediated via their ability to potentiate phasic and tonic inhibitory neurotransmission mediated by ?-aminobutyric acid type A receptors (GABAARs). Although neurosteroids are accepted allosteric modulators of GABAARs, here we reveal they exert sustained effects on GABAergic inhibition by selectively enhancing the trafficking of GABAARs that mediate tonic inhibition. We demonstrate that neurosteroids potentiate the protein kinase C-dependent phosphorylation of S443 within ?4 subunits, a component of GABAAR subtypes that mediate tonic inhibition in many brain regions. This process enhances insertion of ?4 subunit-containing GABAAR subtypes into the membrane, resulting in a selective and sustained elevation in the efficacy of tonic inhibition. Therefore, the ability of neurosteroids to modulate the phosphorylation and membrane insertion of ?4 subunit-containing GABAARs may underlie the profound effects these endogenous signaling molecules have on neuronal excitability and behavior. PMID:24778259
Abramian, Armen M; Comenencia-Ortiz, Eydith; Modgil, Amit; Vien, Thuy N; Nakamura, Yasuko; Moore, Yvonne E; Maguire, Jamie L; Terunuma, Miho; Davies, Paul A; Moss, Stephen J
Phase transitions in insertion electrodes for lithium batteries
Phase transitions that occur during lithium insertion into layered and framework structures are discussed in the context of their application as positive and negative electrodes in lithium-ion batteries. The discussion is focused on the two-dimensional structures of graphite, LiNi{sub 1{minus}x}M{sub x}O{sub 2} (M = Co, Ti and Mg), and Li{sub 1.2}V{sub 3}O{sub 8}; examples of framework structures with a three-dimensional interstitial space for Li{sup +}-ion transport include the spinel oxides and intermetallic compounds with zinc-blende-type structures. The phase transitions are discussed in terms of their tolerance to lithium insertion and extraction and to the chemical stability of the electrodes in the cell environment.
Thackeray, M. M.
Operation of synchrotron light sources with multiple insertion devices
The stability requirements of the next generation of synchrotron radiation facilities have been achieved on insertion device beamlines of existing rings. However, one insertion device (ID) affects the stored beam and hence the performance of all the other beamlines. Since the effects of the undulators are cumulative, higher levels of performance are required of the accelerator in order to meet and exceed present day standards in rings with many undulators. This paper will report experience to date in the areas mentioned above at several multi-undulator facilities and efforts to address these problems at facilities in the planning and construction phase. Section 2 will treat orbit control and feedback. Section 3 will describe work on linear and nonlinear effects of ideal undulators in accelerators. Section 4 will mention undulator imperfections and the demands they make on the accelerator control system. 11 refs., 2 figs., 3 tabs.
Galayda, J. (Argonne National Lab., IL (USA)); Fauchet, A.M. (Brookhaven National Lab., Upton, NY (USA))
A male-sterile insertion in the mouse
Is(7;l)40H was found in the daughter of a male mouse given spermatozoal X-irradiation. It is a non-inverted insertion of about half of chromosome 7 into chromosome 1, generating a long somatic marker chromosome. Breakpoints are in bands IB, 7B1, and 7F1; linkage tests show that these breakpoints are about midway between fz and In on the 1, and 0.2 units
A. G. Searle; C. V. Beechey; P. de Boer; D. G. de Rooij; E. P. Evans; M. Kirk
Selection analyses of insertional mutants using subgenic-resolution arrays
We describe a method of genome-wide analysis of quantitative growth phenotypes using insertional mutagenesis and DNA microarrays. We applied the method to assess the fitness contributions of Escherichia coli gene domains under specific growth conditions. A transposon library was subjected to competitive growth selection in LuriaľBertani (LB) and in glucose minimal media. Transposon-containing genomic DNA fragments from the selected libraries
Vasudeo Badarinarayana; Preston W. Estep III; Jay Shendure; Jeremy Edwards; Saeed Tavazoie; Felix Lam; George M. Church
Developmental Outcomes after Early or Delayed Insertion of Tympanostomy Tubes
BACKGROUND To prevent later developmental impairments, myringotomy with the insertion of tympanostomy tubes has often been undertaken in young children who have persistent otitis media with effusion. We previously reported that prompt as compared with delayed insertion of tympanostomy tubes in children with persistent effusion who were younger than three years of age did not result in improved developmental outcomes at three or four years of age. However, the effect on the outcomes of school-age children is unknown. METHODS We enrolled 6350 healthy infants younger than 62 days of age and evaluated them regularly for middle-ear effusion. Before three years of age, 429 children with persistent middle-ear effusion were randomly assigned to have tympanostomy tubes inserted either promptly or up to nine months later if effusion persisted. We assessed developmental outcomes in 395 of these children at six years of age. RESULTS At six years of age, 85 percent of children in the early-treatment group and 41 percent in the delayed-treatment group had received tympanostomy tubes. There were no significant differences in mean (▒SD) scores favoring early versus delayed treatment on any of 30 measures, including the Wechsler Full-Scale Intelligence Quotient (98▒13 vs. 98▒14); Number of Different Words test, a measure of word diversity (183▒36 vs. 175▒36); Percentage of Consonants CorrectľRevised test, a measure of speech-sound production (96▒2 vs. 96▒3); the SCAN test, a measure of central auditory processing (95▒15 vs. 96▒14); and several measures of behavior and emotion. CONCLUSIONS In otherwise healthy children younger than three years of age who have persistent middle-ear effusion within the duration of effusion that we studied, prompt insertion of tympanostomy tubes does not improve developmental outcomes at six years of age.
Paradise, Jack L.; Campbell, Thomas F.; Dollaghan, Christine A.; Feldman, Heidi M.; Bernard, Beverly S.; Colborn, D. Kathleen; Rockette, Howard E.; Janosky, Janine E.; Pitcairn, Dayna L.; Kurs-Lasky, Marcia; Sabo, Diane L.; Smith, Clyde G.
High Throughput Fingerprint Analysis of Large-Insert Clones
As part of the Human Genome Project, the Washington University Genome Sequencing Center has commenced systematic sequencing of human chromsome 7. To organize and supply the effort, we have undertaken the construction of sequence-ready physical maps for defined chromosomal intervals. Map construction is a serial process composed of three main activities. First, candidate STS-positive large-insert PAC and BAC clones are
Marco A. Marra; Tamara A. Kucaba; Nicole L. Dietrich; Eric D. Green; Buddy Brownstein; Richard K. Wilson; Ken M. McDonald; LaDeana W. Hillier; John D. McPherson; Robert H. Waterston
The inserted piezoelectric sensor method for monitoring thermosets cure
NASA Astrophysics Data System (ADS)
An original method enabling the in-situ and real time monitoring of a polymer structure evolution using an inserted piezoelectric sensor is presented. The validation of this technique is done at constant temperature by comparison to the results obtained on several polymers by classical ultrasonic methods in practically the same conditions. The complete validation is achieved by monitoring the complex cure of a polymer matrix fibre reinforced composite PMR-15 under sever conditions of temperature.
Tabellout, M.; Raquois, A.; Emery, J. R.; Jayet, Y.
Management of subcalcaneal pain and Achilles tendonitis with heel inserts
Soft tissue symptoms in the leg due to sporting activity are commonly associated with the force of heel strike. Conventional training shoes compromise between comfort and performance; few models are suitably designed for both considerations. Using a visco-elastic polymer insert the symptoms of heel pain and Achilles tendonitis have been largely or completely abolished in a preliminary study. Imagesp117-ap117-bp117-cp118-a
Maclellan, G. E.; Vyvyan, Barbara
Forensic implications in self-insertion of urethral foreign bodies
A 54-year-old Caucasian male presented to our emergency department because he had self inserted a transparent plastic and flexible tube into the urethral meatus, during erotic games. A plain pelvic x-ray film and an abdominal ultrasound were executed to determine the size, shape, orientation and location of the foreign body, and any eventually associated visceral injuries or complications. As results,
R. Boscolo-Berto; M. Iafrate; G. Viel
Microstructure and Mechanical Characteristics of DMLS Tool-Inserts
A commercial metal powder mixture was used for the laser sintering experiments. Different substrates in the form of simple blocks and complex testing tool-inserts for pressure die casting of Al-based alloy were prepared at different process conditions and post-surface treatments. Additional improvement of surface quality was obtained with the hard coating. In the present article, the basic principles of laser
B. Sustarsic; S. Dolinsek; M. Jenko; V. LeskovÜek
Minimally Invasive Technique for Insertion of a Unicompartmental Knee Arthroplasty
\\u000a The Oxford meniscal unicompartmental knee arthrop lasty has been used to treat anteromedial arthritis of the knee since 1982.1 The design of the prosthesis employs a spherical femoral component with a flat tibial base-plate, between which a fully congruent\\u000a unconstrained mobile bearing is inserted (Figure 17.1). This provides a large contact area for articulation in all angles of flexion, reducing
A. J. Price; D. W. Murray
Lithium insertion in manganese oxides: A model lithium ion system
A model lithium ion system, based on the manganese oxide, LixMn2O4, has been used to study the reversibility of the lithium insertion processes in both the approximate 3 V and 4 V versus LiLi+ voltages ranges. We have used the half-cell voltage responses for the Li1 ? xMn2O4 and Li1 + xMn2O4 electrodes to predict the voltage profile for the
Jerry Barker; RenÚ Koksbang; M. Yazid Sa´di
Cryptococcus neoformans Virulence Gene Discovery through Insertional Mutagenesis
Insertional mutagenesis was applied to Cryptococcus neoformans to identify genes associated with virulence attributes. Using biolistic transformation, we generated 4,300 nourseothricin (NAT)-resistant strains, of which 590 exhibited stable resistance. We focused on mutants with defects in established virulence factors and identified two with reduced growth at 37░C, four with reduced production of the antioxidant pigment melanin, and two with an
Alexander Idnurm; Jennifer L. Reedy; Jesse C. Nussbaum; Joseph Heitman
The Thomsen model of inserts in sandwich composites: An evaluation
An one-dimensional finite element model of a sandwich panel with insert is derived using the approach used in the Thomsen model. The one-dimensional model produces results that are close to those of a two-dimensional axisysmmetric model. Both models assume that the core is homogeneous. Our results indicate that the one-dimensional model may be well suited for small deformations of sandwich
Biswajit Banerjee; Bryan Smith
Electrochemical Insertion of Lithium into GraphiteľZinc Composites
Metal-based composites are currently under investigation as possible negative-electrode materials in lithium-ion batteries. We present here a new composite material composed of zinc particles deposited mainly onto