Science.gov

Sample records for att minska transportlogistikens

  1. AT&T: "Apologies Are Not Enough."

    ERIC Educational Resources Information Center

    Benoit, William L.; Brinson, Susan L.

    1994-01-01

    Describes AT&T's long distance service interruption in New York on September 17, 1991, as a serious threat to its corporate image. Analyzes the advertising actions taken by AT&T to restore its image. Discusses three primary strategies developed in the advertising: mortification, plans for correction, and bolstering. (HB)

  2. TV via satellite from AT&T

    NASA Astrophysics Data System (ADS)

    Kiely, J.

    1982-04-01

    AT&T's Satellite Television Service, which relays TV feeds by means of the Comstar domestic communications satellite system, is discussed. Advantages include the ability to provide multiple feeds of program and advertising material simultaneously to affiliates, which can help accommodate the trend toward greater program diversity, and the ability to distribute one signal to many points, making their use economically attractive. A proposed radio service from AT&T is also discussed, which could provide radio broadcasters and programmers with a high-quality, versatile means of transmitting radio programs via satellite.

  3. 78 FR 69878 - AT&T Corporation, a Subsidiary of AT&T Inc., Business Billing Customer Care, Pittsburgh...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-21

    ... in the Federal Register on July 2, 2013 (78 FR 39776). Workers at the subject firm were engaged in... Employment and Training Administration AT&T Corporation, a Subsidiary of AT&T Inc., Business Billing Customer... apply for Trade Adjustment Assistance (TAA) applicable to workers and former workers of AT&T...

  4. The Arms Trade Treaty (ATT): a public health imperative.

    PubMed

    Valenti, Maria; Mtonga, Robert; Gould, Robert; Christ, Michael

    2014-02-01

    The United Nations adopted an historic international Arms Trade Treaty (ATT) in April 2013. A 1997 meeting of Nobel Peace Prize laureates who called for an International Code of Conduct to address the 'destructive effects of the unregulated arms trade' initiated discussions that led to the Treaty. Public health institutions, including the World Health Organization and the International Committee of the Red Cross, and nongovernmental health groups such as International Physicians for the Prevention of Nuclear War, made adoption of the ATT a public health imperative. The poorly regulated $70 billion annual trade in conventional arms fuels conflict, with devastating effects on global health. The ATT aims to 'reduce human suffering'. It prohibits arms' sales if there is knowledge that the arms would be used in the commission of genocide, attacks against civilians, or war crimes. The health community has much to contribute to ensuring ratification and implementation of the ATT.

  5. The Unilateral, Self-Deregulation of AT&T

    NASA Astrophysics Data System (ADS)

    Maloff, Joel H.

    1990-01-01

    The provision of telecommunications products and services in the United States has been dominated by AT&T and the Bell System for more than one hundred years. From the early days of telephones, it was clear that some framework of logic was required to provide "universal" telephone services to the vast majority of American citizens. The concept was known as "natural monopoly", and for the most part, has served us well. We are all aware of statements concerning absolute power corrupting absolutely, and AT&T has not been immune to this law of human nature. Several times during the past century, action has been required by the Federal Government to regulate and constrain AT&T and the Bell System from anti-competitive and predatory actions. These have been called consent decrees, Computer Inquiry II, Computer Inquiry III, and the Modified Final Judgment. The regulations and constraints imposed upon AT&T, with good reason, are now becoming eroded. The perception of a more highly competitive marketplace capable of accommodating an unrestrained AT&T, the impact of uncoordinated national and regional government policies, and unilateral actions by AT&T themselves are bringing us to the edge of an abyss. There are substantial reasons to believe that AT&T will use its considerable might to eliminate its competitors, once free to do so. Must we be doomed to repeat history with still another antitrust case against AT&T some years from now? Logic demands that we learn from the past, and that telecommunications policies be based upon that knowledge.

  6. Conjugal transfer using the bacteriophage phiC31 att/int system and properties of the attB site in Streptomyces ambofaciens.

    PubMed

    Kim, Mi-Kyung; Ha, Heon-Su; Choi, Sun-Uk

    2008-04-01

    To facilitate molecular genetic studies of Streptomyces ambofaciens that produces spiramycin, a commercially important macrolide antibiotic used in human medicine against Gram-positive pathogenic bacteria, the conditions for the conjugal transfer of DNA from E. coli to S. ambofaciens were established using a bacteriophage phiC31 att/int system. The transconjugation efficiency of S. ambofaciens varied with the medium used; the highest frequency was obtained on AS-1 medium containing 10 mM MgCl(2) without heat treatment of the spores. In addition, by cloning and sequencing the attB site, we identified that S. ambofaciens contains a single attB site within an ORF coding for a pirin homolog, and its attB site sequence shows 100% nt identity to the sequence of S. coelicolor and S. lividans, which have the highest efficiency in transconjugation using the phiC31 att/int system.

  7. QCD Prediction of ATT for Small QT Dimuon Production in pp and pp-bar Collisions

    SciTech Connect

    Kawamura, Hiroyuki; Kodaira, Jiro; Tanaka, Kazuhiro

    2007-06-13

    We present QCD prediction of double-spin asymmetries (ATT) in transversely polarized Drell-Yan process at small transverse momentum QT of dimuon. Resummation of large logarithmic corrections, relevant in small QT region, is performed up to next-to-leading logarithmic (NLL) accuracy. ATT at RHIC, J-PARC and GSI are studied numerically in the corresponding kinematic regions. We show that the large ATT is obtained for small QT and moderate energies.

  8. 76 FR 5831 - AT&T; Reynoldsburg, OH; Notice of Affirmative Determination Regarding Application for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-02

    ... published in the Federal Register on January 3, 2011 (76 FR 182). The workers supply customer care call... Employment and Training Administration AT&T; Reynoldsburg, OH; Notice of Affirmative Determination Regarding... Adjustment Assistance (TAA) applicable to workers and former workers of AT&T, Reynoldsburg, Ohio ]...

  9. 78 FR 76096 - Request for Comment on Petition Filed by AT&T Services, Inc., Regarding the Provision of Muting...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-16

    ... of the requirement contained in the 2013 STS Order, published at 78 FR 49693, August 15, 2013, and... COMMISSION 47 CFR Part 64 Request for Comment on Petition Filed by AT&T Services, Inc., Regarding the... seeks comment on an AT&T Services, Inc. (AT&T) petition requesting clarification or, in the...

  10. 76 FR 5831 - Amdocs, Inc., Global Support Services, Advertising And Media AT&T Division, New Haven...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-02

    ... Federal Register on November 23, 2010 (75 FR 71461). The negative determination was based on the findings... Employment and Training Administration Amdocs, Inc., Global Support Services, Advertising And Media AT&T...., Global Support Services, Advertising and Media AT&T Division, New Haven, Connecticut (subject firm)....

  11. 76 FR 27666 - AT&T Services, Inc., Reynoldsburg, OH; Notice of Negative Determination on Reconsideration

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-12

    ... Employment and Training Administration AT&T Services, Inc., Reynoldsburg, OH; Notice of Negative... February 2, 2011 (76 FR 5831). Workers supply customer care call services. Pursuant to 29 CFR 90.18(c... reconsideration of the decision. The negative determination was based on the findings that the worker...

  12. An Overview of Research and Development Projects at the AT&T National Teletraining Center.

    ERIC Educational Resources Information Center

    Chute, Alan G.; Balthazar, Lee B.

    Research and development projects at the AT&T (American Telephone and Telegraph Company) National Teletraining Center (NTC) are geared toward increasing understanding of the various psychological, sociological, ergonomic, and environmental factors involved in teletraining, and toward improving the ability to manage the distance learning…

  13. The streptomyces genome contains multiple pseudo-attB sites for the (phi)C31-encoded site-specific recombination system.

    PubMed

    Combes, Patricia; Till, Rob; Bee, Sally; Smith, Margaret C M

    2002-10-01

    The integrase from the Streptomyces phage (phi)C31 is a member of the serine recombinase family of site-specific recombinases and is fundamentally different from that of lambda or its relatives. Moreover, (phi)C31 int/attP is used widely as an essential component of integration vectors (such as pSET152) employed in the genetic analysis of Streptomyces species. phiC31 or integrating plasmids containing int/attP have been shown previously to integrate at a locus, attB, in the chromosome. The DNA sequences of the attB sites of various Streptomyces species revealed nonconserved positions. In particular, the crossover site was narrowed to the sequence 5'TT present in both attP and attB. Strains of Streptomyces coelicolor and S. lividans were constructed with a deletion of the attB site ((Delta)attB), and pSET152 was introduced into these strains by conjugation. Thus, secondary or pseudo-attB sites were identified by Southern blotting and after rescue of plasmids containing DNA flanking the insertion sites from the chromosome. The sequences of the integration sites had similarity to those of attB. Analysis of the insertions of pSET152 into both attB(+) and (Delta)attB strains indicated that this plasmid can integrate at several loci via independent recombination events within a transconjugant.

  14. ATtRACT-a database of RNA-binding proteins and associated motifs.

    PubMed

    Giudice, Girolamo; Sánchez-Cabo, Fátima; Torroja, Carlos; Lara-Pezzi, Enrique

    2016-01-01

    RNA-binding proteins (RBPs) play a crucial role in key cellular processes, including RNA transport, splicing, polyadenylation and stability. Understanding the interaction between RBPs and RNA is key to improve our knowledge of RNA processing, localization and regulation in a global manner. Despite advances in recent years, a unified non-redundant resource that includes information on experimentally validated motifs, RBPs and integrated tools to exploit this information is lacking. Here, we developed a database named ATtRACT (available athttp://attract.cnic.es) that compiles information on 370 RBPs and 1583 RBP consensus binding motifs, 192 of which are not present in any other database. To populate ATtRACT we (i) extracted and hand-curated experimentally validated data from CISBP-RNA, SpliceAid-F, RBPDB databases, (ii) integrated and updated the unavailable ASD database and (iii) extracted information from Protein-RNA complexes present in Protein Data Bank database through computational analyses. ATtRACT provides also efficient algorithms to search a specific motif and scan one or more RNA sequences at a time. It also allows discoveringde novomotifs enriched in a set of related sequences and compare them with the motifs included in the database.Database URL:http:// attract. cnic. es. PMID:27055826

  15. ATtRACT-a database of RNA-binding proteins and associated motifs.

    PubMed

    Giudice, Girolamo; Sánchez-Cabo, Fátima; Torroja, Carlos; Lara-Pezzi, Enrique

    2016-01-01

    RNA-binding proteins (RBPs) play a crucial role in key cellular processes, including RNA transport, splicing, polyadenylation and stability. Understanding the interaction between RBPs and RNA is key to improve our knowledge of RNA processing, localization and regulation in a global manner. Despite advances in recent years, a unified non-redundant resource that includes information on experimentally validated motifs, RBPs and integrated tools to exploit this information is lacking. Here, we developed a database named ATtRACT (available athttp://attract.cnic.es) that compiles information on 370 RBPs and 1583 RBP consensus binding motifs, 192 of which are not present in any other database. To populate ATtRACT we (i) extracted and hand-curated experimentally validated data from CISBP-RNA, SpliceAid-F, RBPDB databases, (ii) integrated and updated the unavailable ASD database and (iii) extracted information from Protein-RNA complexes present in Protein Data Bank database through computational analyses. ATtRACT provides also efficient algorithms to search a specific motif and scan one or more RNA sequences at a time. It also allows discoveringde novomotifs enriched in a set of related sequences and compare them with the motifs included in the database.Database URL:http:// attract. cnic. es.

  16. ATtRACT—a database of RNA-binding proteins and associated motifs

    PubMed Central

    Giudice, Girolamo; Sánchez-Cabo, Fátima; Torroja, Carlos; Lara-Pezzi, Enrique

    2016-01-01

    RNA-binding proteins (RBPs) play a crucial role in key cellular processes, including RNA transport, splicing, polyadenylation and stability. Understanding the interaction between RBPs and RNA is key to improve our knowledge of RNA processing, localization and regulation in a global manner. Despite advances in recent years, a unified non-redundant resource that includes information on experimentally validated motifs, RBPs and integrated tools to exploit this information is lacking. Here, we developed a database named ATtRACT (available at http://attract.cnic.es) that compiles information on 370 RBPs and 1583 RBP consensus binding motifs, 192 of which are not present in any other database. To populate ATtRACT we (i) extracted and hand-curated experimentally validated data from CISBP-RNA, SpliceAid–F, RBPDB databases, (ii) integrated and updated the unavailable ASD database and (iii) extracted information from Protein-RNA complexes present in Protein Data Bank database through computational analyses. ATtRACT provides also efficient algorithms to search a specific motif and scan one or more RNA sequences at a time. It also allows discovering de novo motifs enriched in a set of related sequences and compare them with the motifs included in the database. Database URL: http:// attract. cnic. es PMID:27055826

  17. Site-specific integration of the temperate bacteriophage phi adh into the Lactobacillus gasseri chromosome and molecular characterization of the phage (attP) and bacterial (attB) attachment sites.

    PubMed Central

    Raya, R R; Fremaux, C; De Antoni, G L; Klaenhammer, T R

    1992-01-01

    The temperate bacteriophage phi adh integrates its genome into the chromosomal DNA of Lactobacillus gasseri ADH by a site-specific recombination process. Southern hybridization analysis of BclI-digested genomic DNA from six relysogenized derivatives of the prophage-cured strain NCK102 displayed phage-chromosomal junction fragments identical to those of the lysogenic parent. The phi adh attachment site sequence, attP, was located within a 365-bp EcoRI-HindIII fragment of phage phi adh. This fragment was cloned and sequenced. DNA sequence analysis revealed striking features common to the attachment sites of other site-specific recombination systems: five direct repeats of the sequence TGTCCCTTTT(C/T) and a 14-bp inverted repeat. Oligonucleotides derived from the sequence of the attP-containing fragment enabled us to amplify predicted junction fragment sequences and thus to identify attL, attR, and attB. The core region was defined as the 16-bp sequence TACACTTCTTAGGAGG. Phage-encoded functions essential for site-specific insertion of phage phi adh were located in a 4.5-kb BclI fragment. This fragment was cloned in plasmid pSA34 to generate the insertional vector pTRK182. Plasmid pTRK182 was introduced into L. gasseri NCK102 by electroporation. Hybridization analysis showed that a single copy of pTRK182 had integrated at the attB site of the NCK102 erythromycin-resistant transformants. This is the first site-specific recombination system described in lactobacilli, as well as the first attP-based site-specific integration vector constructed for L. gasseri ADH. Images PMID:1512192

  18. Effects of mastoparan upon the late stages of the ACTH secretory pathway of AtT-20 cells.

    PubMed Central

    McFerran, B. W.; Guild, S. B.

    1995-01-01

    1. The mouse AtT-20/D16-16 anterior pituitary tumour cell line was used as a model system for the study of the effects of mastoparan upon the late stages of the adrenocorticotrophin (ACTH) secretory pathway. 2. Mastoparan (10(-8)-10(-5) M), an activator of heterotrimeric guanosine 5'-triphosphate binding proteins (G-proteins), stimulated ACTH secretion from electrically-permeabilized AtT-20 cells in a concentration-dependent manner in the effective absence of calcium ions with a threshold of 10(-6) M. Guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) (10(-8)-10(-4) M) also stimulated ACTH secretion from electrically-permeabilized AtT-20 cells in a concentration-dependent manner in the effective absence of calcium ions with a threshold of 10(-6) M. This GTP-gamma-S-evoked secretion is consistent with previous studies which demonstrated that a G-protein, termed GE, mediates calcium evoked ACTH secretion from AtT-20 cells. GTP-gamma-S-evoked secretion however was not as great as that obtained in response to mastoparan. 3. Both mastoparan (10(-5) M) and GTP-gamma-S (10(-4) M) stimulated ACTH secretion from electrically-permeabilized AtT20 cells in a time-dependent manner. A time of 30 min was adopted as the standard incubation period for the study of both mastoparan and GTP-gamma-S-stimulated ACTH secretion from permeabilized AtT-20 cells. 4. Mastoparan (10(-8)-10(-5) M) stimulated ACTH secretion from permeabilized AtT-20 cells to the same extent in the presence and absence of the protein kinase C (PKC) inhibitor, chelerythrine chloride (10(-5) M).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7582493

  19. 78 FR 79362 - Request for Comment on Petition Filed by AT&T Services, Inc., Regarding the Provision of Muting...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-30

    ..., the Federal Communications Commission (Commission) corrects a document published at 78 FR 76096... Rights Office, (202) 418-1264, email: Caitlin.vogus@fcc.gov . SUPPLEMENTARY INFORMATION: Correction In FR... COMMISSION 47 CFR Part 64 Request for Comment on Petition Filed by AT&T Services, Inc., Regarding...

  20. 78 FR 2600 - Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-11

    ... CFR Part 69 Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To Reform Regulation of Incumbent Local Exchange Carrier Rates for Interstate Special Access... analytic framework capable of assessing whether the Commission's price cap and pricing flexibility...

  1. 77 FR 57504 - Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-18

    ... COMMISSION 47 CFR Part 1 Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To Reform Regulation of Incumbent Local Exchange Carrier Rates for Interstate Special Access... access prices and deter anticompetitive practices by price cap local exchange carriers. In the Report...

  2. 78 FR 24683 - Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-26

    ... COMMISSION 47 CFR Part 69 Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To Reform Regulation of Incumbent Local Exchange Carrier Rates for Interstate Special Access.... DATES: The effective date of the Report and Order published on January 11, 2013, at 78 FR 2572,...

  3. An Evaluation of the Electronic Classroom: The AT&T Teaching Theatre at the University of Maryland.

    ERIC Educational Resources Information Center

    Norman, Kent L.; Carter, Leslie E.

    1994-01-01

    Describes the AT&T Teaching Theater at the University of Maryland, an electronic classroom outfitted with a high performance workstation at each student's desk and the instructor's podium. Topics addressed include the facility and its objectives; issues and concepts; informal faculty and student reactions; and recommendations and guidance.…

  4. Spontaneous deletion mutants of the Lactococcus lactis temperate bacteriophage BK5-T and localization of the BK5-T attP site.

    PubMed Central

    Boyce, J D; Davidson, B E; Hillier, A J

    1995-01-01

    Spontaneous deletion mutants of the temperate lactococcal bacteriophage BK5-T were obtained when the phage was grown vegetatively on the indicator strain Lactococcus lactis subsp. cremoris H2. One deletion mutant was unable to form stable lysogens, and analysis of this mutant led to the identification of the BK5-T attP site and the integrase gene (int). The core sequences of the BK5-T attP and host attB regions are conserved in a number of lactococcal phages and L. lactis strains. PMID:8526525

  5. Pseudo attP sites in favor of transgene integration and expression in cultured porcine cells identified by Streptomyces phage phiC31 integrase.

    PubMed

    Bi, Yanzhen; Liu, Ximei; Zhang, Long; Shao, Changwei; Ma, Zhuo; Hua, Zaidong; Zhang, Liping; Li, Li; Hua, Wenjun; Xiao, Hongwei; Wei, Qingxin; Zheng, Xinmin

    2013-09-08

    Phage PhiC31 integrase integrates attB-containing plasmid into pseudo attP site in eukaryotic genomes in a unidirectional site-specific manner and maintains robust transgene expression. Few studies, however, explore its potential in livestock. This study aims to discover the molecular basis of PhiC31 integrase-mediated site-specific recombination in pig cells. We show that PhiC31 integrase can mediate site-specific transgene integration into the genome of pig kidney PK15 cells. Intramolecular recombination in pig PK15 cell line occurred at maximum frequency of 82% with transiently transfected attB- and attP-containing plasmids. An optimal molar ratio of pCMV-Int to pEGFP-N1-attB at 5:1 was observed for maximum number of cell clones under drug selection. Four candidate pseudo attP sites were identified by TAIL-PCR from those cell clones with single-copy transgene integration. Two of them gave rise to higher integration frequency occurred at 33%. 5' and 3' junction PCR showed that transgene integration mediated by PhiC31 integrase was mono-allelic. Micro- deletion and insertion were observed by sequencing the integration border, indicating that double strand break was induced by the recombination. We then constructed rescue reporter plasmids by ABI-REC cloning of the four pseudo attP sites into pBCPB + plasmid. Transfection of these rescue plasmids and pCMV-Int resulted in expected intramolecular recombination between attB and pseudo attP sites. This proved that the endogenous pseudo attP sites were functional substrates for PhiC31 integrase-mediated site-specific recombination. Two pseudo attP sites maintained robust extracellular and intracellular EGFP expression. Alamar blue assay showed that transgene integration into these specific sites had little effect on cell proliferation. This is the first report to document the potential use of PhiC31 integrase to mediate site-specific recombination in pig cells. Our work established an ideal model to study the position

  6. Transcription of AAT•ATT Triplet Repeats in Escherichia coli Is Silenced by H-NS and IS1E Transposition

    PubMed Central

    Pan, Xuefeng; Liao, Lingni; Yang, Li; Li, Hongquan

    2010-01-01

    Background The trinucleotide repeats AAT•ATT are simple DNA sequences that potentially form different types of non-B DNA secondary structures and cause genomic instabilities in vivo. Methodology and Principal Findings The molecular mechanism underlying the maintenance of a 24-triplet AAT•ATT repeat was examined in E.coli by cloning the repeats into the EcoRI site in plasmid pUC18 and into the attB site on the E.coli genome. Either the AAT or the ATT strand acted as lagging strand template in a replication fork. Propagations of the repeats in either orientation on plasmids did not affect colony morphology when triplet repeat transcription using the lacZ promoter was repressed either by supplementing LacIQ in trans or by adding glucose into the medium. In contrast, transparent colonies were formed by inducing transcription of the repeats, suggesting that transcription of AAT•ATT repeats was toxic to cell growth. Meanwhile, significant IS1E transposition events were observed both into the triplet repeats region proximal to the promoter side, the promoter region of the lacZ gene, and into the AAT•ATT region itself. Transposition reversed the transparent colony phenotype back into healthy, convex colonies. In contrast, transcription of an 8-triplet AAT•ATT repeat in either orientation on plasmids did not produce significant changes in cell morphology and did not promote IS1E transposition events. We further found that a role of IS1E transposition into plasmids was to inhibit transcription through the repeats, which was influenced by the presence of the H-NS protein, but not of its paralogue StpA. Conclusions and Significance Our findings thus suggest that the longer AAT•ATT triplet repeats in E.coli become vulnerable after transcription. H-NS and its facilitated IS1E transposition can silence long triplet repeats transcription and preserve cell growth and survival. PMID:21151567

  7. Measuring Attitudes About Intimate Partner Violence Against Women: The ATT-IPV Scale

    PubMed Central

    Yount, Kathryn M.; VanderEnde, Kristin; Zureick-Brown, Sarah; Anh, Hoang Tu; Schuler, Sidney Ruth; Minh, Tran Hung

    2014-01-01

    In lower-income settings, women more often than men justify intimate partner violence (IPV). Yet, the role of measurement invariance across gender is unstudied. We developed the ATT-IPV scale to measure attitudes about physical violence against wives in 1,055 married men and women ages 18–50 in My Hao district, Vietnam. Across 10 items about transgressions of the wife, women more often than men agreed that a man had good reason to hit his wife (3 % to 92 %; 0 % to 67 %). In random split-half samples, one-factor exploratory factor analysis (EFA) (N1 = 527) and confirmatory factor analysis (CFA) (N2 = 528) models for nine items with sufficient variability had significant loadings (0.575–0.883; 0.502–0.897) and good fit (RMSEA = 0.068, 0.048; CFI = 0.951, 0.978, TLI = 0.935, 0.970). Three items had significant uniform differential item functioning (DIF) by gender, and adjustment for DIF revealed that measurement noninvariance was partially masking men’s lower propensity than women to justify IPV. A CFA model for the six items without DIF had excellent fit (RMSEA = 0.019, CFI = 0.994, TLI = 0.991) and an attitudinal gender gap similar to the DIF-adjusted nine-item model, suggesting that the six-item scale reliably measures attitudes about IPV across gender. Researchers should validate the scale in urban Vietnam and elsewhere and decompose DIF-adjusted gender attitudinal gaps. PMID:24846070

  8. Involvement of multiple protein kinase C isozymes in the ACTH secretory pathway of AtT-20 cells.

    PubMed Central

    McFerran, B. W.; MacEwan, D. J.; Guild, S. B.

    1995-01-01

    1. The mouse AtT-20/D16-16 anterior pituitary tumour cell line was used as a model system for the study of protein kinase C (PKC)-mediated enhancement of calcium- and guanine nucleotide-evoked adrenocorticotrophin (ACTH) secretion. 2. A profile of the PKC isozymes present in AtT-20 cells was obtained by Western blotting analysis and it was found that AtT-20 cells express the alpha, beta, epsilon and zeta isoforms of PKC. 3. PKC isozymes were activated by the use of substances reported to activate particular isoforms of the enzyme. The effects of these substances were investigated in both intact and electrically-permeabilized cells. Phorbol 12-myristate 13-acetate (PMA, EC50 = 1 +/- 0.05 nM, which activates all isozymes of PKC, except the zeta isozyme), thymeleatoxin (TMX, EC50 = 10 +/- 0.5 nM, which activates the alpha, beta and gamma isozymes) and 12-deoxyphorbol 13-phenylacetate 20-acetate (dPPA, EC50 = 3 +/- 0.5 nM, a beta 1-selective isozyme activator) all stimulated ACTH secretion from intact cells in a concentration-dependent manner. Maximal TMX stimulated ACTH secretion was of a similar degree to that obtained in response to PMA but maximal dPPA-stimulated ACTH secretion was only 60-70% of that obtained in response to PMA or TMX. 4. Calcium stimulated ACTH secretion from electrically-permeabilized cells over the concentration-range of 100 nM to 10 microM. PMA (100 nM), TMX (100 nM) but not dPPA (100 nM) enhanced the amount of ACTH secreted at every concentration of calcium investigated.(ABSTRACT TRUNCATED AT 250 WORDS) Images Figure 1 PMID:7670732

  9. The silent codon change I507-ATC→ATT contributes to the severity of the ΔF508 CFTR channel dysfunction

    PubMed Central

    Lazrak, Ahmed; Fu, Lianwu; Bali, Vedrana; Bartoszewski, Rafal; Rab, Andras; Havasi, Viktoria; Keiles, Steve; Kappes, John; Kumar, Ranjit; Lefkowitz, Elliot; Sorscher, Eric J.; Matalon, Sadis; Collawn, James F.; Bebok, Zsuzsanna

    2013-01-01

    The most common disease-causing mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene is the out-of-frame deletion of 3 nucleotides (CTT). This mutation leads to the loss of phenylalanine-508 (ΔF508) and a silent codon change (SCC) for isoleucine-507 (I507-ATC→ATT). ΔF508 CFTR is misfolded and degraded by endoplasmic reticulum-associated degradation (ERAD). We have demonstrated that the I507-ATC→ATT SCC alters ΔF508 CFTR mRNA structure and translation dynamics. By comparing the biochemical and functional properties of the I507-ATT and I507-ATC ΔF508 CFTR, we establish that the I507-ATC→ATT SCC contributes to the cotranslational misfolding, ERAD, and to the functional defects associated with ΔF508 CFTR. We demonstrate that the I507-ATC ΔF508 CFTR is less susceptible to the ER quality-control machinery during translation than the I507-ATT, although 27°C correction is necessary for sufficient cell-surface expression. Whole-cell patch-clamp recordings indicate sustained, thermally stable cAMP-activated Cl− transport through I507-ATC and unstable function of the I507-ATT ΔF508 CFTR. Single-channel recordings reveal improved gating properties of the I507-ATC compared to I507-ATT ΔF508 CFTR (NPo=0.45±0.037 vs. NPo=0.09±0.002; P<0.001). Our results signify the role of the I507-ATC→ATT SCC in the ΔF508 CFTR defects and support the importance of synonymous codon choices in determining the function of gene products.—Lazrak, A., Fu, L., Bali, V., Bartoszewski, R., Rab, A., Havasi, V., Keiles, S., Kappes, J., Kumar, R., Lefkowitz, E., Sorscher, E. J., Matalon, S., Collawn, J. F., Bebok, Z. The silent codon change I507-ATC→ATT contributes to the severity of the ΔF508 CFTR. PMID:23907436

  10. Unusual site-specific DNA integration into the highly active pseudo-attB of the Streptomyces albus J1074 genome.

    PubMed

    Bilyk, Bohdan; Luzhetskyy, Andriy

    2014-06-01

    The φC31-encoded recombination system has become a widely used tool for genetic analysis of streptomycetes, gene therapy and generation of transgenic animals. However, the application of this system, even in the context of its natural host genus, Streptomyces, may require a specific approach for each species. In this study, we have identified a novel pseudo-attB site, called pseB4, for integration of vectors using the φC31 system. More than 90 % of clones contained two copies of pSET152- or pOJ436-based cosmids, after their introduction into S. albus. The efficiency of the integration of φC31-based vectors into pseB4 is therefore comparable to that of the integration into attB. Moreover, in contrast with integration into the native attB, integration into pseB4 is not polar and does not require a complementary sequence in the TT-core region. Furthermore, an analysis of conjugation frequency revealed mutual inhibition of plasmid integration into either site when both the attB and pseB4 sites were present in the genome.

  11. Expression of multiple somatostatin receptor genes in AtT-20 cells. Evidence for a novel somatostatin-28 selective receptor subtype.

    PubMed

    Patel, Y C; Panetta, R; Escher, E; Greenwood, M; Srikant, C B

    1994-01-14

    The pattern of expression of somatostatin receptor (SSTR) genes and gene products in AtT-20 cells was characterized in an attempt to explain the SST-28 binding selectivity that typifies these cells. AtT-20 cells expressed multiple SSTR mRNAs. Paradoxically, this included mRNA for three of the four SST-14 selective receptors: SSTR2 ( +), SSTR1 (+), SSTR4 (+). The SST-28 selective SSTR5 was expressed as a 3.8-kilobase (kb) transcript of relatively low abundance (+) in contrast to normal mouse pituitary which displayed high levels ( ) of a 2.4-kb SSTR5 mRNA. Immunoblot analysis of solubilized membranes with an antipeptide SSTR2 antibody revealed a single SSTR2 protein of 72 +/- 2 kDa. Preincubation of AtT-20 cell membranes with SSTR2 antibody reduced 125I-[Leu8,D-Trp22,Tyr25]SST-28 binding sites by 38%. Residual binding sites exhibited a 4.9-fold increase in affinity for SST-28, a 2.6-fold decrease in affinity for SST-14, and an SST-28:SST-14 potency ratio of 40:1 compared with a potency ratio of 3.5:1 in control membranes. These results demonstrate the expression of four SSTR genes in AtT-20 cells of which SSTR2 predominates. Blockade of SSTR2 with antibody exposes high affinity SST-28 selective sites with comparable binding characteristics to those reported for cloned SSTR5. These SST-28 binding sites may arise from a SSTR5 variant encoded by a high molecular weight 3.8-kb transcript or more likely from another as yet undiscovered member of the SST-28 selective SSTR subfamily.

  12. Ternary arsenides ATt3As3 (A=K, Rb; Tt=Ge, Sn) with layered structures

    NASA Astrophysics Data System (ADS)

    Khatun, Mansura; Stoyko, Stanislav S.; Mar, Arthur

    2016-06-01

    The four ternary arsenides ATt3As3 (A=K, Rb; Tt=Ge, Sn) were obtained by reaction of the elements at 600-650 °C. They adopt an orthorhombic structure (space group Pnma, Z=4, with cell parameters ranging from a=9.9931(11) Å, b=3.7664(4) Å, c=18.607(2) Å for KGe3As3 to a=10.3211(11) Å, b=4.0917(4) Å, c=19.570(2) Å for RbSn3As3) containing corrugated [Tt3As3] layers built from Tt-centred trigonal pyramids and tetrahedra forming five-membered rings decorated with As handles. They can be considered to be Zintl phases with Tt atoms in +4, +3, and +1 oxidation states. Band structure calculations predict that these compounds are semiconductors with narrow band gaps (0.71 eV in KGe3As3, 0.50 eV in KSn3As3).

  13. Efficiency of integron cassette insertion in correct orientation is ensured by the interplay of the three unpaired features of attC recombination sites.

    PubMed

    Nivina, Aleksandra; Escudero, José Antonio; Vit, Claire; Mazel, Didier; Loot, Céline

    2016-09-19

    The integron is a bacterial recombination system that allows acquisition, stockpiling and expression of cassettes carrying protein-coding sequences, and is responsible for the emergence and rise of multiresistance in Gram-negative bacteria. The functionality of this system depends on the insertion of promoterless cassettes in correct orientation, allowing their expression from the promoter located upstream of the cassette array. Correct orientation is ensured by strand selectivity of integron integrases for the bottom strand of cassette recombination sites (attC), recombined in form of folded single-stranded hairpins. Here, we investigated the basis of such strand selectivity by comparing recombination of wild-type and mutated attC sites with different lengths, sequences and structures. We show that all three unpaired structural features that distinguish the bottom and top strands contribute to strand selectivity. The localization of Extra-Helical Bases (EHBs) directly favors integrase binding to the bottom strand. The Unpaired Central Spacer (UCS) and the Variable Terminal Structure (VTS) influence strand selectivity indirectly, probably through the stabilization of the bottom strand and the resulting synapse due to the nucleotide skew between the two strands. These results underscore the importance of the single-stranded nature of the attC site that allows such tight control over integron cassette orientation. PMID:27496283

  14. Efficiency of integron cassette insertion in correct orientation is ensured by the interplay of the three unpaired features of attC recombination sites

    PubMed Central

    Nivina, Aleksandra; Escudero, José Antonio; Vit, Claire; Mazel, Didier; Loot, Céline

    2016-01-01

    The integron is a bacterial recombination system that allows acquisition, stockpiling and expression of cassettes carrying protein-coding sequences, and is responsible for the emergence and rise of multiresistance in Gram-negative bacteria. The functionality of this system depends on the insertion of promoterless cassettes in correct orientation, allowing their expression from the promoter located upstream of the cassette array. Correct orientation is ensured by strand selectivity of integron integrases for the bottom strand of cassette recombination sites (attC), recombined in form of folded single-stranded hairpins. Here, we investigated the basis of such strand selectivity by comparing recombination of wild-type and mutated attC sites with different lengths, sequences and structures. We show that all three unpaired structural features that distinguish the bottom and top strands contribute to strand selectivity. The localization of Extra-Helical Bases (EHBs) directly favors integrase binding to the bottom strand. The Unpaired Central Spacer (UCS) and the Variable Terminal Structure (VTS) influence strand selectivity indirectly, probably through the stabilization of the bottom strand and the resulting synapse due to the nucleotide skew between the two strands. These results underscore the importance of the single-stranded nature of the attC site that allows such tight control over integron cassette orientation. PMID:27496283

  15. Ionic milieu controls the compartment-specific activation of pro-opiomelanocortin processing in AtT-20 cells.

    PubMed Central

    Schmidt, W K; Moore, H P

    1995-01-01

    Newly synthesized prohormones and their processing enzymes transit through the same compartments before being packaged into regulated secretory granules. Despite this coordinated intracellular transport, prohormone processing does not occur until late in the secretory pathway. In the mouse pituitary AtT-20 cell line, conversion of pro-opiomelanocortin (POMC) to mature adrenocorticotropic hormone involves the prohormone convertase PC1. The mechanism by which this proteolytic processing is restricted to late secretory compartments is unknown; PC1 activity could be regulated by compartment-specific activators/inhibitors, or through changes in the ionic milieu that influence its activity. By arresting transport in a semi-intact cell system, we have addressed whether metabolically labeled POMC trapped in early secretory compartments can be induced to undergo conversion if the ionic milieu in these compartments is experimentally manipulated. Prolonged incubation of labeled POMC trapped in the endoplasmic reticulum or Golgi/trans-Golgi network did not result in processing, thereby supporting the theory that processing is normally a post-Golgi/trans-Golgi network event. However, acidification of these compartments allowed effective processing of POMC to the intermediate and mature forms. The observed processing increased sharply at a pH below 6.0 and required millimolar calcium, regardless of the compartment in which labeled POMC resided. These conditions also resulted in the coordinate conversion of PC1 from the 84/87 kDa into the 74-kDa and 66-kDa forms. We propose that POMC processing is predominantly restricted to acidifying secretory granules, and that a change in pH within these granules is both necessary and sufficient to activate POMC processing. Images PMID:8573786

  16. Marine integrons containing novel integrase genes, attachment sites, attI, and associated gene cassettes in polluted sediments from Suez and Tokyo Bays

    PubMed Central

    Elsaied, Hosam; Stokes, Hatch W; Kitamura, Keiko; Kurusu, Yasurou; Kamagata, Yoichi; Maruyama, Akihiko

    2011-01-01

    In order to understand the structure and biological significance of integrons and associated gene cassettes in marine polluted sediments, metagenomic DNAs were extracted from sites at Suez and Tokyo Bays. PCR amplicons containing new integrase genes, intI, linked with novel gene cassettes, were recovered and had sizes from 1.8 to 2.5 kb. This approach uncovered, for the first time, the structure and diversity of both marine integron attachment site, attI, and the first gene cassette, the most efficiently expressed integron-associated gene cassette. The recovered 13 and 20 intI phylotypes, from Suez and Tokyo Bay samples, respectively, showed a highly divergence, suggesting a difference in integron composition between the sampling sites. Some intI phylotypes showed similarity with that from Geobacter metallireducens, belonging to Deltaproteobacteria, the dominant class in both sampling sites, as determined by 16S rRNA gene analysis. Thirty distinct families of putative attI site, as determined by the presence of an attI-like simple site, were recovered. A total of 146 and 68 gene cassettes represented Suez and Tokyo Bay unsaturated cassette pools, respectively. Gene cassettes, including a first cassette, from both sampling sites encoded two novel families of glyoxalase/bleomycin antibiotic-resistance protein. Gene cassettes from Suez Bay encoded proteins similar to haloacid dehalogenases, protein disulfide isomerases and death-on-curing and plasmid maintenance system killer proteins. First gene cassettes from Tokyo Bay encoded a xenobiotic-degrading protein, cardiolipin synthetase, esterase and WD40-like β propeller protein. Many of the first gene cassettes encoded proteins with no ascribable function but some of them were duplicated and possessed signal functional sites, suggesting efficient adaptive functions to their bacterial sources. Thus, each sampling site had a specific profile of integrons and cassette types consistent with the hypothesis that the

  17. Inhibition of Ubiquitin-specific Peptidase 8 Suppresses Adrenocorticotropic Hormone Production and Tumorous Corticotroph Cell Growth in AtT20 Cells

    PubMed Central

    Jian, Fang-Fang; Li, Yun-Feng; Chen, Yu-Fan; Jiang, Hong; Chen, Xiao; Zheng, Li-Li; Zhao, Yao; Wang, Wei-Qing; Ning, Guang; Bian, Liu-Guan; Sun, Qing-Fang

    2016-01-01

    Background: Two recent whole-exome sequencing researches identifying somatic mutations in the ubiquitin-specific protease 8 (USP8) gene in pituitary corticotroph adenomas provide exciting advances in this field. These mutations drive increased epidermal growth factor receptor (EGFR) signaling and promote adrenocorticotropic hormone (ACTH) production. This study was to investigate whether the inhibition of USP8 activity could be a strategy for the treatment of Cushing's disease (CD). Methods: The anticancer effect of USP8 inhibitor was determined by testing cell viability, colony formation, apoptosis, and ACTH secretion. The immunoblotting and quantitative reverse transcription polymerase chain reaction were conducted to explore the signaling pathway by USP8 inhibition. Results: Inhibition of USP8-induced degradation of receptor tyrosine kinases including EGFR, EGFR-2 (ERBB2), and Met leading to a suppression of AtT20 cell growth and ACTH secretion. Moreover, treatment with USP8 inhibitor markedly induced AtT20 cells apoptosis. Conclusions: Inhibition of USP8 activity could be an effective strategy for CD. It might provide a novel pharmacological approach for the treatment of CD. PMID:27569239

  18. Involvement of calyculin A inhibitable protein phosphatases in the cyclic AMP signal transduction pathway of mouse corticotroph tumour (AtT20) cells

    PubMed Central

    Antaraki, A; Ang, K L; Antoni, F A

    1997-01-01

    The role of non-calcineurin protein phosphatases in the cyclic AMP signal transduction pathway was examined in mouse pituitary corticotroph tumour (AtT20) cells. Blockers of protein phosphatases, calyculin A and okadaic acid, were applied in AtT20 cells depleted of rapidly mobilizable pools of intracellular calcium and activated by various cyclic AMP generating agonists. Inhibitors of cyclic nucleotide phosphodiesterases were present throughout. The accumulation of cyclic AMP was monitored by radioimmunoassay, phosphodiesterase activity in cell homogenates was measured by radiometric assay. Neither calyculin A nor okadaic acid altered basal cyclic AMP levels but cyclic AMP formation induced by 41 amino acid residue corticotrophin releasing-factor (CRF) was strongly inhibited (up to 80%). 1-Norokadaone was inactive. Similar data were also obtained when isoprenaline or pituitary adenylate cyclase activating peptide1–38 were used as agonists. Pertussis toxin did not modify the inhibition of CRF-induced cyclic AMP production by calyculin A. Pretreatment with calyculin A completely prevented the stimulation of cyclic AMP formation by cholera toxin even in the presence of 0.5 mM isobutylmethylxanthine (IBMX) and 0.1 mM rolipram. Cholera toxin mediated ADP-ribosylation of the 45K and 52K molecular weight Gsα isoforms in membranes from calyculin A-pretreated cells was enhanced to 150–200% when compared with controls. Cholera toxin-induced cyclic AMP was reduced by calyculin A within 10 min when calyculin A was applied after a 90 min pretreatment with cholera toxin. Under these conditions the effect of calyculin A could be blocked by the combination of 0.5 mM IBMX and 0.1 mM rolipram, but not by 0.5 mM IBMX alone. Phosphodiesterase activity in AtT20 cell homogenates showed a significant, 2.7 fold increase after treatment with calyculin A. In control cells phosphodiesterase activity was blocked by 80% in the presence of IBMX (0.5 mM), or IBMX plus

  19. Cell cycle dependent induction of apoptosis by somatostatin analog SMS 201-995 in AtT-20 mouse pituitary cells.

    PubMed

    Srikant, C B

    1995-04-17

    Somatostatin (SS) and its analogs exert direct antiproliferative effects in a variety of tumor cells not only by inhibiting the mitogenic signalling of growth factor receptor kinases, but also by inducing apoptosis. In this study, the apoptotic effect of the octreotide SS analog, SMS 201-995, was investigated in phase synchronized AtT-20 mouse pituitary tumor cells. SMS 201-995 added to cells synchronized in G1 or in G2 phases induces apoptosis which is manifested in the subsequent cell cycle. No evidence of apoptosis was seen in cells subjected to cell cycle arrest in G1 by lovastatin or in G2 with 160 nM staurosporine. SMS 201-995 did not induce detectable apoptosis in unsynchronized cells which remained predominantly in G1 phase. These findings provide the first documentation that SS peptides regulate cell growth in G2 to induce apoptosis.

  20. Modulation of CRF signaling through receptor splicing in mouse pituitary cell line AtT-20 - Emerging role of soluble isoforms

    PubMed Central

    Żmijewski, M.A.; Slominski, A.T.

    2009-01-01

    Summary Previously, using cultured human epidermal keratinocytes we have demonstrated that the activity of CRF1 receptor can be modulated by the process of alternative splicing. This phenomenon has been further investigated in the mouse corticotroph AtT-20 cell line. In the cells, transiently transfected with the plasmids coding human CRF1 isoforms, only isoforms α and c have shown expression on the cell membrane. Other isoforms d, e, g and h had intracellular localization with the isoform e also found in the nucleus. Co-expression of the CRF1α (main form of the receptor) with isoforms d, f and g prevented its expression on the cell surface resulting in accumulation of CRF1α inside of the cell. As expected, CRF stimulated time and dose dependent activation of CRE, CARE, AP-1 transcription elements and POMC promoter in AtT-20 cells overexpressing human CRF1α, while having no effect on the AP-1 transcriptional activity in cells transfected with other isoforms (d, f, g and h). However, when cells were co-transfected with CRF1α and CRF1e or h the CRF stimulated transcriptional activity of CRE and AP-1 was amplified in comparison to the cells expressing solely CRF1α; the effect was more pronounced for CRF1h than for CRF1e. In contrast, the conditioned media from the cells overexpressing CRF1e and h inhibited the CRF induced transcriptional activity in cells overexpressing CRF1α. Media from cells expressing CRF1h were significantly more potent that from cells transfected with CRF1e. In summary, we have demonstrated that alternatively spliced CRF1 isoforms can regulate the cellular localization of CRF1α, and that soluble CRF1 isoforms can have a dual effect on CRF1α activity depending on the intracellular vs. extracellular localization. PMID:20083850

  1. Predictors and Timing of ATT Initiation among HIV-TB Patients at ART Centers of Karnataka, India: Two Year Follow-Up

    PubMed Central

    Shastri, Suresh; Nagaraja, Sharath Burugina; Tripathy, Jaya Prasad; Satyanarayana, Srinath; Rewari, Bharat Bhushan

    2015-01-01

    Background In India, TB and HIV co-infection remains as a serious public health problem. From 2006 onwards, the intensified TB-HIV collaborative activities are being jointly implemented by National AIDS Control Programme (NACP) and Revised National TB Control programme (RNTCP) at high HIV burden states. Objectives To determine (a) the predictors of outcome among a cohort of HIV-TB co-infected patients after two years after initiation of ART treatment. (b) prognostic significance of time difference between the initiation of ATT and ART in HIV-TB co-infected patients. Methods Patients registered at sixteen ART centres in Karnataka, from October through December 2009 formed the study cohort and were followed till December 2011. Results A total of 604 HIV-TB patients were registered. Follow-up (a) at the end of one year had shown 63.6% (377)patients with unfavorable TB treatment outcomes (b) at the end of second year, 55.6% (336)patients were alive on ART treatment. The variables male, smear negative TB, CD4 count less than 50cells per cumm and unfavorable TB outcome were significantly associated with unfavorable ART treatment outcome. Conclusions The programmes need to review the existing strategies and strengthen HIV-TB collaborative activities for timely treatment initiation with intensive monitoring of HIV-TB patients on treatment. PMID:26394397

  2. Dual Role of the Saccharomyces cerevisiae TEA/ATTS Family Transcription Factor Tec1p in Regulation of Gene Expression and Cellular Development

    PubMed Central

    Köhler, Tim; Wesche, Stefanie; Taheri, Naimeh; Braus, Gerhard H.; Mösch, Hans-Ulrich

    2002-01-01

    In Saccharomyces cerevisiae, the transcription factors Tec1p and Ste12p are required for haploid invasive and diploid pseudohyphal growth. Tec1p and Ste12p have been postulated to regulate these developmental processes primarily by cooperative binding to filamentous and invasion-responsive elements (FREs), which are combined enhancer elements that consist of a Tec1p-binding site (TCS) and an Ste12p-binding site (PRE). They are present in the promoter regions of target genes, e.g., FLO11. Here, we show that Tec1p efficiently activates target gene expression and cellular development in the absence of Ste12p. We further demonstrate that TCS elements alone are sufficient to mediate Tec1p-driven gene expression by a mechanism termed TCS control that is operative even when Ste12p is absent. Mutational analysis of TEC1 revealed that TCS control, FLO11 expression, and haploid invasive growth require the C terminus of Tec1p. In contrast, the Ste12p-dependent FRE control mechanism is sufficiently executed by the N-terminal portion of Tec1p, which contains the TEA/ATTS DNA-binding domain. Our study suggests that regulation of haploid invasive and diploid pseudohyphal growth by Ste12p and Tec1p is not only executed by combinatorial control but involves additional control mechanisms in which Ste12p activates TEC1 expression via clustered PREs and where Tec1p regulates expression of target genes, e.g., FLO11, by TCS control. PMID:12455687

  3. The BlcC (AttM) Lactonase of Agrobacterium tumefaciens Does Not Quench the Quorum-Sensing System That Regulates Ti Plasmid Conjugative Transfer ▿ †

    PubMed Central

    Khan, Sharik R.; Farrand, Stephen K.

    2009-01-01

    The conjugative transfer of Agrobacterium plasmids is controlled by a quorum-sensing system consisting of TraR and its acyl-homoserine lactone (HSL) ligand. The acyl-HSL is essential for the TraR-mediated activation of the Ti plasmid Tra genes. Strains A6 and C58 of Agrobacterium tumefaciens produce a lactonase, BlcC (AttM), that can degrade the quormone, leading some to conclude that the enzyme quenches the quorum-sensing system. We tested this hypothesis by examining the effects of the mutation, induction, or mutational derepression of blcC on the accumulation of acyl-HSL and on the conjugative competence of strain C58. The induction of blc resulted in an 8- to 10-fold decrease in levels of extracellular acyl-HSL but in only a twofold decrease in intracellular quormone levels, a measure of the amount of active intracellular TraR. The induction or mutational derepression of blc as well as a null mutation in blcC had no significant effect on the induction of or continued transfer of pTiC58 from donors in any stage of growth, including stationary phase. In matings performed in developing tumors, wild-type C58 transferred the Ti plasmid to recipients, yielding transconjugants by 14 to 21 days following infection. blcC-null donors yielded transconjugants 1 week earlier, but by the following week, transconjugants were recovered at numbers indistinguishable from those of the wild type. Donors mutationally derepressed for blcC yielded transconjugants in planta at numbers 10-fold lower than those for the wild type at weeks 2 and 3, but by week 4, the two donors showed no difference in recoverable transconjugants. We conclude that BlcC has no biologically significant effect on Ti plasmid transfer or its regulatory system. PMID:19011037

  4. Muscarinic cholinergic ligand binding to intact mouse pituitary tumor cells (AtT-20/D16-16) coupling with two biochemical effectors: adenylate cyclase and phosphatidylinositol turnover.

    PubMed

    Akiyama, K; Vickroy, T W; Watson, M; Roeske, W R; Reisine, T D; Smith, T L; Yamamura, H I

    1986-03-01

    (-)-[3H]Quinuclidinyl benzilate (QNB) binding to muscarinic receptors on intact mouse pituitary tumor cells (AtT-20/D16-16) was characterized in an attempt to correlate radioligand binding properties with receptor-coupled biochemical responses. Performing rinse time studies for 2 hr produced a remarkably improved ratio of specific/total (+)-[3H]QNB binding (85%). Kinetic experiments yielded association (k+1) and dissociation (k-1) rate constants of 2.2 X 10(8) M-1 min-1 and 6.8 X 10(-3) min-1, respectively. Receptor occupancy curves demonstrated a uniform population of specific, saturable (-)-[3H]QNB binding sites with a Hill coefficient equal to 1.0 and an apparent dissociation constant (Kd) equal to 34 pM under our conditions. Stereoselectivity was observed with the enantiomers (dexetimide and levetimide) of benzetimide (a factor of 4300). Concentrations of carbachol that produced a half-maximal inhibition of cyclic AMP formation and a concentration of carbachol for producing half-maximal stimulation of phosphatidylinositol turnover in the intact cells were 0.45 and 170 microM, respectively. Schild analysis revealed that pirenzepine, a nonclassical muscarinic antagonist, had a 40-fold greater affinity for reversing carbachol-stimulated phosphatidylinositol turnover (inhibition constant or Ki = 7 nM), compared to its antagonism of the carbachol-mediated inhibition of isoproterenol-stimulated cyclic AMP formation (Ki = 280 nM). Interestingly, pirenzepine inhibited (-)-[3H]QNB binding with a Ki value of 72 nM. In contrast, atropine was nearly equipotent (Ki = 0.3-0.5 nM) in binding studies and in both effector systems. PMID:3005550

  5. Parametric analysis of ATT configurations.

    NASA Technical Reports Server (NTRS)

    Lange, R. H.

    1972-01-01

    This paper describes the results of a Lockheed parametric analysis of the performance, environmental factors, and economics of an advanced commercial transport envisioned for operation in the post-1985 time period. The design parameters investigated include cruise speeds from Mach 0.85 to Mach 1.0, passenger capacities from 200 to 500, ranges of 2800 to 5500 nautical miles, and noise level criteria. NASA high performance configurations and alternate configurations are operated over domestic and international route structures. Indirect and direct costs and return on investment are determined for approximately 40 candidate aircraft configurations. The candidate configurations are input to an aircraft sizing and performance program which includes a subroutine for noise criteria. Comparisons are made between preferred configurations on the basis of maximum return on investment as a function of payload, range, and design cruise speed.

  6. Inadequacy of clinical and/or laboratory criteria for the diagnosis of lupus vulgaris, re-infection cutaneous tuberculosis: fallout/implication of 6 weeks of anti-tubular therapy (ATT) as a precise diagnostic supplement to complete the scheduled regimen.

    PubMed

    Sehgal, Virendra N; Sardana, Kabir; Sharma, Sonal

    2008-01-01

    The morphological diagnosis of the plaque form of lupus vulgaris was made on the basis of distinct clinical features. An endeavor to supplement this diagnosis was made through laboratory investigations comprising total differential leukocyte count,erythrocyte sedimentation rate (ESR), immunoglobulin M (IgM), and polymerase chain reaction (PCR); the results of which were largely ambiguous. Regimented anti-tubular therapy (ATT) was utilized to arrive at the diagnosis. A perceptible regression of the lesions over the course of 6 weeks was remarkably beneficial to completing the treatment subsequently. ATT seems to bea feasible and well-conceived tool in the diagnosis of lupus vulgaris tuberculosis verrucosa cutis and pediatric scleroderma.

  7. High performance addition-type thermoplastics (ATTs) - Evidence for the formation of a Diels-Alder adduct in the reaction of an acetylene-terminated material and a bismaleimide

    NASA Technical Reports Server (NTRS)

    Pater, R. H.; Soucek, M. D.; Chang, A. C.; Partos, R. D.

    1991-01-01

    Recently, the concept and demonstration of a new versatile synthetic reaction for making a large number of high-performance addition-type thermoplastics (ATTs) were reported. The synthesis shows promise for providing polymers having an attractive combination of easy processability, good toughness, respectable high temperature mechanical performance, and excellent thermo-oxidative stability. The new chemistry involves the reaction of an acetylene-terminated material with a bismaleimide or benzoquinone. In order to clarify the reaction mechanism, model compound studies were undertaken in solutions as well as in the solid state. The reaction products were purified by flash chromatography and characterized by conventional analytical techniques including NMR, FT-IR, UV-visible, mass spectroscopy, and high pressure liquid chromatography. The results are presented of the model compound studies which strongly support the formation of a Diels-Alder adduct in the reaction of an acetylene-terminated compound and a bismaleimide or benzoquinone.

  8. 78 FR 55103 - Notice of Proposed Exemption involving AT&T Inc. (Together With AT&T Inc.'s Affiliates, AT&T or...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-09

    ... comments and hearing requests may be posted on the Internet and can be retrieved by most Internet search engines. FOR FURTHER INFORMATION CONTACT: Anna Mpras Vaughan, Office of Exemption Determinations, Employee... CFR Part 2570, Subpart B (76 FR 66637, 66644, October 27, 2011). Effective December 31, 1978,...

  9. The life and afterlife of AT&T's HealthSite.

    PubMed

    Campbell, S

    1996-01-01

    What was going to be the richest, most potent healthcare web site died only 30 days after it debuted. Its strategic partners, surprised and feeling a little abandoned, say they learned a critical lesson in the new field of marketing on the Internet.

  10. LIFE CYCLE DESIGN FRAMEWORK AND DEMONSTRATION PROJECTS - PROFILES OF AT&T AND ALLIED SIGNAL

    EPA Science Inventory

    This document offers guidance and practical experience for integrating environmental considerations into product system development. Life cycle design seeks to minimize the environmental burden associated with a product's life cycle from raw materials acquisition through manufact...

  11. Performance of repaired defects and attPSM in EUV multilayer masks

    NASA Astrophysics Data System (ADS)

    Deng, Yunfei; La Fontaine, Bruno; Neureuther, Andrew R.

    2002-12-01

    The imaging performance of non-planar topographies in EUV masks for both partially repaired defects and non-planar attenuating phase-shifting masks made with repair treatments are evaluated using rigorous electromagnetic simulation with TEMPEST. Typical topographies produced by treatment techniques in the literature such as removal of top layers and compaction produced by electron-beam heating are considered. Isolated defects on/near the surface repaired by material removal are shown to result in an image intensity within 5% of the clear field value. Deeply buried defects within the multilayer treated by electron-beam heating can be repaired to 3% of the clear field but over repair can result in some degradation. Compaction from a 6.938 nm period to a 6.312 nm period shows a 540° phase-shift and an intensity reduced to about 6% suggesting such a treatment may be used to create attenuated phase-shifting masks for EUV. The quality of the aerial image for such a mask is studied as a function of the lateral transition distance between treated and untreated regions.

  12. Investing in Quality Child Care: A Report for AT&T.

    ERIC Educational Resources Information Center

    Galinsky, Ellen; Friedman, Dana E.

    More than 50 child care experts were asked (1) What aspects of child care are most likely to ensure high quality? (2) What are the current barriers to achieving quality in child care? and (3) What corporate or labor initiative would make the greatest difference in improving the quality of child care services? Recommendations generated from the…

  13. 78 FR 67053 - Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-08

    ... Collection Order, 78 FR 2571, January 11, 2013, as implemented by this Report and Order, are not effective... questioned whether they can use Uniform Service Order Codes (USOCs) for their unique billing code IDs....

  14. 76 FR 14697 - Amdocs, Inc., Global Support Services, Advertising and Media AT&T Division, New Haven, CT; Notice...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-17

    ... 22, 2010, the petitioner requested administrative reconsideration of the Department's negative... February 22, 2011 (76 FR 5831). The subject workers are engaged in employment related to the supply...

  15. 76 FR 44036 - Proposed Low-Effect Habitat Conservation Plan for the California Tiger Salamander, AT&T Portable...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-22

    ... Fish and Wildlife Service Proposed Low-Effect Habitat Conservation Plan for the California Tiger... potential for ``take'' of one Federally listed animal, the California tiger salamander. The applicant would... for the California tiger salamander into a new storage facility for portable generators within...

  16. 78 FR 2571 - Special Access for Price Cap Local Exchange Carriers; AT&T Corporation Petition for Rulemaking To...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-11

    ...In this document, the Commission continues the process of reviewing its special access rules to ensure that they reflect the state of competition today and promote competition, investment, and access to dedicated communications services businesses across the country rely on every day to deliver their products and services to American consumers. The Report and Order initiates a comprehensive......

  17. Guide to use the data features of the AT&T 7506, 7507, and 8510T ISDN phones

    SciTech Connect

    Eldridge, J.M.

    1994-07-01

    Integrated Service digital Network, ISDN, technology is an integral component of Sandia National Laboratories telecommunications infrastructure. ISDN is a fully digital telephone service that allows simultaneous voice and data communication from the same telephone instrument. Almost all ISDN phones in use at Sandia/New Mexico and most ISDN phones at Sandia/California have a built-in module for data communication. This user guide describes the use and operation of ISDN data module and services as they are installed at Sandia.

  18. Tumor tracking and motion compensation with an adaptive tumor tracking system (ATTS): System description and prototype testing

    SciTech Connect

    Wilbert, Juergen; Meyer, Juergen; Baier, Kurt; Guckenberger, Matthias; Herrmann, Christian; Hess, Robin; Janka, Christian; Ma Lei; Mersebach, Torben; Richter, Anne; Roth, Michael; Schilling, Klaus; Flentje, Michael

    2008-09-15

    A novel system for real-time tumor tracking and motion compensation with a robotic HexaPOD treatment couch is described. The approach is based on continuous tracking of the tumor motion in portal images without implanted fiducial markers, using the therapeutic megavoltage beam, and tracking of abdominal breathing motion with optical markers. Based on the two independently acquired data sets the table movements for motion compensation are calculated. The principle of operation of the entire prototype system is detailed first. In the second part the performance of the HexaPOD couch was investigated with a robotic four-dimensional-phantom capable of simulating real patient tumor trajectories in three-dimensional space. The performance and limitations of the HexaPOD table and the control system were characterized in terms of its dynamic behavior. The maximum speed and acceleration of the HexaPOD were 8 mm/s and 34.5 mm/s{sup 2} in the lateral direction, and 9.5 mm/s and 29.5 mm/s{sup 2} in longitudinal and anterior-posterior direction, respectively. Base line drifts of the mean tumor position of realistic lung tumor trajectories could be fully compensated. For continuous tumor tracking and motion compensation a reduction of tumor motion up to 68% of the original amplitude was achieved. In conclusion, this study demonstrated that it is technically feasible to compensate breathing induced tumor motion in the lung with the adaptive tumor tracking system.

  19. Att Tolka Barns Signaler: Gravt utvecklingsstorda flerhandikappade barns lek och kommunikation (To Interpret Childrens' Signals: Play and Communication in Profoundly Mentally Retarded and Multiply Handicapped Children).

    ERIC Educational Resources Information Center

    Brodin, Jane

    Written in Swedish with an English-language summary, this report describes a study which examined the interaction between mothers or caregivers and their children with profound mental retardation and multiple disabilities, particularly looking at the function of play in communicative interaction. The six children all had five or six handicaps in…

  20. Simulation directe de l'atténuation par contrôle adaptatif du rayonnement d'une cavité affleurée par un écoulement

    NASA Astrophysics Data System (ADS)

    Marsden, Olivier; Gloerfelt, Xavier; Bailly, Christophe

    2003-06-01

    The Large Eddy Simulation of closed-loop active flow control applied to a 3D cavity excited by a compressible airflow with a Mach number of 0.6 is presented. The control actuator is an idealized synthetic jet located at the upstream cavity edge, and the control function is supplied by a feedback LMS-type algorithm whose input is a pressure signal measured inside the cavity. The radiated sound, provided directly by the LES simulation, was shown to decrease substantially when active control was applied. A simultaneous reduction of the vertical velocity fluctuations in the shear layer was observed. The intensity of vortical structures inside the cavity was also reduced, although the general aspect of the recirculation zone was not modified. The direct noise computation technique, which supplies the pressure field by solving the fluid mechanics equations, is shown to constitute a powerful tool for studying active aeroacoustic noise control. To cite this article: O. Marsden et al., C. R. Mecanique 331 (2003).

  1. Measuring Syntactic Complexity in Spontaneous Spoken Swedish

    ERIC Educational Resources Information Center

    Roll, Mikael; Frid, Johan; Horne, Merle

    2007-01-01

    Hesitation disfluencies after phonetically prominent stranded function words are thought to reflect the cognitive coding of complex structures. Speech fragments following the Swedish function word "att" "that" were analyzed syntactically, and divided into two groups: one with "att" in disfluent contexts, and the other with "att" in fluent…

  2. Helical polyurethane@attapulgite nanocomposite: Preparation, characterization and study of optical activity

    NASA Astrophysics Data System (ADS)

    Wang, Zhiqiang; Zhou, Yuming; Sun, Yanqing; Fan, Kai; Guo, Xingxing; Jiang, Xiaolei

    2009-08-01

    Helical polyurethane@attapulgite (BM-ATT) based on R-1,1'-binaphthyl-2',2-diol (R-BINOL) composite was prepared after the surface modification of attapulgite (ATT). BM-ATT was characterized by Fourier-transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high resolution transmission electron microscopy (HTEM) and vibrational circular dichroism (VCD) spectroscopy. FT-IR and XRD analyses indicate that the helical polyurethane has been successfully grafted onto the surfaces of the modified ATT without destroying the original crystalline structure of ATT. BM-ATT exhibits the rod-like structure by SEM, TEM, and HTEM photographs. BM-ATT displays obvious Cotton effect for some absorbance in VCD spectrum, and its optical activity results from the singlehanded conformation of helical polyurethane.

  3. The mechanism of ϕC31 integrase directionality: experimental analysis and computational modelling

    PubMed Central

    Pokhilko, Alexandra; Zhao, Jia; Ebenhöh, Oliver; Smith, Margaret C. M.; Stark, W. Marshall; Colloms, Sean D.

    2016-01-01

    Serine integrases, DNA site-specific recombinases used by bacteriophages for integration and excision of their DNA to and from their host genomes, are increasingly being used as tools for programmed rearrangements of DNA molecules for biotechnology and synthetic biology. A useful feature of serine integrases is the simple regulation and unidirectionality of their reactions. Recombination between the phage attP and host attB sites is promoted by the serine integrase alone, giving recombinant attL and attR sites, whereas the ‘reverse’ reaction (between attL and attR) requires an additional protein, the recombination directionality factor (RDF). Here, we present new experimental data on the kinetics and regulation of recombination reactions mediated by ϕC31 integrase and its RDF, and use these data as the basis for a mathematical model of the reactions. The model accounts for the unidirectionality of the attP × attB and attL × attR reactions by hypothesizing the formation of structurally distinct, kinetically stable integrase–DNA product complexes, dependent on the presence or absence of RDF. The model accounts for all the available experimental data, and predicts how mutations of the proteins or alterations of reaction conditions might increase the conversion efficiency of recombination. PMID:27387286

  4. Control of directionality in Streptomyces phage φBT1 integrase-mediated site-specific recombination.

    PubMed

    Zhang, Lin; Zhu, Binyan; Dai, Ruixue; Zhao, Guoping; Ding, Xiaoming

    2013-01-01

    Streptomyces phage φBT1 integrates its genome into the attB site of the host chromosome with the attP site to generate attL and attR. The φBT1 integrase belongs to the large serine recombinase subfamily which directly binds to target sites to initiate double strand breakage and exchange. A recombination directionality factor (RDF) is commonly required for switching integration to excision. Here we report the characterization of the RDF protein for φBT1 recombination. The RDF, is a phage-encoded gp3 gene product (28 KDa), which allows efficient active excision between attL and attR, and inhibits integration between attB and attP; Gp3 can also catalyze topological relaxation with the integrase of supercoiled plasmids containing a single excision site. Further study showed that Gp3 could form a dimer and interact with the integrase whether it bound to the substrate or not. The synapse formation of attL or attR alone with integrase and Gp3 showed that synapsis did not discriminate between the two sites, indicating that complementarity of central dinucleotides is the sole determinant of outcome in correct excision synapses. Furthermore, both in vitro and in vivo evidence support that the RDFs of φBT1 and φC31 were fully exchangeable, despite the low amino acid sequence identity of the two integrases.

  5. TG1 integrase-based system for site-specific gene integration into bacterial genomes.

    PubMed

    Muroi, Tetsurou; Kokuzawa, Takaaki; Kihara, Yoshihiko; Kobayashi, Ryuichi; Hirano, Nobutaka; Takahashi, Hideo; Haruki, Mitsuru

    2013-05-01

    Serine-type phage integrases catalyze unidirectional site-specific recombination between the attachment sites, attP and attB, in the phage and host bacterial genomes, respectively; these integrases and DNA target sites function efficiently when transferred into heterologous cells. We previously developed an in vivo site-specific genomic integration system based on actinophage TG1 integrase that introduces ∼2-kbp DNA into an att site inserted into a heterologous Escherichia coli genome. Here, we analyzed the TG1 integrase-mediated integrations of att site-containing ∼10-kbp DNA into the corresponding att site pre-inserted into various genomic locations; moreover, we developed a system that introduces ∼10-kbp DNA into the genome with an efficiency of ∼10(4) transformants/μg DNA. Integrations of attB-containing DNA into an attP-containing genome were more efficient than integrations of attP-containing DNA into an attB-containing genome, and integrations targeting attP inserted near the replication origin, oriC, and the E. coli "centromere" analogue, migS, were more efficient than those targeting attP within other regions of the genome. Because the genomic region proximal to the oriC and migS sites is located at the extreme poles of the cell during chromosomal segregation, the oriC-migS region may be more exposed to the cytosol than are other regions of the E. coli chromosome. Thus, accessibility of pre-inserted attP to attB-containing incoming DNA may be crucial for the integration efficiency by serine-type integrases in heterologous cells. These results may be beneficial to the development of serine-type integrases-based genomic integration systems for various bacterial species.

  6. PhiC31 integrase induces efficient site-specific recombination in the Capra hircus genome.

    PubMed

    Ma, Haiyan; Ma, Qingwen; Lu, Yao; Wang, Juan; Hu, Wei; Gong, Zhijuan; Cai, Linlin; Huang, Ying; Huang, Shu-Zhen; Zeng, Fanyi

    2014-08-01

    Streptomyces phage φC31 integrase induces efficient site-specific recombination capable of integrating exogenous genes at pseudo attP sites in human, mouse, rat, rabbit, sheep, Drosophila, and bovine genomes. However, the φC31-mediated recombination between attB and the corresponding pseudo attP sites has not been investigated in Capra hircus. Here, we identified eight pseudo attP sites located in the intron or intergenic regions of the C. hircus genome, and demonstrated different levels of foreign gene expression after φC31 integrase-mediated integration. These pseudo attP sites share similar sequences with each other and with pseudo attP sites in other mammalian genomes, and these are associated with a neighboring consensus motif found in other genomes. The application of the φC31 integrase system in C. hircus provides a new option for genetic engineering of this economically important goat species.

  7. Site-specific integration of bacteriophage VWB genome into Streptomyces venezuelae and construction of a VWB-based integrative vector.

    PubMed

    Van Mellaert, L; Mei, L; Lammertyn, E; Schacht, S; Anné, J

    1998-12-01

    The temperate bacteriophage VWB integrates into the chromosome of Streptomyces venezuelae ETH14630 via site-specific integration. Following recombination of the VWB attP region with the chromosomal attB sequence, the host-phage junctions attL and attR are formed. Nucleotide sequence analysis of attP, attB, attL and attR revealed a 45 bp common core sequence. In attB this 45 bp sequence consists of the 3' end of a putative tRNA Arg(AGG) gene with a 3'-terminal CCA sequence which is typical for prokaryotic tRNAs. Phage DNA integration restores the putative tRNA Arg(AGG) gene in attL. However, following recombination the CCA sequence is missing as is the case for most Streptomyces tRNA genes described so far. Adjacent to VWB attP, an ORF encoding a 427 aa protein was detected. The C-terminal region of this protein shows high similarity to the conserved C-terminal domain of site-specific recombinases belonging to the integrase family. To prove the functionality of this putative integrase gene (int), an integrative vector pKT02 was constructed. This vector consists of a 2.3 kb HindIII-SphI restriction fragment of VWB DNA containing attP and int cloned in a non-replicative Escherichia coli vector carrying a thiostrepton-resistance (tsr) gene. Integration of pKT02 was obtained after transformation of Streptomyces venezuelae ETH14630 and Streptomyces lividans TK24 protoplasts. This vector will thus be useful for a number of additional Streptomyces species in which a suitable tRNA gene can be functional as integration site.

  8. Tough, High-Performance, Thermoplastic Addition Polymers

    NASA Technical Reports Server (NTRS)

    Pater, Ruth H.; Proctor, K. Mason; Gleason, John; Morgan, Cassandra; Partos, Richard

    1991-01-01

    Series of addition-type thermoplastics (ATT's) exhibit useful properties. Because of their addition curing and linear structure, ATT polymers have toughness, like thermoplastics, and easily processed, like thermosets. Work undertaken to develop chemical reaction forming stable aromatic rings in backbone of ATT polymer, combining high-temperature performance and thermo-oxidative stability with toughness and easy processibility, and minimizing or eliminating necessity for tradeoffs among properties often observed in conventional polymer syntheses.

  9. Helical polyurethane-attapulgite nanocomposite: Preparation, characterization and study of optical activity

    SciTech Connect

    Wang Zhiqiang; Zhou Yuming; Sun Yanqing; Fan Kai; Guo Xingxing; Jiang Xiaolei

    2009-08-15

    Helical polyurethane-attapulgite (BM-ATT) based on R-1,1'-binaphthyl-2',2-diol (R-BINOL) composite was prepared after the surface modification of attapulgite (ATT). BM-ATT was characterized by Fourier-transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high resolution transmission electron microscopy (HTEM) and vibrational circular dichroism (VCD) spectroscopy. FT-IR and XRD analyses indicate that the helical polyurethane has been successfully grafted onto the surfaces of the modified ATT without destroying the original crystalline structure of ATT. BM-ATT exhibits the rod-like structure by SEM, TEM, and HTEM photographs. BM-ATT displays obvious Cotton effect for some absorbance in VCD spectrum, and its optical activity results from the singlehanded conformation of helical polyurethane. - Graphical Abstract: Helical polyurethane-attapulgite (BM-ATT) based on R-1,1'-binaphthyl-2',2-diol (R-BINOL) nanocomposite was prepared after surface modification of attapulgite (ATT). This rod-like composite is coated by the optically active polyurethane shell on the surfaces.

  10. The Evolution of Access. ONA Technical and Regulatory Issues.

    ERIC Educational Resources Information Center

    Jackson, Charles L.

    In June 1986, the Federal Communications Commission (FCC) released its "Report and Order" and its "Third Computer Inquiry," which proposed to remove the then-current requirement that AT&T and the Bell Operating Companies (BOCs) provide enhanced services only through separate subsidiaries. Instead, AT&T and the BOCs would be subject to several…

  11. The integrated conjugative plasmid pSAM2 of Streptomyces ambofaciens is related to temperate bacteriophages.

    PubMed

    Boccard, F; Smokvina, T; Pernodet, J L; Friedmann, A; Guérineau, M

    1989-03-01

    Streptomyces ambofaciens ATCC23877 and derivatives contain the 11-kb element pSAM2 present in an integrated state or as a free and integrated plasmid. This element, able to integrate site-specifically in the genome of different Streptomyces species, is conjugative and mobilizes chromosomal markers. Besides these plasmid functions, we have shown that the site-specific recombination system of pSAM2 presents strong similarities with that of several temperate phages. The integration event is promoted by a site-specific recombinase of the integrase family. The int gene encoding this integrase is closely linked to the plasmid attachment site (attP). A small open reading frame (ORF) overlaps the int gene and the predicted protein exhibits similarities with Xis proteins involved in phages excision. The integrated copy of pSAM2 in strain ATCC23877 is flanked by att sequences (attL and attR). Another att sequence (attX) is present in this strain and attX and attL are the boundaries of a 42-kb fragment (xSAM1) absent, as well as pSAM2, from S.ambofaciens DSM40697. Sequences partially similar to pSAM2 int gene are found near the chromosomal integration zone in both S.ambofaciens strains. The possible origin of pSAM2, an element carrying plasmid as well as phage features, is discussed.

  12. A Pilot Study to Assess the Feasibility of Group Exercise and Animal-Assisted Therapy in Older Adults.

    PubMed

    Grubbs, Brandon; Artese, Ashley; Schmitt, Karla; Cormier, Eileen; Panton, Lynn

    2016-04-01

    This pilot study assessed the feasibility of incorporating animal-assisted therapy teams (ATT) into a 6-week group exercise program for older adults (77 ± 6 years). Fifteen participants were randomly assigned to an exercise with ATT (E+ATT; n = 8) or exercise only (E; n = 7) group. Groups exercised 3x/week for 45 min. Feasibility was assessed by three objectives: (1) ATT will not need extensive preparation beyond their original therapy training; (2) the study will require minimal cost; and (3) ATT must not impair the effectiveness of the exercise program. By the study conclusion, all objectives were met. Time and cost were minimal for ATT, and adherence was 93% and 90% for E+ATT and E, respectively. There were significant improvements in both groups (p ≤ .05) for arm curls, get-up and go, and 6-min walk. The results of this pilot study suggest that it is feasible to incorporate ATT into group exercise programming for older adults.

  13. The origin of the curing behavior, mechanical and thermal properties of surface functionalized attapulgite/bismaleimide/diallylbisphenol composites

    NASA Astrophysics Data System (ADS)

    Zhao, Lin; Liu, Ping; Liang, Guozheng; Gu, Aijuan; Yuan, Li; Guan, Qingbao

    2014-01-01

    High curing temperature and big brittleness are two disadvantages of heat resistant thermosetting resins. To simultaneously resolve these problems, surface functionalized fibrous attapulgite (N-ATT) was used to modify 4,4‧-bismaleimidodiphenylmethane (BDM)/2,2‧-diallyl bisphenol A (DBA) resin, and then a new kind of high performance nanocomposites (BD/N-ATT) were developed. The structure and properties of BD/N-ATT nanocomposites with different loadings of N-ATT were intensively studied. Results prove that the surface functionalization of attapulgite endows N-ATT has multi-effects with BD resin, and thus change the chemical and aggregation state (including crosslinking density and free volume) structures of the crosslinked network, and consequently, the composites have obviously different properties from BD resin. For the composite with only 0.5 wt% N-ATT, its impact strength is about 1.6 times the value of BD resin, while the glass transition temperature and initial degradation temperature are about 25 and 20 °C higher than those of BD resin, respectively. The attractive properties of BD/N-ATT composites with a small loading of N-ATTs suggest that the composites have great potential in many cutting-edge fields; besides, interestingly, the method developed herein suggests a new way to develop high performance resins and related composites with simultaneously improved curing feature, toughness and thermal resistance.

  14. Genetic and Biochemical Characterization of Nectria haematococca Strains with Adhesive and Adhesion-Reduced Macroconidia

    PubMed Central

    Epstein, L.; Kwon, Y. H.; Almond, D. E.; Schached, L. M.; Jones, M. J.

    1994-01-01

    A previous study reported the isolation of two mutants (LE1 and LE2) of the plant pathogenic fungus Nectria haematococca (anamorph, Fusarium solani f. sp. cucurbitae) with macroconidia with reduced ability to adhere (Att-) to zucchini fruits and polystyrene. The adhesion-reduced-phenotype in LE1 and LE2 macroconidia is temperature sensitive and dependent on the concentration of nutrients. Classical genetic analysis of progeny derived from LE1 identified a mutation in a genetic locus, named Att1. The 90-kDa glycoprotein and macroconidial tip mucilage which were previously associated with the development of adhesion competence in Att+ macroconidia are specifically associated with macroconidia; neither is produced on microconidia, which are relatively nonadherent. However, macroconidia of both Att+ and Att- strains produce the 90-kDa glycoprotein and the macroconidial tip mucilage. Images PMID:16349181

  15. Anti-tubercular therapy for intraocular tuberculosis: A systematic review and meta-analysis.

    PubMed

    Kee, Ae Ra; Gonzalez-Lopez, Julio J; Al-Hity, Aws; Gupta, Bhaskar; Lee, Cecilia S; Gunasekeran, Dinesh Visva; Jayabalan, Nirmal; Grant, Robert; Kon, Onn Min; Gupta, Vishali; Westcott, Mark; Pavesio, Carlos; Agrawal, Rupesh

    2016-01-01

    Intraocular tuberculosis remains a diagnostic and management conundrum for both ophthalmologists and pulmonologists. We analyze the efficacy and safety of anti-tubercular therapy (ATT) in patients with intraocular tuberculosis and factors associated with favorable outcome. Twenty-eight studies are included in this review, with a total of 1,917 patients. Nonrecurrence of inflammation was observed in pooled estimate of 84% of ATT-treated patients (95% CI 79-89). There was minimal difference in the outcome between patients treated with ATT alone (85% successful outcome; 95% CI 25-100) and those with concomitant systemic corticosteroid (82%; 95% CI 73-90). The use of ATT may be of benefit to patients with suspected intraocular tuberculosis; however, this conclusion is limited by the lack of control group analysis and standardized recruitment and treatment protocols. We propose further prospective studies to better establish the efficacy of ATT and ascertain the factors associated with favorable treatment outcomes. PMID:26970263

  16. Efficient site-specific integration in Plasmodium falciparum chromosomes mediated by mycobacteriophage Bxb1 integrase

    PubMed Central

    Nkrumah, Louis J; Muhle, Rebecca A; Moura, Pedro A; Ghosh, Pallavi; Hatfull, Graham F; Jacobs, William R; Fidock, David A

    2010-01-01

    Here we report an efficient, site-specific system of genetic integration into Plasmodium falciparum malaria parasite chromosomes. This is mediated by mycobacteriophage Bxb1 integrase, which catalyzes recombination between an incoming attP and a chromosomal attB site. We developed P. falciparum lines with the attB site integrated into the glutaredoxin-like cg6 gene. Transfection of these attB+ lines with a dual-plasmid system, expressing a transgene on an attP-containing plasmid together with a drug resistance gene and the integrase on a separate plasmid, produced recombinant parasites within 2 to 4 weeks that were genetically uniform for single-copy plasmid integration. Integrase-mediated recombination resulted in proper targeting of parasite proteins to intra-erythrocytic compartments, including the apicoplast, a plastid-like organelle. Recombinant attB × attP parasites were genetically stable in the absence of drug and were phenotypically homogeneous. This system can be exploited for rapid genetic integration and complementation analyses at any stage of the P. falciparum life cycle, and it illustrates the utility of Bxb1-based integrative recombination for genetic studies of intracellular eukaryotic organisms. PMID:16862136

  17. Anterior Tibial Translation in Collegiate Athletes with Normal Anterior Cruciate Ligament Integrity

    PubMed Central

    Rosene, John M.; Fogarty, Tracey D.

    1999-01-01

    Objective: To examine differences in anterior tibial translation (ATT) among sports, sex, and leg dominance in collegiate athletes with normal anterior cruciate ligament integrity. Design and Setting: Subjects from various athletic teams were measured for ATT in right and left knees. Subjects: Sixty subjects were measured for ATT with a KT-1000 knee arthrometer. Measurements: Statistical analyses were computed for each sex and included a 2 × 3 × 4 mixed-factorial analysis of variance (ANOVA) for anterior cruciate ligament displacement, right and left sides, and force and sport. A 2 × 2 × 3 mixed-factorial ANOVA was computed to compare means for sex and force. A 2 × 3 mixed-factorial ANOVA was computed to compare sex differences across 3 forces. Results: For males and females, no significant interactions were found among leg, force, and sport for mean ATT, for leg and sport or leg and force, or for translation values between dominant and nondominant legs. Males had a significant interaction for force and sport, and a significant difference was found for side of body, since the right side had less translation than the left side. Females had greater ATT than males at all forces. Conclusions: Sex differences exist for ATT, and differences in ATT exist among sports for both sexes. Differences between the right and left sides of the body should be expected when making comparisons of ligamentous laxity. ImagesFigure 2.Figure 3.Figure 5. PMID:16558565

  18. Actinophage R4 integrase-based site-specific chromosomal integration of non-replicative closed circular DNA.

    PubMed

    Miura, Takamasa; Nishizawa, Akito; Nishizawa, Tomoyasu; Asayama, Munehiko; Shirai, Makoto

    2016-06-01

    The actinophage R4 integrase (Sre)-based molecular genetic engineering system was developed for the chromosomal integration of multiple genes in Escherichia coli. A cloned DNA fragment containing two attP sites, green fluorescent protein (gfp) as a first transgene, and an antibiotic resistance gene as a selection marker was self-ligated to generate non-replicative closed circular DNA (nrccDNA) for integration. nrccDNA was introduced into attB-inserted E. coli cells harboring the plasmid expressing Sre by electroporation. The expressed Sre catalyzed site-specific integration between one of the two attP sites on nrccDNA and the attB site on the E. coli chromosome. The integration frequency was affected by the chromosomal location of the target site. A second nrccDNA containing two attB sites, lacZα encoding the alpha fragment of β-galactosidase as a transgene, and another antibiotic resistance gene was integrated into the residual attP site on the gfp-integrated E. coli chromosome via one of the two attB sites according to reiterating site-specific recombination. The integrants clearly exhibited β-galactosidase activity and green fluorescence, suggesting the simultaneous expression of multiple recombinant proteins in E. coli. The results of the present study showed that a step-by-step integration procedure using nrccDNA achieved the chromosomal integration of multiple genes. PMID:26870903

  19. Gene insertion and replacement in Schizosaccharomyces pombe mediated by the Streptomyces bacteriophage phiC31 site-specific recombination system.

    PubMed

    Thomason, L C; Calendar, R; Ow, D W

    2001-08-01

    The site-specific recombination system used by the Streptomyces bacteriophage phiC31 was tested in the fission yeast Schizosaccharomyces pombe. A target strain with the phage attachment site attP inserted at the leu1 locus was co-transformed with one plasmid containing the bacterial attachment site attB linked to a ura4+ marker, and a second plasmid expressing the phiC31 integrase gene. High-efficiency transformation to the Ura+ phenotype occurred when the integrase gene was expressed. Southern analysis revealed that the attB-ura4+ plasmid integrated into the chromosomal attP site. Sequence analysis showed that the attBxattP recombination was precise. In another approach, DNA with a ura4+ marker flanked by two attB sites in direct orientation was used to transform S. pombe cells bearing an attP duplication. The phiC31 integrase catalyzed two reciprocal cross-overs, resulting in a precise gene replacement. The site-specific insertions are stable, as no excision (the reverse reaction) was observed on maintenance of the integrase gene in the integrant lines. The irreversibility of the phiC31 site-specific recombination system sets it apart from other systems currently used in eukaryotic cells, which reverse readily. Deployment of the phiC31 recombination provides new opportunities for directing transgene and chromosome rearrangements in eukaryotic systems.

  20. Timing of antiretroviral therapy and TB treatment outcomes in patients with TB-HIV in Myanmar

    PubMed Central

    Shewade, H. D.; Kyaw, N. T. T.; Oo, M. M.; Aung, T. K.; Aung, S. T.; Oo, H. N.; Win, T.; Harries, A. D.

    2016-01-01

    Setting: Integrated HIV Care programme, Mandalay, Myanmar. Objectives: To determine time to starting antiretroviral treatment (ART) in relation to anti-tuberculosis treatment (ATT) and its association with TB treatment outcomes in patients co-infected with tuberculosis (TB) and the human immunodeficiency virus (HIV) enrolled from 2011 to 2014. Design: Retrospective cohort study. Results: Of 1708 TB-HIV patients, 1565 (92%) started ATT first and 143 (8%) started ART first. Treatment outcomes were missing for 226 patients and were thus not included. In those starting ATT first, the median time to starting ART was 8.6 weeks. ART was initiated after 8 weeks in 830 (53%) patients. Unsuccessful outcome was found in 7%, with anaemia being an independent predictor. In patients starting ART first, the median time to starting ATT was 21.6 weeks. ATT was initiated within 3 months in 56 (39%) patients. Unsuccessful outcome was found in 12%, and in 20% of those starting ATT within 3 months. Patients with CD4 count <100/mm3 had a four times higher risk of an unsuccessful outcome. Conclusions: Timing of ART in relation to ATT was not an independent risk factor for unsuccessful outcome. Extensive screening for TB with rapid and sensitive diagnostic tests in HIV-infected persons and close monitoring of anaemia and immunosuppression are recommended to further improve TB treatment outcomes among patients with TB-HIV. PMID:27358804

  1. The retrovirus pol gene encodes a product required for DNA integration: identification of a retrovirus int locus.

    PubMed Central

    Panganiban, A T; Temin, H M

    1984-01-01

    We mutagenized cloned spleen necrosis virus DNA to identify a region of the retrovirus genome encoding a polypeptide required for integration of viral DNA. Five plasmids bearing different lesions in the 3' end of the pol gene were examined for the ability to integrate or replicate following transfection of chicken embryo fibroblasts. Transfection with one of these DNAs resulted in the generation of mutant virus incapable of integrating but able to replicate at low levels; this phenotype is identical to that of mutants bearing alterations in the cis-acting region, att. To determine whether the 3' end of the pol gene encodes a protein that interacts with att, we did a complementation experiment. Cells were first infected with an att- virus and then superinfected with the integration-deficient virus containing a lesion in the pol gene and a wild-type att site. The results showed that the att- virus provided a transacting function allowing integration of viral DNA derived from the mutant bearing a wild-type att site. Thus, the 3' end of the pol gene serves as an "int" locus and encodes a protein mediating integration of retrovirus DNA through interaction with att. Images PMID:6083562

  2. [Is the abnormal toxicity test still relevant for the safety of vaccines, sera and immunoglobulins?

    PubMed

    Krämer, Beate; Nagel, Margit; Duchow, Karin; Schwanig, Michael; Cussler, Klaus

    1996-01-01

    The German Pharmacopoeia (DAB) requires the abnormal toxicity test (ATT) using mice and guinea pigs as a non-specific safety test for vaccines, sera and immuno-globulines. The purpose of this project was to investigate the relevance of ATT after the introduction of GMP- and GLP-principles in the manufacturing of biological products. A great variability in the test performance became evident for the different test laboratories, involving the animal number as well as the vaccine dosage administration and test duration. The retrospective analysis of ATT results reveals reasons for the incompatibility of particular preparation groups, vaccine components or additives with the animal species used. There were highly significant differences between the manufacturers and the PEI regarding the frequency of deviating test results for identical test batches. Positive ATT"s never resulted from the insufficient quality of a batch. On the other hand vaccines causing adverse reactions in the target species were not identified by the ATT. For these reasons the abnormal toxicity test is unsuitable to detect harmful batches. The results of the analysis of data show that finally the ATT has always been passed, i.e. there was no retention by the vaccine manufacturers and no refusal by the controlling authorities due to the ATT-results. Considering the present animal model and the questionable transferability of the test results to the target species only a poor reliability is evident. Taking into account aspects of drug safety and animal welfare it is recommended to the DAB to omit the ATT.

  3. Adult thymus transplantation with allogeneic intra-bone marrow–bone marrow transplantation from same donor induces high thymopoiesis, mild graft-versus-host reaction and strong graft-versus-tumour effects

    PubMed Central

    Miyake, Takashi; Hosaka, Naoki; Cui, Wenhao; Nishida, Teruhisa; Takaki, Takashi; Inaba, Muneo; Kamiyama, Yasuo; Ikehara, Susumu

    2009-01-01

    Although allogeneic bone marrow transplantation (BMT) plus donor lymphocyte infusion (DLI) is performed for solid tumours to enhance graft-versus-tumour (GVT) effects, a graft-versus-host reaction (GVHR) is also elicited. We carried out intra-bone marrow–bone marrow transplantation (IBM-BMT) plus adult thymus transplantation (ATT) from the same donor to supply alloreactive T cells continually. Normal mice treated with IBM-BMT + ATT survived for a long time with high donor-derived thymopoiesis and mild GVHR. The percentage of CD4+ FoxP3+ regulatory T cells in the spleen of the mice treated with IBM-BMT + ATT was lower than in normal B6 mice or mice treated with IBM-BMT alone, but higher than in mice treated with IBM-BMT + DLI; the mice treated with IBM-BMT + DLI showed severe GVHR. In tumour-bearing mice, tumour growth was more strongly inhibited by IBM-BMT + ATT than by IBM-BMT alone. Mice treated with IBM-BMT + a high dose of DLI also showed tumour regression comparable to that of mice treated with IBM-BMT + ATT but died early of GVHD. By contrast, mice treated with IBM-BMT + a low dose of DLI showed longer survival but less tumour regression than the mice treated with IBM-BMT + ATT. Histologically, significant numbers of CD8+ T cells were found to have infiltrated the tumour in the mice treated with IBM-BMT + ATT. The number of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labelling (TUNEL)-positive apoptotic tumour cells also significantly increased in the mice treated with IBM-BMT + ATT. Allogeneic IBM-BMT + ATT thus can induce high thymopoiesis, preserving strong GVT effects without severe GVHR. PMID:18778285

  4. Attachment sites for bacteriophage P2 on the Escherichia coli chromosome: DNA sequences, localization on the physical map, and detection of a P2-like remnant in E. coli K-12 derivatives.

    PubMed

    Barreiro, V; Haggård-Ljungquist, E

    1992-06-01

    Integration of bacteriophage P2 into the Escherichia coli genome involves recombination between two attachment sites, attP and attB, one on the phage and one on the host genome, respectively. At least 10 different attB sites have been identified over the years. In E. coli C, one site, called locI, is preferred, being occupied before any of the others. In E. coli K-12, no such preference is seen (reviewed in L. E. Bertani and E. W. Six, p. 73-143, in R. Calendar, ed., The Bacteriophages, vol. 2, 1988). The DNA sequence of locI has been determined, and it shows a core sequence of 27 nucleotides identical to attP (A. Yu, L. E. Bertani, and E. Haggård-Ljungquist, Gene 80:1-12, 1989). By inverse polymerase chain reactions, the prophage-host junctions of DNA extracted from P2 lysogenic strains have been amplified, cloned, and sequenced. By combining the attL and attR sequences, the attB sequences of locations II, III, and H have been deduced. The core sequence of location II had 20 matches to the 27-nucleotide core sequence of attP; the sequences of locations III and H had 17 matches. Thus, the P2 integrase accepts at least up to 37% mismatches within the core sequence. The E. coli K-12 strains examined all contain a 639-nucleotide-long cryptic remnant of P2 at a site with a sequence similar to that of locI but that may have a different map position. The P2 remnant consists of the C-terminal part of gene D, all of gene ogr, and attR. Locations II, III, and H have been located on Kohara's physical map to positions 3670, 1570 to 1575, and 2085, respectively.

  5. Libraries in Florida: MedlinePlus

    MedlinePlus

    ... this page: https://medlineplus.gov/libraries/florida.html Libraries in Florida To use the sharing features on ... please enable JavaScript. Atlantis JFK Medical Center MEDICAL LIBRARY 5301 S. Congress Ave. Att: Karin H. Pancake Atlantis, ...

  6. 78 FR 18579 - Combined Notice of Filings #1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-27

    ...: Monongahela Power Company, PJM Interconnection, L.L.C. Description: Mon Power submits revisions to PJM ``OATT... Company, PJM Interconnection, L.L.C. Description: Mon Power submits revisions to PJM OATT Att H-11A &...

  7. 33 CFR 165.1191 - Northern California and Lake Tahoe Area Annual Fireworks Events.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Fireworks Display Sponsor San Francisco Giants Baseball Team. Event Description Fireworks display in conjunction with baseball season home games. Date All season home games at AT&T Park. Location 1,000 feet...

  8. 33 CFR 165.1191 - Northern California and Lake Tahoe Area Annual Fireworks Events.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Fireworks Sponsor San Francisco Giants Baseball Team. Event Description Fireworks display in conjunction with baseball season home games. Date All season home games at AT&T Park. Location 700 feet off of...

  9. 33 CFR 165.1191 - Northern California and Lake Tahoe Area Annual Fireworks Events.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Fireworks Display Sponsor San Francisco Giants Baseball Team. Event Description Fireworks display in conjunction with baseball season home games. Date All season home games at AT&T Park. Location 1,000 feet...

  10. The life cycles of the temperate lactococcal bacteriophage phiLC3 monitored by a quantitative PCR method.

    PubMed

    Lunde, M; Blatny, J M; Kaper, F; Nes, I F; Lillehaug, D

    2000-11-01

    We present here a new and general approach for monitoring the life cycles of temperate bacteriophages which establish lysogeny by inserting their genomes site-specifically into the bacterial host chromosome. The method is based on quantitative amplification of specific DNA sites involved in various cut-and-join events during the life cycles of the phages (i.e. the cos, attP, attB, attL and attR sites) with the use of sequence-specific primers. By comparing the amounts of these specific DNA sites at different intervals, we were able to follow the development of the lytic and lysogenic life cycles of the temperate lactococcal bacteriophage phiLC3 after infection of its bacterial host Lactococcus lactis ssp. cremoris IMN-C18. PMID:11040439

  11. The life cycles of the temperate lactococcal bacteriophage phiLC3 monitored by a quantitative PCR method.

    PubMed

    Lunde, M; Blatny, J M; Kaper, F; Nes, I F; Lillehaug, D

    2000-11-01

    We present here a new and general approach for monitoring the life cycles of temperate bacteriophages which establish lysogeny by inserting their genomes site-specifically into the bacterial host chromosome. The method is based on quantitative amplification of specific DNA sites involved in various cut-and-join events during the life cycles of the phages (i.e. the cos, attP, attB, attL and attR sites) with the use of sequence-specific primers. By comparing the amounts of these specific DNA sites at different intervals, we were able to follow the development of the lytic and lysogenic life cycles of the temperate lactococcal bacteriophage phiLC3 after infection of its bacterial host Lactococcus lactis ssp. cremoris IMN-C18.

  12. NASA Now: The Sun: The Impact of Solar Activity on Earth

    NASA Video Gallery

    Meet Mitzi Adams, an astrophysicist studying the sun. Adams has been working as a solar scientist at NASA's Marshall Space Flight Center in Huntsville, Ala., since 1988. She analyzes data in an att...

  13. [Progress of φC31 integrase system in site-specific integration].

    PubMed

    Ma, Qing-Wen

    2011-06-01

    Integrase of phage fC31 catalyses the homologous recombination between Streptomyces attachment site attB and the phage attachment site attP. Meanwhile, this integrase can mediate integration of attB-containing donor plasmids into the pseudo attP sites in eukaryotic genomes by a site-specific manner and resulting long-term and robust expression of integrated genes. Nowadays, fC31 integrase system is becoming a potential tool for genome modification, gene therapy and transgenic research. Recent progress of fC31 integrase system in integration mode in mammalian genomes, efficiency improvement and researches concerned on transgenic safety were summarized in this review.

  14. Young's procedure for the treatment of valgus flatfoot deformity caused by a posterior tibial tendon dysfunction, stage II.

    PubMed

    Schinca, Nuri; Lasalle, Alicia; Alvarez, Josefina

    2012-06-01

    Young’s procedure contains an action mechanism that works better than other techniques on the pathophysiology of FFD. It respects the anatomy and biomechanics of the foot to reach the necessary muscular balance. The benefits of this technique include that the ATT is not detached, so its function mechanism is still active; the new trajectory of the ATT provides a powerful sling function at the level of the navicular; and the horizontal trajectory of the ATT and the osteoperiosteal flaps constitute a powerful inner capsular–tendinous–ligamentous support. What is more, an insufficiency of the ATT is created, which results in a predominance of the peroneus lateral longus, that descends and prones the forefoot. Additional procedures, such as medial displacement calcaneal osteotomy, should be considered to correct the entire deformity. The combination of these techniques do not sacrifice the joint mobility. PMID:22541522

  15. 78 FR 2306 - Self-Regulatory Organizations; New York Stock Exchange LLC; Notice of Filing and Immediate...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-10

    ... Act Release No. 68021 (October 9, 2012), 77 FR 63406 (October 16, 2012) (SR-NYSE-2012-50). Company... Chase & Co JPM Nokia Corporation NOK PFIZER Inc PFE Sprint Nextel Corporation S AT&T Inc T Wells...

  16. Attracting Water Drops

    NASA Video Gallery

    Astronauts Cady Coleman and Ron Garan perform the Attracting Water Drops experiment from Chabad Hebrew Academy in San Diego, Calif. This research determines if a free-floating water drop can be att...

  17. Assembly and transfer of tripartite integrative and conjugative genetic elements

    PubMed Central

    Haskett, Timothy L.; Terpolilli, Jason J.; Bekuma, Amanuel; O’Hara, Graham W.; Sullivan, John T.; Wang, Penghao; Ronson, Clive W.

    2016-01-01

    Integrative and conjugative elements (ICEs) are ubiquitous mobile genetic elements present as “genomic islands” within bacterial chromosomes. Symbiosis islands are ICEs that convert nonsymbiotic mesorhizobia into symbionts of legumes. Here we report the discovery of symbiosis ICEs that exist as three separate chromosomal regions when integrated in their hosts, but through recombination assemble as a single circular ICE for conjugative transfer. Whole-genome comparisons revealed exconjugants derived from nonsymbiotic mesorhizobia received three separate chromosomal regions from the donor Mesorhizobium ciceri WSM1271. The three regions were each bordered by two nonhomologous integrase attachment (att) sites, which together comprised three homologous pairs of attL and attR sites. Sequential recombination between each attL and attR pair produced corresponding attP and attB sites and joined the three fragments to produce a single circular ICE, ICEMcSym1271. A plasmid carrying the three attP sites was used to recreate the process of tripartite ICE integration and to confirm the role of integrase genes intS, intM, and intG in this process. Nine additional tripartite ICEs were identified in diverse mesorhizobia and transfer was demonstrated for three of them. The transfer of tripartite ICEs to nonsymbiotic mesorhizobia explains the evolution of competitive but suboptimal N2-fixing strains found in Western Australian soils. The unheralded existence of tripartite ICEs raises the possibility that multipartite elements reside in other organisms, but have been overlooked because of their unusual biology. These discoveries reveal mechanisms by which integrases dramatically manipulate bacterial genomes to allow cotransfer of disparate chromosomal regions. PMID:27733511

  18. 78 FR 35826 - Unfair Competitive Advantages; Enhancement of the Formal Complaint Process

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-14

    ... provide AT&T with access to live coverage of San Diego Padres baseball games. See AT&T Services, Inc. v... that raised issues relating to unfair competition: Docket No. C2009-1, GameFly Inc. (GameFly). Although the GameFly complaint did not directly raise issues related to 39 U.S.C. 404a, without changes to...

  19. Treatment paradox in musculo-skeletal tuberculosis in an immunocompetent adult male; a case report from a tertiary care hospital.

    PubMed

    Chaskar, Priyanka; Rana, Geetika; Anuradha; Duggal, Nandini; Arora, Jyoti

    2015-04-01

    Paradoxical reactions like immune reconstitution inflammatory syndrome (IRIS) as seen with patients on retroviral treatment in HIV infection, have also been identified in HIV sero-negative patients with extra pulmonary tuberculosis especially lymph-node tuberculosis. Musculo-skeletal tuberculosis presenting as a cold abscess of the anterior chest wall is a rare entity which poses diagnostic and therapeutic challenge. A 35-year-old immunocompetent male came with complains of painless lump on right side of his chest over 9th and 10th intercostal space which gradually increased and extended upto 11th rib area. Clinically, diagnosis of cold abscess was made and anti-tubercular therapy (ATT) was started. Despite of being on ATT for 3 weeks, patient developed pain and signs of inflammation. Fluid was aspirated and sent for biochemical and microbiological investigations. The aspirated fluid was positive for acid fast bacilli by ZN stain and grew Mycobacterium tuberculosis in culture, sensitive to first line ATT. Pyogenic and fungal culture was negative. This case presented as an anterior chest wall cold abscess which deteriorated on initiation of first line ATT, thus creating a suspicion of resistance to ATT which was cleared on ATT susceptibility testing. Hence, this case underlines the possibility of treatment paradoxes seen in immunocompetent musculo-skeletal tuberculosis. PMID:26046019

  20. Analysis of the site-specific integration system of the Streptomyces aureofaciens phage μ1/6.

    PubMed

    Farkašovská, Jarmila; Godány, Andrej

    2012-03-01

    The bacteriophage μ1/6 integrates its DNA into the chromosome of tetracycline producing strains of Streptomyces aureofaciens by a site-specific recombination process. A bioinformatic analysis of the μ1/6 genome revealed that orf5 encodes a putative integrase, a basic protein of 416 amino acids. The μ1/6 integrase was found to belong to the integrase family of site-specific tyrosine recombinases. The phage attachment site (attP) was localized downstream of the int gene. The attachment junctions (attL and attR) were determined, allowing identification of the bacterial attachment site (attB). All attachment sites shared a 46-bp common core sequence within which a site-specific recombination occurs. This core sequence comprises the 3' end of a putative tRNA(Thr) gene (anticodon TGT) which is completely restored in attL after integration of the phage into the host genome. An integration vector containing μ1/6 int-attP region was inserted stably into the S. aureofaciens B96, S. lividans TK24, and S. coelicolor A3. The μ1/6 integrase was shown to be functional in vivo in heterologous Escherichia coli without any other factors encoded by Streptomyces. In vitro recombination assay using purified μ1/6 integrase demonstrated its ability to catalyze integrative recombination in the presence of a crude extract of E. coli cells.

  1. Streptomyces temperate bacteriophage integration systems for stable genetic engineering of actinomycetes (and other organisms).

    PubMed

    Baltz, Richard H

    2012-05-01

    ϕC31, ϕBT1, R4, and TG1 are temperate bacteriophages with broad host specificity for species of the genus Streptomyces. They form lysogens by integrating site-specifically into diverse attB sites located within individual structural genes that map to the conserved core region of streptomycete linear chromosomes. The target genes containing the ϕC31, ϕBT1, R4, and TG1 attB sites encode a pirin-like protein, an integral membrane protein, an acyl-CoA synthetase, and an aminotransferase, respectively. These genes are highly conserved within the genus Streptomyces, and somewhat conserved within other actinomycetes. In each case, integration is mediated by a large serine recombinase that catalyzes unidirectional recombination between the bacteriophage attP and chromosomal attB sites. The unidirectional nature of the integration mechanism has been exploited in genetic engineering to produce stable recombinants of streptomycetes, other actinomycetes, eucaryotes, and archaea. The ϕC31 attachment/integration (Att/Int) system has been the most widely used, and it has been coupled with the ϕBT1 Att/Int system to facilitate combinatorial biosynthesis of novel lipopeptide antibiotics in Streptomyces fradiae.

  2. DNA cleavage is independent of synapsis during Streptomyces phage phiBT1 integrase-mediated site-specific recombination.

    PubMed

    Zhang, Lin; Wang, Lu; Wang, Jin; Ou, Xijun; Zhao, Guoping; Ding, Xiaoming

    2010-10-01

    Bacteriophage-encoded serine recombinases have great potential in genetic engineering but their catalytic mechanisms have not been adequately studied. Integration of ϕBT1 and ϕC31 via their attachment (att) sites is catalyzed by integrases of the large serine recombinase subtype. Both ϕBT1 and ϕC31 integrases were found to cleave single-substrate att sites without synaptic complex formation, and ϕBT1 integrase relaxed supercoiled DNA containing a single integration site. Systematic mutation of the central att site dinucleotide revealed that cleavage was independent of nucleotide sequence, but rejoining was crucially dependent upon complementarity of the cleavage products. Recombination between att sites containing dinucleotides with antiparallel complementarity led to antiparallel recombination. Integrase-substrate pre-incubation experiments revealed that the enzyme can form an attP-integrase tetramer complex that then captures naked attB DNA, and suggested that two alternative assembly pathways can lead to synaptic complex formation.

  3. The bacteroides NBU1 integrase performs a homology-independent strand exchange to form a holliday junction intermediate.

    PubMed

    Rajeev, Lara; Segall, Anca; Gardner, Jeffrey

    2007-10-26

    The Bacteroides mobilizable transposon NBU1 uses an integrase (IntN1) that is a tyrosine recombinase for its integration and excision from the host chromosome. Previously we showed that IntN1 makes 7-bp staggered cuts within the NBU1 att sites, and certain mismatches within the crossover region of the attN1 site (G(-2)C attN1) or the chromosomal target site (C(-3)G attBT1-1) enhanced the in vivo integration efficiency. Here we describe an in vitro integration system for NBU1. We used nicked substrates and a Holliday junction trapping peptide to show that NBU1 integration proceeds via formation of a Holliday junction intermediate that is formed by exchange of bottom strands. Some mismatches next to the first strand exchange site (in reactions with C(-3)G attBT1-1 or G(-2)C attN1 with their wild-type partner site) not only allowed formation of the Holliday junction intermediate but also increased the rate of recombinant formation. The second strand exchange appears to be homology-dependent. IntN1 is the only tyrosine recombinase known to catalyze a reaction that is more efficient in the presence of mismatches and where the first strand exchange is homology-independent. The possible mechanisms by which the mismatches stimulate recombination are discussed.

  4. Cellular Performance Comparison of Biomimetic Calcium Phosphate Coating and Alkaline-Treated Titanium Surface

    PubMed Central

    Wei, Mei

    2013-01-01

    The influence of biomimetic calcium phosphate coating on osteoblasts behavior in vitro is not well established yet. In this study, we investigated the behavior of osteoblastic rat osteosarcoma 17/2.8 cells (ROS17/2.8) on two groups of biomaterial surfaces: alkaline-treated titanium surface (ATT) and biomimetic calcium phosphate coated ATT (CaP). The cell attachment, proliferation, differentiation, and morphology on these surfaces were extensively evaluated to reveal the impact of substrate surface on osteoblastic cell responses. It was found that the ROS17/2.8 cells cultured on the ATT surface had higher attachment and proliferation rates compared to those on the CaP surface. Our results also showed that the calcium phosphate coatings generated in this work have an inhibiting effect on osteoblast adhesion and further influenced the proliferation and differentiation of osteoblast compared to the ATT surface in vitro. Cells on the ATT surface also exhibited a higher alkaline phosphatase activity than on the CaP surface after two weeks of culture. Immunofluorescence staining and scanning electron microscopy results showed that the cells adhered and spread faster on the ATT surface than on the CaP surface. These results collectively suggested that substrate surface properties directly influence cell adhesion on different biomaterials, which would result in further influence on the cell proliferation and differentiation. PMID:24455730

  5. Identification and analysis of integrons and cassette arrays in bacterial genomes.

    PubMed

    Cury, Jean; Jové, Thomas; Touchon, Marie; Néron, Bertrand; Rocha, Eduardo Pc

    2016-06-01

    Integrons recombine gene arrays and favor the spread of antibiotic resistance. Their broader roles in bacterial adaptation remain mysterious, partly due to lack of computational tools. We made a program - IntegronFinder - to identify integrons with high accuracy and sensitivity. IntegronFinder is available as a standalone program and as a web application. It searches for attC sites using covariance models, for integron-integrases using HMM profiles, and for other features (promoters, attI site) using pattern matching. We searched for integrons, integron-integrases lacking attC sites, and clusters of attC sites lacking a neighboring integron-integrase in bacterial genomes. All these elements are especially frequent in genomes of intermediate size. They are missing in some key phyla, such as α-Proteobacteria, which might reflect selection against cell lineages that acquire integrons. The similarity between attC sites is proportional to the number of cassettes in the integron, and is particularly low in clusters of attC sites lacking integron-integrases. The latter are unexpectedly abundant in genomes lacking integron-integrases or their remains, and have a large novel pool of cassettes lacking homologs in the databases. They might represent an evolutionary step between the acquisition of genes within integrons and their stabilization in the new genome.

  6. Identification and analysis of integrons and cassette arrays in bacterial genomes

    PubMed Central

    Cury, Jean; Jové, Thomas; Touchon, Marie; Néron, Bertrand; Rocha, Eduardo PC

    2016-01-01

    Integrons recombine gene arrays and favor the spread of antibiotic resistance. Their broader roles in bacterial adaptation remain mysterious, partly due to lack of computational tools. We made a program – IntegronFinder – to identify integrons with high accuracy and sensitivity. IntegronFinder is available as a standalone program and as a web application. It searches for attC sites using covariance models, for integron-integrases using HMM profiles, and for other features (promoters, attI site) using pattern matching. We searched for integrons, integron-integrases lacking attC sites, and clusters of attC sites lacking a neighboring integron-integrase in bacterial genomes. All these elements are especially frequent in genomes of intermediate size. They are missing in some key phyla, such as α-Proteobacteria, which might reflect selection against cell lineages that acquire integrons. The similarity between attC sites is proportional to the number of cassettes in the integron, and is particularly low in clusters of attC sites lacking integron-integrases. The latter are unexpectedly abundant in genomes lacking integron-integrases or their remains, and have a large novel pool of cassettes lacking homologs in the databases. They might represent an evolutionary step between the acquisition of genes within integrons and their stabilization in the new genome. PMID:27130947

  7. Phage-encoded Serine Integrases and Other Large Serine Recombinases.

    PubMed

    Smith, Margaret C M

    2015-08-01

    The large serine recombinases (LSRs) are a family of enzymes, encoded in temperate phage genomes or on mobile elements, that precisely cut and recombine DNA in a highly controllable and predictable way. In phage integration, the LSRs act at specific sites, the attP site in the phage and the attB site in the host chromosome, where cleavage and strand exchange leads to the integrated prophage flanked by the recombinant sites attL and attR. The prophage can excise by recombination between attL and attR but this requires a phage-encoded accessory protein, the recombination directionality factor (RDF). Although the LSRs can bind specifically to all the recombination sites, only specific integrase-bound sites can pair in a synaptic complex prior to strand exchange. Recent structural information has led to a breakthrough in our understanding of the mechanism of the LSRs, notably how the LSRs bind to their substrates and how LSRs display this site-selectivity. We also understand that the RDFs exercise control over the LSRs by protein-protein interactions. Other recent work with the LSRs have contributed to our understanding of how all serine recombinases undergo strand exchange subunit rotation, facilitated by surfaces that resemble a molecular bearing.

  8. Identification and analysis of integrons and cassette arrays in bacterial genomes.

    PubMed

    Cury, Jean; Jové, Thomas; Touchon, Marie; Néron, Bertrand; Rocha, Eduardo Pc

    2016-06-01

    Integrons recombine gene arrays and favor the spread of antibiotic resistance. Their broader roles in bacterial adaptation remain mysterious, partly due to lack of computational tools. We made a program - IntegronFinder - to identify integrons with high accuracy and sensitivity. IntegronFinder is available as a standalone program and as a web application. It searches for attC sites using covariance models, for integron-integrases using HMM profiles, and for other features (promoters, attI site) using pattern matching. We searched for integrons, integron-integrases lacking attC sites, and clusters of attC sites lacking a neighboring integron-integrase in bacterial genomes. All these elements are especially frequent in genomes of intermediate size. They are missing in some key phyla, such as α-Proteobacteria, which might reflect selection against cell lineages that acquire integrons. The similarity between attC sites is proportional to the number of cassettes in the integron, and is particularly low in clusters of attC sites lacking integron-integrases. The latter are unexpectedly abundant in genomes lacking integron-integrases or their remains, and have a large novel pool of cassettes lacking homologs in the databases. They might represent an evolutionary step between the acquisition of genes within integrons and their stabilization in the new genome. PMID:27130947

  9. Advanced Technology Training System on Motor-Operated Valves

    NASA Technical Reports Server (NTRS)

    Wiederholt, Bradley J.; Widjaja, T. Kiki; Yasutake, Joseph Y.; Isoda, Hachiro

    1993-01-01

    This paper describes how features from the field of Intelligent Tutoring Systems are applied to the Motor-Operated Valve (MOV) Advanced Technology Training System (ATTS). The MOV ATTS is a training system developed at Galaxy Scientific Corporation for the Central Research Institute of Electric Power Industry in Japan and the Electric Power Research Institute in the United States. The MOV ATTS combines traditional computer-based training approaches with system simulation, integrated expert systems, and student and expert modeling. The primary goal of the MOV ATTS is to reduce human errors that occur during MOV overhaul and repair. The MOV ATTS addresses this goal by providing basic operational information of the MOV, simulating MOV operation, providing troubleshooting practice of MOV failures, and tailoring this training to the needs of each individual student. The MOV ATTS integrates multiple expert models (functional and procedural) to provide advice and feedback to students. The integration also provides expert model validation support to developers. Student modeling is supported by two separate student models: one model registers and updates the student's current knowledge of basic MOV information, while another model logs the student's actions and errors during troubleshooting exercises. These two models are used to provide tailored feedback to the student during the MOV course.

  10. Enhanced correlation between quantitative ultrasound and structural and mechanical properties of bone using combined transmission-reflection measurement

    PubMed Central

    Lin, Liangjun; Lin, Wei; Qin, Yi-Xian

    2015-01-01

    Quantitative ultrasound (QUS) is capable of predicting the principal structural orientation of trabecular bone; this orientation is highly correlated with the mechanical strength of trabecular bone. Irregular shape of bone, however, would increase variation in such a prediction, especially under human in vivo measurement. This study was designed to combine transmission and reflection modes of QUS measurement to improve the prediction for the structural and mechanical properties of trabecular bone. QUS, mechanical testing, and micro computed tomography (μCT) scanning were performed on 24 trabecular bone cubes harvested from a bovine distal femur to obtain the mechanical and structural parameters. Transmission and reflection modes of QUS measurement in the transverse and frontal planes were performed in a confined 60° angle range with 5° increment. The QUS parameters, attenuation (ATT) and velocity (UV), obtained from transmission mode, were normalized to the specimen thickness acquired from reflection mode. Analysis of covariance showed that the combined transmission-reflection modes improved prediction for the structural and Young's modulus of bone in comparison to the traditional QUS measurement performed only in the medial-lateral orientation. In the transverse plane, significant improvement between QUS and μCT was found in ATT vs bone surface density (BS/BV) (p < 0.05), ATT vs trabecular thickness (Tb.Th) (p < 0.01), ATT vs degree of anisotropy (DA) (p < 0.05), UV vs trabecular bone number (Tb.N) (p < 0.05), and UV vs Tb.Th (p < 0.001). In the frontal plane, significant improvement was found in ATT vs structural model index (SMI) (p < 0.01), ATT vs bone volume fraction (BV/TV) (p < 0.01), ATT vs BS/BV (p < 0.001), ATT vs Tb.Th (p < 0.001), ATT vs DA (p < 0.001), and ATT vs modulus (p < 0.001), UV vs SMI (p < 0.01), UV vs BV/TV (p < 0.05), UV vs BS/BV (p < 0.05), UV vs Tb.Th (p < 0.01), UV vs

  11. Gilded and gelded. Hard-won lessons from the PR wars.

    PubMed

    Martin, Dick

    2003-10-01

    A golden statue of a winged youth once perched on the roof of AT&T's old headquarters. But when AT&T lowered the 24-foot-high statue for regilding so that it could be placed in the company's new headquarters, the chairman was shocked to discover that the figure was anatomically correct. So he decreed that it also be gelded. The altered "Golden Boy" thus became a metaphor for AT&T's recent embattled history, and it serves as a cautionary symbol for all companies operating in today's brutal business environment, where perception can be as important as reality. While image consultants and executives work to gild a company's image, special interest groups and the media can geld a company with countless little cuts. The author, a former executive vice president of public relations for AT&T, provides an insider's view of some of the company's most painful public-relations scrapes. They include the collapse of two apparent CEO succession plans, AT&T's inability to meet heightened expectations after Mike Armstrong was appointed CEO, and the racially charged furor over a cartoon in an employee publication. The author offers four lessons: Don't become hypnotized by your own buzz; understand the way the business media think; address the needs of all your stakeholders; and be sensitive to the possible emotional resonance of what appear to be straightforward facts. To illustrate the final point, the author mentions AT&T's elimination of 40,000 jobs in 1996. Wall Street was impressed, but NBC's Tom Brokaw said the workforce reduction might signal "another long, anxious year for the American middle class." No rational argument from AT&T could overcome the layoffs' symbolic impact. Wounded but wiser after numerous public-relations battles, the company eventually learned to stop aggregating job-reduction information for the media. PMID:14521097

  12. SU-D-18C-05: Variable Bolus Arterial Spin Labeling MRI for Accurate Cerebral Blood Flow and Arterial Transit Time Mapping

    SciTech Connect

    Johnston, M; Jung, Y

    2014-06-01

    Purpose: Arterial spin labeling (ASL) is an MRI perfusion imaging method from which quantitative cerebral blood flow (CBF) maps can be calculated. Acquisition with variable post-labeling delays (PLD) and variable TRs allows for arterial transit time (ATT) mapping and leads to more accurate CBF quantification with a scan time saving of 48%. In addition, T1 and M0 maps can be obtained without a separate scan. In order to accurately estimate ATT and T1 of brain tissue from the ASL data, variable labeling durations were invented, entitled variable-bolus ASL. Methods: All images were collected on a healthy subject with a 3T Siemens Skyra scanner. Variable-bolus Psuedo-continuous ASL (PCASL) images were collected with 7 TI times ranging 100-4300ms in increments of 700ms with TR ranging 1000-5200ms. All boluses were 1600ms when the TI allowed, otherwise the bolus duration was 100ms shorter than the TI. All TI times were interleaved to reduce sensitivity to motion. Voxel-wise T1 and M0 maps were estimated using a linear least squares fitting routine from the average singal from each TI time. Then pairwise subtraction of each label/control pair and averaging for each TI time was performed. CBF and ATT maps were created using the standard model by Buxton et al. with a nonlinear fitting routine using the T1 tissue map. Results: CBF maps insensitive to ATT were produced along with ATT maps. Both maps show patterns and averages consistent with literature. The T1 map also shows typical T1 contrast. Conclusion: It has been demonstrated that variablebolus ASL produces CBF maps free from the errors due to ATT and tissue T1 variations and provides M0, T1, and ATT maps which have potential utility. This is accomplished with a single scan in a feasible scan time (under 6 minutes) with low sensivity to motion.

  13. Gilded and gelded. Hard-won lessons from the PR wars.

    PubMed

    Martin, Dick

    2003-10-01

    A golden statue of a winged youth once perched on the roof of AT&T's old headquarters. But when AT&T lowered the 24-foot-high statue for regilding so that it could be placed in the company's new headquarters, the chairman was shocked to discover that the figure was anatomically correct. So he decreed that it also be gelded. The altered "Golden Boy" thus became a metaphor for AT&T's recent embattled history, and it serves as a cautionary symbol for all companies operating in today's brutal business environment, where perception can be as important as reality. While image consultants and executives work to gild a company's image, special interest groups and the media can geld a company with countless little cuts. The author, a former executive vice president of public relations for AT&T, provides an insider's view of some of the company's most painful public-relations scrapes. They include the collapse of two apparent CEO succession plans, AT&T's inability to meet heightened expectations after Mike Armstrong was appointed CEO, and the racially charged furor over a cartoon in an employee publication. The author offers four lessons: Don't become hypnotized by your own buzz; understand the way the business media think; address the needs of all your stakeholders; and be sensitive to the possible emotional resonance of what appear to be straightforward facts. To illustrate the final point, the author mentions AT&T's elimination of 40,000 jobs in 1996. Wall Street was impressed, but NBC's Tom Brokaw said the workforce reduction might signal "another long, anxious year for the American middle class." No rational argument from AT&T could overcome the layoffs' symbolic impact. Wounded but wiser after numerous public-relations battles, the company eventually learned to stop aggregating job-reduction information for the media.

  14. Group IIC Intron with an Unusual Target of Integration in Enterobacter cloacae

    PubMed Central

    Rodríguez-Martínez, José-Manuel; Poirel, Laurent

    2012-01-01

    A potential role of group IIC-attC introns in integron gene cassette formation, that is, the way in which they could provide the attC sequence essential for recombination, has been proposed. Group IIC introns usually target the attC site of gene cassettes and more specifically their inverse core. Here we characterized a novel group IIC intron targeting the core site of the aadA1 gene cassette attC site (aadA1-qacEΔ1 gene cassette junction) from enterobacterial isolates. Intron mobility (retrohoming) was analyzed using a two-plasmid assay performed in Escherichia coli. Intron mobility assays confirmed the mobilization-integration of the group II intron into the core site of the aadA2, blaVIM-2, blaCARB-2, aac(6′)-Ib, dfrXVb, arr2, cmlA4, and aadB gene cassettes but not into the attI site. This mobility was dependent on maturase activity. Reverse transcriptase PCR showed that this intron was transcriptionally active, and an intermediate circular form was detected by inverse PCR. This element was linked to the blaVEB-1 extended-spectrum β-lactamase gene in a high number of enterobacterial isolates. A phylogenetic tree showed that the identified element was located in a branch separate from group IIC-attC introns, being an IIC intron possessing the ability to integrate using the core site of the attC sites as target. PMID:22020643

  15. Construction of transformed, cultured silkworm cells and transgenic silkworm using the site-specific integrase system from phage φC31.

    PubMed

    Yin, Yajuan; Cao, Guangli; Xue, Renyu; Gong, Chengliang

    2014-10-01

    The Streptomyces bacteriophage, φC31, uses a site-specific integrase enzyme to perform efficient recombination. The recombination system uses specific sequences to integrate exogenous DNA from the phage into a host. The sequences are known as the attP site in the phage and the attB site in the host. The system can be used as a genetic manipulation tool. In this study it has been applied to the transformation of cultured BmN cells and the construction of transgenic Bombyx mori individuals. A plasmid, pSK-attB/Pie1-EGFP/Zeo-PASV40, containing a cassette designed to express a egfp-zeocin fusion gene, was co-transfected into cultured BmN cells with a helper plasmid, pSK-Pie1/NLS-Int/NSL. Expression of the egfp-zeocin fusion gene was driven by an ie-1 promoter, downstream of a φC31 attB site. The helper plasmid encoded the φC31 integrase enzyme, which was flanked by two nuclear localization signals. Expression of the egfp-zeocin fusion gene could be observed in transformed cells. The two plasmids were also transferred into silkworm eggs to obtain transgenic silkworms. Successful integration of the fusion gene was indicated by the detection of green fluorescence, which was emitted by the silkworms. Nucleotide sequence analysis demonstrated that the attB site had been cut, to allow recombination between the attB and endogenous pseudo attP sites in the cultured silkworm cells and silkworm individuals.

  16. Characterization of IntA, a bidirectional site-specific recombinase required for conjugative transfer of the symbiotic plasmid of Rhizobium etli CFN42.

    PubMed

    Hernández-Tamayo, Rogelio; Sohlenkamp, Christian; Puente, José Luis; Brom, Susana; Romero, David

    2013-10-01

    Site-specific recombination occurs at short specific sequences, mediated by the cognate recombinases. IntA is a recombinase from Rhizobium etli CFN42 and belongs to the tyrosine recombinase family. It allows cointegration of plasmid p42a and the symbiotic plasmid via site-specific recombination between attachment regions (attA and attD) located in each replicon. Cointegration is needed for conjugative transfer of the symbiotic plasmid. To characterize this system, two plasmids harboring the corresponding attachment sites and intA were constructed. Introduction of these plasmids into R. etli revealed IntA-dependent recombination events occurring at high frequency. Interestingly, IntA promotes not only integration, but also excision events, albeit at a lower frequency. Thus, R. etli IntA appears to be a bidirectional recombinase. IntA was purified and used to set up electrophoretic mobility shift assays with linear fragments containing attA and attD. IntA-dependent retarded complexes were observed only with fragments containing either attA or attD. Specific retarded complexes, as well as normal in vivo recombination abilities, were seen even in derivatives harboring only a minimal attachment region (comprising the 5-bp central region flanked by 9- to 11-bp inverted repeats). DNase I-footprinting assays with IntA revealed specific protection of these zones. Mutations that disrupt the integrity of the 9- to 11-bp inverted repeats abolish both specific binding and recombination ability, while mutations in the 5-bp central region severely reduce both binding and recombination. These results show that IntA is a bidirectional recombinase that binds to att regions without requiring neighboring sequences as enhancers of recombination.

  17. Characterization of IntA, a Bidirectional Site-Specific Recombinase Required for Conjugative Transfer of the Symbiotic Plasmid of Rhizobium etli CFN42

    PubMed Central

    Hernández-Tamayo, Rogelio; Sohlenkamp, Christian; Puente, José Luis; Brom, Susana

    2013-01-01

    Site-specific recombination occurs at short specific sequences, mediated by the cognate recombinases. IntA is a recombinase from Rhizobium etli CFN42 and belongs to the tyrosine recombinase family. It allows cointegration of plasmid p42a and the symbiotic plasmid via site-specific recombination between attachment regions (attA and attD) located in each replicon. Cointegration is needed for conjugative transfer of the symbiotic plasmid. To characterize this system, two plasmids harboring the corresponding attachment sites and intA were constructed. Introduction of these plasmids into R. etli revealed IntA-dependent recombination events occurring at high frequency. Interestingly, IntA promotes not only integration, but also excision events, albeit at a lower frequency. Thus, R. etli IntA appears to be a bidirectional recombinase. IntA was purified and used to set up electrophoretic mobility shift assays with linear fragments containing attA and attD. IntA-dependent retarded complexes were observed only with fragments containing either attA or attD. Specific retarded complexes, as well as normal in vivo recombination abilities, were seen even in derivatives harboring only a minimal attachment region (comprising the 5-bp central region flanked by 9- to 11-bp inverted repeats). DNase I-footprinting assays with IntA revealed specific protection of these zones. Mutations that disrupt the integrity of the 9- to 11-bp inverted repeats abolish both specific binding and recombination ability, while mutations in the 5-bp central region severely reduce both binding and recombination. These results show that IntA is a bidirectional recombinase that binds to att regions without requiring neighboring sequences as enhancers of recombination. PMID:23935046

  18. Succinic Semialdehyde Promotes Prosurvival Capability of Agrobacterium tumefaciens

    PubMed Central

    Wang, Chao; Tang, Desong; Gao, Yong-Gui

    2016-01-01

    ABSTRACT Succinic semialdehyde (SSA), an important metabolite of γ-aminobutyric acid (GABA), is a ligand of the repressor AttJ regulating the expression of the attJ-attKLM gene cluster in the plant pathogen Agrobacterium tumefaciens. While the response of A. tumefaciens to GABA and the function of attKLM have been extensively studied, genetic and physiological responses of A. tumefaciens to SSA remain unknown. In combination with microarray and genetic approaches, this study sets out to explore new roles of the SSA-AttJKLM regulatory mechanism during bacterial infection. The results showed that SSA plays a key role in regulation of several bacterial activities, including C4-dicarboxylate utilization, nitrate assimilation, and resistance to oxidative stress. Interestingly, while the SSA relies heavily on the functional AttKLM in mediating nitrate assimilation and oxidative stress resistance, the compound could regulate utilization of C4-dicarboxylates independent of AttJKLM. We further provide evidence that SSA controls C4-dicarboxylate utilization through induction of an SSA importer and that disruption of attKLM attenuates the tumorigenicity of A. tumefaciens. Taken together, these findings indicate that SSA could be a potent plant signal which, together with AttKLM, plays a vital role in promoting the bacterial prosurvival abilities during infection. IMPORTANCE Agrobacterium tumefaciens is a plant pathogen causing crown gall diseases and has been well known as a powerful tool for plant genetic engineering. During the long history of microbe-host interaction, A. tumefaciens has evolved the capabilities of recognition and response to plant-derived chemical metabolites. Succinic semialdehyde (SSA) is one such metabolite. Previous results have demonstrated that SSA functions to activate a quorum-quenching mechanism and thus to decrease the level of quorum-sensing signals, thereby avoiding the elicitation of a plant defense. Here, we studied the effect of SSA on gene

  19. Mortality in HIV-infected patients with tuberculosis treated with streptomycin and a two-week intensified regimen: data from an HIV cohort study using inverse probability of treatment weighting.

    PubMed

    Alvarez-Uria, Gerardo; Midde, Manoranjan; Naik, Praveen K

    2016-01-01

    Background. Despite the dramatic scale-up of antiretroviral therapy in low- and middle-income countries, tuberculosis (TB) is still the main cause of death among HIV-infected patients in resource-limited settings. Previous studies in patients with TB meningitis suggest that the use of higher doses of common anti-TB drugs could reduce mortality. Methods. Using clinical data from an HIV cohort study in India, we compared the mortality among HIV-infected patients diagnosed with TB according to the regimen received during the first two weeks of treatment: standard anti-tuberculosis therapy (ATT) (N = 847), intensified ATT (N = 322), and intensified ATT with streptomycin (N = 446). The intensified ATT comprised double dose of rifampicin and substitution of ethambutol with levofloxacin. Multivariate analysis was performed using Cox proportional hazard models and inverse probability of treatment weighting (IPTW) based on propensity scores. Patients with TB meningitis were excluded. Results. The use of intensified ATT alone did not improve survival. However, when streptomycin was added, the use intensified ATT was associated with reduced mortality in Cox models (adjusted hazard ratio 0.72, 95% CI [0.57-0.91]) and after IPTW (hazard ratio 0.77, 95% CI [0.67-0.96]). Other factors associated with improved survival were high serum albumin concentration, high CD4 lymphocyte cell-counts, and high glomerular filtration rates. Factors associated with increased mortality were high urea concentrations, being on antiretroviral therapy at the time of ATT initiation and high BUN/creatinine ratio. In an effect modification analysis, the survival benefits of the intensified ATT with streptomycin disappeared in patients with severe hypoalbuminemia. Conclusion. The results of this study are in accordance with a previous study from our cohort involving patients with TB meningitis, and suggest that an intensified 2-week ATT with streptomycin could reduce mortality in HIV infected patients

  20. Inactivation of a GAL4-Like Transcription Factor Improves Cell Fitness and Product Yield in Glycoengineered Pichia pastoris Strains

    PubMed Central

    Argyros, Rebecca; Bukowski, John; Nelson, Stephanie; Sharkey, Nathan; Kim, Sehoon; Copeland, Victoria; Davidson, Robert C.; Chen, Ronghua; Zhuang, Jun; Sethuraman, Natarajan; Stadheim, Terrance A.

    2014-01-01

    With a completely reengineered and humanized glycosylation pathway, glycoengineered Pichia pastoris has emerged as a promising production host for the manufacture of therapeutic glycoproteins. However, the extensive genetic modifications have also negatively affected the overall fitness levels of the glycoengineered host cells. To make glycoengineered Pichia strains more compatible with a scalable industrial fermentation process, we sought to identify genetic solutions to broadly improve cell robustness during fermentation. In this study, we report that mutations within the Pichia pastoris ATT1 (PpATT1) gene (a homolog of the Saccharomyces cerevisiae GAL4 [ScGAL4] transcriptional activator) dramatically increased the cellular fitness levels of glycoengineered Pichia strains. We demonstrate that deletion of the PpATT1 gene enabled glycoengineered Pichia strains to improve their thermal tolerance levels, reduce their cell lysis defects, and greatly improve fermentation robustness. The extension of the duration of fermentation enabled the PpATT1-modified glycoengineered Pichia strains to increase their product yields significantly without any sacrifice in product quality. Because the ATT1 gene could be deleted from any Pichia strains, including empty hosts and protein-expressing production strains alike, we suggest that the findings described in this study are broadly applicable to any Pichia strains used for the production of therapeutic proteins, including monoclonal antibodies, Fc fusions, peptides, hormones, and growth factors. PMID:25344235

  1. High knee valgus in female subjects does not yield higher knee translations during drop landings: a biplane fluoroscopic study.

    PubMed

    Torry, Michael R; Shelburne, Kevin B; Myers, Casey; Giphart, J Erik; Pennington, W Wesley; Krong, Jacob P; Peterson, Daniel S; Steadman, J Richard; Woo, Savio L-Y

    2013-02-01

    The goal of this study was to determine the effects of peak knee valgus angle and peak knee abductor moment on the anterior, medial, and lateral tibial translations (ATT, MTT, LTT) in the "at risk" female knee during drop landing. Fifteen female subjects performed drop landings from 40 cm. Three-dimension knee motion was simultaneously recorded using a high speed, biplane fluoroscopy system, and a video-based motion analysis system. Valgus knee angles and knee abduction moments were stratified into low, intermediate, and high groups and peak ATT, MTT, and LTT were compared between these groups with ANOVA (α = 0.05). Significant differences were observed between stratified groups in peak knee valgus angle (p < 0.0001) and peak knee abduction moment (p < 0.0001). However, no corresponding differences in peak ATT, LTT, and MTT between groups exhibiting low to high-peak knee valgus angles (ATT: p = 0.80; LTT: p = 0.25; MTT: p = 0.72); or, in peak ATT (p = 0.61), LTT (p = 0.26) and MTT (p = 0.96) translations when stratified according to low to high knee abduction moments, were found. We conclude that the healthy female knee is tightly regulated with regard to translations even when motion analysis derived knee valgus angles and abduction moments are high.

  2. A Rare Case of Flare-Up of PID in Infertility Treatment.

    PubMed

    Wadhwa, Leena; Wadhwa, Sanjana N; Jindal, Sunita

    2015-01-01

    Case Presentation. Mrs. X, 35 years old, case of primary infertility, was diagnosed to have genital tuberculosis on the basis of PCR positive and hysterolaparoscopy findings and received category I ATT for 6 months. Following ATT completion, her USG revealed no evidence of tuboovarian mass or hydrosalpinx. Since her tubes were patent, she underwent 3 cycles of ovulation induction and 2 cycles of IUI. The women presented with acute PID, five days after IUI, and was conservatively managed. She again presented 24 days after IUI with persistent low grade fever and abdominal pain. Suspecting relapse of genital tuberculosis, she was started on category II ATT. She had acute episodes of high grade fever with chills 2 weeks after starting ATT and MRI revealed bilateral TO masses suggestive of pyosalpinx. Emergency laparotomy was done, pus was drained, and cyst wall was removed and HPE was suggestive of chronic inflammation with few granulation tissues. ATT was continued for one year and the woman improved. Conclusion. The possibility of flare-up of PID (pelvic inflammatory disease) in treated case of tuberculosis undergoing infertility management should be kept in mind and aggressive management should be done. PMID:26600959

  3. High Knee Valgus in Female Subjects Does Not Yield Higher Knee Translations During Drop Landings: A Biplane Fluoroscopic Study

    PubMed Central

    Torry, Michael R.; Shelburne, Kevin B.; Myers, Casey; Giphart, J. Erik; Pennington, W. Wesley; Krong, Jacob P.; Peterson, Daniel S.; Steadman, J. Richard; Woo, Savio L-Y.

    2012-01-01

    The goal of this study was to determine the effects of peak knee valgus angle and peak knee abductor moment on the anterior, medial, and lateral tibial translations (ATT, MTT, LTT) in the ‘at risk’ female knee during drop landing. Fifteen female subjects performed drop landings from 40 cm. 3D knee motion was simultaneously recorded using a high speed, biplane fluoroscopy system and a video-based motion analysis system. Valgus knee angles and knee abduction moments were stratified into low, intermediate and high groups and peak ATT, MTT and LTT were compared between these groups with ANOVA (α = .05). Significant differences were observed between stratified groups in peak knee valgus angle (p < .0001) and peak knee abduction moment (p < .0001). However, no corresponding differences in peak ATT, LTT and MTT between groups exhibiting low to high peak knee valgus angles (ATT: p = .80; LTT: p = .25; MTT: p = .72); or, in peak ATT (p = .61), LTT (p = .26) and MTT (p = .96) translations when stratified according to low to high knee abduction moments, were found. We conclude that the healthy female knee is tightly regulated with regard to translations even when motion analysis derived knee valgus angles and abduction moments are high. PMID:22968826

  4. Comment on 'Microwave attenuation of hydrogen plasma in carbon nanotubes' [J. Appl. Phys. 104, 124315 (2008)

    SciTech Connect

    Moradi, Afshin

    2010-03-15

    In a recent article, Babaei and Solari [J. Appl. Phys. 104, 124315 (2008)] studied the effects of the electron temperature, and the external static magnetic field on the attenuation (ATT) of the microwave in the hydrogen plasma embedded inside the carbon nanotubes (CNTs), which were grown by iron-catalyzed high-pressure disproportionation (HiPco). They showed that the position of ATT peak shifts significantly toward high frequency with increasing thermal frequency and in the presence of an external magnetic field in the Faraday configuration, for {upsilon}{sub c}<20 GHz, the ATT coefficient increases with increasing cyclotron frequency, and for {upsilon}{sub c}>20 GHz, the ATT level variations extremely increase, where {upsilon}{sub c} is the cyclotron frequency. Here we derive the correct form of the microwave absorption coefficient of the magnetized hydrogen plasma embedded inside the CNTs and show that the absorption band moves from low to high frequencies when the magnetic field strength increases. Also, we show that the ATT of the microwave in the system is not sensitive to the thermal frequency.

  5. Historical data analyses and scientific knowledge suggest complete removal of the abnormal toxicity test as a quality control test.

    PubMed

    Garbe, Joerg H O; Ausborn, Susanne; Beggs, Claire; Bopst, Martin; Joos, Angelika; Kitashova, Alexandra A; Kovbasenco, Olga; Schiller, Claus-Dieter; Schwinger, Martina; Semenova, Natalia; Smirnova, Lilia; Stodart, Fraser; Visalli, Thomas; Vromans, Lisette

    2014-11-01

    In the early 1900s, the abnormal toxicity test (ATT) was developed as an auxiliary means to ensure safe and consistent antiserum production. Today, the ATT is utilized as a quality control (QC) release test according to pharmacopoeial or other regulatory requirements. The study design has not been changed since around 1940. The evidence of abnormal toxicity testing as a prediction for harmful batches is highly questionable and lacks a scientific rationale. Numerous reviews of historical ATT results have revealed that no reliable conclusions can be drawn from this QC measure. Modern pharmaceutical manufacturers have thorough control of the manufacturing process and comply with good manufacturing practice rules. Contaminants are appropriately controlled by complying with the validated manufacturing processes and strict QC batch release confirming batch-to-batch consistency. Recognizing that product safety, efficacy, and stability can be ensured with strict QC measures, nowadays most regulatory authorities do not require the ATT for most product classes. In line with the replacement, reduction, and refinement (3Rs) initiative, the test requirement has been deleted from approximately 80 monographs of the European Pharmacopoeia and for the majority of product classes in the United States. For these reasons, it is recommended that the ATT should be consistently omitted world-wide and be removed from pharmacopoeias and other regulatory requirements.

  6. Reexamining the role of the accessory plasmid pAtC58 in the virulence of Agrobacterium tumefaciens strain C58.

    PubMed

    Nair, Gauri R; Liu, Zhenying; Binns, Andrew N

    2003-11-01

    Isogenic strains of Agrobacterium tumefaciens carrying pTiC58, pAtC58, or both were constructed and assayed semiquantitatively and quantitatively for virulence and vir gene expression to study the effect of the large 542-kb accessory plasmid, pAtC58, on virulence. Earlier studies indicate that the att (attachment) genes of A. tumefaciens are crucial in the ability of this soil phytopathogen to infect susceptible host plants. Mutations in many att genes, notably attR and attD, rendered the strain avirulent. These genes are located on pAtC58. Previous work also has shown that derivatives of the wild-type strain C58 cured of pAtC58 are virulent as determined by qualitative virulence assays and, hence, pAtC58 was described as nonessential for virulence. We show here that the absence of pAtC58 in pTiC58-containing strains results in reduced virulence but that disruption of the attR gene does not result in avirulence or a reduction in virulence. Our studies indicate that pAtC58 has a positive effect on vir gene induction as revealed by immunoblot analysis of Vir proteins and expression of a PvirB::lacZ fusion. PMID:14551325

  7. Can the attention training technique turn one marshmallow into two? Improving children's ability to delay gratification.

    PubMed

    Murray, Joanne; Theakston, Anna; Wells, Adrian

    2016-02-01

    The seminal Marshmallow Test (Mischel & Ebbesen, 1970) has reliably demonstrated that children who can delay gratification are more likely to be emotionally stable and successful later in life. However, this is not good news for those children who can't delay. Therefore, this study aimed to explore whether a metacognitive therapy technique, Attention Training (ATT: Wells, 1990) can improve young children's ability to delay gratification. One hundred children participated. Classes of 5-6 year olds were randomly allocated to either the ATT or a no-intervention condition and were tested pre and post-intervention on ability to delay gratification, verbal inhibition (executive control), and measures of mood. The ATT intervention significantly increased (2.64 times) delay of gratification compared to the no-intervention condition. After controlling for age and months in school, the ATT intervention and verbal inhibition task performance were significant independent predictors of delay of gratification. These results provide evidence that ATT can improve children's self-regulatory abilities with the implication that this might reduce psychological vulnerability later in life. The findings highlight the potential contribution that the Self-Regulatory Executive Function (S-REF) model could make to designing techniques to enhance children's self-regulatory processes.

  8. Anterior Cruciate Ligament Injury: Compensation during Gait using Hamstring Muscle Activity

    PubMed Central

    Catalfamo, Paola Formento; Aguiar, Gerardo; Curi, Jorge; Braidot, Ariel

    2010-01-01

    Previous research has shown that an increase in hamstring activation may compensate for anterior tibial transalation (ATT) in patients with anterior cruciate ligament deficient knee (ACLd); however, the effects of this compensation still remain unclear. The goals of this study were to quantify the activation of the hamstring muscles needed to compensate the ATT in ACLd knee during the complete gait cycle and to evaluate the effect of this compensation on quadriceps activation and joint contact forces. A two dimensional model of the knee was used, which included the tibiofemoral and patellofemoral joints, knee ligaments, the medial capsule and two muscles units. Simulations were conducted to determine the ATT in healthy and ACLd knee and the hamstring activation needed to correct the abnormal ATT to normal levels (100% compensation) and to 50% compensation. Then, the quadriceps activation and the joint contact forces were calculated. Results showed that 100% compensation would require hamstring and quadriceps activations larger than their maximum isometric force, and would generate an increment in the peak contact force at the tibiofemoral (115%) and patellofemoral (48%) joint with respect to the healthy knee. On the other hand, 50% compensation would require less force generated by the muscles (less than 0.85 of maximum isometric force) and smaller contact forces (peak tibiofemoral contact force increased 23% and peak patellofemoral contact force decreased 7.5% with respect to the healthy knee). Total compensation of ATT by means of increased hamstring activity is possible; however, partial compensation represents a less deleterious strategy. PMID:20721326

  9. A Rare Case of Flare-Up of PID in Infertility Treatment

    PubMed Central

    Wadhwa, Leena; Wadhwa, Sanjana N.; Jindal, Sunita

    2015-01-01

    Case Presentation. Mrs. X, 35 years old, case of primary infertility, was diagnosed to have genital tuberculosis on the basis of PCR positive and hysterolaparoscopy findings and received category I ATT for 6 months. Following ATT completion, her USG revealed no evidence of tuboovarian mass or hydrosalpinx. Since her tubes were patent, she underwent 3 cycles of ovulation induction and 2 cycles of IUI. The women presented with acute PID, five days after IUI, and was conservatively managed. She again presented 24 days after IUI with persistent low grade fever and abdominal pain. Suspecting relapse of genital tuberculosis, she was started on category II ATT. She had acute episodes of high grade fever with chills 2 weeks after starting ATT and MRI revealed bilateral TO masses suggestive of pyosalpinx. Emergency laparotomy was done, pus was drained, and cyst wall was removed and HPE was suggestive of chronic inflammation with few granulation tissues. ATT was continued for one year and the woman improved. Conclusion. The possibility of flare-up of PID (pelvic inflammatory disease) in treated case of tuberculosis undergoing infertility management should be kept in mind and aggressive management should be done. PMID:26600959

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

    PubMed Central

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

    2015-01-01

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

  11. Corrosion of copper in aerated acidic pickling solutions and its inhibition by 3-amino-1,2,4-triazole-5-thiol.

    PubMed

    Sherif, El-Sayed M; Erasmus, R M; Comins, J D

    2007-02-01

    Corrosion of copper in aerated acidic chloride pickling (0.5 M HCl) solutions and its inhibition by 3-amino-1,2,4-triazole-5-thiol (ATT) have been investigated using electrochemical techniques and weight-loss measurements, along with Raman spectroscopy. Electrochemical measurements for copper after varied immersion periods of 0, 24, and 48 h showed that the presence of ATT and the increase of its concentration significantly decrease cathodic, anodic, corrosion (j(Corr)) currents and corrosion rates (K(Corr)), as well as the dissolution currents at 300 mV vs Ag/AgCl, while increasing polarization resistance (Rp), degree of surface coverage (theta) and inhibition efficiency (IE%) to a great extent. Weight-loss measurements after different immersion periods of 6 to 48 h revealed that the dissolution of copper decreased to a minimum and the corresponding IE% increased with increasing ATT concentration. The detection of ATT molecules on the copper surface by Raman spectroscopy indicated that inhibition of copper corrosion is achieved by strong adsorption of ATT molecules onto the copper surface. PMID:17084854

  12. Breakdown of the consent process at a quaternary medical center: our full disclosure.

    PubMed

    Fairfax, Lindsay M; Christmas, A Britton; Norton, H James; Jacobs, David G

    2012-08-01

    Circumstances may arise in the intensive care unit (ICU) when the physician is unable to obtain informed consent. We undertook this study to determine the variations in the consent process. An anonymous survey was distributed to all critical care nurses (RN), resident physicians (RES), advanced practitioners (AP), and attending physicians (ATT). Participants were asked to describe the risks of nine common ICU procedures (central venous line, peripherally inserted central catheter, bronchoscopy, tube thoracostomy, tracheostomy, vena cava filter, angioembolization, image-guided drainage, and percutaneous endoscopic gastrostomy tube). Participants were also asked which member of the healthcare team should obtain consent. All groups were compared with ATT responses and RN responses were compared with the remaining groups. The response rate was 134 of 610 participants (22%) with 51 per cent RN (n = 68), 17 per cent RES (n = 23), 7 per cent AP (n = 9), and 25 per cent ATT (n = 34). Compared with ATT, RN assessment of important risks varied significantly for eight of nine procedures. RES responses varied in three procedures. A minority believed that nurses should obtain consent. However, many physicians (34% ATT and 27% RES) denied having informed consent discussions with 50 per cent or more of their patients. This study has exposed a wide variation in consent practices. Future efforts to standardize consent processes are needed to protect patients and physicians.

  13. Myeloid-derived suppressor cell impact on endogenous and adoptively transferred T cells.

    PubMed

    Arina, Ainhoa; Bronte, Vincenzo

    2015-04-01

    Novel models of autochthonous tumorigenesis and adoptive T cell therapy (ATT) are providing new clues regarding the pro-tumorigenic and immunosuppressive effects of myeloid-derived suppressor cells (MDSC), and their interaction with T cells. New findings are shifting the perception of the main level at which MDSC act, from direct cell-to-cell suppression to others, such as limiting T cell infiltration. Adoptively transferred, high-avidity T cells recognizing peptides with high-affinity for MHC-I eliminated large tumors. However, low-avidity T cells or low-affinity peptides resulted in failure to eradicate tumors. Manipulation of intratumoral myeloid cells improved the outcome of otherwise unsuccessful ATT. Therefore, therapeutic intervention directed at the tumor stroma might be required when using suboptimal T cells for ATT.

  14. Tn7 transposition in vitro proceeds through an excised transposon intermediate generated by staggered breaks in DNA.

    PubMed

    Bainton, R; Gamas, P; Craig, N L

    1991-05-31

    We have developed a cell-free system in which the bacterial transposon Tn7 inserts at high frequency into its preferred target site in the Escherichia coli chromosome, attTn7; Tn7 transposition in vitro requires ATP and Tn7-encoded proteins. Tn7 transposes via a cut and paste mechanism in which the element is excised from the donor DNA by staggered double-strand breaks and then inserted into attTn7 by the joining of 3' transposon ends to 5' target ends. Neither recombination intermediates nor products are observed in the absence of any protein component or DNA substrate. Thus, we suggest that Tn7 transposition occurs in a nucleoprotein complex containing several proteins and the substrate DNAs and that recognition of attTn7 within this complex provokes strand cleavages at the Tn7 ends.

  15. Aggressive Therapy for Patients with Non-small Cell Lung Carcinoma and Synchronous Brain-only Oligometastatic Disease is Associated with Long-term Survival

    PubMed Central

    Gray, Phillip J.; Mak, Raymond H.; Yeap, Beow Y.; Cryer, Sarah K.; Pinnell, Nancy E.; Christianson, Laura W.; Sher, David J.; Arvold, Nils D.; Baldini, Elizabeth H.; Chen, Aileen B.; Kozono, David E.; Swanson, Scott J.; Jackman, David M.; Alexander, Brian M.

    2015-01-01

    Objectives Optimal therapy for patients with non-small cell lung carcinoma (NSCLC) presenting with synchronous brain-only oligometastases (SBO) is not well defined. We sought to analyze the effect of differing therapeutic paradigms in this subpopulation. Materials and Methods We retrospectively analyzed NSCLC patients with 1-4 SBO diagnosed between 1/2000 and 1/2011 at our institution. Patients with T0 tumors or documented Karnofsky Performance Status <70 were excluded. Aggressive thoracic therapy (ATT) was defined as resection of the primary disease or chemoradiotherapy whose total radiation dose exceeded 45 Gy. Cox proportional hazards and competing risks models were used to analyze factors affecting survival and first recurrence in the brain. Results Sixty-six patients were included. Median follow-up was 31.9 months. Intrathoracic disease extent included 9 stage I, 10 stage II and 47 stage III patients. Thirty-eight patients received ATT, 28 did not. Patients receiving ATT were younger (median age 55 vs. 60.5 years, p=0.027) but were otherwise similar to those who did not. Receipt of ATT was associated with prolonged median overall survival (OS) (26.4 vs. 10.5 months; p<0.001) with actuarial 2-year rates of 54% vs. 26%. ATT remained associated with OS after controlling for age, thoracic stage, performance status and initial brain therapy (HR 0.40, p=0.009). On multivariate analysis, the risk of first failure in the brain was associated with receipt of ATT (HR 3.62, p=0.032) and initial combined modality brain therapy (HR 0.34, p=0.046). Conclusion Aggressive management of thoracic disease in NSCLC patients with SBO is associated with improved survival. Careful management of brain disease remains important, especially for those treated aggressively. PMID:24974152

  16. A New Generation of Modified Live-Attenuated Avian Influenza Viruses Using a Two-Strategy Combination as Potential Vaccine Candidates▿

    PubMed Central

    Song, Haichen; Nieto, Gloria Ramirez; Perez, Daniel R.

    2007-01-01

    In light of the recurrent outbreaks of low pathogenic avian influenza (LPAI) and highly pathogenic avian influenza (HPAI), there is a pressing need for the development of vaccines that allow rapid mass vaccination. In this study, we introduced by reverse genetics temperature-sensitive mutations in the PB1 and PB2 genes of an avian influenza virus, A/Guinea Fowl/Hong Kong/WF10/99 (H9N2) (WF10). Further genetic modifications were introduced into the PB1 gene to enhance the attenuated (att) phenotype of the virus in vivo. Using the att WF10 as a backbone, we substituted neuraminidase (NA) for hemagglutinin (HA) for vaccine purposes. In chickens, a vaccination scheme consisting of a single dose of an att H7N2 vaccine virus at 2 weeks of age and subsequent challenge with the wild-type H7N2 LPAI virus resulted in complete protection. We further extended our vaccination strategy against the HPAI H5N1. In this case, we reconstituted an att H5N1 vaccine virus, whose HA and NA genes were derived from an Asian H5N1 virus. A single-dose immunization in ovo with the att H5N1 vaccine virus in 18-day-old chicken embryos resulted in more than 60% protection for 4-week-old chickens and 100% protection for 9- to 12-week-old chickens. Boosting at 2 weeks posthatching provided 100% protection against challenge with the HPAI H5N1 virus for chickens as young as 4 weeks old, with undetectable virus shedding postchallenge. Our results highlight the potential of live att avian influenza vaccines for mass vaccination in poultry. PMID:17596317

  17. Arterial Transit Time Mapping Obtained by Pulsed Continuous 3D ASL Imaging with Multiple Post-Label Delay Acquisitions: Comparative Study with PET-CBF in Patients with Chronic Occlusive Cerebrovascular Disease

    PubMed Central

    Tsujikawa, Tetsuya; Kimura, Hirohiko; Matsuda, Tsuyoshi; Fujiwara, Yasuhiro; Isozaki, Makoto; Kikuta, Ken-ichiro; Okazawa, Hidehiko

    2016-01-01

    Arterial transit time (ATT) is most crucial for measuring absolute cerebral blood flow (CBF) by arterial spin labeling (ASL), a noninvasive magnetic resonance (MR) perfusion assessment technique, in patients with chronic occlusive cerebrovascular disease. We validated ASL-CBF and ASL-ATT maps calculated by pulsed continuous ASL (pCASL) with multiple post-label delay acquisitions in patients with occlusive cerebrovascular disease. Fifteen patients underwent MR scans, including pCASL, and positron emission tomography (PET) scans with 15O-water to obtain PET-CBF. MR acquisitions with different post-label delays (1.0, 1.5, 2.0, 2.5 and 3.0 sec) were also obtained for ATT correction. The theoretical framework of 2-compartmental model (2CM) was also used for the delay compensation. ASL-CBF and ASL-ATT were calculated based on the proposed 2CM, and the effect on the CBF values and the ATT correction characteristics were discussed. Linear regression analyses were performed both on pixel-by-pixel and region-of-interest bases in the middle cerebral artery (MCA) territory. There were significant correlations between ASL-CBF and PET-CBF both for voxel values (r = 0.74 ± 0.08, slope: 0.87 ± 0.22, intercept: 6.1 ± 4.9) and for the MCA territorial comparison in both affected (R2 = 0.67, y = 0.83x + 6.3) and contralateral sides (R2 = 0.66, y = 0.74x + 6.3). ASL-ATTs in the affected side were significantly longer than those in the contralateral side (1.51 ± 0.41 sec and 1.12 ± 0.30 sec, respectively, p <0.0005). CBF measurement using pCASL with delay compensation was feasible and fairly accurate even in altered hemodynamic states. PMID:27275779

  18. Adverse Drug Reaction Profile in Patients on Anti-tubercular Treatment Alone and in Combination with Highly Active Antiretroviral Therapy

    PubMed Central

    Sadiq, Shamiya; Khajuria, Vijay; Mahajan, Annil; Singh, Jang B.

    2015-01-01

    Background and Objectives Adverse drug reactions are very common among patients on anti-tubercular treatment alone or in combination with highly active antiretroviral therapy but comparatively studied very less. Hence, the current study was done to evalaute the adverse drug reaction (ADR) profile in patients receiving anti-tubercular treatment (ATT) and ATT with highly active antiretroviral therapy (HAART). Materials and Methods A one year prospective, cross-sectional observational study was undertaken using suspected adverse drug data collection form available under Pharmacovigilance Programme of India. Results Seventy four patients receiving ATT & 32 patients on both ATT & HAART presented with 74 and 45 adverse drug events (ADE) respectively. Males were more affected than females in both the groups. DOTS category- 1 regimen was mostly responsible for ADE in both the groups. Epigastric pain was the most common ADE in TB patients, while anaemia was the most common presentation in TB with HIV group. On comparison, ADE rate of TB with HIV co-morbid patients was more (55.8%) than TB patients (0.36%) (p < 0.001). Urban population presented more with ADR in TB/HIV group unlike rural population in TB group (p<0.0001). Whereas, illiterate were more involved in TB group unlike literate in TB/HIV group (p<0.05). Type A reactions were more common in TB group (p < 0.001). Addition of drugs for the management of ADR events was more in TB/HIV group (p < 0.001) as compared to TB group. Rest all the parameters were comparable. Conclusion The study underscores that concomitant HAART and ATT, result in more ADRs in comparison to ATT alone demanding collaboration & integration of National AIDS Control programme and PvPI to enhance drug safety in this field. PMID:26557538

  19. EEG oscillatory activity associated to monetary gain and loss signals in a learning task: effects of attentional impulsivity and learning ability.

    PubMed

    De Pascalis, Vilfredo; Varriale, Vincenzo; Rotonda, Marco

    2012-07-01

    This study investigated the influence of individual differences in attentional impulsivity (Att-Imp), learning ability, and learning practice on oscillatory activity and phase synchrony responses to monetary gain and loss signals during an instrumental-learning task in healthy women. We used a trial-by-trial wavelet-based time-frequency analysis of the electroencephalographic (EEG) signal to provide amplitude and inter-site phase synchrony measures from 30 electrodes in theta (4-8 Hz, 350-500 ms), alpha (8-12 Hz, 100-200 ms), beta (13-25 Hz, 100-200 ms), and gamma (30-40 Hz, 350-450 ms) time-frequency ranges. Oscillatory amplitude and inter-site phase synchrony were both greater following loss signals as compared to gain signals in theta, beta, and gamma frequency bands. Low Att-Imp subjects had higher theta activity within a 350-500 ms time window over frontocentral, and centroparietal sites than high Att-Imp subjects. Monetary gain signals elicited higher theta and gamma activities in high Att-Imp individuals and loss signals elicited higher activities to loss signals in low Att-Imp individuals. Good learners showed enhanced intrahemispheric theta synchrony between frontoparietal, and fronto-occipital sites to monetary loss compared to gain signals. In good learners, monetary loss produced an increase of gamma synchrony that enhanced in the second stage of learning. In low Att-Imp individuals, there was a reduction of theta synchrony during the second stage, as compared with the first stage of learning, between temporal, parietal and fronto-parietal brain regions. These findings may offer valuable clues to understand outcome processing, attentional impulsivity, and learning ability. We propose that the punishment-related theta and gamma waves play a leading role in learning process.

  20. Multicopy integration of mini-Tn7 transposons into selected chromosomal sites of a Salmonella vaccine strain

    PubMed Central

    Roos, Karen; Werner, Esther; Loessner, Holger

    2015-01-01

    Chromosomal integration of expression modules for transgenes is an important aspect for the development of novel Salmonella vectors. Mini-Tn7 transposons have been used for the insertion of one such module into the chromosomal site attTn7, present only once in most Gram-negative bacteria. However, integration of multiple mini-Tn7 copies might be suitable for expression of appropriate amounts of antigen or combination of different modules. Here we demonstrate that integration of a 9.6 kb mini-Tn7 harbouring the luciferase luxCDABE (lux) occurs at the natural attTn7 site and simultaneously other locations of the Salmonella chromosome, which were engineered using λ-Red recombinase to contain one or two additional artificial attTn7 sites (a-attTn7). Multicopy integration even at closely spaced attTn7 sites was unexpected in light of the previously reported distance-dependent Tn7 target immunity. Integration of multiple copies of a mini-Tn7 containing a gfp cassette resulted in increasing green fluorescence of bacteria. Stable consecutive integration of two mini-Tn7 encoding lacZ and lux was achieved by initial transposition of lacZ-mini-Tn7, subsequent chromosomal insertion of a-attTn7 and a second round of transposition with lux-mini-Tn7. Mini-Tn7 thus constitutes a versatile method for multicopy integration of expression cassettes into the chromosome of Salmonella and possibly other bacteria. PMID:25488129

  1. A Case of Mycobacterium riyadhense in an Acquired Immune Deficiency Syndrome (AIDS) Patient with a Suspected Paradoxical Response to Antituberculosis Therapy

    PubMed Central

    Badreddine, Samar Assem

    2016-01-01

    A 30-year-old male patient with acquired immune deficiency syndrome (AIDS) on highly active antiretroviral therapy (HAART) presented with clinical picture suggestive of pulmonary tuberculosis. He was commenced on antituberculosis therapy (ATT) with signs of improvement. Then he developed cervical lymph node abscess which was drained. Steroid was started for presumed paradoxical response to ATT which results in clinical regression. The culture result revealed Mycobacterium riyadhense. This report addresses the rarity of this bacteria in medical literature. It reviews clinical presentations and medical treatment particularly in the setting of coinfections. PMID:27703819

  2. A Case of Tuberculous Meningitis with Atypical Multiple Lesions

    PubMed Central

    Zhang, W; Hu, Z; Li, T

    2014-01-01

    ABSTRACT This case describes a rare case of tuberculous meningitis. A 50-year old female presented with seven days of numbness on the left side, fatigue and a three-day headache. Magnetic resonance imaging (MRI) showed multiple lesions in the dorsal medulla and upper cervical spine. After admission, she developed a long-term fever, cranial nerves palsy and showed little response to corticosteroid, antibacterial and antiviral therapy. She received a diagnostic anti-tuberculous therapy (ATT); despite that, all examinations for tuberculosis were negative. After ATT lasting 16 days, she recovered and was discharged from hospital with slight asthenia and hypoesthesia. PMID:25867567

  3. Bot, Cyborg and Automated Turing Test

    NASA Astrophysics Data System (ADS)

    Yan, Jeff

    The Automated Turing test (ATT) is almost a standard security technique for addressing the threat of undesirable or malicious bot programs. In this paper, we motivate an interesting adversary model, cyborgs, which are either humans assisted by bots or bots assisted by humans. Since there is always a human behind these bots, or a human can always be available on demand, ATT fails to differentiate such cyborgs from humans. The notion of “telling humans and cyborgs apart” is novel, and it can be of practical relevance in network security. Although it is a challenging task, we have had some success in telling cyborgs and humans apart automatically.

  4. Intramedullary Tuberculoma of Dorsal Spinal Cord: A Case Report with Review of Literature

    PubMed Central

    Sonawane, Dhiraj Vithal; Jagtap, Sanjay Anand; Patil, Hitendra Gulabrao; Biraris, Sandeep Ramesh; Chandanwale, Ajay S

    2015-01-01

    Introduction: Intramedullary tuberculoma (IMT)are extremely rare lesions should be distinguished from other space occupying lesions like neoplasms. Although it is a treatable condition delaying diagnosis leads to significant morbidity. Case Report: We report a case of intramedullary thoracic tuberculoma with paraparesis showing deterioration of neurological status during medical treatment. Surgical resection had shown the excellent result. Conclusion: The possibility of IMT should be seriously considered when an intraspinal mass is found with evidence of tuberculosis. Anti-tuberculous therapy (ATT) is the mainstay of treatment and had shown good results. Surgical removal is to be considered in cases of worsening neurological status while the patient is on ATT. PMID:27299042

  5. Effect of antitubercular treatment on ovarian function in female genital tuberculosis with infertility

    PubMed Central

    Sharma, Jai Bhagwan; Sneha, J.; Singh, U. B.; Kumar, Sunesh; Roy, K. K.; Singh, Neeta; Dharmendra, Sona

    2016-01-01

    AIM: To evaluate the effect of antitubercular therapy (ATT) on an ovarian function such as ovarian reserve, ovarian dimensions, and ovarian stromal blood flow. SETTINGS AND DESIGN: Prospective study design. MATERIALS AND METHODS: Fifty infertile women with female genital tuberculosis (FGTB) without tubo-ovarian masses diagnosed by positive acid-fast bacilli culture or epithelioid granuloma on endometrial aspirate or positive polymerase chain reaction with positive findings on laparoscopy or hysteroscopy were recruited. The ovarian function tests were performed on day 2/3 as follicle-stimulating hormone (FSH) levels and anti-Mullerian hormone (AMH) levels. Ovarian dimensions (length, width, and depth) were measured using a transvaginal ultrasound. Mean antral follicle count (AFC) and ovarian stromal blood flow (peak systolic velocity [PSV], pulsatility index (PI), and resistive index [RI]) were measured using a transvaginal ultrasound. All women were started on ATT for 6 months by directly observed treatment strategy. After completion of ATT, all the parameters were repeated. RESULTS: There was a significant increase in AMH (2.68 ± 0.97 ng/ml to 2.8 ± 1.03 ng/ml) pre- to post-ATT, nonsignificant increase in FSH (7.16 ± 2.34 mIU/ml to 7.26 ± 2.33 mIU/ml) post-ATT, significant increase in mean AFC (7.40 ± 2.12–8.14 ± 2.17), PSV in the right ovary (6.015–6.11 cm/s) and left ovary (6.05–6.08 cm/s), PI in the right ovary (0.935–0.951 cm/s) and left ovary (0.936–0.957 cm/s), and RI in the right ovary (0.62 ± 0.01–0.79 ± 0.02) and left ovary (0.65 ± 0.02–0.84 ± 0.01) with ATT. There was no significant change in mean ovarian dimensions (ovarian length, breadth, and width) and summed ovarian volume with ATT. On laparoscopy, tubercles were seen in 27 (54%) women. Caseous nodules and encysted ascites were seen in 8% cases each. CONCLUSION: ATT improves the ovarian function (AMH and AFC) and ovarian blood flow in women with FGTB. PMID:27803581

  6. Bacteriophage φC31 Integrase Mediated Transgenesis in Xenopus laevis for Protein Expression at Endogenous Levels

    NASA Astrophysics Data System (ADS)

    Allen, Bryan G.; Weeks, Daniel L.

    Bacteriophage φC31 inserts its genome into that of its host bacterium via the integrase enzyme which catalyzes recombination between a phage attachment site (attP) and a bacterial attachment site (attB). Integrase requires no accessory factors, has a high efficiency of recombination, and does not need perfect sequence fidelity for recognition and recombination between these attachment sites. These imperfect attachment sites, or pseudo-attachment sites, are present in many organisms and have been used to insert transgenes in a variety of species. Here we describe the φC31 integrase approach to make transgenic Xenopus laevis embryos.

  7. Pituitary tumor evaluation

    SciTech Connect

    Albertson, B.; Binney, S.

    1995-11-01

    This paper describes research on the following: the structure of {sup 10}B{sub 10}-ovine corticotropin releasing hormone and {sup 10}B{sub 10}-growth hormone releasing hormone; the BNCT effect on AtT-20 cell {sup 10}B{sub 10}-CRH incubations in vitro; BNCT effects on GH{sub 4}C{sub 1} cell {sup 10}B{sub 10} growth hormone releasing factor incubation in vitro; and competitive inhibition of AtT-20 cell BNCT effect.

  8. Studies on attitude toward teaching science and anxiety about teaching science in preservice elementary teachers

    NASA Astrophysics Data System (ADS)

    Westerback, Mary E.

    These studies examined attitude toward teaching science (ATTS) using an adaptation of the Bratt Attitude Test (M-BAT); anxiety about teaching science (ANX-TS), as measured by the State-Trait Anxiety Inventory (STAI A-State); and selected demographic variables in preservice elementary teachers for the 1977-1978 and 1978-1979 academic years and a follow-up of those students who completed their student teaching in May 1979. The M-BAT and STAI were administered in September at the beginning of Science 6 (earth science and biology course), in December on the next to last day of Science 6, in May on the next to the last day of Science 5 (physical science), and in May 1979 after student teaching. In the two academic years, both ATTS and ANX-TS became more positive during the sequence Science 6-5. Both changes in ATTS and ANX-TS continued to change in a positive direction after completion of Science 6-5, after student teaching. There were differences in the times that the greatest changes in ATTS and ANX-TS occurred. In both studies, the greatest change in ATTS took place between September and December, during Science 6. The greatest change in ANX-TS, however, took place during Science 5 between December and May in the 1977-1978 study. In the 1978-1979 study, the greatest changes in ANX-TS occurred in Science 6, between September and December. The delayed reduction of ANX-TS in the 1977-1978 study may be explained by differences in teaching patterns. In 1977-1978, two teachers taught only their academic specialty, biology or earth science, to students who switched teachers midsemester. In 1978-1979, the same two instructors taught both biology and earth science to the same students. Correlation coefficients for successive and corresponding administrations of both the M-BAT and STAI suggest these variables are related. Students with more positive ATTS tended to have reduced ANX-TS. Neither the number of high school or college science and math courses completed nor the level

  9. Proceedings of the Fifth Annual Participants' Information Meeting: DOE Low-Level Waste Management Program

    SciTech Connect

    Not Available

    1983-12-01

    The meeting consisted of the following six sessions: (1) plenary session I; (2) disposal technology; (3) characteristics and treatment of low-level waste; (4) environmental aspects and performance prediction; (5) overall summary sessions; and (6) plenary session II. Fifty two papers of the papers presented were processed for inclusion in the Energy Data Base. (ATT)

  10. 32 CFR Attachment 5 to Part 855 - Sample Temporary Agreement

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 6 2012-07-01 2012-07-01 false Sample Temporary Agreement 5 Attachment 5 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 5 Attachment 5 to Part 855—Sample Temporary Agreement Letter of Agreement...

  11. 32 CFR Attachment 5 to Part 855 - Sample Temporary Agreement

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 6 2014-07-01 2014-07-01 false Sample Temporary Agreement 5 Attachment 5 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 5 Attachment 5 to Part 855—Sample Temporary Agreement Letter of Agreement...

  12. 32 CFR Attachment D to Subpart B... - Decision Tables

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 1 2011-07-01 2011-07-01 false Decision Tables D Attachment D to Subpart B of Part 147 National Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE PERSONNEL, MILITARY... Investigative Standards Pt. 147, Subpt. B, Att. D Attachment D to Subpart B of Part 147—Decision Tables Table...

  13. Stanniocalcin-1 regulates endothelial gene expression and modulates trans-endothelial migration of leukocytes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The mammalian counterpart of the fish calcium-regulating hormone stanniocalcin-1 (STC1) inhibits monocyte chemotactic protein-1- and stromal-derived factor-1alpha (SDF-1alpha)-mediated chemotaxis and diminishes chemokinesis in macrophage-like RAW264.7 and U937 cells in a manner that may involve atte...

  14. Rehabilitation after ACL Injury: A Fluoroscopic Study on the Effects of Type of Exercise on the Knee Sagittal Plane Arthrokinematics

    PubMed Central

    Norouzi, Sadegh; Esfandiarpour, Fateme; Shakourirad, Ali; Salehi, Reza; Akbar, Mohammad; Farahmand, Farzam

    2013-01-01

    A safe rehabilitation exercise for anterior cruciate ligament (ACL) injuries needs to be compatible with the normal knee arthrokinematics to avoid abnormal loading on the joint structures. The objective of this study was to measure the amount of the anterior tibial translation (ATT) of the ACL-deficient knees during selective open and closed kinetic chain exercises. The intact and injured knees of fourteen male subjects with unilateral ACL injury were imaged using uniplanar fluoroscopy, while the subjects performed forward lunge and unloaded/loaded open kinetic knee extension exercises. The ATTs were measured from fluoroscopic images, as the distance between the tibial and femoral reference points, at seven knee flexion angles, from 0° to 90°. No significant differences were found between the ATTs of the ACL-deficient and intact knees at all flexion angles during forward lunge and unloaded open kinetic knee extension (P < 0.05). During loaded open kinetic knee extension, however, the ATTs of the ACL deficient knees were significantly larger than those of the intact knees at 0° (P = 0.002) and 15° (P = 0.012). It was suggested that the forward lunge, as a weight-bearing closed kinetic chain exercise, provides a safer approach for developing muscle strength and functional stability in rehabilitation program of ACL-deficient knees, in comparison with open kinetic knee extension exercise. PMID:24066288

  15. APPLICATION OF COMPUTER-AIDED TOMOGRAPHY (CAT) AS A POTENTIAL INDICATOR OF MARINE MARCO BENTHIC ACTIVITY ALONG POLLUTION GRADIENTS

    EPA Science Inventory

    Sediment cores were imaged using a local hospital CAT scanner. These image data were transferred to a personal computer at our laboratory using specially developed software. Previously, we reported an inverse correlation (r2 = 0.98, P<0.01) between the average sediment x-ray atte...

  16. Just-in-Time Knowledge Performance Support: A Test of Concept.

    ERIC Educational Resources Information Center

    Dorsey, Laura T.; And Others

    1993-01-01

    Describes a project developed by AT&T (American Telephone and Telegraph) and Indiana University that focused on the needs and tasks of sales personnel by building prototypes of components of a performance support system. Combining innovative technology to support a changing work environment is discussed, and the evaluation process is reported.…

  17. Rehabilitation after ACL injury: a fluoroscopic study on the effects of type of exercise on the knee sagittal plane arthrokinematics.

    PubMed

    Norouzi, Sadegh; Esfandiarpour, Fateme; Shakourirad, Ali; Salehi, Reza; Akbar, Mohammad; Farahmand, Farzam

    2013-01-01

    A safe rehabilitation exercise for anterior cruciate ligament (ACL) injuries needs to be compatible with the normal knee arthrokinematics to avoid abnormal loading on the joint structures. The objective of this study was to measure the amount of the anterior tibial translation (ATT) of the ACL-deficient knees during selective open and closed kinetic chain exercises. The intact and injured knees of fourteen male subjects with unilateral ACL injury were imaged using uniplanar fluoroscopy, while the subjects performed forward lunge and unloaded/loaded open kinetic knee extension exercises. The ATTs were measured from fluoroscopic images, as the distance between the tibial and femoral reference points, at seven knee flexion angles, from 0° to 90°. No significant differences were found between the ATTs of the ACL-deficient and intact knees at all flexion angles during forward lunge and unloaded open kinetic knee extension (P < 0.05). During loaded open kinetic knee extension, however, the ATTs of the ACL deficient knees were significantly larger than those of the intact knees at 0° (P = 0.002) and 15° (P = 0.012). It was suggested that the forward lunge, as a weight-bearing closed kinetic chain exercise, provides a safer approach for developing muscle strength and functional stability in rehabilitation program of ACL-deficient knees, in comparison with open kinetic knee extension exercise. PMID:24066288

  18. Contracts for field projects and supporting research on enhanced oil recovery and improved drilling technology. Progress review No. 34, quarter ending March 31, 1983

    SciTech Connect

    Linville, B.

    1983-07-01

    Progress achieved for the quarter ending March 1983 are presented for field projects and supporting research for the following: chemical flooding; carbon dioxide injection; and thermal/heavy oil. In addition, progress reports are presented for: resource assessment technology; extraction technology; environmental and safety; microbial enhanced oil recovery; oil recovered by gravity mining; improved drilling technology; and general supporting research. (ATT)

  19. Heating poultry houses with an attic ventilation system

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fuel use constitutes a large percentage of the energy costs incurred by growers during winter flocks and increasing fuel costs have illustrated the need for alternative methods to reduce energy usage in broiler production. The objective of this study was to determine the feasibility of using the att...

  20. 78 FR 76710 - Notice of Buy America Waiver for a Video Ready Access Device Cabinet

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-18

    ... cabinet needed for an AT&T utility relocation associated with the Charlotte Area Transit System's (CATS... Transit System's (CATS or City of Charlotte) LYNX Blue Line Extension (BLE) project, which is an FTA... interpretation of FTA's Buy America rules with respect to the utility relocation performed for the CATS LYNX...

  1. Software process assessments

    NASA Technical Reports Server (NTRS)

    Miller, Sharon E.; Tucker, George T.; Verducci, Anthony J., Jr.

    1992-01-01

    Software process assessments (SPA's) are part of an ongoing program of continuous quality improvements in AT&T. Their use was found to be very beneficial by software development organizations in identifying the issues facing the organization and the actions required to increase both quality and productivity in the organization.

  2. Root assays to study pattern-triggered immunity in plant-nematode interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plants employ extracellular immune receptors to perceive conserved pathogen-associated molecular patterns (PAMPs), triggering the first layer of defense known as pattern-triggered immunity (PTI). The understanding of PTI is mainly based on the studies focusing on leaves. Plants are vulnerable to att...

  3. Multiple staphylococcal cassette chromosomes and allelic variants of cassette chromosome recombinases in Staphylococcus aureus and coagulase-negative staphylococci from Norway.

    PubMed

    Hanssen, Anne-Merethe; Sollid, Johanna U Ericson

    2007-05-01

    We investigated the nature of the staphylococcal cassette chromosome mec (SCCmec) elements and cognate insertion sites in a collection of 42 clinical staphylococcal isolates of various species from Norway. The ccr and mec genes and the attachment sites (attL/attR) were identified by PCR, Southern blot hybridization, and DNA sequencing. We found 10 possibly new SCCmec types and one previously unreported variant of SCCmec type III (mec complex A, ccrAB3, and ccrC7) in Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis. Eleven of 42 strains contained multiple copies of ccr, suggesting the presence of mosaic structures composed of multiple SCC elements. S. haemolyticus contained ccrAB2 genes identical to those in S. aureus SCCmec type IV but lacked IS1272 and mec regulators. Two new allelic ccr variants, ccrC6 and ccrC7, were identified. Also, the presumed functional version of ccrB1 was found in a mecA-positive S. hominis strain and in mecA-negative S. epidermidis and S. hominis strains. Only minor differences in direct repeats in the left and right boundaries (attR/attL) were observed, while there was more variation in the inverted repeats. Coagulase-negative staphylococci (CoNS) contained several representatives of different ccr complexes and thus seemed to harbor multiple or composite new types of SCCmec. The enormous diversity observed in the SCCmec elements implies a large SCCmec reservoir in CoNS.

  4. Integrated Communications at America's Leading Total Quality Management Corporations.

    ERIC Educational Resources Information Center

    Gronstedt, Anders

    1996-01-01

    Examines how to create organizational processes that allow communication professionals with a variety of expertise to support each other through coordination and integration. Studies eight of America's leading total quality management corporations, including AT&T, Federal Express, Saturn, and Xerox. Explores how various total quality management…

  5. Campaign Policies, Broadcasters, and the Presidential Election of 1924.

    ERIC Educational Resources Information Center

    Benjamin, Louise M.

    The policies initiated by broadcasters for campaign coverage during the 1924 presidential election influenced the treatment of political candidates on the air and helped set the stage for inclusion of the equal opportunities clause in the Radio Act of 1927. AT&T, RCA, and General Electric formed policies to treat candidates fairly in allocating…

  6. Workplace Education Initiative: Year Two Observations.

    ERIC Educational Resources Information Center

    Sperazi, Laura; Astrein, Bruce

    Site visits to 17 workplace education projects funded in the second year of the Massachusetts Workplace Education Initiative are reported for year 1 and year 2 projects. The year one projects are as follows: EASCO Handtool Company (Springfield), T.J. Maxx Distribution Center (Worcester), laundry workers (Boston/Lynn), AT&T (Lawrence), and Columbia…

  7. Cutting out the φC31 prophage.

    PubMed

    Stark, W Marshall

    2011-06-01

    To establish a lysogenic lifestyle, the temperate bacteriophage φC31 integrates its genome into the chromosome of its Streptomyces host, by site-specific recombination between attP (the attachment site in the phage DNA) and attB (the chromosomal attachment site). This reaction is promoted by a phage-encoded serine recombinase Int. To return to the lytic lifestyle, the prophage excises its DNA by a similar Int-mediated reaction between the recombinant sites flanking the prophage, attL and attR. φC31 Int has been developed into a popular experimental tool for integration of transgenic DNA into the genomes of eukaryotic organisms. However, until now it has not been possible to use Int to promote the reverse reaction, excision. In many other phages, the presence of a recombination directionality factor (RDF) protein biases the phage-encoded integrase towards prophage excision, whereas absence of the RDF favours integration; but the φC31 RDF had proved elusive. In this issue of Molecular Microbiology, Khaleel et al. (2011) report the identification and purification of the φC31 RDF, and show that it both promotes excision and inhibits integration by direct protein-protein interactions with Int itself. PMID:21564340

  8. Proceedings of the Symposium on Training of Nuclear Facility Personnel (8th, Gatlinburg, Tennessee, April 24-27, 1989).

    ERIC Educational Resources Information Center

    Oak Ridge National Lab., TN.

    Education-related titles among the 56 papers are "Panel Discussion: Plant Expectation of Training" (Sellman, Zach, Cross); "Managing Training AT&T" (Solomon); "Training Management Systems" (Waylett); "Managing the Training Function" (Wiggin); "Training Management" (Newton); Three Alternative Simulation Systems for Training Nuclear Power Plant…

  9. A SEARCH FOR RESIDUAL BEHAVIORAL EFFECTS OF TRICHLOROETHYLENE (TCE) IN RATS EXPOSED AS YOUNG ADULTS

    EPA Science Inventory

    Trichloroethylene (TCE) is a solvent of concern to the EPA due to its extensive use in industry, its prevalence in urban air, and its appearance in water supplies. Human clinical studies have associated short and long-termsolvent exposures with cognitive dysfunction including att...

  10. Attention Training with Auditory Hallucinations: A Case Study

    ERIC Educational Resources Information Center

    Valmaggia, Lucia R.; Bouman, Theo K.; Schuurman, Laura

    2007-01-01

    The case presented in this paper illustrates how Attention Training (ATT; [Wells, A. (1990). "Panic disorder in association with relaxation induced anxiety: An attentional training approach to treatment." "Behavior Therapy," 21, 273-280.]) can be applied in an outpatient setting in the treatment of auditory hallucinations. The 25-year-old male…

  11. Installing the Future. Fiber Optics Program Readies Students for Lucrative Jobs.

    ERIC Educational Resources Information Center

    Serrano, Kenneth M.

    1995-01-01

    A fiber optics program at Somerset County Technical Institute (SCTI) prepares college students and trades workers for telecommunication's new wave of installation. The program was born of a partnership among an electricians' union, AT&T, and SCTI to meet the expected need for fiber optic technicians. (JOW)

  12. 47 CFR 64.1201 - Restrictions on billing name and address disclosure.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... due; (ii) Any purpose associated with the “equal access” requirement of United States v. AT&T 552 F... provided to a local exchange company by each of its local exchange customers to which the local exchange company directs bills for its services. (2) The term “telecommunications service provider”...

  13. The Impact of the AT&T-EEO Consent Decree. Labor Relations and Public Rolicy Series No. 20.

    ERIC Educational Resources Information Center

    Northrup, Herbert R.; Larson, John A.

    The consent decree agreed to in January 1973 by the American Telephone and Telegraph Company (AT&T), the United States Equal Employment Opportunity Commission (EEOC), the Department of Justice, and the Department of Labor was a milestone of EEOC enforcement. Large amounts of back pay were paid to women and minorities, and goals were set which, as…

  14. 47 CFR 53.201 - Services for which a section 272 affiliate is required.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... provide previously authorized interLATA information services and manufacturing activities through a... the AT&T Consent Decree that authorized such services. (b) InterLATA information services. A BOC shall provide an interLATA information service through a section 272 affiliate when it provides the...

  15. The membrane IgM-associated proteins MB-1 and Ig-beta are sufficient to promote surface expression of a partially functional B-cell antigen receptor in a nonlymphoid cell line.

    PubMed

    Matsuuchi, L; Gold, M R; Travis, A; Grosschedl, R; DeFranco, A L; Kelly, R B

    1992-04-15

    The B-cell antigen receptors consist of membrane immunoglobulins (mIgs) noncovalently associated with two accessory proteins, MB-1 and Ig-beta. We used transfection into a nonlymphoid cell line to test whether MB-1 and Ig-beta were sufficient to promote cell surface expression of mIgM capable of signal transduction. Expression of MB-1 and Ig-beta, but not MB-1 alone, allowed high-level surface expression of mIgM in the AtT20 endocrine cell line, which presumably lacks other B-cell-specific components. The reconstituted antigen receptor was capable of mediating some of the signaling reactions characteristic of mIgM in B lymphocytes. Crosslinking mIgM on transfected AtT20 cells stimulated tyrosine phosphorylation of MB-1 and Ig-beta and also increased the amount of phosphatidylinositol 3-kinase activity that could be precipitated with anti-phosphotyrosine antibodies. When total cell lysates were analyzed by anti-phosphotyrosine immunoblotting, however, no induced phosphorylation of more abundant proteins was detected. Moreover, crosslinking of the receptor in AtT20 cells did not stimulate inositol phospholipid breakdown. Thus, the transfected B-cell antigen receptor could initiate some signal transduction events but AtT20 cells may lack components required for other signaling events associated with mIgM.

  16. UNIX Systems in Higher Education: A Paradoxical Success.

    ERIC Educational Resources Information Center

    McCredie, John W.

    1983-01-01

    Bell Laboratories' much acclaimed UNIX operating system is widely used in educational computing environments. Discusses history of the system, system features and weaknesses, and policy issues. Also discusses some ways UNIX systems are used and recent developments at American Telephone and Telegraph (AT&T) impacting UNIX systems. (JN)

  17. Research on Teletraining: Student Acceptance, Learning Effectiveness and Cost Benefits in the Corporate Environment.

    ERIC Educational Resources Information Center

    Chute, Alan G.

    The Sales and Marketing Education organization of AT&T Communications conducts ongoing research to monitor the impact of their National Teletraining Network (NTN) programs for professional sales personnel on: (1) learning, (2) student acceptance of courses and instructors, and (3) student willingness to take additional teletrained programs. During…

  18. Lupus vulgaris of external nose with septal perforation--a rarity in antibiotic era.

    PubMed

    Garg, Ajay; Wadhera, Raman; Gulati, S P; Singh, Jagjit

    2010-07-01

    Lupus vulgaris (LV) is the commonest morphological variant of cutaneous tuberculosis. Case of LV of external nose extending to internal nose causing septal perforation is documented here. Histopathology of biopsy taken confirmed the diagnosis of LV. Patient responded well to Anti-tubercular therapy (ATT).

  19. 32 CFR Attachment A to Subpart B... - Standard A-National Agency Check With Local Agency Checks and Credit Check (NACLC)

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Access Programs by sect. 4.4 of Executive Order 12958) (60 FR 19825, 3 CFR 1995 Comp., p. 333); (2) “L... Agency Checks and Credit Check (NACLC) A Attachment A to Subpart B of Part 147 National Defense..., Subpt. B, Att. A Attachment A to Subpart B of Part 147—Standard A—National Agency Check With...

  20. 32 CFR Attachment 3 to Part 2800 - Sample

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Sample 3 Attachment 3 to Part 2800 National Defense Other Regulations Relating to National Defense OFFICE OF THE VICE PRESIDENT OF THE UNITED STATES SECURITY PROCEDURES Pt. 2800, Att. 3 Attachment 3 to Part 2800—Sample EC25OC91.012...

  1. 32 CFR Attachment 5 to Part 855 - Sample Temporary Agreement

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Sample Temporary Agreement 5 Attachment 5 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 5 Attachment 5 to Part 855—Sample...

  2. 32 CFR Attachment 2 to Part 2800 - Security Termination Statement

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Security Termination Statement 2 Attachment 2 to Part 2800 National Defense Other Regulations Relating to National Defense OFFICE OF THE VICE PRESIDENT OF THE UNITED STATES SECURITY PROCEDURES Pt. 2800, Att. 2 Attachment 2 to Part...

  3. 32 CFR Attachment 3 to Part 855 - Landing Permit Application Instructions

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Landing Permit Application Instructions 3 Attachment 3 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 3 Attachment 3 to Part...

  4. 32 CFR Attachment 4 to Part 855 - Sample Joint-Use Agreement

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Sample Joint-Use Agreement 4 Attachment 4 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 4 Attachment 4 to Part 855—Sample...

  5. 32 CFR Attachment 1 to Part 2800 - Employment Agreement & Indoctrination Statement

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Employment Agreement & Indoctrination Statement 1 Attachment 1 to Part 2800 National Defense Other Regulations Relating to National Defense OFFICE OF THE VICE PRESIDENT OF THE UNITED STATES SECURITY PROCEDURES Pt. 2800, Att. 1 Attachment 1 to...

  6. 32 CFR Attachment B to Subpart B... - Standard B-Single Scope Background Investigation (SSBI)

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 1 2010-07-01 2010-07-01 false Standard B-Single Scope Background Investigation (SSBI) B Attachment B to Subpart B of Part 147 National Defense Department of Defense OFFICE OF THE... FOR ACCESS TO CLASSIFIED INFORMATION Investigative Standards Pt. 147, Subpt. B, Att. B Attachment B...

  7. 32 CFR Attachment D to Subpart B... - Decision Tables

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 1 2010-07-01 2010-07-01 false Decision Tables D Attachment D to Subpart B of Part 147 National Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE PERSONNEL, MILITARY... Investigative Standards Pt. 147, Subpt. B, Att. D Attachment D to Subpart B of Part 147—Decision Tables Table...

  8. Developmental and Behavioral Performance of Internationally Adopted Preschoolers: A Pilot Study

    ERIC Educational Resources Information Center

    Jacobs, Emma; Miller, Laurie C.; Tirella, Linda G.

    2010-01-01

    Most international adoptees (IA) have rapid catch-up of the delays common at arrival. However, it is not known whether development at arrival predicts later abilities or school readiness. Therefore, we comprehensively evaluated language, fine motor, visual reception (VR), executive function (EF), attention (ATT), and sensory skills (SS) in IA…

  9. Percentage of Protected Area Amounts within each Watershed Boundary for the Conterminous US

    EPA Science Inventory

    Abstract: This dataset uses spatial information from the Watershed Boundary Dataset (WBD, March 2011) and the Protected Areas Database of the United States (PAD-US Version 1.0). The resulting data layer, with percentages of protected areas by category, was created using the ATtI...

  10. 77 FR 27221 - Combined Notice of Filings #2

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-09

    ... following electric rate filings: Docket Numbers: ER12-1695-000. Applicants: Liberty Electric Power, LLC.... Comments Due: 5 p.m. ET 5/23/12. Docket Numbers: ER12-1696-000. Applicants: Michigan Electric Transmission...: Louisville Gas and Electric Company. Description: Att M LGIA LGIP Rev to be effective 5/3/2012. Filed Date:...

  11. End-User Searching in a Large Library Network: A Case Study of Patent Attorneys.

    ERIC Educational Resources Information Center

    Vollaro, Alice J.; Hawkins, Donald T.

    1986-01-01

    Reports results of study of a group of end users (patent attorneys) doing their own online searching at AT&T Bell Laboratories. Highlights include DIALOG databases used by the attorneys, locations and searching modes, characteristics of patent attorney searchers, and problem areas. Questionnaire is appended. (5 references) (EJS)

  12. Electronic Baby Steps.

    ERIC Educational Resources Information Center

    Froman, Ann D.

    1992-01-01

    Describes the participation of a small Catholic school near Philadelphia in the AT&T Learning Circles Project, which involved networking with schools in several states. The integration of the telecommunications project with the language arts curriculum is discussed, including creative writing about local food and historic places to share with…

  13. A theoretical analysis in the time-domain of wave reflection on a bone plate

    NASA Astrophysics Data System (ADS)

    Grimal, Quentin; Naïli, Salah

    2006-11-01

    This paper presents an original method for simulating ultrasonic wave reflection on a fluid-loaded plate. Geometrical and material parameters used are relevant to the "axial transmission technique" (ATT) setup. Devices based on the ATT are used for the assessment of cortical bone strength (estimate of a fracture risk). In this work, the cortical bone layer is represented as a plate of infinite extent surrounded by fluid (soft tissues). A line source and a receiver are placed in the fluid. Transient waves generated upon reflection at the plate-fluid interfaces are addressed. Analytic Green's functions are derived with the generalized ray/Cagniard-de Hoop method. The acoustic response is obtained upon convolution of Green's functions with a given source pulse. The method associates each wave amplitude in the time-domain (lateral waves, reflected body waves, etc.) to a specific term of the final solution. The method is ideally suited to a detailed analysis of the ultrasonic signal for various geometrical and mechanical parameters. The results presented highlight the potential of the method for the understanding of wave phenomena involved in the ATT and similar setups. They also bring new elements that reinforce our theoretical knowledge of the ATT.

  14. ANALYTICAL TOOLS INTERFACE FOR LANDSCAPE ASSESSMENTS (ATTILA) USER MANUAL

    EPA Science Inventory

    ATtlLA is an ArcView extension that allows users to easily calculate many common landscape metrics. GIS expertise is not required, but some experience with ArcView is recommended. Four metric groups are currently included in ATtILA: landscape characteristics, riparian characteris...

  15. Proprioception in the ACL-ruptured knee: the contribution of the medial collateral ligament and patellar ligament. An in vivo experimental study in the cat.

    PubMed

    Bonsfills, N; Raygoza, J J; Boemo, E; Garrido, J; Núñez, A; Gómez-Barrena, E

    2007-01-01

    In the absence of anterior cruciate ligament (ACL), secondary restraints such as menisci, ligaments, and tendons restrict anterior knee laxity. Strain detection at these sites could define the contribution of this alternative signalling system to knee proprioception after ACL injury. The hypothesis in this study questions if measurements of anterior tibial translation (ATT) from surface strain gauges on the insertions of the medial collateral ligament (MCL) and the patellar tendon (PT) are sufficiently sensitive and specific to differentiate normal, stable knees from acutely unstable knees due to ACL section. Twelve cats received miniaturized strain gauges on the surface of MCL and PT distal insertions. A purpose-made receiver transformed into measurements any voltage variation obtained during passive knee flexion-extension and anterior tibial translation manoeuvres. Variables under evaluation included first peak latency, normalized amplitude, and slope of voltage along time. Femorotibial displacements were video recorded, digitized, and used as the ATT reference. The proposed system detected significant changes in the slope of the voltage/time signal, with higher specificity and sensitivity during ATT after experimental ACL section. Changes were not significant during flexion or extension. It was found that a pattern of earlier and more intense strain in MCL and PT distal insertions was found during ATT in the ACL deficient knee. Enhanced pattern recognition learning from these structures could be a future target for proprioceptive training after ACL injury.

  16. Aspirin

    MedlinePlus

    ... Guidelines . Circulation . 2011 ; 123 : 2022 – 2060 . OpenUrl FREE Full Text 4. ↵ Antithrombotic Trialists' (ATT) Collaboration , Baigent C , Blackwell ... BMJ . 2002 ; 324 : 71 – 86 . OpenUrl Abstract / FREE Full Text 6. ↵ Furie KL , Kasner SE , Adams RJ , Albers ...

  17. Low level hydrogen peroxide generation from a nonwoven fibrous pectin-cellulose blend

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fibrous pectic-cellulose (FPC) (cellulose blended with primary cell wall pectin at 2 percent by weight of pectin) is product made from naturally occurring plant fibers. FPC is a fibrous mixture of polysaccharides with a low percent by weight of pectin-based primary cell wall and lipid components att...

  18. 75 FR 61716 - Combined Notice of Filings #2

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-06

    ... per 35.13(a)(2)(iii): 2010-09-24--Att-O--PSCo--SmartGrid--Tbls2- 3--chngs to be effective 8/1/2010.... Applicants: Farmers City Wind, LLC. Description: Farmers City Wind, LLC submits its Baseline Filing of...

  19. 32 CFR Attachment 1 to Part 855 - Glossary of References, Abbreviations, Acronyms, and Terms

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 10-10, Civil Aircraft Use of United States Air Force Airfields AFI 10-1001, Civil Aircraft Landing Permits AFI 13-201, Air Force Airspace Management AFI 32-7061(32 CFR part 989), Environmental Impact... THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att....

  20. [Construction of the oocyte-specific expressing phiC31 integrase vector pZP3-INT and its expression in mouse oocytes].

    PubMed

    Xu, Huan-Yu; Gong, Xiu-Li; Guo, Xin-Bing; Ma, Qing-Wen; Zeng, Yi-Tao

    2009-06-01

    Streptomyces phage phiC31 integrase is a site-specific recombinase, which can catalyze site-specific, unidirectional recombination between the attP site and attB site. To explore whether it can be used to mediate the recombination of specific gene in oocytes, GV-stage oocytes were collected from 3-week-old Kunming White mice by puncturing antral follocles with a sharp needle, and micro-injected with oocyte-specific expressing phiC31 integrase vector pZP3-INT and site -specific recombination detection vector pBCPB+. phiC31 integrase mRNA were detected by RT-PCR and the recombination of pBCPB+ was evaluated by PCR in mouse oocytes at 48 h after injection. Both can get corresponding bands. These results indicated that the expression of phiC31 integrase can be driven by ZP3 promoter efficiently and phiC31 integrase can mediate the site-specific recombination between attP site and attB site in mouse GV-stage oocytes. It could be a powerful tool for the study of recombination of specific gene in mouse oocytes and would provide an alternative way for the mouse oocyte genome manipulation.

  1. Characterizing the molecular basis of attenuation of Marek’s disease virus via in vitro serial passage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Marek’s disease (MD) is a lymphoproliferative disease of chickens caused by the oncogenic Gallid herpesvirus 2, commonly known as Marek’s disease virus (MDV). MD vaccines, the primary control method, are often generated by repeated in vitro serial passage of this highly cell-associated virus to atte...

  2. A novel approach to investigate the uptake and internalization of Escherichia coli O157:H7 in spinach cultivated in soil and hydroponic media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Internalization of E. coli O157:H7 into spinach plants through root uptake is a potential route of contamination. A Tn7-based plasmid vector was used to insert the green fluorescent protein (gfp) gene into the attTn7 site in the E. coli chromosome. Three gfp-labeled E. coli inocula, O157:H7 strains ...

  3. Two-stage, low noise advanced technology fan. 5: Acoustic final report

    NASA Technical Reports Server (NTRS)

    Sofrin, T. G.; Riloff, N., Jr.

    1975-01-01

    The NASA Q2S(quiet two-stage) fan is a 0.836m (32.9 in.) diameter model of the STF 433 engine fan, selected in a 1972 study for an Advanced Technology Transport (ATT) airplane. Noise-control features include: low tip speed, moderate stage pressure rise, large blade-vane spacings, no inlet guide vanes, and optimum blade and vane numbers. Tests were run on the baseline Q2S fan with standard inlet and discharge ducts. Further tests were made of a translating centerbody sonic inlet device and treated discharge ducts. Results were scaled to JT8D and JT3D engine fan size for comparison with current two-stage fans, and were also scaled to STF 433 fan size to compare calculated ATT flyover noise with FAR 36 limits. Baseline Q2S results scaled to JT8D and JT3D engine fan sizes showed substantial noise reductions. Calculated unsuppressed baseline ATT flyovers averaged about 2.5 EPNdB below FAR 36 limits. Using measured sonic inlet results, scaled baseline Q2S fan results, and calculated attenuations for a 1975 technology duct liner, projected flyover noise calculations for the ATT averaged about FAR 36 limits minus 10 EPNdB. Advances in suppression technology required to meet the 1985 goal of FAR 36 limits minus 20 EPNdB are discussed.

  4. Technology and the Research Environment of the Future. The Impact of the Information Science Revolution on the Research Environment of the Future. Proceedings of Three Seminars (Albany, New York, September 15, 1987, February 2, 1988, and May 6, 1988).

    ERIC Educational Resources Information Center

    New York State Library, Albany.

    In a series of three half-day seminars, scientists from AT&T Bell Laboratories, IBM (International Business Machines), and the Corporation for National Research Initiatives described current research in materials science, computing, and networking with implications for scholarly communication in the future. The audience for the seminars included…

  5. Isolation and characterization of a proteinase K sensitive PrPSc fraction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent studies have shown that a sizeable fraction of PrPSc present in prion-infected tissues is,contrary to previous conceptions, sensitive to digestion by proteinase K (PK). This finding has important implications in the context of diagnosis of prion disease, as PK has been extensively used in att...

  6. Using Technology in School Supervision.

    ERIC Educational Resources Information Center

    Bercik, Janet T.; Blair-Larsen, Susan M.

    2000-01-01

    Notes that few college faculty use technology in their teaching despite rapid growth in technology-based instruction in K-12 education. Describes two projects using AT&T's PersonaLink Service and Sony's Magic Link PIC 1000 in field experience supervision. (SG)

  7. Disparities in provision of transplant information affect access to kidney transplantation.

    PubMed

    Kucirka, L M; Grams, M E; Balhara, K S; Jaar, B G; Segev, D L

    2012-02-01

    Recently Centers for Medicare and Medicaid Services (CMS) began asking providers on Form-2728 whether they informed patients about transplantation, and if not, to select a reason. The goals of this study were to describe national transplant education practices and analyze associations between practices and access to transplantation (ATT), based on United States Renal Data System (USRDS) data from 2005 to 2007. Multinomial logistic regression was used to examine factors associated with not being informed about transplantation, and modified Poisson regression to examine associations between not being informed and ATT (all models adjusted for demographics/comorbidities). Of 236,079 incident end-stage renal disease (ESRD) patients, 30.1% were not informed at time of 2728 filing, for reasons reported by providers as follows: 42.1% unassessed, 30.4% medically unfit, 16.9% unsuitable due to age, 3.1% psychologically unfit and 1.5% declined counsel. Older, obese, uninsured, Medicaid-insured and patients at for-profit centers were more likely to be unassessed. Women were more likely to be reported as unsuitable due to age, medically unfit and declined, and African Americans as psychologically unfit. Uninformed patients had a 53% lower rate of ATT, a disparity persisting in the subgroup of uninformed patients who were unassessed. Disparities in ATT may be partially explained by disparities in provision of transplant information; dialysis centers should ensure this critical intervention is offered equitably.

  8. Synthesis of formyl esters of vegetable oils: Milkweed, Pennycress and Soy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a previous study of the characteristics of acyl derivatives of polyhydroxy milkweed oil (PHMWO), it was observed that the densities and viscosities of the respective derivatives decreased with increased chain length of the substituent acyl group. Thus, from the polyhydroxyl starting material, att...

  9. Strategic Human Resources Management of Employer Cooperative Education Programs.

    ERIC Educational Resources Information Center

    Nielsen, Richard P.; Porter, Ralph C.

    1982-01-01

    This article discusses effective strategic planning and management of employer cooperative education programs. It examines types of planning, the need for such programs, and implementation strategies. Several case studies are considered: Dayton-Hudson, IBM, AT&T, Delta Airlines, and Rockwell International. (CT)

  10. 32 CFR Attachment D to Subpart B... - Decision Tables

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 1 2013-07-01 2013-07-01 false Decision Tables D Attachment D to Subpart B of Part 147 National Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE PERSONNEL, MILITARY... Investigative Standards Pt. 147, Subpt. B, Att. D Attachment D to Subpart B of Part 147—Decision Tables Table...

  11. Heat Stress Screening of Peanut Seedlings for Acquired Thermotolerance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this research was to develop a user-friendly and medium throughput laboratory protocol using acquired thermotolerance (ATT) in peanut seedlings as a measure of one mechanism of heat stress tolerance. Sixteen genotypes, including selected accessions of the U.S. peanut min...

  12. 32 CFR Attachment 2 to Part 855 - Weather Alternate List

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 6 2011-07-01 2011-07-01 false Weather Alternate List 2 Attachment 2 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 2 Attachment 2 to Part...

  13. 32 CFR Attachment 2 to Part 855 - Weather Alternate List

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 6 2014-07-01 2014-07-01 false Weather Alternate List 2 Attachment 2 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 2 Attachment 2 to Part...

  14. 32 CFR Attachment 2 to Part 855 - Weather Alternate List

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false Weather Alternate List 2 Attachment 2 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 2 Attachment 2 to Part...

  15. 32 CFR Attachment 2 to Part 855 - Weather Alternate List

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 6 2012-07-01 2012-07-01 false Weather Alternate List 2 Attachment 2 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 2 Attachment 2 to Part...

  16. 32 CFR Attachment 2 to Part 855 - Weather Alternate List

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 6 2013-07-01 2013-07-01 false Weather Alternate List 2 Attachment 2 to Part 855 National Defense Department of Defense (Continued) DEPARTMENT OF THE AIR FORCE AIRCRAFT CIVIL AIRCRAFT USE OF UNITED STATES AIR FORCE AIRFIELDS Pt. 855, Att. 2 Attachment 2 to Part...

  17. The effects of QEEG-informed neurofeedback in ADHD: an open-label pilot study.

    PubMed

    Arns, Martijn; Drinkenburg, Wilhelmus; Leon Kenemans, J

    2012-09-01

    In ADHD several EEG biomarkers have been described before, with relevance to treatment outcome to stimulant medication. This pilot-study aimed at personalizing neurofeedback treatment to these specific sub-groups to investigate if such an approach leads to improved clinical outcomes. Furthermore, pre- and post-treatment EEG and ERP changes were investigated in a sub-group to study the neurophysiological effects of neurofeedback. Twenty-one patients with ADHD were treated with QEEG-informed neurofeedback and post-treatment effects on inattention (ATT), hyperactivity/impulsivity (HI) and comorbid depressive symptoms were investigated. There was a significant improvement for both ATT, HI and comorbid depressive complaints after QEEG-informed neurofeedback. The effect size for ATT was 1.78 and for HI was 1.22. Furthermore, anterior individual alpha peak frequency (iAPF) demonstrated a strong relation to improvement on comorbid depressive complaints. Pre- and post-treatment effects for the SMR neurofeedback sub-group exhibited increased N200 and P300 amplitudes and decreased SMR EEG power post-treatment. This pilot study is the first study demonstrating that it is possible to select neurofeedback protocols based on individual EEG biomarkers and suggests this results in improved treatment outcome specifically for ATT, however these results should be replicated in further controlled studies. A slow anterior iAPF at baseline predicts poor treatment response on comorbid depressive complaints in line with studies in depression. The effects of SMR neurofeedback resulted in specific ERP and EEG changes.

  18. Airborne gamma-ray spectrometer and magnetometer survey: Christmas Mountains, Texas Detail Area. Final report

    SciTech Connect

    Not Available

    1980-01-01

    Volume IIA contains the following: geology of Christmas Mountain; radioactive mineral occurrences in Texas; geophysical data interpretation; references; and appendices for stacked profiles, geologic histograms; geochemical histograms; speed and altitude histograms; geologic statistical tables; geochemical statistical tables; magnetic and ancillary profiles, and test line data. (ATT)

  19. Integrated circuit with dissipative layer for photogenerated carriers

    DOEpatents

    Myers, D.R.

    1989-09-12

    The sensitivity of an integrated circuit to single-event upsets is decreased by providing a dissi The U.S. Government has rights in this invention pursuant to Contract No. DE-ACO4-76DP00789 between the Department of Energy and AT&T Technologies, Inc.

  20. Integrated circuit with dissipative layer for photogenerated carriers

    DOEpatents

    Myers, David R.

    1989-01-01

    The sensitivity of an integrated circuit to single-event upsets is decreased by providing a dissi The U.S. Government has rights in this invention pursuant to Contract No. DE-ACO4-76DP00789 between the Department of Energy and AT&T Technologies, Inc.

  1. Erratum.

    PubMed

    1984-07-20

    In the credit for the photograph on page 1086 of the issue of 8 June accompanying the Research News article "Crystal anisotropy directs solidification" by Arthur L. Robinson (p. 1085), Kurt Nassau's affiliation is incorrectly given as Western Electric. Nassau is at AT&T Bell Laboratories, Murray Hill, New Jersey.

  2. Flood stress as a technicque to assess preventive insecticide and fungicide treatments for protecting trees against ambrosia beetles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The exotic ambrosia beetles Xylosandrus germanus, Xylosandrus crassiusculus, and Xylosandrus compactus tunnel into the heartwood of trees where they cultivate and feed upon a symbiotic fungus. We assessed the effectiveness of flood stress as a tactic for making trees attractive and vulnerable to att...

  3. New developments in food-based synthetic attractants for pest Tephritidae.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Optimal capture of several species of Anastrepha and Bactrocera spp. tephritid fruit flies is in traps baited with the aqueous protein bait Nulure combined with borax. Nulure is produced by acid hydrolysis of corn and has an acidic pH. Addition of borax makes the solution more alkaline and more att...

  4. The Slick Hair Coat Locus Maps to Bovine Chromosome 20 in Senepol Derived Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability to maintain normal temperatures during heat stress is an important attribute for cattle in the subtropics and tropics. Previous studies have shown that Senepol cattle and their crosses with the Holstein, Charolais and Angus breeds are as heat tolerant as Brahman cattle. This has been att...

  5. Telecommunications Policy Research Conference. Impacts of Telephone Deregulation Section. Papers.

    ERIC Educational Resources Information Center

    Telecommunications Policy Research Conference, Inc., Washington, DC.

    Three papers consider various aspects of the AT&T (American Telephone and Telegraph Company) court-ordered divestiture. The first paper, "InterLATA Toll Alternatives for the Bell Regional Holding Companies" (Elizabeth A. La Blanc, Ann M. Wolf, and Richard M. Wolf), examines six options available to the RHCs (regional holding companies) for…

  6. Coal slurry combustion and technology. Volume 2

    SciTech Connect

    Not Available

    1983-01-01

    Volume II contains papers presented at the following sessions of the Coal Slurry Combustion and Technology Symposium: (1) bench-scale testing; (2) pilot testing; (3) combustion; and (4) rheology and characterization. Thirty-three papers have been processed for inclusion in the Energy Data Base. (ATT)

  7. Probiotics and colostrum/milk differentially affect neonatal humoral immune responses to oral rotavirus vaccine.

    PubMed

    Chattha, Kuldeep S; Vlasova, Anastasia N; Kandasamy, Sukumar; Esseili, Malak A; Siegismund, Christine; Rajashekara, Gireesh; Saif, Linda J

    2013-04-01

    Breast milk (colostrum [col]/milk) components and gut commensals play important roles in neonatal immune maturation, establishment of gut homeostasis and immune responses to enteric pathogens and oral vaccines. We investigated the impact of colonization by probiotics, Lactobacillus rhamnosus GG (LGG) and Bifidobacterium lactis Bb12 (Bb12) with/without col/milk (mimicking breast/formula fed infants) on B lymphocyte responses to an attenuated (Att) human rotavirus (HRV) Wa strain vaccine in a neonatal gnotobiotic pig model. Col/milk did not affect probiotic colonization in AttHRV vaccinated pigs. However, unvaccinated pigs fed col/milk shed higher numbers of probiotic bacteria in feces than non-col/milk fed colonized controls. In AttHRV vaccinated pigs, col/milk feeding with probiotic treatment resulted in higher mean serum IgA HRV antibody titers and intestinal IgA antibody secreting cell (ASC) numbers compared to col/milk fed, non-colonized vaccinated pigs. In vaccinated pigs without col/milk, probiotic colonization did not affect IgA HRV antibody titers, but serum IgG HRV antibody titers and gut IgG ASC numbers were lower, suggesting that certain probiotics differentially impact HRV vaccine responses. Our findings suggest that col/milk components (soluble mediators) affect initial probiotic colonization, and together, they modulate neonatal antibody responses to oral AttHRV vaccine in complex ways. PMID:23453730

  8. Making Education and Training Work: Case Studies of Good Practice.

    ERIC Educational Resources Information Center

    Pollitt, David. Ed.

    2000-01-01

    Includes 12 case studies of education and training in various settings and countries: AT&T, Microsoft, and IBM; Maytag; Vauxhall; St. John Ambulance; McDonald's; Bayer; Newcastle (England) City Council; corporate universities; European management program; distance learning at Leuven University (Belgium); learning organizations; and technology…

  9. The UNIX Operating System in Libraries: Advantages of LAN's and Bulletin Board Systems in Your Library.

    ERIC Educational Resources Information Center

    Rose, Phillip E.

    1988-01-01

    Describes the experiences of the AT&T Technical Library in installing a local area network (LAN) and bulletin board using the UNIX operating system. Reasons why a LAN was needed, how the system works, and hardware and software used are discussed. (1 reference) (MES)

  10. Teachers Learning in Networked Communities. Phase I Evaluation Report

    ERIC Educational Resources Information Center

    Carroll, Tom; Fulton, Kathleen; Yoon, Irene

    2005-01-01

    In 2003 the National Commission on Teaching and America's Future convened a design team to launch the Teachers Learning in Networked Communities (TLINC) project. The initial one-year phase, funded by AT&T, involved a TLINC design team partnered with four communities, Pueblo, Colorado; Seattle, Washington; Portland, Maine; and Socorro, Texas. The…

  11. Schooling for Survival.

    ERIC Educational Resources Information Center

    Bowen, Ezra

    1985-01-01

    U.S. corporations are moving en masse into the learning business. A Carnegie Foundation study, "Corporate Classrooms: The Learning Business," is discussed, and programs run by AT&T, Wang, Bell and Howell, and the National Technological University are cited. (MLW)

  12. 32 CFR Attachment A to Subpart B... - Standard A-National Agency Check With Local Agency Checks and Credit Check (NACLC)

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Access Programs by sect. 4.4 of Executive Order 12958) (60 FR 19825, 3 CFR 1995 Comp., p. 333); (2) “L... Agency Checks and Credit Check (NACLC) A Attachment A to Subpart B of Part 147 National Defense..., Subpt. B, Att. A Attachment A to Subpart B of Part 147—Standard A—National Agency Check With...

  13. 32 CFR Attachment A to Subpart B... - Standard A-National Agency Check With Local Agency Checks and Credit Check (NACLC)

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Access Programs by sect. 4.4 of Executive Order 12958) (60 FR 19825, 3 CFR 1995 Comp., p. 333); (2) “L... Agency Checks and Credit Check (NACLC) A Attachment A to Subpart B of Part 147 National Defense..., Subpt. B, Att. A Attachment A to Subpart B of Part 147—Standard A—National Agency Check With...

  14. Changes Come as Old Hiring Practices Go.

    ERIC Educational Resources Information Center

    Journal of Career Planning & Employment, 1995

    1995-01-01

    Assembles a number of short takes that describe how some human resource professionals have changed their operations to upgrade their bottom line. Includes: Deloitte & Touche, Andersen Consulting, Baxter Healthcare Corporation, Brown & Root, AT&T, Ford Motor Company, and The Monsanto Company. Provides advice and tips. (JBJ)

  15. 75 FR 71464 - Metlife Technology, Operations, and Information Technology Groups Including On-Site Leased...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-23

    ... Workers From At&T Solutions, Chimes, Cognizant, Patni, Siemens, Xerox Clarks Summit, PA; Notice of Revised...,210) and MetLife, Technology, Operations, and Information Technology Groups, Clarks Summit... workers (TA-W-73,210) and the Clarks Summit, Pennsylvania facility, including on-site leased workers...

  16. 77 FR 3470 - Combined Notice of Filings #1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-24

    ... Energy Regulatory Commission Combined Notice of Filings 1 Take notice that the Commission received the... filing per 35: Att. L--MVP Compliance to be effective 1/ 1/2012. Filed Date: 1/12/12. Accession Number... effective 12/31/9998. Filed Date: 1/11/12. Accession Number: 20120111-5204. Comments Due: 5 p.m. ET...

  17. 77 FR 30272 - Combined Notice of Filings #1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-22

    ... Energy Regulatory Commission Combined Notice of Filings 1 Take notice that the Commission received the.... Filed Date: 5/1/12. Accession Number: 20120501-5447. Comments Due: 5 p.m. ET 5/22/12. Docket Numbers... of the PJM OATT Att K Appx & OA Schedule 1 to be effective 4/1/2012. Filed Date: 5/14/12....

  18. New Frontiers in Educational Technology: Trends and Issues. Proceedings of the Conference of the University/Urban Schools National Task Force (8th, San Francisco, California, November 14-15, 1986).

    ERIC Educational Resources Information Center

    Bossone, Richard M., Ed.; Polishook, Irwin H., Ed.

    The papers in this collection of presentations at a conference of the University/Urban Schools National Task Force address topics on trends in educational technology and issues in teaching with educational technology. The following papers are included: (1) "The Appropriate Use of Technology in Education" (James E. Dezell, Jr.); (2) AT&T and the…

  19. Estimation of Ligament Loading and Anterior Tibial Translation in Healthy and ACL-Deficient Knees During Gait and the Influence of Increasing Tibial Slope Using EMG-Driven Approach

    PubMed Central

    Shao, Qi; MacLeod, Toran D.; Manal, Kurt

    2010-01-01

    The purpose of this study was to develop a biomechanical model to estimate anterior tibial translation (ATT), anterior shear forces, and ligament loading in the healthy and anterior cruciate ligament (ACL)-deficient knee joint during gait. This model used electromyography (EMG), joint position, and force plate data as inputs to calculate ligament loading during stance phase. First, an EMG-driven model was used to calculate forces for the major muscles crossing the knee joint. The calculated muscle forces were used as inputs to a knee model that incorporated a knee–ligament model in order to solve for ATT and ligament forces. The model took advantage of using EMGs as inputs, and could account for the abnormal muscle activation patterns of ACL-deficient gait. We validated our model by comparing the calculated results with previous in vitro, in vivo, and numerical studies of healthy and ACL-deficient knees, and this gave us confidence on the accuracy of our model calculations. Our model predicted that ATT increased throughout stance phase for the ACL-deficient knee compared with the healthy knee. The medial collateral ligament functioned as the main passive restraint to anterior shear force in the ACL-deficient knee. Although strong co-contraction of knee flexors was found to help restrain ATT in the ACL-deficient knee, it did not counteract the effect of ACL rupture. Posterior inclination angle of the tibial plateau was found to be a crucial parameter in determining knee mechanics, and increasing the tibial slope inclination in our model would increase the resulting ATT and ligament forces in both healthy and ACL-deficient knees. PMID:20683675

  20. Abundance and length polymorphism of microsatellite repeats in Beta vulgaris L.

    PubMed

    Mörchen, M; Cuguen, J; Michaelis, G; Hänni, C; Saumitou-Laprade, P

    1996-03-01

    Simple sequence repeats (SSRs) are known to exhibit high degrees of variability even among closely related individuals. Their usage as nuclear genetic markers requires their conversion into sequence-tagged sites (STSs). In this paper we present the development of simple sequences as STSs for Beta vulgaris. This species comprises wild, cultivated, and weedy forms; the latter are thought to originate from accidental hybridisation between the other two. Two partial genomic libraries were screened with simple sequence motifs (AT, CA, CT, ATT, GTG, and CA, CT, respectively). Clones of 22 CA, nine CT, eight ATT, and one GTG sequence were obtained. AT micro satellites were present in compound motifs, not recognised by the probe. Sequence comparisons revealed that 20 CA clones containing short motifs (<16 bp) were variants of a previously described approximately 320-bp satellite DNA (Schmidt et al. 1991), and hence did not correspond to unique loci. Polymorphism of one (ATT)15 and three (CT)n, with n=15, 17 and 26, was detected by PCR on a sample of 64 plants from the different forms of B. vulgaris. 13 (ATT), 13 (CT), nine (CT) alleles and one (CT) allele were detected. One of the ATT alleles was much larger than the others (>800 bp). Genetic variability was high among wild beets, lower among cultivated beets, and intermediate among weed beets. One allele of each locus was found at high frequencies in cultivated beets and, to a lower extent, in weed beets. The combination of three polymorphic loci allowed the individual identification of 17/17 wild and 15/15 weed beets, and 21/32, mostly homozygous, cultivated beets.

  1. Relationship of anterior knee laxity to knee translations during drop landings: a bi-plane fluoroscopy study

    PubMed Central

    Myers, C.; Pennington, W. W.; Shelburne, K. B.; Krong, J. P.; Giphart, J. E.; Steadman, J. R.; Woo, Savio L-Y

    2014-01-01

    Purpose Passive anterior knee laxity has been linked to non-contact ACL injury risk. High deceleration movements have been implicated in the non-contact ACL injury mechanism, and evidence suggests that greater anterior tibial translations (ATT) may occur in healthy knees that are lax compared to a tight knee. The purpose of this study was to determine the relationship between anterior knee laxity scores and ATT during drop landings using biplane fluoroscopy. Methods Sixteen healthy adults (10 women; 6 men) performed stiff drop landings (40 cm) while being filmed using a high-speed, biplane fluoroscopy system. Initial, peak and excursions for rotations and translations were calculated and regression analysis used to determine the 6DoF kinematic relationships with KT1000 scores with peak ATT occurring during the landing. Results KT1000 values were (+) correlated with peak ATT values for group (r = 0.89; P < 0.0001) and both genders (males, r = 0.97; P = 0.0003; females, r = 0.93; P = < 0.0001). Regression analysis yielded a significant linear fit for the group (r2 = 0.80; YATT-group = −0.516 + 1.2 × XKT1000-group) and for each gender (females: r2 = 0.86; YATT-females = 0.074 + 1.2 × XKT1000-females and males: r2 = 0.94; YATT-males = −0.79 + 1.2 × XKT1000-males). Conclusion A strong relationship was observed between passive anterior knee laxity measured via KT1000 and peak ATT experienced during dynamic activity in otherwise healthy persons performing a stiff drop-landing motion. PMID:21153545

  2. Tuberculous Drug-induced Liver Injury and Treatment Re-challenge in Human Immunodeficiency Virus Co-infection

    PubMed Central

    Costiniuk, Cecilia T.; Gosnell, Bernadett I.; Manzini, Thandekile C.; Du Plessis, Camille N.; Moosa, Mahomed Yunus S.

    2015-01-01

    Background: Tuberculosis drug-induced liver injury (TB-DILI) is the most common adverse event necessitating therapy interruption. The optimal re-challenge strategy for antituberculous therapy (ATT) remains unclear, especially in human immunodeficiency virus (HIV) co-infected individuals in high-prevalence settings such as South Africa. Objective: To determine the incidence of and risk factors for the recurrence of TB-DILI with different ATT re-challenge strategies. Materials and Methods: We conducted a retrospective chart review of patients managed for TB-DILI from 2005 to 2013 at King Edward VIII Hospital in Durban, South Africa. Relevant clinical and laboratory data at the presentation of TB-DILI, time to recovery of liver function, method of ATT re-challenge and outcome of re-challenge were documented. Results: 1016 charts were reviewed, and 53 individuals with TB-DILI (48 HIV-co-infected) were identified. Following discontinuation of ATT, the median time to alanine aminotransferase normalization was 28 days (interquartile range 13-43). Forty-two subjects were re-challenged (30 regimen re-challenges and 12 step-wise re-challenges). 5 (12%) cases of recurrent TB-DILI were noted. Recurrences were not associated with the method of re-challenge. Conclusion: Based on the data available, it appears that full ATT can be safely restarted in the majority of subjects with a recurrence of DILI occurring in about 12% of subjects. The method of re-challenge did not appear to impact on the risk of recurrence. Ideally, a prospective randomized trial is needed to determine the best method of re-challenge. PMID:26752869

  3. ACL mismatch reconstructions: influence of different tunnel placement strategies in single-bundle ACL reconstructions on the knee kinematics.

    PubMed

    Herbort, Mirco; Lenschow, Simon; Fu, Freddie H; Petersen, Wolf; Zantop, Thore

    2010-11-01

    To evaluate the influence of tibial and femoral tunnel position in ACL reconstruction on knee kinematics, we compared ACL reconstruction with a tibial and femoral tunnel in anteromedial (AM-AM reconstruction) and in posterolateral footprint (PL-PL reconstruction) with a reconstruction technique with tibial posterolateral and femoral anteromedial tunnel placement (PL-AM reconstruction). In 9 fresh-frozen human cadaveric knees, the knee kinematics under simulated Lachman (134 N anterior tibial load) and a simulated pivot shift test (10 N/m valgus and 4 N/m internal tibial torque) were determined at 0°, 30°, 60°, and 90° of flexion. Kinematics were recorded for intact, ACL-deficient, and single-bundle ACL reconstructed knees using three different reconstruction strategies in randomized order: (1) PL-AM, (2) AM-AM and (3) PL-PL reconstructions. Under simulated Lachman test, single-bundle PL-AM reconstruction and PL-PL reconstructions both showed significantly increased anterior tibial translation (ATT) at 60° and 90° when compared to the intact knee. At all flexion angles, AM-AM reconstruction did not show any statistical significant differences in ATT compared to the intact knee. Under simulated pivot shift, PL-AM reconstruction resulted in significantly higher ATT at 0°, 30°, and 60° knee flexion and AM-AM reconstructions showed significantly higher ATT at 30° compared to the intact knee. PL-PL reconstructions did not show any significant differences to the intact knee. AM-AM reconstructions restore the intact knee kinematics more closely when compared to a PL-AM technique resembling a transtibial approach. PL-PL reconstructions showed increased ATT at higher flexion angles, however, secured the rotational stability at all flexion angles. Due to the independent tibial and femoral tunnel location, a medial portal technique may be superior to a transtibial approach. PMID:20461359

  4. Tuberculous optochiasmatic arachnoiditis.

    PubMed

    Anupriya, A; Sunithi, M; Maya, T; Goel, M; Alexander, M; Aaron, S; Mathew, V

    2010-01-01

    Arachnoiditis involving optic nerve and the optic chiasm can occur as a complication of tuberculous meningitis (TBM). This study evaluates the clinical features, cerebrospinal fluid (CSF) and laboratory parameters and imaging findings of optochiasmatic arachnoiditis (OCA) and also tried to identify any factors which can predict this complication in patients with TBM. Patients admitted with TBM in the neurology wards of a tertiary care teaching hospital over a period of 6 years formed the material for this study. Student's "t" test and univariate analysis were done to identify any predictors for this complication and the variables found to be significant were further analyzed by multivariate logistic regression analysis. One hundred sixty-three patients with TBM, admitted over a 6-year period, were studied. Twenty-three (14%) patients developed OCA. Eighteen out of 23 (78%) developed this complication while on antituberculous treatment (ATT) and 5/23 (22%) were newly diagnosed cases of TBM. Of those already on treatment, 12/23 (52%) were receiving only ATT, the remaining 6/23 (26%) had received steroids along with ATT in varying doses and duration. The average period from diagnosis of TBM to visual symptoms was 6.4 months. On the multivariate logistic regression analysis, female sex (P < 0.037), age less than 27 (P < 0.008) years and protein content in the CSF > 260 mg% (P < 0.021) were the factors predisposing toward this complication. At 6 months follow-up, on treatment with steroids and ATT, 17% had improvement and no further deterioration was noted in visual acuity in 52%. OCA can develop even while on treatment with ATT. Young women with a high CSF protein content seem to be more prone for this complication.

  5. Dual functions of Lactobacillus acidophilus NCFM™ at the intermediate dose in protection against rotavirus diarrhea in gnotobiotic pigs vaccinated with a human rotavirus vaccine

    PubMed Central

    Liu, Fangning; Wen, Ke; Li, Guohua; Yang, Xingdong; Kocher, Jacob; Bui, Tammy; Jones, Dorothy; Pelzer, Kevin; Clark-Deener, Sherrie; Yuan, Lijuan

    2014-01-01

    Objectives To examine dose effects of Lactobacillus acidophilus NCFM (LA) ™ strain on rotavirus-specific antibody and B cell responses in gnotobiotic pigs vaccinated with an oral attenuated human rotavirus (AttHRV). Methods Pigs were inoculated with AttHRV vaccine in conjunction with high dose LA (14 doses, total 2.2×109 colony forming units [CFU]), intermediate dose LA (9 doses, total 3.2×106 CFU), low dose LA (5 doses, total 2.1×106 CFU) or without LA feeding. Protection against rotavirus shedding and diarrhea was assessed upon challenge with a virulent HRV. Rotavirus-specific IgA and IgG antibodies in serum and rotavirus-specific IgA and IgG antibody-secreting cells (ASC) and memory B cells in ileum, spleen and blood of the pigs were measured and compared among treatment groups. Results The intermediate dose LA (MidLA), but not high or low dose LA, significantly reduced rotavirus diarrhea (MidLA only group) and significantly improved the protection conferred by AttHRV vaccine (MidLA+AttHRV group). Associated with the increased protection, MidLA significantly enhanced rotavirus-specific antibody, ASC and memory B cell responses to AttHRV vaccine. High or low dose LA did not enhance virus-specific antibody and ASC responses, hence did not improve the vaccine efficacy. Conclusions These findings highlight the importance of dose selection and indicate that certain specific lactobacilli strains at the appropriate dose have the dual function of reducing rotavirus diarrhea and enhancing the immunogenicity and protective efficacy of rotavirus vaccines. PMID:24126832

  6. Prophage, phiPV83-pro, carrying panton-valentine leukocidin genes, on the Staphylococcus aureus P83 chromosome: comparative analysis of the genome structures of phiPV83-pro, phiPVL, phi11, and other phages.

    PubMed

    Zou, D; Kaneko, J; Narita, S; Kamio, Y

    2000-12-01

    Staphylococcus aureus P83 has Panton-Valentine leukocidin (PVL)-like genes, lukM and lukF-PV. Here, lukM and lukF-PV genes were found on the genome of a prophage, which was designated as phiPV83-pro. The precise genome size was 45,636 bp with att core sequences of 10 base pairs. Sixty-four ORFs were identified on the phiPV83-pro genome, including two extra operons, lukM-lukF-PV and orfs63-64. The lukM-lukF-PV cluster was located 2.1 kb upstream of the attL site. The most striking feature of the phiPV83-pro genome was a constituent of at least 4 regions from phi11, phiPVL, and other phages, i.e., (i) att sites identical with those of phi11, (ii) a cos sequence and the genes encoding packaging and head proteins of phiPVL (occupied half region of phiPV83-pro), and (iii) the other two regions which showed no significant similarity with known phages (occupied about 40% of phiPV83-pro). Furthermore, two insertion sequences, ISSA1 and ISSA2 were integrated into attL site and orf44, respectively. PhiPV83-pro was not induced as phage particles from S. aureus P83 regardless of its treatment with mitomycin C. The insertion of ISSA1 into the attL site was one of the reasons of the failure of the induction of the phage particles by mitomycin C treatment of the strain P83.

  7. Lactobacilli and Bifidobacteria enhance mucosal B cell responses and differentially modulate systemic antibody responses to an oral human rotavirus vaccine in a neonatal gnotobiotic pig disease model

    PubMed Central

    Kandasamy, Sukumar; Chattha, Kuldeep S; Vlasova, Anastasia N; Rajashekara, Gireesh; Saif, Linda J

    2014-01-01

    B cells play a key role in generation of protective immunity against rotavirus infection, a major cause of gastroenteritis in children. Current RV vaccines are less effective in developing countries compared to developed countries. Commensals/probiotics influence mucosal immunity, but the role of early gut colonizing bacteria in modulating intestinal B cell responses to RV vaccines is largely unknown. We co-colonized neonatal gnotobiotic pigs, the only animal model susceptible to HRV diarrhea, with 2 dominant bacterial species present in the gut of breastfed infants, Lactobacillus rhamnosus strain GG and Bifidobacterium animalis lactis Bb12 to evaluate their impact on B cell responses to an attenuated (Att) human rotavirus (HRV) Wa strain vaccine. Following HRV challenge, probiotic-colonized, AttHRV vaccinated piglets had significantly lower fecal scores and reduced HRV shedding titers compared to uncolonized, AttHRV vaccinated pigs. The reduction in HRV diarrhea was significantly correlated with higher intestinal IgA HRV antibody titers and intestinal HRV-specific IgA antibody secreting cell (ASC) numbers in probiotic-colonized, AttHRV vaccinated pigs compared to uncolonized, vaccinated pigs. The significantly higher small intestinal HRV IgA antibody responses coincided with higher IL-6, IL-10 and APRIL responses of ileal mononuclear cells (MNCs) and the immunomodulatory effects of probiotics genomic DNA on TGF-β and IL-10 responses. However, serum RV IgG antibody titers and total IgG titers were significantly lower in probiotic-colonized, AttHRV vaccinated pigs compared to uncolonized, vaccinated pigs, both pre- and post-challenge. In summary, LGG and Bb12 beneficially modulated intestinal B cell responses to HRV vaccine. PMID:25483333

  8. Lactobacilli and Bifidobacteria enhance mucosal B cell responses and differentially modulate systemic antibody responses to an oral human rotavirus vaccine in a neonatal gnotobiotic pig disease model.

    PubMed

    Kandasamy, Sukumar; Chattha, Kuldeep S; Vlasova, Anastasia N; Rajashekara, Gireesh; Saif, Linda J

    2014-01-01

    B cells play a key role in generation of protective immunity against rotavirus infection, a major cause of gastroenteritis in children. Current RV vaccines are less effective in developing countries compared to developed countries. Commensals/probiotics influence mucosal immunity, but the role of early gut colonizing bacteria in modulating intestinal B cell responses to RV vaccines is largely unknown. We co-colonized neonatal gnotobiotic pigs, the only animal model susceptible to HRV diarrhea, with 2 dominant bacterial species present in the gut of breastfed infants, Lactobacillus rhamnosus strain GG and Bifidobacterium animalis lactis Bb12 to evaluate their impact on B cell responses to an attenuated (Att) human rotavirus (HRV) Wa strain vaccine. Following HRV challenge, probiotic-colonized, AttHRV vaccinated piglets had significantly lower fecal scores and reduced HRV shedding titers compared to uncolonized, AttHRV vaccinated pigs. The reduction in HRV diarrhea was significantly correlated with higher intestinal IgA HRV antibody titers and intestinal HRV-specific IgA antibody secreting cell (ASC) numbers in probiotic-colonized, AttHRV vaccinated pigs compared to uncolonized, vaccinated pigs. The significantly higher small intestinal HRV IgA antibody responses coincided with higher IL-6, IL-10 and APRIL responses of ileal mononuclear cells (MNCs) and the immunomodulatory effects of probiotics genomic DNA on TGF-β and IL-10 responses. However, serum RV IgG antibody titers and total IgG titers were significantly lower in probiotic-colonized, AttHRV vaccinated pigs compared to uncolonized, vaccinated pigs, both pre- and post-challenge. In summary, LGG and Bb12 beneficially modulated intestinal B cell responses to HRV vaccine. PMID:25483333

  9. 6-trifluoromethyl-2-thiouracil possesses anti-Toxoplasma gondii effect in vitro and in vivo with low hepatotoxicity.

    PubMed

    Choi, Hwa-Jung; Yu, Seung-Taek; Lee, Kee-In; Choi, Joong-Kwon; Chang, Bo-Yoon; Kim, Sung-Yeon; Ko, Mi-Hwa; Song, Hyun-Ok; Park, Hyun

    2014-08-01

    Since pyrimethamine, the general therapeutic drug for toxoplasmosis, presents several adverse side effects, the need to develop and evaluate new drugs for the condition is critical. In this study, anti-Toxoplasma gondii activities of 3-[{2-((E)-furan-2-ylmethylene)hydrazinyl}methylene]-1,3-dihydroindol-2-one (ATT-5126) and 6-trifluoromethyl-2-thiouracil (KH-0562) were evaluated in vitro using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and in vivo by measuring amount of the tachyzoites in mice ascites. Biochemical parameters such as lipid peroxidation (LPO), glutathione (GSH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were also evaluated in livers of mice at 4 days post-infection. As a result, the ATT-5126 and KH-0562 showed anti-T. gondii activity in vitro. Treatment of ATT-5126 or KH-0562 decreased the amount of tachyzoites in T. gondii infected ICR mice. The relative weight of liver and spleen increased by T. gondii infection were decreased by treatment of ATT-5126 or KH-0562. The levels of LPO, ALT and AST, which are biochemical parameters involved in liver injury, were also significantly recovered by treatment of ATT-5126 or KH-0562 (p<0.05). In particular, the recovered levels by KH-0562 were similar to those of pyrimethamine-treated group (p<0.05). However, the level of GSH, which is an antioxidant indicator, showed insignificant statistics. The results suggest that KH-0562 show anti-T. gondii activities in vitro and in vivo with low hepatotoxicity. Therefore, KH-0562 may be a useful candidate for treating T. gondii infection. PMID:24830752

  10. [Expression, purification of recombinant cationic peptide AIK in Escherichia coli and its antitumor activity].

    PubMed

    Fan, Fangfang; Sun, Huiying; Xu, Hui; Liu, Jiawei; Zhang, Haiyuan; Li, Yilan; Ning, Xuelian; Sun, Yue; Bai, Jing; Fu, Songbin; Zhou, Chunshui

    2015-12-01

    AIK is a novel cationic peptide with potential antitumor activity. In order to construct the AIK expression vector by Gateway technology, and establish an optimal expression and purification method for recombinant AIK, a set of primers containing AttB sites were designed and used to create the AttB-TEV-FLAG-AIR fusion gene by overlapping PCR. The resulting fusion gene was cloned into the donor vector pDONR223 by attB and attP mediated recombination (BP reaction), then, transferred into the destination vector pDESTl 5 by attL and attR mediated recombination (LR reaction). All the cloning was verified by both colony PCR and DNA sequencing. The BL21 F. coli transformed by the GST-AIR expression plasmid was used to express the GST-AIK fusion protein with IPTG induction and the induction conditions were optimized. GST-AIR fusion protein was purified by glutathione magnetic beads, followed by rTEV cleavage to remove GST tag and MTS assay to test the growth inhibition activity of the recombinant AIR on human leukemia HL-60 cells. We found that a high level of soluble expression of GST-AIK protein (more than 30% out of the total bacterial proteins) was achieved upon 0.1 mmol/L ITPG induction for 4 h at 37 °C in the transformed BL21 F. coli with starting OD₆₀₀ at 1.0. Through GST affinity purification and rTEV cleavage, the purity of the resulting recombinant AIK was greater than 95%. And the MTS assays on HL-60 cells confirmed that the recombinant AIK retains an antitumor activity at a level similar to the chemically synthesized AIK. Taken together, we have established a method for expression and purification of recombinant AIK with a potent activity against tumor cells, which will be beneficial for the large-scale production and application of recombinant AIK in the future. PMID:27093838

  11. Cold inducible RNA binding protein upregulation in pituitary corticotroph adenoma induces corticotroph cell proliferation via Erk signaling pathway

    PubMed Central

    Fu, Wei; Tang, Hao; Chen, Xiao; Zhao, Yao; Zheng, Lili; Pan, Sijian; Wang, Weiqing; Bian, Liuguan; Sun, Qingfang

    2016-01-01

    Cushing's disease is caused by pituitary corticotroph adenoma, and the pathogenesis of it has remained obscure. Here, we showed that cold inducible RNA binding protein (CIRP) was markedly elevated in corticotroph tumors. Forced overexpression of CIRP in murine AtT20 pituitary corticotroph cell line increased corticotroph precursor hormone proopiomelanocortin (POMC) transcription, ACTH secretion and cellular proliferation. In vivo, CIRP overexpression promotes murine corticotroph tumor growth and enhances ACTH production. Mechanistically, we show that CIRP could promote AtT20 cells proliferation by inducing cyclinD1 and decreasing p27 expression via Erk1/2 signaling pathway. Clinically, CIRP overexpression is significantly correlated with Cushing's disease recurrence. CIRP appears to play a critical tumorigenesis function in Cushing's disease and its expression might be a useful biomarker for tumor recurrence. PMID:26824322

  12. A signal sequence is sufficient for green fluorescent protein to be routed to regulated secretory granules.

    PubMed

    El Meskini, R; Jin, L; Marx, R; Bruzzaniti, A; Lee, J; Emeson, R; Mains, R

    2001-02-01

    To investigate trafficking in neuroendocrine cells, green fluorescent protein (GFP) tags were fused to various portions of the preproneuropeptide Y (NPY) precursor. Two neuroendocrine cell lines, AtT-20 corticotrope tumor cells and PC-12 pheochromocytoma cells, along with primary anterior pituitary cells, were examined. Expression of chimeric constructs did not disrupt trafficking or regulated secretion of endogenous ACTH and prohormone convertase 1 in AtT-20 cells. Western blot and immunocytochemical analyses demonstrated that the chimeric constructs remained intact, as long as the Lys-Arg cleavage site within preproNPY was deleted. GFP was stored in, and released from, regulated granules in cells expressing half of the NPY precursor fused to GFP, and also in cells in which only the signal sequence of preproNPY was fused to GFP. Thus, in neuroendocrine cells, entering the lumen of the secretory pathway is sufficient to target GFP to regulated secretory granules.

  13. Timing of antiretroviral therapy and TB treatment outcomes in patients with TB-HIV in Myanmar.

    PubMed

    Thi, A M; Shewade, H D; Kyaw, N T T; Oo, M M; Aung, T K; Aung, S T; Oo, H N; Win, T; Harries, A D

    2016-06-21

    Contexte : Programme intégré de prise en charge du virus de l'immunodéficience humaine (VIH), Mandalay, Myanmar.Objectifs : Chez les patients atteints de tuberculose (TB) et VIH enrôlés entre 2011 et 2014, déterminer la date du début du traitement antirétroviral (TAR) en relation avec le traitement antituberculeux (ATT) et son association avec le résultat d'ATT.Schéma : Etude rétrospective de cohorte.Résultats : Sur 1708 patients TB-VIH, 1565 (92%) ont débuté l'ATT en premier et 143 (8%) ont commencé le TAR en premier. Le résultat du traitement a été manquant pour 226 patients qui n'ont pas été inclus. Chez les patients ayant débuté l'ATT en premier, le délai médian de mise en route du TAR a été de 8,6 semaines. L'initiation du TAR a été retardée d'un délai médian de 8 semaines chez 830 (53%) patients. Parmi ces patients, 7% ont eu un résultat médiocre, avec une anémie qui a constitué un facteur de risque indépendant. Chez les patients ayant débuté le TAR en premier, le délai médian de mise en route de l'ATT a été de 21,6 semaines. L'ATT a été initié au cours des 3 mois chez 56 (39%) patients. Le traitement a échoué chez 12% des patients et chez 20% de ceux qui ont débuté l'ATT dans les 3 mois. Les patients ayant des CD4 <100/mm(3) ont eu un risque quatre fois plus élevé d'échec.Conclusions: La chronologie du TAR en rapport avec l'ATT n'a pas été un facteur de risque indépendant d'échec du traitement. Un dépistage extensif de la TB avec des tests de diagnostic rapides et sensibles chez les personnes infectées par le VIH et un suivi étroit de l'anémie et de l'immunosuppression sont recommandés afin d'améliorer encore le résultat du traitement de TB parmi les patients TB-VIH.

  14. Determining average path length and average trapping time on generalized dual dendrimer

    NASA Astrophysics Data System (ADS)

    Li, Ling; Guan, Jihong

    2015-03-01

    Dendrimer has wide number of important applications in various fields. In some cases during transport or diffusion process, it transforms into its dual structure named Husimi cactus. In this paper, we study the structure properties and trapping problem on a family of generalized dual dendrimer with arbitrary coordination numbers. We first calculate exactly the average path length (APL) of the networks. The APL increases logarithmically with the network size, indicating that the networks exhibit a small-world effect. Then we determine the average trapping time (ATT) of the trapping process in two cases, i.e., the trap placed on a central node and the trap is uniformly distributed in all the nodes of the network. In both case, we obtain explicit solutions of ATT and show how they vary with the networks size. Besides, we also discuss the influence of the coordination number on trapping efficiency.

  15. Restructuring the US telecommunications industry - Impact on innovation

    NASA Technical Reports Server (NTRS)

    Agnew, C. E.; Romeo, A. A.

    1981-01-01

    The Communications Act of 1934, which regulates the United States telecommunications industry, is becoming less able to deal with the changes brought by modern technology. Therefore, proposals are being made to restructure the industry. Breaking up AT&T into smaller firms would have only minor effects, since its ratio of R and D to sales has been near the median for large telephone companies. Restructuring AT&T into subsidiaries dealing at arms' length might cause a reordering of priorities toward more directly marketable and useful products, but too extensive a break-up could endanger Bell Labs' commitment to basic research. Regulation leads a firm to ignore true social value of possible products, expand into low profit markets, and favor capital-intensive over labor-intensive innovation. If regulatory lag occurs, a firm will produce cost-reducing innovation in order to increase its profit.

  16. [Construction of RNAi vectors for SmNAC1 transcription factors of Salvia miltiorrhiza using Gateway cloning technology].

    PubMed

    Zhao, Rong; Rong, Qi-Xian; Liu, Yu-Zhong; Shen, Ye; Huang, Lu-Qi

    2014-05-01

    NAC transcription factors involved in plant growth and development, as well as responses to biotic and abiotic stress. RNAi Vectors for SmNAC transcription factors of Salvia miltiorrhiza was constructed by using Gateway cloning technology, in order to further study the function of SmNAC1 transcription factor. According to Gateway cloning technology, the specific fragments of SmNAC1 containing attB adapter was amplified by PCR using ultra-fideling phusion polymerase of NEB. By the BP recombination reaction, the PCR product containing attB was transferred to an donor vector (pENTR/SD/D-TOPO). Finally, SmNACi specific gene was cloned into pK7GWIWG2D plant expression vectors by LR recombination reaction. Experimental results showed that Gateway cloning technology provide a rapid and highly efficient way to clone the interested gene. PMID:25095362

  17. Bilateral Presumed Tuberculous Choroiditis

    PubMed Central

    Mohammadi, Naseh; Ghassemi, Fariba; Shojaei, Esfandiar; Moradnejad, Pardis

    2016-01-01

    Purpose: To describe a case of bilateral presumed tuberculous choroiditis which resolved promptly with a short course of antituberculous therapy. Case Report: A 44-year-old lady presented with bilateral visual loss of 8 months’duration. Considering the presence of pan-uveitis, severe macular edema, multifocal leakage on fluorescein angiography, positive tuberculin skin test, and after excluding other diseases, she underwent anti-tuberculous therapy (ATT) for disseminated choroiditis due to presumed ocular tuberculosis. She improved promptly and completely. There were no signs of relapse 18 months after completion of the treatment course. In a young patient with disseminated choroiditis and relatively short duration of symptoms, a course of ATT for 6 months may effectively treat the disease preventing relapse for a considerable period of time. Conclusion: In Presumed Tuberculous chroiditis early diagnosis is crucial for prompt and complete improvement. PMID:27413507

  18. Permanent genetic memory with >1 byte capacity

    PubMed Central

    Yang, Lei; Nielsen, Alec A.K.; Fernandez-Rodriguez, Jesus; McClune, Conor J.; Laub, Michael T.; Lu, Timothy K.; Voigt, Christopher A.

    2014-01-01

    Genetic memory enables the recording of information in the DNA of living cells. Memory can record a transient environmental signal or cell state that is then recalled at a later time. Permanent memory is implemented using irreversible recombinases that invert the orientation of a unit of DNA, corresponding to the [0,1] state of a bit. To expand the memory capacity, we have applied bioinformatics to identify 34 phage integrases (and their cognate attB and attP recognition sites), from which we build 11 memory switches that are perfectly orthogonal to each other and the FimE and HbiF bacterial invertases. Using these switches, a memory array is constructed in Escherichia coli that can record 1.375 bytes of information. It is demonstrated that the recombinases can be layered and used to permanently record the transient state of a transcriptional logic gate. PMID:25344638

  19. Determination of genome size of Pseudomonas aeruginosa by PFGE: analysis of restriction fragments.

    PubMed Central

    Hector, J S; Johnson, A R

    1990-01-01

    Genomic DNA size was measured in three strains of Pseudomonas aeruginosa, ATCC 29260 (exotoxin A), ATCC 33467 (type I smooth) and ATCC 33468 (type 2 mucoid) by transverse alternating field electrophoresis of restriction fragments. Because of the high (67%) G + C content of Pseudomonas aeruginosa, restriction enzymes that recognize sequences with at least 4 AT base pairs were expected to be rare cutters. Eight enzymes produced fragments greater than 200 kb in size: Dral (TTT/AAA), Asnl (ATT/AAT), Hpal (GTT/AAC), AfIII (C/TTAAG), Xbal (T/CTAGA), Spel (A/CTAGT), Sspl (AAT/ATT) and Ndel (CA/TATG). All eight enzymes recognized one of three rare tetranucleotide sequences, TTAA, CTAG or ATAT. Pseudomonas aeruginosa strain 29260 has a genomic DNA size of 5573 kb. Strains 33467 and 33468 have identical restriction patterns and a possible deletion with a genomic size of 5407 kb. Images PMID:1972559

  20. Quantitative assessment of cerebral hemodynamic parameters by QUASAR arterial spin labeling in Alzheimer's disease and cognitively normal Elderly adults at 3-tesla.

    PubMed

    Mak, Henry K F; Chan, Queenie; Zhang, Zhipeng; Petersen, Esben T; Qiu, Deqiang; Zhang, Linda; Yau, Kelvin K W; Chu, Leung-Wing; Golay, Xavier

    2012-01-01

    QUASAR arterial spin labeling (ASL) was used to investigate the role of vascular impairment in Alzheimer's disease (AD). We hypothesized that the hemodynamic parameters monitoring cerebrovascular integrity, i.e., cerebral blood flow (CBF), arterial blood volume (aBV), and arterial transit time (aTT), would be affected. 13 AD patients and 15 healthy control (HC) subjects underwent 3T MRI scanning. Two separate blood flow acquisitions were obtained with 1 slice overlap for whole brain coverage. CBF, aBV, and aTT maps were calculated using in-house software. Preprocessing and statistical analyses were performed on SPM5. Region-of-interest (ROI) studies of ten selected cerebral regions were also conducted. There were significant differences in mini mental status exam (MMSE) (AD: 16.3 ± 4.55, HC: 28.5 ± 2.00) and Alzheimer's disease assessment scale-cognitive subscale (ADAS-cog) scores (AD: 25.25 ± 9.64, HC: 5.51 ± 2.62) between the 2 groups (p < 0.001) but none in age (p = 0.068). CBF decreased significantly (p < 0.01) in AD compared to controls in the right middle cingulate, left cuneus, left inferior and middle frontal, right superior frontal, left inferior parietal, and right supramarginal gyri. ROI studies confirmed significant hemodynamic impairments in AD compared to HC (p < 0.05): CBF in middle and posterior cingulate, aBV in left superior temporal, right inferior parietal, and posterior cingulate, and aTT in left inferior frontal and middle cingulate gyri. CBF correlated positively while aTT correlated negatively to MMSE, and vice versa for ADAS-cog. Using QUASAR ASL, we found patterns of regional hemodynamic impairment typical of moderate AD, suggesting underlying vascular abnormality. As potential biomarkers, these hemodynamic parameters could differentiate patients from volunteers, and possibly indicate the conversion from healthy aging to mild cognitive impairment to AD.

  1. Mouse embryonic stem cells with a multi-integrase mouse artificial chromosome for transchromosomic mouse generation.

    PubMed

    Yoshimura, Yuki; Nakamura, Kazuomi; Endo, Takeshi; Kajitani, Naoyo; Kazuki, Kanako; Kazuki, Yasuhiro; Kugoh, Hiroyuki; Oshimura, Mitsuo; Ohbayashi, Tetsuya

    2015-08-01

    The mouse artificial chromosome (MAC) has several advantages as a gene delivery vector, including stable episomal maintenance of the exogenous genetic material and the ability to carry large and/or multiple gene inserts including their regulatory elements. Previously, a MAC containing multi-integration site (MI-MAC) was generated to facilitate transfer of multiple genes into desired cells. To generate transchromosomic (Tc) mice containing a MI-MAC with genes of interest, the desired genes were inserted into MI-MAC in CHO cells, and then the MI-MAC was transferred to mouse embryonic stem (mES) cells via microcell-mediated chromosome transfer (MMCT). However, the efficiency of MMCT from CHO to mES cells is very low (<10(-6)). In this study, we constructed mES cell lines containing a MI-MAC vector to directly insert a gene of interest into the MI-MAC in mES cells via a simple transfection method for Tc mouse generation. The recombination rate of the GFP gene at each attachment site (FRT, PhiC31attP, R4attP, TP901-1attP and Bxb1attP) on MI-MAC was greater than 50% in MI-MAC mES cells. Chimeric mice with high coat colour chimerism were generated from the MI-MAC mES cell lines and germline transmission from the chimera was observed. As an example for the generation of Tc mice with a desired gene by the MI-MAC mES approach, a Tc mouse strain ubiquitously expressing Emerald luciferase was efficiently established. Thus, the findings suggest that this new Tc strategy employing mES cells and a MI-MAC vector is efficient and useful for animal transgenesis.

  2. Mouse embryonic stem cells with a multi-integrase mouse artificial chromosome for transchromosomic mouse generation.

    PubMed

    Yoshimura, Yuki; Nakamura, Kazuomi; Endo, Takeshi; Kajitani, Naoyo; Kazuki, Kanako; Kazuki, Yasuhiro; Kugoh, Hiroyuki; Oshimura, Mitsuo; Ohbayashi, Tetsuya

    2015-08-01

    The mouse artificial chromosome (MAC) has several advantages as a gene delivery vector, including stable episomal maintenance of the exogenous genetic material and the ability to carry large and/or multiple gene inserts including their regulatory elements. Previously, a MAC containing multi-integration site (MI-MAC) was generated to facilitate transfer of multiple genes into desired cells. To generate transchromosomic (Tc) mice containing a MI-MAC with genes of interest, the desired genes were inserted into MI-MAC in CHO cells, and then the MI-MAC was transferred to mouse embryonic stem (mES) cells via microcell-mediated chromosome transfer (MMCT). However, the efficiency of MMCT from CHO to mES cells is very low (<10(-6)). In this study, we constructed mES cell lines containing a MI-MAC vector to directly insert a gene of interest into the MI-MAC in mES cells via a simple transfection method for Tc mouse generation. The recombination rate of the GFP gene at each attachment site (FRT, PhiC31attP, R4attP, TP901-1attP and Bxb1attP) on MI-MAC was greater than 50% in MI-MAC mES cells. Chimeric mice with high coat colour chimerism were generated from the MI-MAC mES cell lines and germline transmission from the chimera was observed. As an example for the generation of Tc mice with a desired gene by the MI-MAC mES approach, a Tc mouse strain ubiquitously expressing Emerald luciferase was efficiently established. Thus, the findings suggest that this new Tc strategy employing mES cells and a MI-MAC vector is efficient and useful for animal transgenesis. PMID:26055730

  3. Versatile P(acman) BAC Libraries for Transgenesis Studies in Drosophila melanogaster

    SciTech Connect

    Venken, Koen J.T.; Carlson, Joseph W.; Schulze, Karen L.; Pan, Hongling; He, Yuchun; Spokony, Rebecca; Wan, Kenneth H.; Koriabine, Maxim; de Jong, Pieter J.; White, Kevin P.; Bellen, Hugo J.; Hoskins, Roger A.

    2009-04-21

    We constructed Drosophila melanogaster BAC libraries with 21-kb and 83-kb inserts in the P(acman) system. Clones representing 12-fold coverage and encompassing more than 95percent of annotated genes were mapped onto the reference genome. These clones can be integrated into predetermined attP sites in the genome using Phi C31 integrase to rescue mutations. They can be modified through recombineering, for example to incorporate protein tags and assess expression patterns.

  4. Rare Offshoot of a Common Malady Anaemia and Tuberculosis

    PubMed Central

    Acharya, Vishak; Balanthimogru, Prashantha; Mani, Arun; Ruman, Shehzad

    2016-01-01

    Haematological manifestations are one of the rarer presentations of tuberculosis and are usually of normocytic normochromic type. An association of Autoimmune Haemolytic Anaemia (AIHA) with active pulmonary tuberculosis is an exceeding rare entity, though anaemia and tuberculosis commonly co-exist. We report a patient with sputum negative pulmonary tuberculosis with associated Coomb’s positive AIHA. The patient responded well to Anti- Tubercular Therapy (ATT) and low dose steroids tapered over a month. PMID:27656489

  5. A comparative study of efficacy and safety of febuxostat and allopurinol in pyrazinamide-induced hyperuricemic tubercular patients

    PubMed Central

    Pichholiya, Meenu; Yadav, Arvind Kumar; Luhadia, S. K.; Tahashildar, Jameela; Aseri, M. L.

    2016-01-01

    Objectives: To compare the efficacy and safety of febuxostat and allopurinol in pyrazinamide (PZA)-induced hyperuricemia in patients taking antitubercular therapy (ATT). Methods: This randomized controlled study was conducted at a tertiary care teaching institute of Rajasthan in all the sputum-positive tubercular patients aged between 18 and 65 years of either sex. Serum uric acid level was monitored at 0th, 2nd, 4th, 6th, and 8th week of ATT. Patients whose uric acid level was found to be increased at 2nd week were finally recruited in the study. Ninety patients who developed hyperuricemia due to ATT were divided randomly into three groups (Group A - febuxostat, Group B - allopurinol, and Group C - control) of thirty patients each. Mean serum uric acid levels were calculated at all the weeks in all the groups, and serum uric acid levels were compared by applying student's t-test and ANOVA. Results: Mean serum uric acid level decreased from 10.698 mg/dl (at 2nd week) to 7.846 mg/dl (at 8th week) in Group A and from 11.34 mg/dl (at 2nd week) to 7.280 mg/dl (at 8th week) in Group B. Numbers of adverse events encountered across both the treatment groups were same with both the drugs. Conclusion: Allopurinol and febuxostat were equally efficacious in lowering PZA induced raised serum uric acid level in tubercular patients, and it was possible to continue ATT without withdrawing PZA. PMID:27721537

  6. Multibody correlations in the hydrophobic solvation of glycine peptides

    SciTech Connect

    Harris, Robert C.; Drake, Justin A.; Pettitt, B. Montgomery

    2014-12-14

    Protein collapse during folding is often assumed to be driven by a hydrophobic solvation energy (ΔG{sub vdw}) that scales linearly with solvent-accessible surface area (A). In a previous study, we argued that ΔG{sub vdw}, as well as its attractive (ΔG{sub att}) and repulsive (ΔG{sub rep}) components, was not simply a linear function of A. We found that the surface tensions, γ{sub rep}, γ{sub att}, and γ{sub vdw}, gotten from ΔG{sub rep}, ΔG{sub att}, and ΔG{sub vdw} against A for four configurations of deca-alanine differed from those obtained for a set of alkanes. In the present study, we extend our analysis to fifty decaglycine structures and atomic decompositions. We find that different configurations of decaglycine generate different estimates of γ{sub rep}. Additionally, we considered the reconstruction of the solvation free energy from scaling the free energy of solvation of each atom type, free in solution. The free energy of the isolated atoms, scaled by the inverse surface area the atom would expose in the molecule does not reproduce the γ{sub rep} for the intact decaglycines. Finally, γ{sub att} for the decaglycine conformations is much larger in magnitude than those for deca-alanine or the alkanes, leading to large negative values of γ{sub vdw} (−74 and −56 cal/mol/Å{sup 2} for CHARMM27 and AMBER ff12sb force fields, respectively). These findings imply that ΔG{sub vdw} favors extended rather than compact structures for decaglycine. We find that ΔG{sub rep} and ΔG{sub vdw} have complicated dependencies on multibody correlations between solute atoms, on the geometry of the molecular surface, and on the chemical identities of the atoms.

  7. Abdominal Lymphonodular Cryptococcosis in an Immunocompetent Child

    PubMed Central

    Zaidi, Mehjabeen; Qureshi, Sonia; Shakoor, Sadia; Fatima, Saira; Mir, Fatima

    2015-01-01

    We describe our experience with an apparently immunocompetent child presenting with pyrexia of unknown origin without focal signs. Investigations revealed lymphadenopathy at lung hila, mesentery, and porta hepatis. The child had received at least two months of empiric antituberculous therapy (ATT) before she came to us. A CT-guided biopsy revealed granulomatous inflammation. PAS stain showed yeasts which stained blue with Alcian blue, suggesting C. neoformans. PMID:26649217

  8. Switching the polarity of a bacteriophage integration system.

    PubMed

    Smith, Matthew C A; Till, Rob; Smith, Margaret C M

    2004-03-01

    During lysogenic growth many temperate bacteriophage genomes are integrated into the host's chromosome and efficient integration and excision are therefore an essential part of the phage life cycle. The Streptomyces phage phiC31 encodes an integrase related to the resolvase/invertases and is evolutionarily and mechanistically distinct from the integrase of phage lambda. We show that during phiC31 integration the polarity of the recombination sites, attB and attP, is dependent on the sequences of the two base pairs (bp) where crossover occurs. A loss or switch in polarity of the recombination sites can occur by mutation of this dinucleotide, leading to incorrectly joined products. The properties of the mutant sites implies that phiC31 integrase interacts symmetrically with the substrates, which during synapsis can align apparently freely in either of two alternative forms that lead to correct or incorrect joining of products. Analysis of the topologies of the reaction products provided evidence that integrase can synapse and activate strand exchange even when recombinant products cannot form due to mismatches at the crossover site. The topologies of the recombination products are complex and indicative of multiple pathways to product formation. The efficiency of integration of a phiC31 derivative, KC859, into an attB site with switched polarity was assayed in vivo and shown to be no different from integration into a wild-type attB. Thus neither the host nor KC859 express a factor that influences the alignment of the recombination sites at synapsis.

  9. Fitness Cost Implications of PhiC31-Mediated Site-Specific Integrations in Target-Site Strains of the Mexican Fruit Fly, Anastrepha ludens (Diptera: Tephritidae)

    PubMed Central

    Meza, José S.; Díaz-Fleischer, Francisco; Sánchez-Velásquez, Lázaro R.; Zepeda-Cisneros, Cristina Silvia; Handler, Alfred M.; Schetelig, Marc F.

    2014-01-01

    Site-specific recombination technologies are powerful new tools for the manipulation of genomic DNA in insects that can improve transgenesis strategies such as targeting transgene insertions, allowing transgene cassette exchange and DNA mobilization for transgene stabilization. However, understanding the fitness cost implications of these manipulations for transgenic strain applications is critical. In this study independent piggyBac-mediated attP target-sites marked with DsRed were created in several genomic positions in the Mexican fruit fly, Anastrepha ludens. Two of these strains, one having an autosomal (attP_F7) and the other a Y-linked (attP_2-M6y) integration, exhibited fitness parameters (dynamic demography and sexual competitiveness) similar to wild type flies. These strains were thus selected for targeted insertion using, for the first time in mexfly, the phiC31-integrase recombination system to insert an additional EGFP-marked transgene to determine its effect on host strain fitness. Fitness tests showed that the integration event in the int_2-M6y recombinant strain had no significant effect, while the int_F7 recombinant strain exhibited significantly lower fitness relative to the original attP_F7 target-site host strain. These results indicate that while targeted transgene integrations can be achieved without an additional fitness cost, at some genomic positions insertion of additional DNA into a previously integrated transgene can have a significant negative effect. Thus, for targeted transgene insertions fitness costs must be evaluated both previous to and subsequent to new site-specific insertions in the target-site strain. PMID:25303238

  10. 78 FR 16491 - Combined Notice of Filings #1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-15

    ... Tariff Att H-14B Part II per Order in ER08-1329 to be effective 5/6/2013. Filed Date: 3/5/13. Accession... effective 3/7/2013. Filed Date: 3/6/13. Accession Number: 20130306-5015. Comments Due: 5 p.m. e.t. 3/28/13...: 20130306-5065. Comments Due: 5 p.m. e.t. 3/28/13. Docket Numbers: ER13-1044-000. Applicants:...

  11. Time-efficient determination of spin compartments by time-encoded pCASL T2-relaxation-under-spin-tagging and its application in hemodynamic characterization of the cerebral border zones.

    PubMed

    Schmid, Sophie; Teeuwisse, Wouter M; Lu, Hanzhang; van Osch, Matthias J P

    2015-12-01

    Information on water-transport across the blood-brain barrier can be determined from the T2 of the arterial spin labeling (ASL) signal. However, the current approach of using separate acquisitions of multiple inversion times is too time-consuming for clinical (research) applications. The aim of this study was to improve the time-efficiency of this method by combining it with time-encoded pseudo-continuous ASL (te-pCASL). Furthermore, the hemodynamic properties of the border zone regions in the brains of healthy, young volunteers were characterized as an example application. The use of te-pCASL instead of multi-TI pCASL significantly reduced the total scan duration, while providing a higher temporal resolution. A significantly lower cerebral blood flow (CBF) was found in the border zone regions compared with the central regions in both the posterior and the middle cerebral artery (MCA) flow territory. The arterial transit time (ATT) was almost two times longer in the border zone regions than in the central regions (p<0.05), with an average delay in ATT of 382ms in the posterior and 539ms in the MCA flow territory. When corrected for the ATT, the change in T2 over time was not significantly different for the border zones as compared to the central regions. In conclusion, te-pCASL-TRUST provided a time-efficient method to distinguish spin compartments based on their T2. The ATT in the border zone is significantly longer than in the central region. However, the exchange of the label from the arterial to the tissue compartment appears to be at a similar rate.

  12. Simultaneous measurement of forward-backward asymmetry and top polarization in dilepton final states from $$t\\bar t$$ production at the Tevatron

    DOE PAGESBeta

    Abazov, Victor Mukhamedovich

    2015-09-22

    We present a simultaneous measurement of the forward-backward asymmetry and the top-quark polarization in tt¯ production in dilepton final states using 9.7 fb–1 of proton-antiproton collisions at √s=1.96 TeV with the D0 detector. To reconstruct the distributions of kinematic observables we employ a matrix element technique that calculates the likelihood of the possible tt¯ kinematic configurations. After accounting for the presence of background events and for calibration effects, we obtain a forward-backward asymmetry of Att¯=(15.0±6.4(stat)±4.9(syst))% and a top-quark polarization times spin analyzing power in the beam basis of κP=(7.2±10.5(stat)±4.2(syst))%, with a correlation of –56% between the measurements. If we constrainmore » the forward-backward asymmetry to its expected standard model value, we obtain a measurement of the top polarization of κP=(11.3±9.1(stat)±1.9(syst))%. If we constrain the top polarization to its expected standard model value, we measure a forward-backward asymmetry of Att¯=(17.5±5.6(stat)±3.1(syst))%. A combination with the D0 Att¯ measurement in the lepton+jets final state yields an asymmetry of Att¯=(11.8±2.5(stat)±1.3(syst))%. Within their respective uncertainties, all these results are consistent with the standard model expectations.« less

  13. A method for producing transgenic cells using a multi-integrase system on a human artificial chromosome vector.

    PubMed

    Yamaguchi, Shigeyuki; Kazuki, Yasuhiro; Nakayama, Yuji; Nanba, Eiji; Oshimura, Mitsuo; Ohbayashi, Tetsuya

    2011-01-01

    The production of cells capable of expressing gene(s) of interest is important for a variety of applications in biomedicine and biotechnology, including gene therapy and animal transgenesis. The ability to insert transgenes at a precise location in the genome, using site-specific recombinases such as Cre, FLP, and ΦC31, has major benefits for the efficiency of transgenesis. Recent work on integrases from ΦC31, R4, TP901-1 and Bxb1 phages demonstrated that these recombinases catalyze site-specific recombination in mammalian cells. In the present study, we examined the activities of integrases on site-specific recombination and gene expression in mammalian cells. We designed a human artificial chromosome (HAC) vector containing five recombination sites (ΦC31 attP, R4 attP, TP901-1 attP, Bxb1 attP and FRT; multi-integrase HAC vector) and de novo mammalian codon-optimized integrases. The multi-integrase HAC vector has several functions, including gene integration in a precise locus and avoiding genomic position effects; therefore, it was used as a platform to investigate integrase activities. Integrases carried out site-specific recombination at frequencies ranging from 39.3-96.8%. Additionally, we observed homogenous gene expression in 77.3-87.5% of colonies obtained using the multi-integrase HAC vector. This vector is also transferable to another cell line, and is capable of accepting genes of interest in this environment. These data suggest that integrases have high DNA recombination efficiencies in mammalian cells. The multi-integrase HAC vector enables us to produce transgene-expressing cells efficiently and create platform cell lines for gene expression. PMID:21390305

  14. Rare Offshoot of a Common Malady Anaemia and Tuberculosis.

    PubMed

    Kolla, Gautham; Acharya, Vishak; Balanthimogru, Prashantha; Mani, Arun; Ruman, Shehzad

    2016-08-01

    Haematological manifestations are one of the rarer presentations of tuberculosis and are usually of normocytic normochromic type. An association of Autoimmune Haemolytic Anaemia (AIHA) with active pulmonary tuberculosis is an exceeding rare entity, though anaemia and tuberculosis commonly co-exist. We report a patient with sputum negative pulmonary tuberculosis with associated Coomb's positive AIHA. The patient responded well to Anti- Tubercular Therapy (ATT) and low dose steroids tapered over a month. PMID:27656489

  15. An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori.

    PubMed

    Long, Dingpei; Lu, Weijian; Zhang, Yuli; Bi, Lihui; Xiang, Zhonghuai; Zhao, Aichun

    2015-01-01

    We developed an efficient strategy that combines a method for the post-integration elimination of all transposon sequences, a site-specific recombination system, and an optimized fibroin H-chain expression system to produce a stable, replaceable, highly efficient transgene expression system in the silkworm (Bombyx mori) that overcomes the disadvantages of random insertion and post-integration instability of transposons. Here, we generated four different transgenic silkworm strains, and of one the transgenic strains, designated TS1-RgG2, with up to 16% (w/w) of the target protein in the cocoons, was selected. The subsequent elimination of all the transposon sequences from TS1-RgG2 was completed by the heat-shock-induced expression of the transposase in vivo. The resulting transgenic silkworm strain was designated TS3-g2 and contained only the attP-flanked optimized fibroin H-chain expression cassette in its genome. A phiC31/att-system-based recombinase-mediated cassette exchange (RMCE) method could be used to integrate other genes of interest into the same genome locus between the attP sites in TS3-g2. Controlling for position effects with phiC31-mediated RMCE will also allow the optimization of exogenous protein expression and fine gene function analyses in the silkworm. The strategy developed here is also applicable to other lepidopteran insects, to improve the ecological safety of transgenic strains in biocontrol programs.

  16. Pyrococcus furiosus-immobilized anodized tubular titania cathode in a hydrogen production system

    NASA Astrophysics Data System (ADS)

    Yoon, Jaekyung; Bae, Sanghyun; Shim, Eunjung; Joo, Hyunku

    Anodized tubular titania (TiO 2) electrodes (ATTEs) are prepared and used as both the photoanode and the cathode substrate in a photoelectrochemical system designed to split water into hydrogen with the assistance of an enzyme and an external bias of 1.5 V. In particular, the ATTE used as the cathode substrate for the immobilization of the enzyme is prepared by two methods-adsorption and crosslinking. Results show that the optimized amount of enzyme is 10.98 units for the slurried enzyme, 3.66 units for the adsorbed one and 7.32 units for the crosslinked one, and the corresponding hydrogen evolution rates are 33.04, 148.58 and 234.88 μmol h -1, respectively. The immobilized enzyme, specifically the chemically crosslinked one, seems to be much superior to the slurried enzyme, due to the enhanced charge-transfer process that is caused by the lower electrical resistance between the enzyme and the ATTE. This results in a greater number of accepted electrons and a larger amount of enzymes able to deal with the electrons.

  17. Photoanodic and cathodic role of anodized tubular titania in light-sensitized enzymatic hydrogen production

    NASA Astrophysics Data System (ADS)

    Bae, Sanghyun; Shim, Eunjung; Yoon, Jaekyung; Joo, Hyunku

    An anodized tubular titania (TiO 2) electrode (ATTE) is prepared and utilized as both a photoanode and a cathode in a photoelectrochemical system designed to split water into hydrogen (for use in fuel cells) with the assistance of a hydrogenase enzyme and an external bias of 1.5 V. In particular, the cathodic ATTE acts as a substrate for the immobilization of the enzyme due to its large surface area that results from the tubular oxides. The optimum molar concentration of KOH in anode and cathode compartments is 1.0 M and the optimum amount of enzyme for the cathode is ca. 3.66 units per geometrical unit area (1 cm × 1 cm) of the cathodic ATTE. After exposure to air for three weeks, the enzyme shows a hydrogen evolution rate that is 85.8% of that of an argon-purged enzyme. The rate of hydrogen evolution is increased from ca. 65 (in a slurry system) to more than 140 μmol cm -2 h -1, even after eliminating the electron relay (methyl viologen) and costly platinum counter electrode.

  18. Vitamin E deficiency reduced lumbar bone calcium content in female rats.

    PubMed

    Norazlina, M; Chua, C W; Ima-Nirwana, S

    2004-12-01

    Vitamin E deficiency has been found to impair bone calcification. This study was done to determine the effects of vitamin E deficiency and supplementation on parathyroid hormone, i.e. the hormone involved in bone regulation. Female Sprague-Dawley rats were divided into 4 groups: 1) normal rat chow (RC), 2) vitamin E deficiency (VED), vitamin E deficient rats supplemented with 3) 60 mg/kg alpha-tocotrienol (ATT) and 4) 60 mg/kg (alpha-tocopherol (ATF). Treatment was carried out for 3 months. Vitamin E deficiency caused hypocalcaemia during the first month of the treatment period, increased the parathyroid hormone level in the second month and decreased the bone calcium content in the 4th lumbar bone at the end of the treatment. Vitamin E supplementation (ATT and ATF) failed to improve these conditions. The bone formation marker, osteocalcin, and the bone resorption marker, deoxypyridinoline did not change throughout the study period. In conclusion vitamin E deficiency impaired bone calcium homeostasis with subsequent secondary hyperparathyroidism and vertebral bone loss. Replacing the vitamin E with pure ATF or pure ATT alone failed to correct the changes seen. PMID:15889565

  19. Physic-mathematical modeling of atmospheric tides influence on background circulation and background temperature of lower Earth thermosphere.

    NASA Astrophysics Data System (ADS)

    Gavrilov, Anatoliy; Kapitsa, Andrey

    Nonstationary semiempirical model of: 1) atmospheric thermal tides (ATT) in middle Earth atmosphere conditional on ozone and water vapor absorption; 2) tidal disturbances (TD) gen-erated by global ozone anomalies. Geophysical phenomenon -distant wave action (teleconnec-tion) of Antarctic ozone anomaly on thermal tidal wind structure, background circulation and background temperature in middle-latitude and polar lower thermosphere of northern hemi-sphere, which was not known earlier, and which was found by means of numerical experiments on constructed model and confirmed by observations, is described. Mean zonal numerical cor-recting models of background circulation and background temperature in lower thermosphere due to semidiurnal and diurnal ATT dissipation at these heights are given. It is noted that background temperature corrections reach maximum value of 40-50 degrees in polar lower ther-mosphere of both hemispheres at 110-metricconverterProductID120 km120 km height due to "heating" caused by semidiurnal ATT, both during equinox and solstice.

  20. Mycobacteria Infection in Incomplete Transverse Myelitis Is Refractory to Steroids: A Pilot Study

    PubMed Central

    Feng, Yanqing; Guo, Ning; Liu, Junxiu; Chen, Xi; Sun, Qiaosong; Lai, Rong; Huang, Fan

    2011-01-01

    Incomplete transverse myelitis (ITM) of unknown origin is associated with high rates of morbidity and mortality. This prospective, open-label study was undertaken to determine whether antituberculous treatment (ATT) might help patients with ITM whose condition continues to deteriorate despite receiving IV methylprednisolone treatment. The study consisted of 67 patients with steroid-refractory ITM in whom Mycobacterium tuberculosis (MTB) was suspected clinically and in whom other known causes of myelopathy were excluded. The study occurred from January 2003 to June 2010. Patients underwent trial chemotherapy with ATT. Efficacy was assessed by the American Spinal Injury Association (ASIA) scoring system, the Barthel Index (BI) and the Hauser Ambulation Index (AI) at baseline, 12 months, and 24 months, using magnetic resonance imaging (MRI). Of the 67 patients enrolled, 51 were assessed and 16 withdrew. At 24 months, 49 patients experienced benefits as indicated by significantly increased ASIA and BI scores. The Hauser AI index also improved with markedly decreased abnormal signals in spinal cord MRI over time. The results from this prospective study provide beneficial clinical and MRI data on the efficacy of ATT in ITM patients and suggests mycobacteria may be an important and neglected cause of myelitis. PMID:21331384

  1. Heat regeneration of hydroxyapatite/attapulgite composite beads for defluoridation of drinking water.

    PubMed

    Feng, Li; Xu, Weihua; Liu, Tengfei; Liu, Jason

    2012-06-30

    Regeneration is one of the key factors in evaluating an adsorbent. A novel heat regeneration method for hydroxyapatite/attapulgite (HAP/ATT) composite beads was studied. The investigation included heat regeneration temperature, regeneration time, and regeneration effects. A possible mechanism for the heat regeneration is described that explains the results of XPS, and SEM with EDAX. Exhausted HAP/ATT composite beads can be regenerated for more than 10 cycles using boiling water or steam. The total capacity increases by 10 times compared to a single defluoridation cycle. The regeneration process involves F(-) ions adsorbed on the surface of the beads to move quickly into the bulk of the HAP through the effect of heating this composite material. The surface active sites are thus re-exposed and the beads recover their fluoride sequestration properties. HAP/ATT composite beads were successfully used for the removal of fluoride from field water taken from a nearby village where fluoride contamination is endemic. Defluoridation and regeneration cycles performed in the same container provide a high efficient and simple operation. No chemical agents are used and no waste products are produced during the heat regeneration process, so this is a nearly zero emission process. This method can easily be up-scaled to a large throughput application.

  2. Effect of ultrasonic treatment on the total phenolic and antioxidant activity of extracts from citrus peel.

    PubMed

    Ma, Y-Q; Chen, J-C; Liu, D-H; Ye, X-Q

    2008-10-01

    Application of ultrasound to extract a variety of biologically active compounds from plant materials has been widely investigated. However, there are few reports on the local effect of ultrasonic irradiation on the yields of these compounds. In the present article, the local effect of ultrasonic treatment on total phenolic content (TPC) and antioxidant activities (ATT) of extracts from citrus peels was investigated. To optimize the extraction process, a response surface methodology (RSM) was used to evaluate the effects of ultrasonic variables including ultrasonic power, ultrasonic time, and extraction temperature on extracts from penggan (Citrus reticulata) peel. The results showed that TPC and ATT increased on increasing ultrasonic time and temperature. The maximum of TPC and ATT by ultrasonic treatment was observed in near ultrasonic irradiation surface, in which ultrasonic power appeared to be positive effect. Furthermore, when the effect of the 3 independent variables was evaluated simultaneously using RSM, the optimal ultrasonic conditions for responses were determined as: 42 to 45 W, 23 to 25 min, 31 to 34 degrees C. The results presented here emphasized that application of ultrasound should be considered both the optimization of ultrasonic variables and available ultrasonic device. PMID:19019132

  3. Structural analysis of glycoconjugates by on-target enzymatic digestion and MALDI-TOF-MS.

    PubMed

    Geyer, H; Schmitt, S; Wuhrer, M; Geyer, R

    1999-01-15

    Exoglycosidase digestion combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has been demonstrated to be an effective method for the structural characterization of glycoconjugates and oligosaccharides in picomolar amounts. A sample preparation method is described, in which 6-aza-2-thiothymine (ATT) in water is used as matrix and enzymes are dialyzed before use against a low concentration of volatile buffer such as ammonium acetate. Under these conditions, a series of sequential on-target exoglycosidase treatments was carried out in one single analyte spot in the presence of ATT matrix. Subsequent mass spectrometric analysis of the resulting products yielded information on both the completeness of the reaction and structural features of the glycoconjugates such as monosaccharide sequence, branching pattern, and anomeric configurations of the corresponding glycosidic linkages. The results show that all exoglycosidases used retain their activity in the presence of ATT matrix. Hence, structural analysis of carbohydrates or mixtures thereof can be performed very fast, without intermediate desalting steps or sample splitting. This approach is illustrated by the analysis of underivatized glycans, oligosaccharide derivatives, glycopeptides, and glycolipids. Depending on the analyte, amounts of sample required could be limited to a few picomoles. PMID:9949734

  4. Targeted gene integration using the combination of a sequence-specific DNA-binding protein and phiC31 integrase.

    PubMed

    Nakanishi, Hideyuki; Higuchi, Yuriko; Yamashita, Fumiyoshi; Hashida, Mitsuru

    2014-09-30

    PhiC31 integrase-based vectors can integrate therapeutic genes selectively into attP or pseudo-attP sites in genomes, but considerable numbers of pseudo-attP sites in human genomes exist inside endogenous gene-coding regions. To avoid endogenous gene disruptions, we aimed to enhance the integration site-specificity of the phiC31 integrase-based vector using a sequence-specific DNA-binding protein containing Gal4 and LexA DNA-binding motifs. The dual DNA-binding protein was designed to tether the UAS-containing donor vector to the target sequence, the LexA operator, and restrict integration to sites close to the LexA operator. To analyze the site-specificity in chromosomal integration, a human cell line having LexA operators on the genome was established, and the cell line was transfected with donor vectors expressing the DNA-binding protein and the phiC31 integrase expression vector (helper vector). Quantitative PCR indicated that integration around the LexA operator was 26-fold higher with the UAS-containing donor vector than with the control. Sequence analysis confirmed that the integration occurred around the LexA operator. The dual DNA-binding protein-based targeted integration strategy developed herein would allow safer and more reliable genetic manipulations for various applications, including gene and cell therapies.

  5. Infrared Attenuation Spectrum of Bulk High-Resistivity CdZnTe Single Crystal in Transparent Wavelength Region Between Electronic and Lattice Absorptions

    NASA Astrophysics Data System (ADS)

    Sarugaku, Yuki; Kaji, Sayumi; Ikeda, Yuji; Kobayashi, Naoto; Sukegawa, Takashi; Nakagawa, Takao; Kataza, Hirokazu; Kondo, Sohei; Yasui, Chikako; Nakanishi, Kenshi; Kawakita, Hideyo

    2016-09-01

    We report measurement of the internal attenuation coefficient, α _{att} , of a bulk high-resistivity cadmium zinc telluride (CdZnTe) single crystal at wavelength, λ = 0.84-26 μ m, to the unprecedentedly low level of α _{att} ˜ 0.001 cm^{-1} . This measurement reveals the spectral behavior for small attenuation in the infrared transparent region between the electronic and lattice absorptions. This result is essential for application of CdZnTe as an infrared transmitting material. Comparing the attenuation spectrum with model spectra obtained on the basis of Mie theory, we find that sub-micrometer-sized Te particles (inclusions) with a number density of approximately 10^{7.5-9} cm^{-3} are the principal source of the small attenuation observed at λ = 0.9-13 μ m. In addition, we determine α _{att} = (7.7 ± 1.9) × 10^{-4} cm^{-1} at λ = 10.6 μ m, which is valuable for CO_2 laser applications. Higher transparency can be achieved by reducing the number of inclusions rather than the number of precipitates. This study also demonstrates that high-accuracy measurement of CdZnTe infrared transmittance is a useful approach to investigating the number density of sub-micrometer-sized Te particles that cannot be identified via infrared microscopy.

  6. Tuberculosis in a renal allograft recipient presenting with intussusception.

    PubMed

    Mohapatra, A; Basu, G; Sen, I; Asirvatham, R; Michael, J S; Pulimood, A B; John, G T

    2012-01-01

    Extra-pulmonary tuberculosis (TB) is more common in renal allograft recipients and may present with dissemination or an atypical features. We report a renal allograft recipient with intestinal TB presenting 3 years after transplantation with persistent fever, weight loss, diarrhea, abdominal pain and mass in the abdomen with intestinal obstruction. He was diagnosed to be having an ileocolic intussusception which on resection showed a granulomatous inflammation with presence of acid-fast bacilli (AFB) typical of Mycobacterium tuberculosis. In addition, AFB was detected in the tracheal aspirate, indicating dissemination. He received anti-TB therapy (ATT) from the fourth postoperative day. However, he developed a probable immune reconstitution inflammatory syndrome (IRIS) with multiorgan failure and died on 11(th) postoperative day. This is the first report of intestinal TB presenting as intussusception in a renal allograft recipient. The development of IRIS after starting ATT is rare in renal allograft recipients. This report highlights the need for a high index of suspicion for diagnosing TB early among renal transplant recipients and the therapeutic dilemma with overwhelming infection and development of IRIS upon reduction of immunosuppression and starting ATT.

  7. Characterization of repairs to KrF 300mm wafer printability for 0.13μm design rule with attenuated phase-shifting mask

    NASA Astrophysics Data System (ADS)

    Chou, William; Chen, Tsung; Tseng, Will; Huang, Peter; Tseng, Chin C.; Chung, Mars; Wang, Dick; Huang, Norman

    2002-12-01

    For sub-0.13um lithography, attenuated phase shifting mask (AttPSM) with optical proximity correction (OPC) is reported as one of the potential methods to achieve manufacturable process by using 248nm exposure wavelength. Unfortunately, the low-k1 imaging process in 130nm lithography imposes much more stringent requirements on defect repair, especially on AttPSM reticle. Therefore, the imperfect repairs will have a significant impact on wafer process window due to quartz damage and phase distortion caused by Ga+ ion stain removal and added carbon material, respectively. In this paper, we have prepared AttPSM test masks having programmed defects with various opaque defects. Each defect area was inspected with KLA-Tencor's SLF27 inspection system to acquire defect coordinates and image, simulated with AIMS to assess the intensity and transmission loss induced by repair process. All of masks were made by DuPont Photomasks Taiwan (DPT) by using the Jbx9000MV2 E-beam writer and dry Chrome etch process. All lithographic experiments were performed on 300mm wafer using high NA ASML AT750S scanner and high contrast CAR resist. In this study, we have focused on the impact of quartz damage and phase error on wafer process window by comparing the wafer CD and pattern profile through focus. In order to establish a efficient way to perform effective judgement on repair defect between mask shop and wafer fab, both AIMS and wafer results will be compared and correlated.

  8. [Integrons: gene collectors].

    PubMed

    Di Conza, J A; Gutkind, G O

    2010-01-01

    Integrons gained great interest due to their participation in resistance gene recruitment and expression. Their basic structure includes a fragment that encodes an integrase (intI) followed by a recognition sequence (attI) into which they may incorporate gene cassettes (encoding resistance mechanisms). A promoter (Pc) embedded within the integrase gene controls the transcription of integrated resistance markers, as these genes do not have their own promoters. When in cassettes, resistance genes are flanked by specific sequences (attC), which are recognized by the integrase that, by site specific recombination, incorporates them after attI in proper orientation for their expression. In the past, integrons were classified according to their sequence homology; currently they are classified according to their location. In general, they are divided into "mobile" integrons (those associated with insertion sequences, transposons and/or plasmids, being most of them associated with resistance mechanisms), and chromosomally-located "super" integrons with large arrangements of cassette genes. "Mobile" class 1 integrons are the most abundant in clinical isolates and are generally associated with Tn21 subgroup transposons, followed by class 2, derived primarily from Tn7. These elements are not mobile themselves, but their association with mobile platforms that facilitate horizontal transfer, explains their wide distribution among bacteria. This review also attempts to describe the mobile integrons described so far in Argentina.

  9. The expression of Streptomyces and Escherichia coli drug-resistance determinants cloned into the Streptomyces phage phi C31.

    PubMed

    Chater, K F; Bruton, C J; King, A A; Suarez, J E

    1982-01-01

    Lysogens obtained by infecting Streptomyces albus G with a phi C31-pBR322 chimaeric prophage or its delta W12 deletion derivative had increased tetracycline resistance. The ability of the delta W12 derivative to transduce tetracycline resistance was inactivated by inserting a viomycin resistance determinant (vph) into the BamHI site of the pBR322 tet gene, and restored by excising the vph gene. Another deletion mutant (delta W17) of the chimaera, carrying an intact tet gene, was normally unable to transduce tetracycline resistance. This inability was correlated with the finding, by Southern hybridisation analysis, that the att site required for insertion of phi C31 prophage into the host chromosome was located within the delta W17 deletion. Use of phi C31 lysogenic recipient permitted the integration of the att-deleted phage, presumably by homologous recombination, giving tetracycline-resistant double lysogens. This technique was extended to S. coelicolor A3(2) in the detection of derivatives of the att-deleted phage into which a thiostrepton-resistance determinant (tsr) had been inserted in vitro. Phage released from double lysogens were mainly recombinants. One such recombinant is a PstI vector for DNA cloning, able to accommodate up to 6 kb of introduced DNA.

  10. An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori

    PubMed Central

    Long, Dingpei; Lu, Weijian; Zhang, Yuli; Bi, Lihui; Xiang, Zhonghuai; Zhao, Aichun

    2015-01-01

    We developed an efficient strategy that combines a method for the post-integration elimination of all transposon sequences, a site-specific recombination system, and an optimized fibroin H-chain expression system to produce a stable, replaceable, highly efficient transgene expression system in the silkworm (Bombyx mori) that overcomes the disadvantages of random insertion and post-integration instability of transposons. Here, we generated four different transgenic silkworm strains, and of one the transgenic strains, designated TS1-RgG2, with up to 16% (w/w) of the target protein in the cocoons, was selected. The subsequent elimination of all the transposon sequences from TS1-RgG2 was completed by the heat-shock-induced expression of the transposase in vivo. The resulting transgenic silkworm strain was designated TS3-g2 and contained only the attP-flanked optimized fibroin H-chain expression cassette in its genome. A phiC31/att-system-based recombinase-mediated cassette exchange (RMCE) method could be used to integrate other genes of interest into the same genome locus between the attP sites in TS3-g2. Controlling for position effects with phiC31-mediated RMCE will also allow the optimization of exogenous protein expression and fine gene function analyses in the silkworm. The strategy developed here is also applicable to other lepidopteran insects, to improve the ecological safety of transgenic strains in biocontrol programs. PMID:25739894

  11. Characterization of the chromosomal integration of Saccharopolyspora plasmid pCM32 and its application to improve production of spinosyn in Saccharopolyspora spinosa.

    PubMed

    Chen, Jian; Xia, Haiyang; Dang, Fujun; Xu, Qingyu; Li, Wenjun; Qin, Zhongjun

    2015-12-01

    Saccharopolyspora spinosa produces tetra-cyclic macrolide spinosyns, a group of highly efficient pesticidal agents. However, this species lacks efficient vectors for genetic manipulation. In this study, the circular plasmid pCM32 was newly isolated from Saccharopolyspora endophytica YIM 61095. The complete nucleotide sequence of pCM32 consists of 14,611 bp and is predicted to encode 17 open reading frames (ORFs). Interestingly, a putative int gene in pCM32 was predicted by homologous alignment to encode an integrase belonging to the tyrosine family of integrases/recombinases. Plasmid pCM238 containing this int locus derived from pCM32 could be transferred by conjugation from Escherichia coli into Sa. spinosa at a high frequency. Integration of pCM238 in the host chromosome was demonstrated as site-specific recombination (at the tRNA (Ser) gene) via a 56-bp core sequence within the attP/attB sites. Plasmid pCM265, a shuttle vector containing the int and attP sequences of pCM32, was constructed to introduce foreign genes into Sa. spinosa. The production of spinosad approximately doubled in Sa. spinosa NRRL18395 after introducing pCM265-derived plasmids carrying the genes for phosphofructokinase (PFK) or anthranilate synthase. These results indicate that plasmid pCM32 is an actinomycete integrative and conjugative element (AICE) and that its derived integrative vectors are useful for efficiently introducing foreign DNA into Sa. spinosa.

  12. Measurements of the charge asymmetry in top-quark pair production in the dilepton final state at s=8TeV with the ATLAS detector

    DOE PAGESBeta

    Aad, G.; Abbott, B.; Abdallah, J.; Abdinov, O.; Abeloos, B.; Aben, R.; Abolins, M.; AbouZeid, O. S.; Abraham, N. L.; Abramowicz, H.; et al

    2016-08-22

    Measurements of the top-antitop quark pair production charge asymmetry in the dilepton channel, characterized by two high-pT leptons (electrons or muons), are presented using data corresponding to an integrated luminosity of 20.3 fb–1 from pp collisions at a center-of-mass energy √s = 8 TeV collected with the ATLAS detector at the Large Hadron Collider at CERN. Inclusive and differential measurements as a function of the invariant mass, transverse momentum, and longitudinal boost of the tt¯ system are performed both in the full phase space and in a fiducial phase space closely matching the detector acceptance. Two observables are studied: AℓℓCmore » based on the selected leptons and Att¯C based on the reconstructed tt¯ final state. As a result, the inclusive asymmetries are measured in the full phase space to be AℓℓC=0.008±0.006 and Att¯C=0.021±0.016, which are in agreement with the Standard Model predictions of AℓℓC=0.0064±0.0003 and Att¯C=0.0111±0.0004.« less

  13. Cardiovascular physiology of androgens and androgen testosterone therapy in postmenopausal women.

    PubMed

    Ling, Shanhong; Komesaroff, Paul A; Sudhir, Krishnankutty

    2009-03-01

    Women before menopause are at relatively lower risk of cardiovascular disease (CVD) compared with age-matched men and after menopause this gender advantage disappears. Androgen has been known to be an independent factor contributing to the higher male susceptibility to CVD, through adverse effects on lipids, blood pressure, and glucose metabolism. High androgen levels also contribute to CVD development in women with polycystic ovary syndrome as well as androgen abusing athletes and body builders. On the other hand, decline in androgen levels, as a result of ageing in men, is associated with hypertension, diabetes and atherosclerosis. Postmenopausal women, particularly those with oophorectomy are generally in low levels of sex hormones and androgen insufficiency is independently associated with the higher incidence of atherosclerosis in postmenopausal women. Androgen testosterone therapy (ATT) has been commonly used to improve well-being and libido in aging men with low androgen levels. The therapy has been demonstrated also to effectively reduce atherogenesis in these people. The use of ATT in postmenopausal women has increased in recent years and to date, however, the cardiovascular benefits of such therapy in these women remain uncertain. This review focuses on research regarding the impact of endogenous androgens and ATT on the cardiovascular physiology and CVD development in postmenopausal women.

  14. Optical measurement of adipose tissue thickness and comparison with ultrasound, magnetic resonance imging, and callipers

    NASA Astrophysics Data System (ADS)

    Geraskin, Dmitri; Boeth, Heide; Kohl-Bareis, Matthias

    2009-07-01

    Near-infrared spectroscopy is used to quantify the subcutaneous adipose tissue thickness (ATT) over five muscle groups (vastus medialis, vastus lateralis, gastrocnemius, ventral forearm and biceps brachii muscle) of healthy volunteers (n=20). The optical lipid signal (OLS) was obtained from the second derivative of broad band attenuation spectra and the lipid absorption peak (λ=930 nm). Ultrasound and MR imaging as well as mechanical calliper readings were taken as reference methods. The data show that the OLS is a good predictor for ATT (<16 mm) with absolute and relative errors of <0.8 mm and <24%, respectively. The optical method compares favourably with calliper reading. The finding of a non-linear relationship of optical signal vs. ultrasound is explained by a theoretical two-layer model based on the diffusion approximation for the transport of photons. The crosstalk between the OLS and tissue hemoglobin concentration changes during an incremental cycling exercise was found to be small, indicating the robustness of OLS. Furthermore, the effect of ATT on spatially-resolved spectroscopy measurements is shown to decrease the calculated muscle hemoglobin concentration and to increase oxygen saturation.

  15. Fragile X mice have robust mGluR5-dependent alterations of social behaviour in the Automated Tube Test.

    PubMed

    de Esch, C E F; van den Berg, W E; Buijsen, R A M; Jaafar, I A; Nieuwenhuizen-Bakker, I M; Gasparini, F; Kushner, S A; Willemsen, R

    2015-03-01

    Fragile X syndrome is the most common monogenetic form of intellectual disability and autism. Although the Fmr1 knockout mouse model recapitulates many aspects of the human FXS condition, the establishment of robust social behavioural phenotypes suitable for drug screening has been difficult. Here, we describe a novel social behavioural paradigm, the Automated Tube Test (ATT), for which Fmr1 knockout mice demonstrate a highly reliable and robust phenotype. Fmr1 KO mice show highly dominant behaviour over wild-type littermates in the ATT. Consistent with previous findings, we observed a highly significant, albeit partial, rescue of the altered social behaviour of Fmr1 knockout mice in the ATT, using genetic (mGluR5 deletion) or pharmacological inhibition (mGluR5 antagonist) of mGluR5 signalling independently. Together, our results validate the Automated Tube Test as a robust outcome measure for social behaviour in preclinical research for FXS, and confirm the pathophysiological relevance of mGluR5 signalling. Moreover, our findings highlight the strategy of initiating pharmacological intervention in adulthood as holding significant clinical potential.

  16. Integration and excision by the large serine recombinase phiRv1 integrase.

    PubMed

    Bibb, Lori A; Hancox, Maria I; Hatfull, Graham F

    2005-03-01

    The Mycobacterium tuberculosis prophage-like element phiRv1 encodes a site-specific recombination system utilizing an integrase of the serine recombinase family. Recombination occurs between a putative attP site and the host chromosome, but is unusual in that the attB site lies within a redundant repetitive element (REP13E12) of which there are seven copies in the M. tuberculosis genome; four of these elements contain attB sites suitable for phiRv1 integration in vivo. Although the mechanism of directional control of large serine integrases is poorly understood, a recombination directionality factor (RDF) has been identified that is required for phiRv1 integrase-mediated excisive recombination in vivo. Here we describe defined in vitro recombination reactions for both phiRv1 integrase-mediated integration and excision and show that the phiRv1 RDF is not only required for excision but inhibits integrative recombination; neither reaction requires DNA supercoiling, host factors, or high-energy cofactors. Integration, excision and excise-mediated inhibition of integration require simple substrates sites, indicating that the control of directionality does not involve the manipulation of higher-order protein-DNA architectures as described for the tyrosine integrases.

  17. Serpinin: A Novel Chromogranin A-Derived, Secreted Peptide Up-Regulates Protease Nexin-1 Expression and Granule Biogenesis in Endocrine Cells

    PubMed Central

    Koshimizu, Hisatsugu; Cawley, Niamh X.; Kim, Taeyoon; Yergey, Alfred L.

    2011-01-01

    Previously we demonstrated that chromogranin A (CgA) promoted secretory granule biogenesis in endocrine cells by stabilizing and preventing granule protein degradation in the Golgi, through up-regulation of expression of the protease inhibitor, protease nexin-1 (PN-1). However, the mechanism by which CgA signals the increase of PN-1 expression is unknown. Here we identified a 2.9-kDa CgA-C-terminus peptide, which we named serpinin, in conditioned media from AtT-20 cells, a corticotroph cell line, which up-regulated PN-1 mRNA expression. Serpinin was secreted from AtT-20 cells upon high potassium stimulation and increased PN-1 mRNA transcription in these cells, in an actinomycin D-inhibitable manner. CgA itself and other CgA-derived peptides, when added to AtT-20 cell media, had no effect on PN-1 expression. Treatment of AtT-20 cells with 10 nm serpinin elevated cAMP levels and PN-1 mRNA expression, and this effect was inhibited by a protein kinase A inhibitor, 6–22 amide. Serpinin and a cAMP analog, 8-bromo-cAMP, promoted the translocation of the transcription factor Sp1 into the nucleus, which is known to drive PN-1 expression. Additionally, an Sp1 inhibitor, mithramycin A inhibited the serpinin-induced PN-1 mRNA up-regulation. Furthermore, a luciferase reporter assay demonstrated serpinin-induced up-regulation of PN-1 promoter activity in an Sp1-dependent manner. When added to CgB-transfected 6T3 cells, a mutant AtT20 cell line, serpinin induced granule biogenesis as evidenced by the presence of CgB puncta accumulation in the processes and tips. Our findings taken together show that serpinin, a novel CgA-derived peptide, is secreted upon stimulation of corticotrophs and plays an important autocrine role in up-regulating PN-1-dependent granule biogenesis via a cAMP-protein kinase A-Sp1 pathway to replenish released granules. PMID:21436258

  18. Survival and Persistence of Nonpathogenic Escherichia coli and Attenuated Escherichia coli O157:H7 in Soils Amended with Animal Manure in a Greenhouse Environment.

    PubMed

    Sharma, Manan; Millner, Patricia D; Hashem, Fawzy; Camp, Mary; Whyte, Celia; Graham, Lorna; Cotton, Corrie P

    2016-06-01

    Animal manure provides benefits to agriculture but may contain pathogens that contaminate ready-to-eat produce. U.S. Food and Drug Administration standards include 90- or 120-day intervals between application of manure and harvest of crop to minimize risks of pathogen contamination of fresh produce. Data on factors affecting survival of Escherichia coli in soils under greenhouse conditions are needed. Three separate studies were conducted to evaluate survival of nonpathogenic E. coli (gEc) and attenuated E. coli O157:H7 (attO157) inoculated at either low (4 log CFU/ml) or high (6 log CFU/ml) populations over 56 days. Studies involved two pot sizes (small, 398 cm(3); large, 89 liters), three soil types (sandy loam, SL; clay loam, CL; silt loam, SIL), and four amendments (poultry litter, PL; dairy manure liquids, DML; horse manure, HM; unamended). Amendments were applied to the surface of the soil in either small or large containers. Study 1, conducted in regularly irrigated small containers, showed that populations of gEc and attO157 (2.84 to 2.88 log CFU/g) in PL-amended soils were significantly (P < 0.05) greater than those in DML-amended (0.29 to 0.32 log CFU/g [dry weight] [gdw]) or unamended (0.25 to 0.28 log CFU/gdw) soils; soil type did not affect E. coli survival. Results from study 2, in large pots with CL and SIL, showed that PL-amended soils supported significantly higher attO157 and gEc populations compared with HM-amended or unamended soils. Study 3 compared results from small and large containers that received high inoculum simultaneously. Overall, in both small and large containers, PLamended soils supported higher gEc and attO157 populations compared with HM-amended and unamended soils. Populations of attO157 were significantly greater in small containers (1.83 log CFU/gdw) than in large containers (0.65 log CFU/gdw) at week 8, perhaps because small containers received more regular irrigation than large pots. Regular irrigation of small pots may have

  19. Hybrid Lentivirus-phiC31-int-NLS Vector Allows Site-Specific Recombination in Murine and Human Cells but Induces DNA Damage

    PubMed Central

    Grandchamp, Nicolas; Altémir, Dorothée; Philippe, Stéphanie; Ursulet, Suzanna; Pilet, Héloïse; Serre, Marie-Claude; Lenain, Aude; Serguera, Che; Mallet, Jacques; Sarkis, Chamsy

    2014-01-01

    Gene transfer allows transient or permanent genetic modifications of cells for experimental or therapeutic purposes. Gene delivery by HIV-derived lentiviral vector (LV) is highly effective but the risk of insertional mutagenesis is important and the random/uncontrollable integration of the DNA vector can deregulate the cell transcriptional activity. Non Integrative Lentiviral Vectors (NILVs) solve this issue in non-dividing cells, but they do not allow long term expression in dividing cells. In this context, obtaining stable expression while avoiding the problems inherent to unpredictable DNA vector integration requires the ability to control the integration site. One possibility is to use the integrase of phage phiC31 (phiC31-int) which catalyzes efficient site-specific recombination between the attP site in the phage genome and the chromosomal attB site of its Streptomyces host. Previous studies showed that phiC31-int is active in many eukaryotic cells, such as murine or human cells, and directs the integration of a DNA substrate into pseudo attP sites (pattP) which are homologous to the native attP site. In this study, we combined the efficiency of NILV for gene delivery and the specificity of phiC31-int for DNA substrate integration to engineer a hybrid tool for gene transfer with the aim of allowing long term expression in dividing and non-dividing cells preventing genotoxicity. We demonstrated the feasibility to target NILV integration in human and murine pattP sites with a dual NILV vectors system: one which delivers phiC31-int, the other which constitute the substrate containing an attB site in its DNA sequence. These promising results are however alleviated by the occurrence of significant DNA damages. Further improvements are thus required to prevent chromosomal rearrangements for a therapeutic use of the system. However, its use as a tool for experimental applications such as transgenesis is already applicable. PMID:24956106

  20. Hybrid lentivirus-phiC31-int-NLS vector allows site-specific recombination in murine and human cells but induces DNA damage.

    PubMed

    Grandchamp, Nicolas; Altémir, Dorothée; Philippe, Stéphanie; Ursulet, Suzanna; Pilet, Héloïse; Serre, Marie-Claude; Lenain, Aude; Serguera, Che; Mallet, Jacques; Sarkis, Chamsy

    2014-01-01

    Gene transfer allows transient or permanent genetic modifications of cells for experimental or therapeutic purposes. Gene delivery by HIV-derived lentiviral vector (LV) is highly effective but the risk of insertional mutagenesis is important and the random/uncontrollable integration of the DNA vector can deregulate the cell transcriptional activity. Non Integrative Lentiviral Vectors (NILVs) solve this issue in non-dividing cells, but they do not allow long term expression in dividing cells. In this context, obtaining stable expression while avoiding the problems inherent to unpredictable DNA vector integration requires the ability to control the integration site. One possibility is to use the integrase of phage phiC31 (phiC31-int) which catalyzes efficient site-specific recombination between the attP site in the phage genome and the chromosomal attB site of its Streptomyces host. Previous studies showed that phiC31-int is active in many eukaryotic cells, such as murine or human cells, and directs the integration of a DNA substrate into pseudo attP sites (pattP) which are homologous to the native attP site. In this study, we combined the efficiency of NILV for gene delivery and the specificity of phiC31-int for DNA substrate integration to engineer a hybrid tool for gene transfer with the aim of allowing long term expression in dividing and non-dividing cells preventing genotoxicity. We demonstrated the feasibility to target NILV integration in human and murine pattP sites with a dual NILV vectors system: one which delivers phiC31-int, the other which constitute the substrate containing an attB site in its DNA sequence. These promising results are however alleviated by the occurrence of significant DNA damages. Further improvements are thus required to prevent chromosomal rearrangements for a therapeutic use of the system. However, its use as a tool for experimental applications such as transgenesis is already applicable. PMID:24956106

  1. Site-specific bacterial chromosome engineering: ΦC31 integrase mediated cassette exchange (IMCE).

    PubMed

    Heil, John R; Cheng, Jiujun; Charles, Trevor C

    2012-03-16

    The bacterial chromosome may be used to stably maintain foreign DNA in the mega-base range. Integration into the chromosome circumvents issues such as plasmid replication, plasmid stability, plasmid incompatibility, and plasmid copy number variance. This method uses the site-specific integrase from the Streptomyces phage (Φ) C31. The ΦC31 integrase catalyzes a direct recombination between two specific DNA sites: attB and attP (34 and 39 bp, respectively). This recombination is stable and does not revert. A "landing pad" (LP) sequence consisting of a spectinomycin-resistance gene, aadA (SpR), and the E. coli ß-glucuronidase gene (uidA) flanked by attP sites has been integrated into the chromosomes of Sinorhizobium meliloti, Ochrobactrum anthropi, and Agrobacterium tumefaciens in an intergenic region, the ampC locus, and the tetA locus, respectively. S. meliloti is used in this protocol. Mobilizable donor vectors containing attB sites flanking a stuffer red fluorescent protein (rfp) gene and an antibiotic resistance gene have also been constructed. In this example the gentamicin resistant plasmid pJH110 is used. The rfp gene may be replaced with a desired construct using SphI and PstI. Alternatively a synthetic construct flanked by attB sites may be sub-cloned into a mobilizable vector such as pK19mob. The expression of the ΦC31 integrase gene (cloned from pHS62) is driven by the lac promoter, on a mobilizable broad host range plasmid pRK7813. A tetraparental mating protocol is used to transfer the donor cassette into the LP strain thereby replacing the markers in the LP sequence with the donor cassette. These cells are trans-integrants. Trans-integrants are formed with a typical efficiency of 0.5%. Trans-integrants are typically found within the first 500-1,000 colonies screened by antibiotic sensitivity or blue-white screening using 5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid (X-gluc). This protocol contains the mating and selection procedures for

  2. Production of transgenic cattle highly expressing human serum albumin in milk by phiC31 integrase-mediated gene delivery.

    PubMed

    Luo, Yan; Wang, Yongsheng; Liu, Jun; Lan, Hui; Shao, Minghao; Yu, Yuan; Quan, Fusheng; Zhang, Yong

    2015-10-01

    Transgenic cattle expressing high levels of recombinant human serum albumin (HSA) in their milk may as an alternative source for commercial production. Our objective was to produce transgenic cattle highly expressing HSA in milk by using phiC31 integrase system and somatic cell nuclear transfer (SCNT). The mammary-specific expression plasmid pIACH(-), containing the attB recognition site for phiC31 integrase, were co-transfected with integrase expression plasmid pCMVInt into bovine fetal fibroblast cells (BFFs). PhiC31 integrase-mediated integrations in genome of BFFs were screened by nested inverse PCR. After analysis of sequence of the PCR products, 46.0% (23/50) of the both attB-genome junction sites (attL and attR) were confirmed, and four pseudo attP sites were identified. The integration rates in BF3, BF11, BF19 and BF4 sites were 4.0% (2/50), 6.0% (3/50), 16.0% (8/50) and 20.0% (10/50), respectively. BF3 is located in the bovine chromosome 3 collagen alpha-3 (VI) chain isomer 2 gene, while the other three sites are located in the non-coding region. The transgenic cell lines from BF11, BF19 and BF4 sites were used as donors for SCNT. Two calves from transgenic cells BF19 were born, one died within a few hours after birth, and another calf survived healthy. PCR and Southern blot analysis revealed integration of the transgene in the genome of cloned calves. The nested reverse PCR confirmed that the integration site in cloned calves was identical to the donor cells. The western blotting assessment indicated that recombinant HSA was expressed in the milk of transgenic cattle and the expression level was about 4-8 mg/mL. The present study demonstrated that phiC31 integrase system was an efficient and safety gene delivery tool for producing HSA transgenic cattle. The production of recombinant HSA in the milk of cattle may provide a large-scale and cost-effective resource.

  3. Modifications in the polymerase genes of a swine-like triple-reassortant influenza virus to generate live attenuated vaccines against 2009 pandemic H1N1 viruses.

    PubMed

    Pena, Lindomar; Vincent, Amy L; Ye, Jianqiang; Ciacci-Zanella, Janice R; Angel, Matthew; Lorusso, Alessio; Gauger, Philip C; Janke, Bruce H; Loving, Crystal L; Perez, Daniel R

    2011-01-01

    On 11 June 2009, the World Health Organization (WHO) declared that the outbreaks caused by novel swine-origin influenza A (H1N1) virus had reached pandemic proportions. The pandemic H1N1 (H1N1pdm) virus is the predominant influenza virus strain in the human population. It has also crossed the species barriers and infected turkeys and swine in several countries. Thus, the development of a vaccine that is effective in multiple animal species is urgently needed. We have previously demonstrated that the introduction of temperature-sensitive mutations into the PB2 and PB1 genes of an avian H9N2 virus, combined with the insertion of a hemagglutinin (HA) tag in PB1, resulted in an attenuated (att) vaccine backbone for both chickens and mice. Because the new pandemic strain is a triple-reassortant (TR) virus, we chose to introduce the double attenuating modifications into a swine-like TR virus isolate, A/turkey/OH/313053/04 (H3N2) (ty/04), with the goal of producing live attenuated influenza vaccines (LAIV). This genetically modified backbone had impaired polymerase activity and restricted virus growth at elevated temperatures. In vivo characterization of two H1N1 vaccine candidates generated using the ty/04 att backbone demonstrated that this vaccine is highly attenuated in mice, as indicated by the absence of signs of disease, limited replication, and minimum histopathological alterations in the respiratory tract. A single immunization with the ty/04 att-based vaccines conferred complete protection against a lethal H1N1pdm virus infection in mice. More importantly, vaccination of pigs with a ty/04 att-H1N1 vaccine candidate resulted in sterilizing immunity upon an aggressive intratracheal challenge with the 2009 H1N1 pandemic virus. Our studies highlight the safety of the ty/04 att vaccine platform and its potential as a master donor strain for the generation of live attenuated vaccines for humans and livestock.

  4. Detrital Zircon U-Pb Age Populations in Time and Space in the Arctic Alaska Terrane

    NASA Astrophysics Data System (ADS)

    Moore, T. E.

    2010-12-01

    The Arctic Alaska Terrane (ATT) occupies the only margin of the Ameriasia Basin whose origin and position since Paleozoic time is incompletely known. To better understand its tectonic history, detrital zircon (DZ) U-Pb ages from about 75 samples of clastic strata were obtained from representative parts of the ATT in northern Alaska. The oldest known strata of the AAT are Neoproterozoic clastic rocks exposed in the northeastern Brooks Range. DZ dating of these rocks show that they contain abundant ~1.8 Ga zircons and subordinate populations that indicate derivation from the northwest part of Laurentia. Upper Neoproterozoic strata in the Brooks Range, in contrast, contain populations dominated by ~600 Ma zircons. The latter ages are similar to those in parts of the ATT outside of northern Alaska, including the Seward Peninsula, Chukotka, and Wrangel Island that are thought to have been derived from the Timanian orogen of northern Baltica. Similar DZ populations have also been obtained from Silurian sandstones of the Lisburne Peninsula, suggesting that much of the western and southern parts of AAT may have formed in or near northern Baltica. A third group of DZ ages were found in deformed clastic rocks that were deposited across large parts of the North Slope in the Silurian and/or Devonian and are also present in parautochthonous settings in the Brooks Range. These rocks typically are dominated by DZ ages of 390-470 Ma, and sometimes contain subordinate non-Laurentian populations of ~1.5 Ga. These DZ ages, the underlying rocks of probable Baltic and Laurentian affinity, and evidence of significant deformation indicate that the ATT may have been constructed by Caledonian tectonism in the Silurian and Devonian. Following Devonian deformation, Mississippian to Triassic platform strata of the Ellesmerian Sequence were deposited on a regional unconformity. DZ ages from these rocks appear to reflect the compositions of the sub-unconformity units and indicate that Timanian

  5. Regulation of pro-adrenocorticotropin-endorphin synthesis and secretion in cultured neonatal rat anterior pituitary

    SciTech Connect

    Sato, S.M.; Mains, R.E. )

    1987-08-01

    Previous work demonstrated that newborn rat anterior pituitary corticotropes display processing patterns for pro-ACTH/endorphin that are different from the adult. The synthesis and release of beta-endorphin-related peptides was examined in dispersed cell and explant cultures of newborn anterior pituitary to investigate corticotrope development further. The temporal pattern of pro-ACTH/endorphin processing differed significantly from adult rat melanotropes and AtT-20 cells. While pro-ACTH/endorphin processing begins within 30 min of synthesis in adult melanotropes and AtT-20 cells, pulse-labeling of newborn corticotropes in culture indicated that pro-ACTH/endorphin remained uncleaved for at least 90 min after synthesis. With further incubation, there was a decrease in radioactivity associated with the precursor and an equivalent rise in the radioactivity associated with beta-endorphin and beta-lipotropin. However, unprocessed precursor still remained in the cultured newborn anterior pituitary cells after a 25-h chase. Although intact pro-ACTH/endorphin from newborn corticotropes was very long-lived, the precursor did undergo oligosaccharide maturation and became endoglycosidase H resistant within 1 h after synthesis. Similar to the adult, pro-ACTH/endorphin synthesis was doubled in cultures of newborn anterior pituitary chronically treated with 10 nM CRF resulting in a 3- to 4-fold stimulation of secretion over the basal rate. However, unlike the AtT-20 cell or adult rat corticotrope, the proteolytic processing of pro-ACTH/endorphin in the newborn corticotrope was altered by chronic secretagogue treatment; less pro-ACTH/endorphin was converted to beta-endorphin in secretagogue-treated corticotropes than in controls. Thus processing of pro-ACTH/endorphin in the corticotrope is not mature by birth and can be regulated by chronic CRF treatment.

  6. Gain-of-Function Mutations in Tnsc, an Atp-Dependent Transposition Protein That Activates the Bacterial Transposon Tn7

    PubMed Central

    Stellwagen, A. E.; Craig, N. L.

    1997-01-01

    The bacterial transposon Tn7 encodes five genes whose protein products are used in different combinations to direct transposition to different types of target sites. TnsABC+D directs transposition to a specific site in the Escherichia coli chromosome called attTn7, whereas TnsABC+E directs transposition to non-attTn7 sites. These transposition reactions can also recognize and avoid ``immune'' targets that already contain a copy of Tn7. TnsD and TnsE are required to activate TnsABC as well as to select a target site; no transposition occurs with wild-type TnsABC alone. Here, we describe the isolation of TnsC gain-of-function mutants that activate the TnsA+B transposase in the absence of TnsD or TnsE. Some of these TnsC mutants enable the TnsABC machinery to execute transposition without sacrificing its ability to discriminate between different types of targets. Other TnsC mutants appear to constitutively activate the TnsABC machinery so that it bypasses target signals. We also present experiments that suggest that target selection occurs early in the Tn7 transposition pathway in vivo: favorable attTn7 targets appear to promote the excision of Tn7 from the chromosome, whereas immune targets do not allow transposon excision to occur. This work supports the view that TnsC plays a central role in the evaluation and utilization of target DNAs. PMID:9055068

  7. Interleukin-7 Modulates Anti-Tumor CD8+ T Cell Responses via Its Action on Host Cells.

    PubMed

    Deiser, Katrin; Stoycheva, Diana; Bank, Ute; Blankenstein, Thomas; Schüler, Thomas

    2016-01-01

    The adoptive transfer of antigen-specific CD8+ T cells is a promising approach for the treatment of chronic viral and malignant diseases. In order to improve adoptive T cell therapy (ATT) of cancer, recent strategies aim at the antibody-based blockade of immunosuppressive signaling pathways in CD8+ T cells. Alternatively, adjuvant effects of immunostimulatory cytokines might be exploited to improve therapeutic CD8+ T cell responses. For example, Interleukin-7 (IL-7) is a potent growth, activation and survival factor for CD8+ T cells that can be used to improve virus- and tumor-specific CD8+ T cell responses. Although direct IL-7 effects on CD8+ T cells were studied extensively in numerous models, the contribution of IL-7 receptor-competent (IL-7R+) host cells remained unclear. In the current study we provide evidence that CD8+ T cell-mediated tumor rejection in response to recombinant IL-7 (rIL-7) therapy is strictly dependent on IL-7R+ host cells. On the contrary, CD8+ T cell expansion is independent of host IL-7R expression. If, however, rIL-7 therapy and peptide vaccination are combined, host IL-7R signaling is crucial for CD8+ T cell expansion. Unexpectedly, maximum CD8+ T cell expansion relies mainly on IL-7R signaling in non-hematopoietic host cells, similar to the massive accumulation of dendritic cells and granulocytes. In summary, we provide evidence that IL-7R+ host cells are major targets of rIL-7 that modulate therapeutic CD8+ T cell responses and the outcome of rIL-7-assisted ATT. This knowledge may have important implications for the design and optimization of clinical ATT protocols.

  8. Diabetes Is Associated with Worse Clinical Presentation in Tuberculosis Patients from Brazil: A Retrospective Cohort Study

    PubMed Central

    Hickson, Lucas S.; Daltro, Carla; Castro, Simone; Kornfeld, Hardy; Netto, Eduardo M.; Andrade, Bruno B.

    2016-01-01

    Background The rising prevalence of diabetes mellitus (DM) worldwide, especially in developing countries, and the persistence of tuberculosis (TB) as a major public health issue in these same regions, emphasize the importance of investigating this association. Here, we compared the clinical profile and disease outcomes of TB patients with or without coincident DM in a TB reference center in Brazil. Methods We performed a retrospective analysis of a TB patient cohort (treatment naïve) of 408 individuals recruited at a TB primary care center in Brazil between 2004 and 2010. Data on diagnosis of TB and DM were used to define the groups. The study groups were compared with regard to TB disease presentation at diagnosis as well as to clinical outcomes such as cure and mortality rates upon anti-tuberculosis therapy (ATT) initiation. A composite score utilizing clinical, radiological and microbiological parameters was used to compare TB severity between the groups. Results DM patients were older than non-diabetic TB patients. In addition, diabetic individuals more frequently presented with cough, night sweats, hemoptysis and malaise than those without DM. The overall pattern of lung lesions assessed by chest radiographic examination was similar between the groups. Compared to non-diabetic patients, those with TB-diabetes exhibited positive acid-fast bacilli in sputum samples more frequently at diagnosis and at 30 days after ATT initiation. Notably, higher values of the TB severity score were significantly associated with TB-diabetes comorbidity after adjustment for confounding factors. Moreover, during ATT, diabetic patients required more frequent transfers to TB reference hospitals for complex clinical management. Nevertheless, overall mortality and cure rates were indistinguishable between the study groups. Conclusions These findings reinforce the idea that diabetes negatively impacts pulmonary TB severity. Our study argues for the systematic screening for DM in TB

  9. An open-source system for in planta gene stacking by Bxb1 and Cre recombinases.

    PubMed

    Hou, Lili; Yau, Yuan-Yeu; Wei, Junjie; Han, Zhiguo; Dong, Zhicheng; Ow, David W

    2014-12-01

    The rapid development of crops with multiple transgenic traits arouses the need for an efficient system for creating stacked cultivars. Most major crops rely on classical breeding to introgress the transgene from a laboratory variety to the numerous cultivars adapted to different growing regions. Even with vegetative propagated crops, genetic crosses are conducted during varietal improvement prior to vegetative cloning. The probability to assort the 'x' number of transgenic loci into a single genome may seem trivial, (¼) (x) for a diploid species, but given the 'y' number of other nontransgenic traits that breeders also need to assemble into the same genome, the (¼) (x+y) probability for a 'breeding stack' could quickly make the line conversion process unmanageable. Adding new transgenes onto existing transgenic varieties without creating a new segregating locus would require site-specific integration of new DNA at the existing transgenic locus. Here, we tested a recombinase-mediated gene-stacking scheme in tobacco. Sequential site-specific integration was mediated by the mycobacteriophage Bxb1 integrase-catalyzed recombination between attP and attB sites. Transgenic DNA no longer needed after integration was excised by Cre recombinase-mediated recombination of lox sites. Site-specific integration occurred in ~10% of the integration events, with half of those events usable as substrates for a next round of gene stacking. Among the site-specific integrants, however, a third experienced gene silencing. Overall, precise structure and reproducible expression of the sequentially added triple traits were obtained at an overall rate of ~3% of the transformed clones--a workable frequency for the development of commercial cultivars. Moreover, since neither the Bxb1-att nor the Cre-lox system is under patent, there is freedom to operate.

  10. Lactic acid does not directly activate hypothalamic-pituitary corticotroph function.

    PubMed

    Petrides, J S; Deuster, P A; Mueller, G P

    1999-02-01

    The role that metabolic products play in regulating the hypothalamic-pituitary-adrenal (HPA) axis during strenuous exercise is speculative. This investigation examined the extent to which lactic acid, a major metabolite of anaerobic exercise, directly affects hypothalamic-pituitary function. Specifically, beta-endorphin secretion was measured from AtT-20 (D-16) mouse corticotroph tumor cells treated either acutely (15 min - 180 min) or chronically (1 day - 3 day) with physiologic levels of lactate (0. 5 x 10-3 M to 5 x 10-2 M) or lactate in combination with the corticotroph releasing factors: corticotroph releasing hormone (CRH), arginine vasopressin (AVP), norepinephrine and/or epinephrine. Findings with AtT-20 cell cultures were shown to be representative of responses in primary cultures of rat anterior pituitary. Lactic acid did not alter the spontaneous release of beta-endorphin by AtT-20 cells under either acute or chronic conditions. While CRH, norepinephrine, and epinephrine evoked significant increases in beta-endorphin release, lactate, in combination with these secretagogues did not alter their effects. Similarly, lactic acid failed to alter basal or stimulated release of beta-endorphin by primary cultures of rat anterior pituitary. The addition of lactate (3 x 103 M) to rat hypothalamic explants did, however, produce a modest but significant reduction in spontaneous CRH release, suggesting that lactate may facilitate the return to basal secretion following exercise. The present findings show that physiologic concentrations of lactate have no effect, either alone or in combination with other pituitary secretagogues, on corticotroph secretion. Whereas a physiologic action for lactate within the hypothalamus is possible, the present findings indicate that lactate is an inhibitor of CRH release. Thus, lactate does not appear to play a direct role in the profound activation of the HPA axis that occurs in response to strenuous exercise.

  11. A6V polymorphism of the human μ-opioid receptor decreases signalling of morphine and endogenous opioids in vitro

    PubMed Central

    Knapman, Alisa; Santiago, Marina; Connor, Mark

    2015-01-01

    Background and Purpose Polymorphisms of the μ opioid receptor (MOPr) may contribute to the variation in responses to opioid drugs in clinical and unregulated situations. The A6V variant of MOPr (MOPr-A6V) is present in up to 20% of individuals in some populations, and may be associated with heightened susceptibility to drug abuse. There are no functional studies examining the acute signalling of MOPr-A6V in vitro, so we investigated potential functional differences between MOPr and MOPr-A6V at several signalling pathways using structurally distinct opioid ligands. Experimental Approach CHO and AtT-20 cells stably expressing MOPr and MOPr-A6V were used. AC inhibition and ERK1/2 phosphorylation were assayed in CHO cells; K channel activation was assayed in AtT-20 cells. Key Results Buprenorphine did not inhibit AC or stimulate ERK1/2 phosphorylation in CHO cells expressing MOPr-A6V, but buprenorphine activation of K channels in AtT-20 cells was preserved. [D-Ala2, N-MePhe4, Gly-ol]-enkephalin, morphine and β-endorphin inhibition of AC was significantly reduced via MOPr-A6V, as was signalling of all opioids to ERK1/2. However, there was little effect of the A6V variant on K channel activation. Conclusions and Implications Signalling to AC and ERK via the mutant MOPr-A6V was decreased for many opioids, including the clinically significant drugs morphine, buprenorphine and fentanyl, as well endogenous opioids. The MOPr-A6V variant is common and this compromised signalling may affect individual responses to opioid therapy, while the possible disruption of the endogenous opioid system may contribute to susceptibility to substance abuse. PMID:25521224

  12. The pro region is not required for the expression or intracellular routeing of carboxypeptidase E.

    PubMed Central

    Song, L; Fricker, L D

    1997-01-01

    Carboxypeptidase E (CPE) is initially synthesized as a larger precursor containing an additional 14-residue propeptide that is highly conserved between human and rat. Previous studies have established that the proenzyme is enzymically active and that deletion of the pro region does not affect the expression of the active enzyme. In the present study the function of the pro region was examined both by deleting this region from CPE and by attaching this region to the N-terminus of albumin. CPE lacking the pro region is sorted into the regulated secretory pathway in AtT-20 cells, based on confocal microscopy and examination of the stimulated secretion of the protein. Stimulation of AtT-20 cells with either forskolin or phorbol 12-myristate 13-acetate induces the secretion of wild-type CPE and of CPE lacking the pro region to similar extents, indicating a similar efficiency of sorting of the mutant. When the pro region of proalbumin is replaced with the pro region of CPE followed by expression in AtT-20 cells, the protein is not sorted into the regulated pathway, based on the lack of stimulated secretion. Confocal microscopy suggests that the proCPE/albumin protein is retained in the endoplasmic reticulum to a greater extent than is proalbumin. Pulse-chase analysis indicates that the pro region of CPE is not efficiently removed from the N-terminus of albumin, and the small amount of propeptide cleavage that does occur takes place soon before secretion of the protein. In contrast, confocal microscopy indicates that the majority of the propeptide is removed from CPE, and that this cleavage occurs in the trans-Golgi network or soon after sorting into the secretory vesicles. Taken together, these results suggest that the pro region of CPE is not required for the expression or intracellular routeing of this protein. PMID:9173892

  13. Transfected human neuropeptide Y cDNA expression in mouse pituitary cells. Inducible high expression, peptide characterization, and secretion.

    PubMed

    Dickerson, I M; Dixon, J E; Mains, R E

    1987-10-01

    An expression vector was constructed that placed the cDNA for human neuropeptide Y (NPY) under the control of the mouse metallothionein promoter and was used to transfect the AtT-20 mouse anterior pituitary corticotrope cell line. AtT-20 cells normally process the pro-ACTH/endorphin precursor but do not produce detectable levels of NPY. The resulting AtT-20/NPY cell line (Mt.NPY1a) was used to study the ability of the corticotrope cells to synthesize, process, and secrete the foreign proNPY-related peptide products. The stable cell line created contains approximately 40 copies of proNPY cDNA per cell. NPY mRNA levels and proNPY synthesis were increased at least 35-fold when maximally induced with cadmium; proNPY synthesis was also induced by glucocorticoids. Upon induction the NPY secretion rate was equimolar to that of the endogenous peptides. ProNPY, NPY, and the COOH-terminal peptide produced by this cell line had molecular weight and amino acid-labeling pattern predicted from cDNA sequence data and from previous isolation of NPY-related molecules from NPY-producing cells. The structures of secreted proNPY, NPY, and COOH-terminal peptide, as well as determination of the site of proteolytic cleavage between NPY and the COOH-terminal peptide, were determined by tryptic mapping and Edman degradation of secreted biosynthetically labeled peptide products. The proNPY molecule appears to be processed in the same pathway responsible for cleavage of the endogenous pro-ACTH/endorphin precursor. Secretion of proNPY-derived peptides paralleled secretion of endogenous pro-ACTH/endorphin-derived products, under both basal and stimulated conditions. With induction proNPY expression there is a dose-dependent inhibition of both proNPY and pro-ACTH/endorphin proteolytic processing.

  14. Malpractice issues in radiology: medicare compliance versus standard of care conformance--real or imaginary conflict?

    PubMed

    Duszak, Richard; Berlin, Leonard

    2010-06-01

    Plaintiff's Attorney (Pl Att:: Doctor, the record shows that the patient was referred to the hospital's radiology department by her gynecologist for a screening mammogram. The record also shows that when completing the mammography information form, the patient wrote that she had a lump in her left breast, correct? Defendant Radiologist (Df Ra:): Yes. Pl Att: But your technologist performed, and you interpreted, a screening mammogram. Doesn't the radiology standard of care require you to do a diagnostic mammogram when the patient has a breast lump? Df Ra:: Well, normally yes, but if it's going to be a diagnostic mammogram, then the referring physician has to order it. In this case our tech called the gynecologist and asked him whether he wanted to order a diagnostic study, and he said no, he didn't feel the lump, and that we should only do a plain screening mammogram. Pl Att:: Please explain something. You're agreeing that a woman with a breast lump should have a diagnostic mammogram, but you are saying that you didn't do one because the patient's physician wouldn't order it? Don't you have a duty to do the diagnostic mammogram in a case like this on your own, without having to ask permission from the patient's gynecologist? Df Ra:: Only the treating physician can change a screening mammogram into a diagnostic mammogram, and I am not the treating physician. If I went ahead and did a diagnostic mammography examination on my own, it would be Medicare fraud, and our hospital's compliance officer says it could result in our hospital being fined and thrown out of the Medicare program. Pl Atty: What prevents you then from recommending-not ordering, but just recommending-a diagnostic mammogram in your report, because the patient says she's got a lump? Df Rad: Well, according to our hospital's compliance officer, that would also be fraud.

  15. Loss of neuromuscular control related to motion in the acutely ACL-injured knee: an experimental study.

    PubMed

    Bonsfills, N; Gómez-Barrena, E; Raygoza, J J; Núñez, A

    2008-10-01

    Ligamentomuscular and muscular stretch reflexes are known to contribute to knee joint stability. After anterior cruciate ligament (ACL) injury, a more intense and adjusted muscular response is required to maintain joint stability, but this neuromuscular control of the knee has not been clearly proved. The aim of the study is to record electromyography (EMG) signal and muscular fibre length variations in quadriceps and hamstrings of the knee with and without ACL, and to analyze and integrate the ligament strain and the muscular reaction to forced anterior tibial translation (ATT). In 17 knees from 12 cats, EMG electrodes and ultrasonomicrometry crystals were inserted into four main periarticular muscles, with strain gauges on periarticular ligament insertions. Their output signal was compared before and after ACL surgical section in series of ATT (at 90 degrees and 30 degrees knee flexion), and also during knee flexion and extension. Linear regression analysis was performed between the EMG signal and muscular fibre length variations, and between the EMG signal and the strain on ligament insertions, in the search of this reflex neuromuscular response. In the ACL deficient knees, the studied muscles showed a poor adjustment to motion of EMG firing, inversely to controls. The muscle stretch reflexes showed poorer correlation with post-peak EMG activity than the ligaments. ATT control depended mainly on hamstrings activity in control knees, whereas in unstable knees, quadriceps activity was associated with more tibial translation. Acute ACL-deficient knees showed poor neuromuscular control with weak ligamentomuscular reflexes and no muscular stretch reflexes, suggesting the ineffectiveness of acute muscular reaction to provide early mechanical knee stabilization after injury.

  16. Interleukin-7 Modulates Anti-Tumor CD8+ T Cell Responses via Its Action on Host Cells

    PubMed Central

    Deiser, Katrin; Stoycheva, Diana; Bank, Ute; Blankenstein, Thomas; Schüler, Thomas

    2016-01-01

    The adoptive transfer of antigen-specific CD8+ T cells is a promising approach for the treatment of chronic viral and malignant diseases. In order to improve adoptive T cell therapy (ATT) of cancer, recent strategies aim at the antibody-based blockade of immunosuppressive signaling pathways in CD8+ T cells. Alternatively, adjuvant effects of immunostimulatory cytokines might be exploited to improve therapeutic CD8+ T cell responses. For example, Interleukin-7 (IL-7) is a potent growth, activation and survival factor for CD8+ T cells that can be used to improve virus- and tumor-specific CD8+ T cell responses. Although direct IL-7 effects on CD8+ T cells were studied extensively in numerous models, the contribution of IL-7 receptor-competent (IL-7R+) host cells remained unclear. In the current study we provide evidence that CD8+ T cell-mediated tumor rejection in response to recombinant IL-7 (rIL-7) therapy is strictly dependent on IL-7R+ host cells. On the contrary, CD8+ T cell expansion is independent of host IL-7R expression. If, however, rIL-7 therapy and peptide vaccination are combined, host IL-7R signaling is crucial for CD8+ T cell expansion. Unexpectedly, maximum CD8+ T cell expansion relies mainly on IL-7R signaling in non-hematopoietic host cells, similar to the massive accumulation of dendritic cells and granulocytes. In summary, we provide evidence that IL-7R+ host cells are major targets of rIL-7 that modulate therapeutic CD8+ T cell responses and the outcome of rIL-7-assisted ATT. This knowledge may have important implications for the design and optimization of clinical ATT protocols. PMID:27447484

  17. A randomized Phase 2 trial of telavancin versus standard therapy in patients with uncomplicated Staphylococcus aureus bacteremia: the ASSURE study

    PubMed Central

    2014-01-01

    Background Staphylococcus aureus bacteremia is a common infection associated with significant morbidity and mortality. Telavancin is a bactericidal lipoglycopeptide active against Gram-positive pathogens, including methicillin-resistant S. aureus (MRSA). We conducted a randomized, double-blind, Phase 2 trial in patients with uncomplicated S. aureus bacteremia. Methods Patients were randomized to either telavancin or standard therapy (vancomycin or anti-staphylococcal penicillin) for 14 days. Continuation criteria were set to avoid complicated S. aureus bacteremia. The primary end point was clinical cure at 84 days. Results In total, 60 patients were randomized and 58 received ≥1 study medication dose (all-treated), 31 patients fulfilled inclusion/exclusion and continuation criteria (all-treated target [ATT]) (telavancin 15, standard therapy 16), and 17 patients were clinically evaluable (CE) (telavancin 8, standard therapy 9). Mean age (ATT) was 60 years. Intravenous catheters were the most common source of S. aureus bacteremia and ~50% of patients had MRSA. A similar proportion of CE patients were cured in the telavancin (88%) and standard therapy (89%) groups. All patients with MRSA bacteremia were cured and one patient with MSSA bacteremia failed study treatment in each group. Although adverse events (AEs) were more common in the telavancin ATT group (90% vs. 72%), AEs leading to drug discontinuation were similar (7%) in both treatment arms. Potentially clinically significant increases in serum creatinine (≥1.5 mg/dl and at least 50% greater than baseline) were more common in the telavancin group (20% vs. 7%). Conclusions This study suggests that telavancin may have utility for treatment of uncomplicated S. aureus bacteremia; additional studies are warranted. (Telavancin for Treatment of Uncomplicated Staphylococcus Aureus Bacteremia (ASSURE); NCT00062647). PMID:24884578

  18. Delayed initiation of anti-retroviral therapy in TB/HIV co-infected patients, Sanyati District, Zimbabwe, 2011-2012

    PubMed Central

    Maponga, Brian Abel; Chirundu, Daniel; Gombe, Notion Tafara; Tshimanga, Mufuta; Bangure, Donewell; Takundwa, Lucia

    2015-01-01

    Introduction Tuberculosis (TB) remains a public health problem and is driven by HIV. Recent studies indicate that anti-retroviral therapy (ART) initiated during the first two months of anti-TB treatment (ATT) reduces risk of HIV morbidity and mortality. In Sanyati district, 14% of TB/HIV co-infected patients were initiated on ART during TB treatment, in 2010. The study was conducted to determine the magnitude and determinants of delay in ART initiation, in TB/HIV co-infected patients. Methods An analytic cross sectional study was conducted at three study sites in Sanyati district. The outcome was delayed ART initiation, being failure to be initiated on ART during the first two months of ATT. Respondents were interviewed using pre-tested questionnaires. Epi-Info™ was used to generate frequencies, means, odds ratios and 95% confidence intervals. Stratified and logistic regression analysis was done. Results Of the 186 respondents, 63% had delayed ART initiation. Median delay from initiation of ATT to ART was 48 days (Q1=20; Q3=82). Risk factors for delayed ART initiation were: being treated for TB first time, AOR=2.23 (p=0.03); initially registered for HIV care outside Sanyati, AOR=3.08 (p<0.01); staying more than 5km from a clinic, AOR=3.29 (p<0.01). Enabling factors for early ART initiation was having a family member on ART, AOR=0.23 (p<0.01). Conclusion Significant delay and barriers to ART initiation were identified. Decentralization of ART initiation should be expedited. ART initiation should be expedited in patients with identified risk factors for delaying ART initiation. Peer support should be strengthened in families and community. Periodic evaluation of magnitude of delay and impact of early ART initiation in TB/HIV patients is recommended. PMID:26401222

  19. Multicopy Integration and Expression of Heterologous Genes in Methylobacterium extorquens ATCC 55366†

    PubMed Central

    Choi, Young J.; Bourque, Denis; Morel, Lyne; Groleau, Denis; Míguez, Carlos B.

    2006-01-01

    High-level expression of chromosomally integrated genes in Methylobacterium extorquens ATCC 55366 was achieved under the control of the strong M. extorquens AM1 methanol dehydrogenase promoter (PmxaF) using the mini-Tn7 transposon system. Stable maintenance and expression of the integrated genes were obtained in the absence of antibiotic selective pressure. Furthermore, using this technology, a multicopy integration protocol for M. extorquens was also developed. Chromosomal integration of one to five copies of the gene encoding the green fluorescent protein (gfp) was achieved. The multicopy-based expression system permitted expression of a preset number of gene copies. A unique specific Tn7 integration locus in the chromosome of M. extorquens, known as the Tn7 attachment site (attTn7 site), was identified. This single attTn7 site was identified in an intergenic region between glmS, which encodes the essential enzyme glucosamine-6-phosphate synthetase, and dhaT, which encodes 1,3-propanediol dehydrogenase. The fact that the integration event is site specific and the fact that the attTn7 site is a noncoding region of the chromosome make the mini-Tn7 transposon system very useful for insertion of target genes and subsequent expression. In all transformants tested, expression and segregation of the transforming gene were stable without generation of secondary mutations in the host. In this paper, we describe single and multicopy chromosome integration and stable expression of heterologous genes (bgl [β-galactosidase], est [esterase], and gfp [green fluorescent protein]) in M. extorquens. PMID:16391115

  20. A Look into the Melting Pot: The mecC-Harboring Region Is a Recombination Hot Spot in Staphylococcus stepanovicii

    PubMed Central

    Semmler, Torsten; Harrison, Ewan M.; Lübke-Becker, Antina; Ulrich, Rainer G.; Wieler, Lothar H.; Guenther, Sebastian; Stamm, Ivonne; Hanssen, Anne-Merethe; Holmes, Mark A.; Vincze, Szilvia; Walther, Birgit

    2016-01-01

    Introduction Horizontal gene transfer (HGT) is an important driver for resistance- and virulence factor accumulation in pathogenic bacteria such as Staphylococcus aureus. Methods Here, we have investigated the downstream region of the bacterial chromosomal attachment site (attB) for the staphylococcal cassette chromosome mec (SCCmec) element of a commensal mecC-positive Staphylococcus stepanovicii strain (IMT28705; ODD4) with respect to genetic composition and indications of HGT. S. stepanovicii IMT28705 was isolated from a fecal sample of a trapped wild bank vole (Myodes glareolus) during a screening study (National Network on “Rodent-Borne Pathogens”) in Germany. Whole genome sequencing (WGS) of IMT28705 together with the mecC-negative type strain CM7717 was conducted in order to comparatively investigate the genomic region downstream of attB (GenBank accession no. KR732654 and KR732653). Results The bank vole isolate (IMT28705) harbors a mecC gene which shares 99.2% nucleotide (and 98.5% amino acid) sequence identity with mecC of MRSA_LGA251. In addition, the mecC-encoding region harbors the typical blaZ-mecC-mecR1-mecI structure, corresponding with the class E mec complex. While the sequences downstream of attB in both S. stepanovicii isolates (IMT28705 and CM7717) are partitioned by 15 bp direct repeats, further comparison revealed a remarkable low concordance of gene content, indicating a chromosomal “hot spot” for foreign DNA integration and exchange. Conclusion Our data highlight the necessity for further research on transmission routes of resistance encoding factors from the environmental and wildlife resistome. PMID:26799070

  1. The patella ligament insertion angle influences quadriceps usage during walking of anterior cruciate ligament deficient patients.

    PubMed

    Shin, Choongsoo S; Chaudhari, Ajit M; Dyrby, Chris O; Andriacchi, Thomas P

    2007-12-01

    Following ACL injury a reduction in the peak knee flexion moment during walking (thought to be created by a decrease of quadriceps contraction) has been described as an adaptation to reduce anterior tibial translation (ATT) relative to the femur. However, the amount of ATT caused by quadriceps contraction is influenced by the patellar ligament insertion angle (PLIA). The purpose of this study was to test the hypothesis that quadriceps usage during walking correlates to individual anatomical variations in the extensor mechanism as defined by PLIA. PLIA and gait were measured for ACL-deficient knees, using subjects' contralateral knees as controls. In ACL-deficient knees, PLIA was negatively correlated (R2 = 0.59) to peak knee flexion moment (balanced by net quadriceps moment), while no correlation was found in contralateral knees. Reduction in peak flexion moment in ACL-deficient knees compared to their contralateral knees was distinctive in subjects with large PLIA, possibly to avoid excessive ATT. These results suggest that subject-specific anatomic variability of knee extensor mechanism may account for the individual variability previously observed in adaptation to a quadriceps reduction strategy following ACL injury. The average (+/-1 SD) PLIA of ACL-deficient knees (21.1 +/- 3.4 degrees) was less than the average PLIA of contralateral knees (23.9 +/- 3.1 degrees). This altered equilibrium position of the tibiofemoral joint associated with reduced PLIA and adaptations of gait patterns following ACL injury may be associated with degenerative changes in the articular cartilage. In the future, individually tailored treatment and rehabilitation considering individuals' specific extensor anatomy may improve clinical outcomes.

  2. Transformation of Mo and W thiosalts into unsupported sulfide catalysts: A temperature dependent in-situ spectroscopic investigation

    SciTech Connect

    Yi, Yanjiao; Williams, Christopher T.; Glascock, Mary; Xiong, Guang; Lauterbach, Jochen; Liang, Changhai

    2014-08-15

    Highlights: • TPD–MS results suggest that the majority of S remains in the solid sample after decomposition. • Decomposition of the (CH{sub 3}){sub 4}N is not directly correlated with that of the MS{sub 4}{sup 2−} species. • Surface of materials does not track directly with the bulk at various treatment temperatures. • ATT-derived materials retained a significant amount of +6 state consistent with the precursor. - Abstract: The thermal decomposition of ammonium thiomolybdate (ATM), ammonium thiotungstate (ATT), tetramethylammonium thiomolybdate (TMATM) and tetramethylammonium thiotungstate (TMATT) are investigated as a function of temperature by using thermogravimetric analysis (TGA), temperature-programmed decomposition with mass spectroscopy (TPS–MS), in-situ Fourier transform infrared (FTIR) and Raman spectroscopies, and X-ray photoelectron spectroscopy (XPS). The results allow for correlations to be made between the changes in the bulk and surface structures of the materials, and the evolution of gas-phase decomposition products. The major difference between the ammonium and tetramethylammonium precursors is the complexity of the thermal decomposition profile, which is found to follow two steps over a wide temperature range for the former, but one step over a relatively narrow range for the latter materials. Raman and FTIR spectra reveal the decomposition of the ammonium and tetramethylammonium groups, along with the decomposition of the initial sulfide structures to metal disulfides. For both sets of precursors, XPS results show that the surface of the resulting materials at various temperatures of treatment does not track directly with the state of the bulk material. While the ATM, TMATM, and TMATT-derived material surfaces are reduced to the 4{sup +} state at the highest temperature, indicating disulfides, the ATT-derived materials still retained a significant amount of W{sup 6+} state consistent with the starting precursor.

  3. Mapping Long-Term Functional Changes in Cerebral Blood Flow by Arterial Spin Labeling

    PubMed Central

    Ssali, Tracy; Anazodo, Udunna C.; Bureau, Yves; MacIntosh, Bradley J.; Günther, Matthias; St. Lawrence, Keith

    2016-01-01

    Although arterial spin labeling (ASL) is appealing for mapping long-term changes in functional activity, inter-sessional variations in basal blood flow, arterial transit times (ATTs), and alignment errors, can result in significant false activation when comparing images from separate sessions. By taking steps to reduce these sources of noise, this study assessed the ability of ASL to detect functional CBF changes between sessions. ASL data were collected in three sessions to image ATT, resting CBF and CBF changes associated with motor activation (7 participants). Activation maps were generated using rest and task images acquired in the same session and from sessions separated by up to a month. Good agreement was found when comparing between-session activation maps to within-session activation maps with only a 16% decrease in precision (within-session: 90 ± 7%) and a 13% decrease in the Dice similarity (within-session: 0.75 ± 0.07) coefficient after a month. In addition, voxel-wise reproducibility (within-session: 4.7 ± 4.5%) and reliability (within-session: 0.89 ± 0.20) of resting grey-matter CBF decreased by less than 18% for the between-session analysis relative to within-session values. ATT variability between sessions (5.0 ± 2.7%) was roughly half the between-subject variability, indicating that its effects on longitudinal CBF were minimal. These results demonstrate that conducting voxel-wise analysis on CBF images acquired on different days is feasible with only modest loss in precision, highlighting the potential of ASL for longitudinal studies. PMID:27706218

  4. The systemic effects of sclerostin overexpression using ΦC31 integrase in mice.

    PubMed

    Zhang, Dongdong; Park, Bo Mi; Kang, Myengmo; Nam, HeeJin; Kim, Eun Jin; Bae, ChuHyun; Lim, Sung Kil

    2016-04-01

    Sclerostin, encoded by the Sost gene, is mainly produced by osteocytes in bone and antagonizes the Wnt/β-catenin signaling pathway, which is a requisite for bone formation. Currently, human anti-sclerostin antibodies are being tested in phase III clinical trials. In addition, serum sclerostin levels are reported to be associated with bone mineral density and fracture risk in normal individuals; however, the correlation between serum sclerostin and bone mass remains controversial. To study the effects of the continuous exposure of exogenous sclerostin on bone, a ΦC31 integrase system, which has the characteristics of site-specificity and efficiency, was applied for the delivery of the Sost gene in this study. We injected Sost-attB plasmid with or without ΦC31 integrase plasmid into the mouse tail vein using a hydrodynamic-based method. The site-specific integration of the Sost gene into the mouse genome was confirmed by examining a pseudo-attP site on the hepatic genomic DNA. Sclerostin was expressed in the hepatocytes, secreted into the blood flow, and maintained at high concentrations in the mice with both Sost-attB plasmid and ΦC31 integrase plasmid injections, which was observed by serial measurement. Moreover, the mice with long-term high levels of serum sclerostin showed trabecular bone loss on micro-CT analysis. Peripheral B cell populations were not affected. Our results suggested that sclerostin could be expressed in the liver and sustained successfully at high levels in the blood by using the ΦC31 integrase system, leading to trabecular bone loss. These findings may help to further ascertain the effects of sclerostin introduced exogenously on the skeleton.

  5. Lack of association between plasma levels of non-nucleoside reverse transcriptase inhibitors & virological outcomes during rifampicin co-administration in HIV-infected TB patients

    PubMed Central

    Ramachandran, Geetha; Kumar, A.K. Hemanth; Ponnuraja, C.; Ramesh, K.; Rajesh, Lakshmi; Chandrasekharan, C.; Swaminathan, Soumya

    2013-01-01

    Background & objectives: Among patients with HIV-associated tuberculosis (TB), reduced plasma non-nucleoside reverse transcriptase inhibitors (NNRTI) concentrations during rifampicin (RMP) co-administration could lead to HIV treatment failure. This study was undertaken to examine the association between plasma nevirapine (NVP) and efavirenz (EFV) concentrations and virological outcomes in patients infected with HIV-1 and TB. Methods: This was a nested study undertaken in a clinical trial of patients with HIV-1 and TB, randomized to two different once-daily antiretroviral treatment (ART) regimens along with anti-TB treatment (ATT). Trough concentrations of plasma NVP and EFV were estimated at months 1 (during ATT and ART) and 6 months (ART only) by HPLC. Plasma HIV-1 RNA level >400 copies/ml or death within 6 months of ART were considered as unfavourable outcomes. Genotyping of CYP2B6 516G>T polymorphism was performed. Results: Twenty nine per cent of patients in NVP arm had an unfavourable outcome at 6 months compared to 9 per cent in EFV arm (P<0.08). The mean NVP and EFV levels estimated at 1 and 6 months did not significantly differ between favourable and unfavourable responders. Logistic regression analysis showed CYP2B6 516G>T polymorphism significantly associated with virologic outcome in patients receiving EFV–based regimen. Interpretation & conclusions: Trough plasma concentrations of NVP and EFV did not show any association with response to ART in patients on ATT and once-daily ART. CYP2B6 516G>T polymorphism was associated with virologic outcome among patients on EFV. PMID:24521642

  6. Kappa Opioid Receptor Activation of p38 MAPK Is GRK3- and Arrestin-dependent in Neurons and Astrocytes*

    PubMed Central

    Bruchas, Michael R.; Macey, Tara A.; Lowe, Janet D.; Chavkin, Charles

    2007-01-01

    AtT-20 cells expressing the wild-type kappa opioid receptor (KOR) increased phospho-p38 MAPK following treatment with the kappa agonist U50,488. The increase was blocked by the kappa antagonist norbinaltorphimine and not evident in untransfected cells. In contrast, U50,488 treatment of AtT-20 cells expressing KOR having alanine substituted for serine-369 (KSA) did not increase phospho-p38. Phosphorylation of serine 369 in the KOR carboxyl terminus by G-protein receptor kinase 3 (GRK3) was previously shown to be required for receptor desensitization, and the results suggest that p38 MAPK activation by KOR may require arrestin recruitment. This hypothesis was tested by transfecting arrestin3-(R170E), a dominant positive form of arrestin that does not require receptor phosphorylation for activation. AtT-20 cells expressing both KSA and arrestin3-(R170E) responded to U50,488 treatment with an increase in phospho-p38 consistent with the hypothesis. Primary cultured astrocytes (glial fibrillary acidic protein-positive) and neurons (γ-aminobutyric acid-positive) isolated from mouse striata also responded to U50,488 by increasing phospho-p38 immunolabeling. p38 activation was not evident in either striatal astrocytes or neurons isolated from KOR knock-out mice or GRK3 knock-out mice. Astrocytes pretreated with small interfering RNA for arrestin3 were also unable to activate p38 in response to U50,488 treatment. Furthermore, in striatal neurons, the kappa-mediated phospho-p38 labeling was colocalized with arrestin3. These findings suggest that KOR may activate p38 MAPK in brain by a GRK3 and arrestin-dependent mechanism. PMID:16648139

  7. Lycopene and Beta-Carotene Induce Growth Inhibition and Proapoptotic Effects on ACTH-Secreting Pituitary Adenoma Cells

    PubMed Central

    Leite de Oliveira, Felipe; Soares, Nathália; de Mattos, Rômulo Medina; Hecht, Fábio; Dezonne, Rômulo Sperduto; Vairo, Leandro; Goldenberg, Regina Coeli dos Santos; Gomes, Flávia Carvalho Alcântara; de Carvalho, Denise Pires; Gadelha, Mônica R.; Nasciutti, Luiz Eurico; Miranda-Alves, Leandro

    2013-01-01

    Pituitary adenomas comprise approximately 10–15% of intracranial tumors and result in morbidity associated with altered hormonal patterns, therapy and compression of adjacent sella turcica structures. The use of functional foods containing carotenoids contributes to reduce the risk of chronic diseases such as cancer and vascular disorders. In this study, we evaluated the influence of different concentrations of beta-carotene and lycopene on cell viability, colony formation, cell cycle, apoptosis, hormone secretion, intercellular communication and expression of connexin 43, Skp2 and p27kip1 in ACTH-secreting pituitary adenoma cells, the AtT20 cells, incubated for 48 and 96 h with these carotenoids. We observed a decrease in cell viability caused by the lycopene and beta-carotene treatments; in these conditions, the clonogenic ability of the cells was also significantly decreased. Cell cycle analysis revealed that beta-carotene induced an increase of the cells in S and G2/M phases; furthermore, lycopene increased the proportion of these cells in G0/G1 while decreasing the S and G2/M phases. Also, carotenoids induced apoptosis after 96 h. Lycopene and beta-carotene decreased the secretion of ACTH in AtT20 cells in a dose-dependent manner. Carotenoids blocked the gap junction intercellular communication. In addition, the treatments increased the expression of phosphorylated connexin43. Finally, we also demonstrate decreased expression of S-phase kinase-associated protein 2 (Skp2) and increased expression of p27kip1 in carotenoid-treated cells. These results show that lycopene and beta-carotene were able to negatively modulate events related to the malignant phenotype of AtT-20 cells, through a mechanism that could involve changes in the expression of connexin 43, Skp2 and p27kip1; and suggest that these compounds might provide a novel pharmacological approach to the treatment of Cushing’s disease. PMID:23667519

  8. 5-ALA Fluorescence in Native Pituitary Adenoma Cell Lines: Resection Control and Basis for Photodynamic Therapy (PDT)?

    PubMed Central

    Poeschke, Stephan; Greve, Burkhard; Prevedello, Daniel; Santacroce, Antonio; Stummer, Walter; Senner, Volker

    2016-01-01

    Objective: Pituitary adenomas (PA), especially invasive ones, are often not completely resectable. Usage of 5-aminolevulinic acid (5-ALA) for fluorescence guided surgery could improve the rate of total resection and, additionally, open the doors for photodynamic therapy (PDT) in case of unresectable or partially resected PAs. The aim of this study was to investigate the uptake of 5-ALA and the effect of 5-ALA based PDT in cell lines. Methods: GH3 and AtT-20 cell lines were incubated with different concentrations of 5-ALA, protoporphyrin IX (PPIX) fluorescence was measured by flow cytometry and fluorescencespectrometry. WST-1 assays were performed to determine the surviving fraction of cells after PDT. PPIX fluorescence intensities and PDT effect of the pituitary adenoma cells were compared to U373MG, a well-known glioblastoma cell line. Results: Both cell lines showed a 5-ALA dependent intracellular PPIX fluorescence. Significant differences after 24hrs of incubation were observed in AtT-20 cells in comparison to GH3. Regardless of the incubation or metabolism time, there was a proliferation inhibiting effect after PDT, with no statistical significance. Conclusion: Since GH3 cells showed a heterogenous uptake of 5-ALA in the flow cytometry profile, but not constantly high concentrations they might have a 5-ALA efflux mechanism, which still needs to be determined. In the case of AtT-20, the cells might need a longer time for the uptake due to their size or slow metabolism. We showed that the different cell lines have different uptake and metabolism mechanisms, which needs to be further investigated. The general uptake of 5-ALA allows the possibility of resection control and PDT for pituitary adenomas. But, the role of PDT for unresectable pituitary adenomas deserves further investigations. PMID:27583461

  9. Divergent immunomodulating effects of probiotics on T cell responses to oral attenuated human rotavirus vaccine and virulent human rotavirus infection in a neonatal gnotobiotic piglet disease model.

    PubMed

    Chattha, Kuldeep S; Vlasova, Anastasia N; Kandasamy, Sukumar; Rajashekara, Gireesh; Saif, Linda J

    2013-09-01

    Rotaviruses (RVs) are a leading cause of childhood diarrhea. Current oral vaccines are not effective in impoverished countries where the vaccine is needed most. Therefore, alternative affordable strategies are urgently needed. Probiotics can alleviate diarrhea in children and enhance specific systemic and mucosal Ab responses, but the T cell responses are undefined. In this study, we elucidated the T cell and cytokine responses to attenuated human RV (AttHRV) and virulent human RV (HRV) in gnotobiotic pigs colonized with probiotics (Lactobacillus rhamnosus strain GG [LGG] and Bifidobacterium lactis Bb12 [Bb12]), mimicking gut commensals in breastfed infants. Neonatal gnotobiotic pigs are the only animal model susceptible to HRV diarrhea. Probiotic colonized and nonvaccinated (Probiotic) pigs had lower diarrhea and reduced virus shedding postchallenge compared with noncolonized and nonvaccinated pigs (Control). Higher protection in the Probiotic group coincided with higher ileal T regulatory cells (Tregs) before and after challenge, and higher serum TGF-β and lower serum and biliary proinflammatory cytokines postchallenge. Probiotic colonization in vaccinated pigs enhanced innate serum IFN-α, splenic and circulatory IFN-γ-producing T cells, and serum Th1 cytokines, but reduced serum Th2 cytokines compared with noncolonized vaccinated pigs (Vac). Thus, LGG+Bb12 induced systemic Th1 immunostimulatory effects on oral AttHRV vaccine that coincided with lower diarrhea severity and reduced virus shedding postchallenge in Vac+Pro compared with Vac pigs. Previously unreported intestinal CD8 Tregs were induced in vaccinated groups postchallenge. Thus, probiotics LGG+Bb12 exert divergent immunomodulating effects, with enhanced Th1 responses to oral AttHRV vaccine, whereas inducing Treg responses to virulent HRV. PMID:23918983

  10. Liquid fossil-fuel technology. Quarterly technical progress report, January-March 1983

    SciTech Connect

    Linville, B.

    1983-07-01

    Accomplishments for the quarter ending March 1983 are presented under the following headings: liquid fossil fuel cycle, processing, utilization, and project integration and technology transfer. Feature articles for this quarter are: (1) abandoned oil field reports issued; (2) oilfield water data bank report published; (3) microbial enhanced recovery report issued; (4) polymer-augmented project could be economic today; (5) carbon dioxide EOR estimates given; (6) BETC passes 65th milestone; and (7) fifty achievements for fifty years (1918-1968). BETC publications are also listed. (ATT)

  11. In vivo analysis of the pivot shift phenomenon during computer navigated ACL reconstruction.

    PubMed

    Lane, Clayton G; Warren, Russell F; Stanford, Fatima C; Kendoff, Daniel; Pearle, Andrew D

    2008-05-01

    ACL insufficiency can be documented clinically with the pivot shift maneuver, but the specific pathologic kinematics of the pivot shift is difficult to quantify. Navigation provides an opportunity to analyze in vivo the motions that comprise the pivot shift and the kinematic changes that are inherent after ACL reconstruction. We hypothesized that tibial rotation, anterior tibial translation (ATT), acceleration of posterior translation (APT) and the newly described angle of P, quantified during navigated pivot shift examination, correlate with clinical grading of the pivot shift phenomena. Navigation data from 12 patients who underwent navigated ACL surgery were retrospectively reviewed. A characteristic P-shaped track of motion is recorded by the navigation software during the pivot-shift examination. The "angle of P" was developed as a means characterizing this track of motion and was measured in all cases. The tibial rotation, maximum anterior tibial translation and acceleration of posterior translation during the pivot shift were also measured. The charts of these patients were reviewed to obtain information on the clinical grading of the pivot-shift before and after ACL reconstruction. Spearman correlation analysis was then used to identify significant correlations between clinical grade of the pivot shift and the angle of p measured with computer navigation. After reconstruction, the clinical grade of the pivot shift was zero in all patients. The tibial rotation, maximum ATT, APT and the angle of p also decreased. On analysis of 24 EUAs, 12 before reconstruction and 12 after, there was excellent and significant correlation between the clinical grade of pivot shift examination and the angle of P (R2 = 0.97, p < 0.001). Only good correlation was noted between the clinical pivot shift and the rotation (R2 = 0.77, p < 0.0001), maximum ATT (R2 = 0.87, p < 0.0001) and APT (R2 = 0.81, p < 0.0001). There was a stepwise increase of 6-7 mm of translation and 5-6 degrees

  12. Participation in the Juntos Conditional Cash Transfer Program in Peru Is Associated with Changes in Child Anthropometric Status but Not Language Development or School Achievement1234

    PubMed Central

    Andersen, Christopher T; Reynolds, Sarah A; Behrman, Jere R; Crookston, Benjamin T; Dearden, Kirk A; Escobal, Javier; Mani, Subha; Sánchez, Alan; Stein, Aryeh D; Fernald, Lia CH

    2015-01-01

    Background: It is unclear what effects a conditional cash transfer (CCT) program would have on child anthropometry, language development, or school achievement in the context of the nutrition transition experienced by many low- and middle-income countries. Objective: We estimated the association of participation in Peru’s Juntos CCT with anthropometry, language development, and school achievement among children aged 7–8 y. Methods: We used data from the Young Lives Study of a cohort born between 2001 and 2002. We estimated associations of the Juntos program with height-for-age z score (HAZ), body mass index–for–age z score (BAZ), stunting, and overweight at age 7–8 y separately for children participating in the program for ≥2 y (n = 169) and children participating for <2 y (n = 188). We then estimated associations with receptive vocabulary and grade achievement among children who had been assessed at age 4–6 y before enrollment in Juntos (n = 243). We identified control subjects using propensity score matching and conducted difference-in-differences comparisons. Results: Juntos participation was associated with increases in HAZ among boys participating for ≥2 y [average effect of treatment among the treated (ATT): 0.43; 95% CI: 0.09, 0.77; P = 0.01] and for boys participating for <2 y (ATT: 0.52; 95% CI: 0.23, 0.80; P < 0.01). Among girls participating in the program for ≥2 y, BAZ declined (ATT: –0.60; 95% CI: –1.00, –0.21; P < 0.01) as did the prevalence of overweight (ATT: –22.0 percentage points; 95% CI: –42.5, –2.7 percentage points; P = 0.03). We observed no significant associations of Juntos participation with receptive vocabulary or grade attainment. Conclusions: CCT program participation in Peru was associated with better linear growth among boys and decreased BAZ among girls, highlighting that a large-scale poverty-alleviation intervention may influence anthropometric outcomes in the context of the nutrition transition. PMID

  13. Advanced subsonic long-haul transport terminal area compatibility study. Volume 2: Research and technology recommendations

    NASA Technical Reports Server (NTRS)

    1974-01-01

    The Terminal Area Compatibility (TAC) study is briefly summarized for background information. The most important research items for the areas of noise congestion, and emissions are identified. Other key research areas are also discussed. The 50 recommended research items are categorized by flight phase, technology, and compatibility benefits. The relationship of the TAC recommendations to the previous ATT recommendations is discussed. The bulk of the document contains the 50 recommended research items. For each item, the potential payoff, state of readiness, recommended action and estimated cost and schedule are given.

  14. Development and validation of the Thai version of the 4 ‘A’s Test for delirium screening in hospitalized elderly patients with acute medical illnesses

    PubMed Central

    Kuladee, Sanchai; Prachason, Thanavadee

    2016-01-01

    Background The English version of the 4 ‘A’s Test (4AT) is a rapid screening tool for delirium with a high sensitivity and specificity among hospitalized elderly patients. Objective To develop the Thai version of the 4AT (4AT-T) and assess its validity. Subjects and setting A total of 97 elderly patients aged 60 years or above who were admitted to the general medical wards were included. Methods Both authors independently translated the English version of the 4AT into Thai and thereafter developed a single reconciled forward translation by consensus. Back translation was performed by a bilingual native English speaker and it was then reviewed to ensure its agreement with the original one. After 24 hours of admission, subjects were enrolled and clinical data collected. Definite diagnosis of delirium was made by a psychiatrist using the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, Text-Revision criteria and the 6-item Thai Delirium Rating Scale; the 4AT was then administered to participants by nurses within 30 minutes. A 4AT score ≥4 was considered positive for delirium screening. The optimal cut-off point of the 4AT-T was identified by Youden’s index. Results In all, 24 out of 97 participants met the Diagnostic and Statistical Manual of Mental Disorders, 4th edition, Text-Revision criteria for delirium. At a cut-off score of 4 or greater, the 4AT-T exhibited satisfactory diagnostic performance with a sensitivity of 83.3% (95% confidence interval (CI): 62.6%–95.3%) and specificity of 86.3% (95% CI: 76.3%–93.2%). The area under the receiver operating characteristic curve was 0.92. The specified score provided maximal Youden’s index, suggesting an optimal criterion value for delirium screening. Conclusion The 4AT-T is a valid delirium-screening instrument for hospitalized elderly patients with acute medical illnesses. PMID:26966365

  15. Characterization of western coals and coal macerals by solid-state nuclear magnetic resonance spectroscopy. Progress report, August 1, 1981-January 31, 1982

    SciTech Connect

    Pugmire, R.J.; Grant, D.M.

    1982-01-01

    Two bituminous coals were selected on the basis of petrographic analysis as having high concentrations of vitrinite, inertinite and liptinite. These coal samples were ground up to about 10-3 ..mu..m, treated with HCl and HF to remove the mineral matter, and the macerals were separated by density gradient centrifugation. Structural features of the three different macerals were obtained by NMR analysis based on conventional CP/MAS and multiple pulse multi-dimensional /sup 13/C NMR spectroscopy. Results are presented in tabular form. (ATT)

  16. Thermal protein denaturation and protein aggregation in cells made thermotolerant by various chemicals: role of heat shock proteins.

    PubMed

    Kampinga, H H; Brunsting, J F; Stege, G J; Burgman, P W; Konings, A W

    1995-08-01

    Thermotolerance (TT) induced by sodium arsenite (A-TT: 100 microM, 1 h, 37 degrees C) was compared to heat-induced thermotolerance (H-TT: 15 min, 44 degrees C) using HeLa S3 cells. All four pretreatments led to comparable levels of thermotolerance and also induced resistance to arsenite-, ethanol-, and diamide-induced toxicity (clonogenic ability). Stress-induced expression of the major heat shock proteins (hsp27, hsc70(p73), hsp70(p72), and hsp90) was generally highest in H-TT cells and lowest in A-TT cells. Interestingly, the four types of TT cells showed distinct differences in certain aspects of resistance against thermal protein damage. Thermal protein denaturation and aggregation determined in isolated cellular membrane fractions was found to be attenuated when they were isolated from H-TT and A-TT cells but not when isolated from E-TT and D-TT cells. The heat resistance in the proteins of the membrane fraction corresponded with elevated levels of hsp70(p72) associated with the isolated membrane fractions. In the nuclear fraction, only marginal (not significant) attenuation of the formation of protein aggregates (as determined by TX-100 (in)solubility) was observed. However, the postheat recovery from heat-induced protein aggregation in the nucleus was faster in H-TT, E-TT, and D-TT cells, but not in A-TT cells. Despite the fact that elevated levels of hsp27, hsp70(p73), and hsp70(p72) were found in the TX-100 insoluble nuclear fraction derived from all TT cells, no correlation was found with the degree of resistance in terms of the accelerated recovery from nuclear protein aggregation. The only correlation between accelerated recovery from nuclear protein aggregates was that with total cellular levels of hsp27. The data indicate that heat-induced loss of clonogenic ability may be a multitarget rather than a single target event. A threshold of damage may exist in cells after exposure to heat; multiple sets of proteins in (different compartments of) the cell

  17. Prevalence of Pulmonary Tuberculosis among Adults in a Rural Sub-District of South India

    PubMed Central

    Chadha, Vineet K.; Kumar, Prahlad; Anjinappa, Sharada M.; Singh, Sanjay; Narasimhaiah, Somashekar; Joshi, Malathi V.; Gupta, Joydev; Lakshminarayana; Ramchandra, Jitendra; Velu, Magesh; Papkianathan, Suganthi; Babu, Suseendra; Krishna, Hemalatha

    2012-01-01

    Background We conducted a survey to estimate point prevalence of bacteriologically positive pulmonary TB (PTB) in a rural area in South India, implementing TB program DOTS strategy since 2002. Methods Survey was conducted among persons ≥15 years of age in fifteen clusters selected by simple random sampling; each consisting of 5–12 villages. Persons having symptoms suggestive of PTB or history of anti-TB treatment (ATT) were eligible for sputum examination by smear microscopy for Acid Fast Bacilli and culture for Mycobacterium tuberculosis; two sputum samples were collected from each eligible person. Persons with one or both sputum specimen positive on microscopy and/or culture were labeled suffering from PTB. Prevalence was estimated after imputing missing values to correct for bias introduced by incompleteness of data. In six clusters, registered persons were also screened by X-ray chest. Persons with any abnormal shadow on X-ray were eligible for sputum examination in addition to those with symptoms and ATT. Multiplication factor calculated as ratio of prevalence while using both screening tools to prevalence using symptoms screening alone was applied to entire study population to estimate prevalence corrected for non-screening by X-ray. Results Of 71,874 residents ≥15 years of age, 63,362 (88.2%) were screened for symptoms and ATT. Of them, 5120 (8.1%) - 4681 (7.4%) with symptoms and an additional 439 (0.7%) with ATT were eligible for sputum examination. Spot specimen were collected from 4850 (94.7%) and early morning sputum specimens from 4719 (92.2%). Using symptom screening alone, prevalence of smear, culture and bacteriologically positive PTB in persons ≥15 years of age was 83 (CI: 57–109), 152 (CI: 108–197) and 196 (CI :145–246) per 100,000 population respectively. Prevalence corrected for non-screening by X-ray was 108 (CI: 82–134), 198 (CI: 153–243) and 254 (CI: 204–301) respectively. Conclusion Observed prevalence suggests further

  18. USGS Telecommunications Responding to Change

    USGS Publications Warehouse

    Hott, James L.

    1985-01-01

    The telecommunications industry is undergoing tremendous change due to the court ordered breakup of the monopoly once enjoyed by American Telephone & Telegraph (AT&T). This action has resulted in a plethora of new services and products in all of the communications fields, including traditional voice and data. The new products are making extensive use of computer technology. At the same time, costs of telecommunications services have risen dramatically over the past three years. This article reviews some of the major actions that the Geological Survey has taken in response to these changes.

  19. STS-51G Crew Portrait

    NASA Technical Reports Server (NTRS)

    1985-01-01

    The crew assigned to the STS-51G mission included (kneeling front left to right) Daniel C. Brandenstein, commander; and John O. Creighton, pilot. Standing, left to right, are mission specialists Shannon W. Lucid, Steven R. Nagel, and John M. Fabian; and payload specialists Sultan Salman Al-Saud, and Patrick Baudrey. Launched aboard the Space Shuttle Discovery on June 17, 1985 at 7:33:00 am (EDT), the STS-51G mission's primary payloads were three communications satellites: MORELOS-A for Mexico; ARABSAT-A , for Arab Satellite communications; and TELSTAR-3D, for ATT.

  20. Limitations on High Data Rate Optical Fiber Transmission Systems Due to Transmission Impairment

    SciTech Connect

    Menyuk, Curtis R.

    2002-03-15

    This project supplemented our regular DOE grant from the Basic Energy Sciences organization with the goal of fostering industrial partnerships and student internships. During the project period, we have interacted with between 15 and 20 companies in the optical fiber telecommunications equipment industry, and our students have participated in a number of highly visible projects with companies such as Ciena, Science Applications International Corporation, KDD, ATT, Virtual Photonics, Inc., Phaethon Telecommunications, PhotonEx, and others. The project led to many successful interactions and numerous job offers for our students.

  1. Sandia National Laboratories: The First Fifty Years

    SciTech Connect

    MORA,CARL J.

    1999-11-03

    On Nov. 1, 1999, Sandia National Laboratories celebrates its 50th birthday. Although Sandia has its roots in the World War II-era Manhattan Project, Sandia began operating as a separate nuclear weapons engineering laboratory under the management of AT&T on Nov. 1, 1949. Today the lab employs more than 7,000 people at its two sites in Albuquerque and Livermore, California, and has research and development missions in national security, energy and environmental technologies, and U.S. economic competitiveness. Lockheed Martin Corporation operates Sandia for the US. Department of Energy.

  2. A PC-interactive stereonet plotting program

    NASA Astrophysics Data System (ADS)

    Pilant, Walter L.

    The computer program here described allows the structural geologist to rotate and revolve structural data interactively (strike dip: trend plunge). These actions can remove the effects of dip and plunge. Once a final screen plot is obtained, it may be printed on an associated graphics printer. This program requires an IBM PC/XT AT or compatible equipped with the color graphics (CGA) card. It also works satisfactorily on compatible PCs such as the COMPAQ or AT&T 6300 (which use a combination mono CGA screen). The addition of a math coprocessor (8087 or 80287) greatly speeds up the response time but is not required.

  3. Spent-fuel-storage alternatives

    SciTech Connect

    Not Available

    1980-01-01

    The Spent Fuel Storage Alternatives meeting was a technical forum in which 37 experts from 12 states discussed storage alternatives that are available or are under development. The subject matter was divided into the following five areas: techniques for increasing fuel storage density; dry storage of spent fuel; fuel characterization and conditioning; fuel storage operating experience; and storage and transport economics. Nineteen of the 21 papers which were presented at this meeting are included in this Proceedings. These have been abstracted and indexed. (ATT)

  4. The precise measurement of the attenuation coefficients of various IR optical materials applicable to immersion grating

    NASA Astrophysics Data System (ADS)

    Kaji, Sayumi; Sarugaku, Yuki; Ikeda, Yuji; Kobayashi, Naoto; Nakanishi, Kenshi; Kondo, Sohei; Yasui, Chikako; Kawakita, Hideyo

    2014-07-01

    Immersion grating is a next-generation diffraction grating which has the immersed the diffraction surface in an optical material with high refractive index of n > 2, and can provide higher spectral resolution than a classical reflective grating. Our group is developing various immersion gratings from the near- to mid-infrared region (Ikeda et al.1, 2, 3, 4, Sarugaku et al.5, and Sukegawa et al.6). The internal attenuation αatt of the candidate materials is especially very important to achieve the high efficiency immersion gratings used for astronomical applications. Nevertheless, because there are few available data as αatt < 0.01cm-1 in the infrared region, except for measurements of CVD-ZnSe, CVD-ZnS, and single-crystal Si in the short near-infrared region reported by Ikeda et al.7, we cannot select suitable materials as an immersion grating in an aimed wavelength range. Therefore, we measure the attenuation coefficients of CdTe, CdZnTe, Ge, Si, ZnSe, and ZnS that could be applicable to immersion gratings. We used an originally developed optical unit attached to a commercial FTIR which covers the wide wavelength range from 1.3μm to 28μm. This measurement system achieves the high accuracy of (triangle)αatt ~ 0.01cm-1. As a result, high-resistivity single-crystal CdZnTe, single-crystal Ge, single-crystal Si, CVD-ZnSe, and CVD-ZnS show αatt < 0.01cm-1 at the wavelength range of 5.5 - 19.0μm, 2.0 - 10.5μm, 1.3 - 5.4μm, 1.7 - 13.2μm, and 1.9 - 9.2μm, respectively. This indicates that these materials are good candidates for high efficiency immersion grating covering those wavelength ranges. We plan to make similar measurement under the cryogenic condition as T <= 10K for the infrared, especially mid-infrared applications.

  5. Children's Deviant Behavior in Primary Education: Comparing Physical Educator's Implicit Theory With Diagnostic Criteria.

    PubMed

    Efstratopoulou, Maria A; Janssen, Rianne; Simons, Johan

    2012-06-11

    Objective: Physical educators' implicit theory of children's deviant behavior in primary education was investigated and compared with diagnostic criteria. Method: A total of 60 physical education (PE) teachers reported deviant behaviors during lessons. Experts sorted these behaviors together with the official diagnostic criteria into categories based on perceived similarity in content. Results: Hierarchical cluster analysis on the derived similarity matrix among the behaviors suggested that PE teachers focus more on attention problems, disobedience, and aggressiveness when internalizing behaviors, such as anxiety and low energy, were less reported. Conclusion: PE teachers may be important and useful informants on children's behavior in school settings. (J. of Att. Dis. 2012; XX(X) 1-XX).

  6. Energies of organic compounds. Technical progress report. [Ketals, orthoesters, norbonyl ketals

    SciTech Connect

    Wiberg, K. B.

    1981-02-19

    Two automated reaction calorimetry systems were built and tested. The first made use of a quartz thermometer probe as the temperature measuring element. The second was constructed using a thermister which was part of a Wheatstone bridge. The off-balance voltage from the bridge was measured using the 6 1/2 digit digital voltmeter, and transferred to the microprocessor. The one disadvantage of the first system is the relatively large size of the quartz sensor. The calorimeter systems were used to measure: the enthalpies of hydrolysis of ketals, acetal, orthoesters, enthalpies of hydration of alkenes; enthalpies of formation of small ring hydrocarbons. (ATT)

  7. Cloud Computing: An Overview

    NASA Astrophysics Data System (ADS)

    Qian, Ling; Luo, Zhiguo; Du, Yujian; Guo, Leitao

    In order to support the maximum number of user and elastic service with the minimum resource, the Internet service provider invented the cloud computing. within a few years, emerging cloud computing has became the hottest technology. From the publication of core papers by Google since 2003 to the commercialization of Amazon EC2 in 2006, and to the service offering of AT&T Synaptic Hosting, the cloud computing has been evolved from internal IT system to public service, from cost-saving tools to revenue generator, and from ISP to telecom. This paper introduces the concept, history, pros and cons of cloud computing as well as the value chain and standardization effort.

  8. Operation and performance of a longitudinal feedback system using digital signal processing

    SciTech Connect

    Teytelman, D.; Fox, J.; Hindi, H.

    1994-11-22

    A programmable longitudinal feedback system using a parallel array of AT&T 1610 digital signal processors has been developed as a component of the PEP-II R&D program. This system has been installed at the Advanced Light Source (LBL) and implements full speed bunch by bunch signal processing for storage rings with bunch spacing of 4ns. Open and closed loop results showing the action of the feedback system are presented, and the system is shown to damp coupled-bunch instabilities in the ALS. A unified PC-based software environment for the feedback system operation is also described.

  9. Bunch-by-bunch longitudinal feedback system for PEP-II

    SciTech Connect

    Oxoby, G.; Claus, R.; Fox, J.

    1994-06-01

    This paper describes the implementation of the bunch-by-bunch longitudinal feedback system for the PEP-II B Factory. Bunch spacing down to 2 ns is achieved using 500 Megasamples per second A/D and D/A converters, and AT&T 1610 Digital Signal Processors are integrated to run a downsampled feedback algorithm for each bunch in parallel. This general purpose programmable system, packaged in VXI and VME, is modular and scalable to offer portability to other accelerator rings. The control and monitoring hardware and software architecture have been developed to provide ease of operation as well as diagnostic tools for machine physics.

  10. Effect of length of anodized TiO 2 tubes on photoreactivity: Photocurrent, Cr(VI) reduction and H 2 evolution

    NASA Astrophysics Data System (ADS)

    Park, Minsung; Heo, Ahyoung; Shim, Eunjung; Yoon, Jaekyung; Kim, Hansung; Joo, Hyunku

    Anodized tubular TiO 2 electrodes (ATTEs) are prepared using an organic additive consisting of either (i) ethylene glycol (EG) or (ii) glycerol (Gly) to make various photoanodes with different length of TiO 2 tubes and thereby to investigate the effect of their length on the photo-driven activity for hydrogen evolution and Cr(VI) reduction, as well as on the photocurrent. The ATTEs with EG have longer TiO 2 tubes (3.42-15.6 μm) than those with Gly (0.26-1.95, 6.82 μm). The former samples exhibit higher photocurrent densities (22.8-32.8 mA cm -2) than the latter (8.0-19.4, 20.3 mA cm -2). The latter samples (tube length of less than 7 μm) clearly exhibit a change of the rate-determining step from electron migration to photohole capture as the scanned applied bias increases, since the photocurrent shows a plateau for tube lengths above 2 μm. Meanwhile, the samples with EG remain in the electron migration step up to a tube length of 16 μm and is due to the difference of the morphology, crystal phase and crystallinity. This favourable characteristic is also applied to and well matched with the results from the reactions of Cr(VI) reduction and hydrogen evolution (up to ca. 250 μmol h -1).

  11. AltPSM contact hole application at DRAM 4xnm nodes with dry 193nm lithography

    NASA Astrophysics Data System (ADS)

    Noelscher, Christoph; Henkel, Thomas; Jauzion-Graverolle, Franck; Hennig, Mario; Morgana, Nicolo; Schlief, Ralph; Moukara, Molela; Koehle, Roderick; Neubauer, Ralf

    2008-03-01

    To avoid expensive immersion lithography and to further use existing dry tools for critical contact layer lithography at 4Xnm DRAM nodes the application of altPSM is investigated and compared to attPSM. Simulations and experiments with several test masks showed that by use of altPSM with suitable 0°/180° coloring and assist placement 30nm smaller contacts can be resolved through pitch with sufficient process windows (PW). This holds for arrays of contacts with variable lengths through short and long side pitches. A further benefit is the lower mask error enhancement factor (MEEF). Nevertheless 3D mask errors (ME) consume benefits in the PW and the assist placement and coloring of the main features (MF) put some constraints on the chip design. An altPSM compatible 4Xnm full-chip layout was realized without loss of chip area. Mask making showed very convincing results with respect to CDU, etch depth uniformity and defectiveness. The printed intra-field CD uniformity was comparable to attPSM despite the smaller target CDs. Room for improvement is identified in OPC accuracy and in automatic assist placement and sizing.

  12. Nonlinear Reduced Order Random Response Analysis of Structures With Shallow Curvature

    NASA Technical Reports Server (NTRS)

    Przekop, Adam; Rizzi, Stephen A.

    2005-01-01

    The goal of this investigation is to further develop nonlinear modal numerical simulation methods for application to geometrically nonlinear response of structures with shallow curvature under random loadings. For reduced order analysis, the modal basis selection must be capable of reflecting the coupling in both the linear and nonlinear stiffness. For the symmetric shallow arch under consideration, four categories of modal basis functions are defined. Those having symmetric transverse displacements (ST modes) can be designated as transverse dominated (ST-T) modes and in-plane dominated (ST-I) modes. Those having anti-symmetric transverse displacements (AT modes) can similarly be designated as transverse dominated (AT-T) modes and in-plane dominated (AT-I) modes. The response of an aluminum arch under a uniformly distributed transverse random loading is investigated. Results from nonlinear modal simulations made using various modal bases are compared with those obtained from a numerical simulation in physical degrees-of-freedom. While inclusion of ST-T modes is important for all response regimes, it is found that the ST-I modes become increasingly important in the nonlinear response regime, and that AT-T and AT-I modes are critical in the autoparametric regime.

  13. Nonlinear Reduced Order Random Response Analysis of Structures with Shallow Curvature

    NASA Technical Reports Server (NTRS)

    Przekop, Adam; Rizzi, Stephen A.

    2006-01-01

    The goal of this investigation is to further develop nonlinear modal numerical simulation methods for application to geometrically nonlinear response of structures with shallow curvature under random loadings. For reduced order analysis, the modal basis selection must be capable of reflecting the coupling in both the linear and nonlinear stiffness. For the symmetric shallow arch under consideration, four categories of modal basis functions are defined. Those having symmetric transverse displacements (ST modes) can be designated as transverse dominated (ST-T) modes and in-plane dominated (ST-I) modes. Those having anti-symmetric transverse displacements (AT modes) can similarly be designated as transverse dominated (AT-T) modes and in-plane dominated (AT-I) modes. The response of an aluminum arch under a uniformly distributed transverse random loading is investigated. Results from nonlinear modal simulations made using various modal bases are compared with those obtained from a numerical simulation in physical degrees-of-freedom. While inclusion of ST-T modes is important for all response regimes, it is found that the ST-I modes become increasingly important in the nonlinear response regime, and that AT-T and AT-I modes are critical in the autoparametric regime.

  14. [A Simple and Efficient Method of Inducing Targeted Deletions in the Drosophila Genome].

    PubMed

    Kravchuk, O I; Mikhailov, V S; Savitsky, M Yu

    2015-11-01

    Deletion mutagenesis is one of the most efficient approaches to studying gene function. However, conventional methods of inducing targeted mutations in the drosophila genome are time- and labor-consuming. This work proposes a new, simple, and effective method of producing drosophila mutants with gene deletions. The method involves the insertion of I-Scel and I-CreI recognition sites and a fragment homologous to the target sequence into the chromosome region of interest by means of an attB-containing construct, the induction of double-strand DNA breaks by the appropriate meganuclease, and their repair by homologous recombination. The procedure results in a deletion extending from the attP-site to the target locus. A cassette was designed to enable single-step construct production for the deletion of any given genomic region. A set of markers facilitates the selection of recombination events. The efficacy of the proposed technique was confirmed by the induction of a 47-kb deletion containing the qtc gene.

  15. On my association with Bell Labs

    NASA Astrophysics Data System (ADS)

    Sondhi, M. Mohan

    2001-05-01

    I joined the Acoustics Research department at Bell Labs in 1962, just eight days before AT&T launched the first communications satellite, Telstar. During the 39 years between 1962 and my retirement in 2001, I worked on several problems related in one way or another to the processing of speech signals. Schroeder and Flanagan are presenting talks from a broad perspective in this session, so I will confine this talk to just my own contributions and collaborations for some of the topics on which I worked, e.g., echo cancellation, inverse problems in acoustics, speech analysis, synthesis, and recognition. I will tell you about one of these contributions that fortunately turned out to yield considerable profits to AT&T. To give you a flavor of the spirit of free inquiry at Bell Labs during that period, I will tell you about the contribution that I am most proud of (which was supported for several years even though it had no monetary value). And I will also mention the contribution that is most often cited of all my papers (which was in collaboration with two mathematicians, and had nothing at all to do with acoustics).

  16. An Efficient Procedure for Marker-Free Mutagenesis of S. coelicolor by Site-Specific Recombination for Secondary Metabolite Overproduction

    PubMed Central

    Dai, Ruixue; Yu, Meiying; Zhao, Guoping; Ding, Xiaoming

    2013-01-01

    Streptomyces bacteria are known for producing important natural compounds by secondary metabolism, especially antibiotics with novel biological activities. Functional studies of antibiotic-biosynthesizing gene clusters are generally through homologous genomic recombination by gene-targeting vectors. Here, we present a rapid and efficient method for construction of gene-targeting vectors. This approach is based on Streptomyces phage φBT1 integrase-mediated multisite in vitro site-specific recombination. Four ‘entry clones’ were assembled into a circular plasmid to generate the destination gene-targeting vector by a one-step reaction. The four ‘entry clones’ contained two clones of the upstream and downstream flanks of the target gene, a selectable marker and an E. coli-Streptomyces shuttle vector. After targeted modification of the genome, the selectable markers were removed by φC31 integrase-mediated in vivo site-specific recombination between pre-placed attB and attP sites. Using this method, part of the calcium-dependent antibiotic (CDA) and actinorhodin (Act) biosynthetic gene clusters were deleted, and the rrdA encoding RrdA, a negative regulator of Red production, was also deleted. The final prodiginine production of the engineered strain was over five times that of the wild-type strain. This straightforward φBT1 and φC31 integrase-based strategy provides an alternative approach for rapid gene-targeting vector construction and marker removal in streptomycetes. PMID:23409083

  17. Extension of 193 nm dry lithography to 45-nm half-pitch node: double exposure and double processing technique

    NASA Astrophysics Data System (ADS)

    Biswas, Abani M.; Li, Jianliang; Hiserote, Jay A.; Melvin, Lawrence S., III

    2006-10-01

    Immersion lithography and multiple exposure techniques are the most promising methods to extend lithography manufacturing to the 45nm node. Although immersion lithography has attracted much attention recently as a promising optical lithography extension, it will not solve all the problems at the 45-nm node. The 'dry' option, (i.e. double exposure/etch) which can be realized with standard processing practice, will extend 193-nm lithography to the end of the current industry roadmap. Double exposure/etch lithography is expensive in terms of cost, throughput time, and overlay registration accuracy. However, it is less challenging compared to other possible alternatives and has the ability to break through the κ I barrier (0.25). This process, in combination with attenuated PSM (att-PSM) mask, is a good imaging solution that can reach, and most likely go beyond, the 45-nm node. Mask making requirements in a double exposure scheme will be reduced significantly. This can be appreciated by the fact that the separation of tightly-pitched mask into two less demanding pitch patterns will reduce the stringent specifications for each mask. In this study, modeling of double exposure lithography (DEL) with att-PSM masks to target 45-nm node is described. In addition, mask separation and implementation issues of optical proximity corrections (OPC) to improve process window are studied. To understand the impact of OPC on the process window, Fourier analysis of the masks has been carried out as well.

  18. Foldback intercoil DNA and the mechanism of DNA transposition.

    PubMed

    Kim, Byung-Dong

    2014-09-01

    Foldback intercoil (FBI) DNA is formed by the folding back at one point of a non-helical parallel track of double-stranded DNA at as sharp as 180° and the intertwining of two double helixes within each other's major groove to form an intercoil with a diameter of 2.2 nm. FBI DNA has been suggested to mediate intra-molecular homologous recombination of a deletion and inversion. Inter-molecular homologous recombination, known as site-specific insertion, on the other hand, is mediated by the direct perpendicular approach of the FBI DNA tip, as the attP site, onto the target DNA, as the attB site. Transposition of DNA transposons involves the pairing of terminal inverted repeats and 5-7-bp tandem target duplication. FBI DNA configuration effectively explains simple as well as replicative transposition, along with the involvement of an enhancer element. The majority of diverse retrotransposable elements that employ a target site duplication mechanism is also suggested to follow the FBI DNA-mediated perpendicular insertion of the paired intercoil ends by non-homologous end-joining, together with gap filling. A genome-wide perspective of transposable elements in light of FBI DNA is discussed.

  19. Analysis of the First Temperate Broad Host Range Brucellaphage (BiPBO1) Isolated from B. inopinata

    PubMed Central

    Hammerl, Jens A.; Göllner, Cornelia; Al Dahouk, Sascha; Nöckler, Karsten; Reetz, Jochen; Hertwig, Stefan

    2016-01-01

    Brucella species are important human and animal pathogens. Though, only little is known about mobile genetic elements of these highly pathogenic bacteria. To date, neither plasmids nor temperate phages have been described in brucellae. We analyzed genomic sequences of various reference and type strains and identified a number of putative prophages residing within the Brucella chromosomes. By induction, phage BiPBO1 was isolated from Brucella inopinata. BiPBO1 is a siphovirus that infects several Brucella species including Brucella abortus and Brucella melitensis. Integration of the phage genome occurs adjacent to a tRNA gene in chromosome 1 (chr 1). The bacterial (attB) and phage (attP) attachment sites comprise an identical sequence of 46 bp. This sequence exists in many Brucella and Ochrobactrum species. The BiPBO1 genome is composed of a 46,877 bp double-stranded DNA. Eighty-seven putative gene products were determined, of which 32 could be functionally assigned. Strongest similarities were found to a temperate phage residing in the chromosome of Ochrobactrum anthropi ATCC 49188 and to prophages identified in several families belonging to the order rhizobiales. The data suggest that horizontal gene transfer may occur between Brucella and Ochrobactrum and underpin the close relationship of these environmental and pathogenic bacteria. PMID:26858702

  20. Rapid Optimization of Engineered Metabolic Pathways with Serine Integrase Recombinational Assembly (SIRA).

    PubMed

    Merrick, C A; Wardrope, C; Paget, J E; Colloms, S D; Rosser, S J

    2016-01-01

    Metabolic pathway engineering in microbial hosts for heterologous biosynthesis of commodity compounds and fine chemicals offers a cheaper, greener, and more reliable method of production than does chemical synthesis. However, engineering metabolic pathways within a microbe is a complicated process: levels of gene expression, protein stability, enzyme activity, and metabolic flux must be balanced for high productivity without compromising host cell viability. A major rate-limiting step in engineering microbes for optimum biosynthesis of a target compound is DNA assembly, as current methods can be cumbersome and costly. Serine integrase recombinational assembly (SIRA) is a rapid DNA assembly method that utilizes serine integrases, and is particularly applicable to rapid optimization of engineered metabolic pathways. Using six pairs of orthogonal attP and attB sites with different central dinucleotide sequences that follow SIRA design principles, we have demonstrated that ΦC31 integrase can be used to (1) insert a single piece of DNA into a substrate plasmid; (2) assemble three, four, and five DNA parts encoding the enzymes for functional metabolic pathways in a one-pot reaction; (3) generate combinatorial libraries of metabolic pathway constructs with varied ribosome binding site strengths or gene orders in a one-pot reaction; and (4) replace and add DNA parts within a construct through targeted postassembly modification. We explain the mechanism of SIRA and the principles behind designing a SIRA reaction. We also provide protocols for making SIRA reaction components and practical methods for applying SIRA to rapid optimization of metabolic pathways.

  1. BioSYNTHESIS: access to a knowledge network of health sciences databases.

    PubMed

    Broering, N C; Hylton, J S; Guttmann, R; Eskridge, D

    1991-04-01

    Users of the IAIMS Knowledge Network at the Georgetown University Medical Center have access to multiple in-house and external databases from a single point of entry through BioSYNTHESIS. The IAIMS project has developed a rich environment of biomedical information resources that represent a medical decision support system for campus physicians and students. The BioSYNTHESIS system is an information navigator that provides transparent access to a Knowledge Network of over a dozen databases. These multiple health sciences databases consist of bibliographic, informational, diagnostic, and research systems which reside on diverse computers such as DEC VAXs, SUN 490, AT&T 3B2s, Macintoshes, IBM PC/PS2s and the AT&T ISN and SYTEK network systems. Ethernet and TCP/IP protocols are used in the network architecture. BioSYNTHESIS also provides network links to the other campus libraries and to external institutions. As additional knowledge resources and technological advances have become available. BioSYNTHESIS has evolved from a two phase to a three phase program. Major components of the system including recent achievements and future plans are described.

  2. Mechanism of action of somatostatin: an overview of receptor function and studies of the molecular characterization and purification of somatostatin receptor proteins.

    PubMed

    Patel, Y C; Murthy, K K; Escher, E E; Banville, D; Spiess, J; Srikant, C B

    1990-09-01

    To determine whether somatostatin receptor subtypes arise from molecular heterogeneity of the receptor protein, we have cross-linked the putative receptor in normal rat tissues and in AtT-20 and GH3 cells, both chemically with SS-14, SS-28 and Tyr3 SMS ligands, as well as by photoaffinity labeling with an azido derivative of Tyr3 SMS (EE 581). Three prominent somatostatin receptor proteins of 58-kDa, 32-kDa, and 27-kDa size have been identified. These proteins exhibit a tissue-specific distribution, ligand selectivity, and relative preference for SS-14 and SS-28 binding, and thus qualify as somatostatin receptor subtypes. Using EE 581 as a photoaffinity probe, the 58-kDa and 32-kDa proteins have been purified to homogeneity from brain and AtT-20 cells by successive SDS-PAGE. The 58-kDa form has been trypsinized and amino acid sequence data obtained from four tryptic fragments. With the help of synthetic oligonucleotides derived from these sequences, work is currently in progress to clone the 58-kDa protein to elucidate its complete sequence, its expression, and its functional relationship to the somatostatin receptor and its pharmacological subtypes.

  3. Multiple gene transcripts of the somatostatin receptor SSTR2: tissue selective distribution and cAMP regulation.

    PubMed

    Patel, Y C; Greenwood, M; Kent, G; Panetta, R; Srikant, C B

    1993-04-15

    The rodent SSTR2 mRNA has been reported to be alternatively spliced to generate long (SSTR2A) and short (SSTR2B) receptor isoforms which differ in sequence at their C-terminal regulatory domains. By extending the 3' nucleotide sequence of the human gene (hSSTR2) we show highly conserved intron/exon boundaries suggesting that hSSTR2 is also capable of generating spliced variants. mRNA blots of rat tissues reveal 2 transcripts of 2.8 and 2.3 kb that are differentially expressed in brain regions and multiple peripheral organs. The 2.3 kb mRNA is preferentially expressed in pituitary tumor cells (AtT-20 mouse, GH3 rat, human prolactinoma, human somatotroph adenoma), but not in rat or human insulinoma cells. This transcript shows 4 fold induction by forskolin in AtT-20 cells suggesting cAMP dependent control of SSTR2 gene expression. The abundant expression of SSTR2 gene, the occurrence of 2 isoforms and evidence of extensive regulation at both gene and peptide levels, suggests that SSTR2 is the principal SST-14 selective subtype.

  4. Conservative site-specific and single-copy transgenesis in human LINE-1 elements

    PubMed Central

    Vijaya Chandra, Shree Harsha; Makhija, Harshyaa; Peter, Sabrina; Myint Wai, Cho Mar; Li, Jinming; Zhu, Jindong; Ren, Zhonglu; D'Alcontres, Martina Stagno; Siau, Jia Wei; Chee, Sharon; Ghadessy, Farid John; Dröge, Peter

    2016-01-01

    Genome engineering of human cells plays an important role in biotechnology and molecular medicine. In particular, insertions of functional multi-transgene cassettes into suitable endogenous sequences will lead to novel applications. Although several tools have been exploited in this context, safety issues such as cytotoxicity, insertional mutagenesis and off-target cleavage together with limitations in cargo size/expression often compromise utility. Phage λ integrase (Int) is a transgenesis tool that mediates conservative site-specific integration of 48 kb DNA into a safe harbor site of the bacterial genome. Here, we show that an Int variant precisely recombines large episomes into a sequence, termed attH4X, found in 1000 human Long INterspersed Elements-1 (LINE-1). We demonstrate single-copy transgenesis through attH4X-targeting in various cell lines including hESCs, with the flexibility of selecting clones according to transgene performance and downstream applications. This is exemplified with pluripotency reporter cassettes and constitutively expressed payloads that remain functional in LINE1-targeted hESCs and differentiated progenies. Furthermore, LINE-1 targeting does not induce DNA damage-response or chromosomal aberrations, and neither global nor localized endogenous gene expression is substantially affected. Hence, this simple transgene addition tool should become particularly useful for applications that require engineering of the human genome with multi-transgenes. PMID:26673710

  5. Patterns of postictal cerebral perfusion in idiopathic generalized epilepsy: a multi-delay multi-parametric arterial spin labelling perfusion MRI study

    PubMed Central

    Chen, Guangxiang; Lei, Du; Ren, Jiechuan; Zuo, Panli; Suo, Xueling; Wang, Danny J. J.; Wang, Meiyun; Zhou, Dong; Gong, Qiyong

    2016-01-01

    The cerebral haemodynamic status of idiopathic generalized epilepsy (IGE) is a very complicated process. Little attention has been paid to cerebral blood flow (CBF) alterations in IGE detected by arterial spin labelling (ASL) perfusion magnetic resonance imaging (MRI). However, the selection of an optimal delay time is difficult for single-delay ASL. Multi-delay multi-parametric ASL perfusion MRI overcomes the limitations of single-delay ASL. We applied multi-delay multi-parametric ASL perfusion MRI to investigate the patterns of postictal cerebral perfusion in IGE patients with absence seizures. A total of 21 IGE patients with absence seizures and 24 healthy control subjects were enrolled. IGE patients exhibited prolonged arterial transit time (ATT) in the left superior temporal gyrus. The mean CBF of IGE patients was significantly increased in the left middle temporal gyrus, left parahippocampal gyrus and left fusiform gyrus. Prolonged ATT in the left superior temporal gyrus was negatively correlated with the age at onset in IGE patients. This study demonstrated that cortical dysfunction in the temporal lobe and fusiform gyrus may be related to epileptic activity in IGE patients with absence seizures. This information can play an important role in elucidating the pathophysiological mechanism of IGE from a cerebral haemodynamic perspective. PMID:27374369

  6. Rapid and efficient introduction of a foreign gene into bacterial artificial chromosome-cloned varicella vaccine by Tn7-mediated site-specific transposition

    SciTech Connect

    Somboonthum, Pranee; Koshizuka, Tetsuo; Okamoto, Shigefumi; Matsuura, Masaaki; Gomi, Yasuyuki; Takahashi, Michiaki; Yamanishi, Koichi; Mori, Yasuko

    2010-06-20

    Using a rapid and reliable system based on Tn7-mediated site-specific transposition, we have successfully constructed a recombinant Oka varicella vaccine (vOka) expressing the mumps virus (MuV) fusion protein (F). The backbone of the vector was our previously reported vOka-BAC (bacterial artificial chromosome) genome. We inserted the transposon Tn7 attachment sequence, LacZ{alpha}-mini-attTn7, into the region between ORF12 and ORF13 to generate a vOka-BAC-Tn genome. The MuV-F expressing cassette was transposed into the vOka-BAC genome at the mini-attTn7 transposition site. MuV-F protein was expressed in recombinant virus, rvOka-F infected cells. In addition, the MuV-F protein was cleaved in the rvOka-F infected cells as in MuV-infected cells. The growth of rvOka-F was similar to that of the original recombinant vOka without the F gene. Thus, we show that Tn7-mediated transposition is an efficient method for introducing a foreign gene expression cassette into the vOka-BAC genome as a live virus vector.

  7. Tuberculous gastric perforation: report of a case.

    PubMed

    Sharma, Deborshi; Gupta, Arun; Jain, Bhupendra Kumar; Agrawal, Vivek; Dargan, Puneet; Upreti, Lalendra; Arora, Vinod

    2004-01-01

    A 21-year-old woman presented with a 2-day history of acute abdominal pain. Contrast-enhanced computed tomography (CT) showed a perforation in the lesser curve of the stomach. The patient suffered a bout of hematemesis, following which an endoscopy showed a bleeding blood vessel at the edge of the perforation. We performed an emergency distal gastrectomy, including the ulcer site. Histopathological examination revealed tuberculous granulation tissue and acid-fast bacilli in the ulcer. The patient was given antituberculosis therapy (ATT) postoperatively, and was well when last seen 1 year 5 months after surgery. We analyzed the clinical data of five cases of tuberculous gastric perforation (TGP), reported between 1948 and 2003, including our patient. The patients ranged in age from 21 to 45 years, with a mean age of 36.8 years (SD +/- 10.21), and a male to female ratio of 3 : 2. The diagnosis was confirmed by surgery or autopsy. Abdominal lymphadenopathy was present in all patients. Gastrectomy was performed in four patients, and two were given ATT. All four patients in the previous reports died of their disease.

  8. Tn5406, a New Staphylococcal Transposon Conferring Resistance to Streptogramin A and Related Compounds Including Dalfopristin

    PubMed Central

    Haroche, Julien; Allignet, Jeanine; El Solh, Névine

    2002-01-01

    We characterized a new transposon, Tn5406 (5,467 bp), in a clinical isolate of Staphylococcus aureus (BM3327). It carries a variant of vgaA, which encodes a putative ABC protein conferring resistance to streptogramin A but not to mixtures of streptogramins A and B. It also carries three putative genes, the products of which exhibit significant similarities (61 to 73% amino acid identity) to the three transposases of the staphylococcal transposon Tn554. Like Tn554, Tn5406 failed to generate target repeats. In BM3327, the single copy of Tn5406 was inserted into the chromosomal att554 site, which is the preferential insertion site of Tn554. In three other independent S. aureus clinical isolates, Tn5406 was either present as a single plasmid copy (BM3318), as two chromosomal copies (BM3252), or both in the chromosome and on a plasmid (BM3385). The Tn5406-carrying plasmids also contain two other genes, vgaB and vatB. The insertion sites of Tn5406 in BM3252 were studied: one copy was in att554, and one copy was in the additional SCCmec element. Amplification experiments revealed circular forms of Tn5406, indicating that this transposon might be active. To our knowledge, a transposon conferring resistance to streptogramin A and related compounds has not been previously described. PMID:12121902

  9. Improved site-specific recombinase-based method to produce selectable marker- and vector-backbone-free transgenic cells

    NASA Astrophysics Data System (ADS)

    Yu, Yuan; Tong, Qi; Li, Zhongxia; Tian, Jinhai; Wang, Yizhi; Su, Feng; Wang, Yongsheng; Liu, Jun; Zhang, Yong

    2014-02-01

    PhiC31 integrase-mediated gene delivery has been extensively used in gene therapy and animal transgenesis. However, random integration events are observed in phiC31-mediated integration in different types of mammalian cells; as a result, the efficiencies of pseudo attP site integration and evaluation of site-specific integration are compromised. To improve this system, we used an attB-TK fusion gene as a negative selection marker, thereby eliminating random integration during phiC31-mediated transfection. We also excised the selection system and plasmid bacterial backbone by using two other site-specific recombinases, Cre and Dre. Thus, we generated clean transgenic bovine fetal fibroblast cells free of selectable marker and plasmid bacterial backbone. These clean cells were used as donor nuclei for somatic cell nuclear transfer (SCNT), indicating a similar developmental competence of SCNT embryos to that of non-transgenic cells. Therefore, the present gene delivery system facilitated the development of gene therapy and agricultural biotechnology.

  10. The filamentous phage XacF1 causes loss of virulence in Xanthomonas axonopodis pv. citri, the causative agent of citrus canker disease.

    PubMed

    Ahmad, Abdelmonim Ali; Askora, Ahmed; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

    2014-01-01

    In this study, filamentous phage XacF1, which can infect Xanthomonas axonopodis pv. citri (Xac) strains, was isolated and characterized. Electron microscopy showed that XacF1 is a member of the family Inoviridae and is about 600 nm long. The genome of XacF1 is 7325 nucleotides in size, containing 13 predicted open reading frames (ORFs), some of which showed significant homology to Ff-like phage proteins such as ORF1 (pII), ORF2 (pV), ORF6 (pIII), and ORF8 (pVI). XacF1 showed a relatively wide host range, infecting seven out of 11 strains tested in this study. Frequently, XacF1 was found to be integrated into the genome of Xac strains. This integration occurred at the host dif site (attB) and was mediated by the host XerC/D recombination system. The attP sequence was identical to that of Xanthomonas phage Cf1c. Interestingly, infection by XacF1 phage caused several physiological changes to the bacterial host cells, including lower levels of extracellular polysaccharide production, reduced motility, slower growth rate, and a dramatic reduction in virulence. In particular, the reduction in virulence suggested possible utilization of XacF1 as a biological control agent against citrus canker disease. PMID:25071734

  11. Deviation from the Maxwell-Cattaneo law: Role of asymmetric interparticle interactions.

    PubMed

    Zhao, Huizhu; Wang, Lei

    2015-10-01

    Nonstationary heat conduction in a few one-dimensional nonlinear lattices is studied numerically based on the Maxwell-Cattaneo (MC) law. We simulate the relaxation process and calculate the magnitudes of the temperature oscillation A(T)(t) and the local heat current oscillation A(j)(t). A phase difference between A(T)(t) and A(j)(t) is observed, which not only verifies the existence of the time lag τ in the MC law but also provides a better way of determining the critical wavelength L(*) that separates between oscillatory and diffusive relaxation modes. However, clear deviations from the MC law are observed. Not only do the decay exponents differ from the theoretical expectations, but, more importantly, suboscillation in the diffusive regime, which is not expected by the MC law, is found in the lattices with asymmetric interactions as well. These findings imply that higher-order effects must be considered in order to well describe the nonstationary heat conduction process in these systems. PMID:26565197

  12. Identification of genetic and environmental factors stimulating excision from Streptomyces scabiei chromosome of the toxicogenic region responsible for pathogenicity.

    PubMed

    Chapleau, Mélanie; Guertin, Julien F; Farrokhi, Ali; Lerat, Sylvain; Burrus, Vincent; Beaulieu, Carole

    2016-05-01

    The genes conferring pathogenicity in Streptomyces turgidiscabies, a pathogen causing common scab of potato, are grouped together on a pathogenicity island (PAI), which has been found to be mobile and appears to transfer and disseminate like an integrative and conjugative element (ICE). However, in Streptomyces scabiei, another common scab-inducing species, the pathogenicity genes are clustered in two regions: the toxicogenic region (TR) and the colonization region. The S. scabiei 87.22 genome was analysed to investigate the potential mobility of the TR. Attachment sites (att), short homologous sequences that delineate ICEs, were identified at both extremities of the TR. An internal att site was also found, suggesting that the TR has a composite structure (TR1 and TR2). Thaxtomin biosynthetic genes, essential for pathogenicity, were found in TR1, whereas candidate genes with known functions in recombination, replication and conjugal transfer were found in TR2. Excision of the TR1 or TR2 subregions alone, or of the entire TR region, was observed, although the excision frequency of TR was low. However, the excision frequency was considerably increased in the presence of either mitomycin C or Streptomyces coelicolor cells. A composite TR structure was not observed in all S. scabiei and Streptomyces acidiscabies strains tested. Of the ten strains analysed, seven lacked TR2 and no TR excision event could be detected in these strains, thus suggesting the implication of TR2 in the mobilization of S. scabiei TR. PMID:26177341

  13. Fluorescent protein-based detection of φC31 integrase activity in mammalian cells.

    PubMed

    Liu, Taian; Fang, Yongxiang; Jia, Huaijie; Chen, Guohua; Guan, Qisai; He, Xiaobing; Yao, Wenjuan; Zeng, Shuang; Jing, Zhizhong

    2013-10-15

    The enzyme φC31 integrase from Streptomyces phage has been documented as functional in mammalian cells and, therefore, has the potential to be a powerful gene manipulation tool. However, the activity of this enzyme is cell-type dependent. The more active mutant forms of φC31 integrase are required. Therefore, a rapid and effective method should be developed to detect the intracellular activity of φC31 integrase. We devised in this study an integrase-inversion cassette that contains the enhanced green fluorescent protein (EGFP) gene and the reverse complementary DsRed gene, which are flanked by attB and reverse complementary attP. This cassette can be inverted by φC31 integrase, thereby altering the fluorescent protein expression. Thus, φC31 integrase activity can be qualitatively or quantitatively evaluated based on the detected fluorescence. Furthermore, this cassette-based method was applied to several cell types, demonstrating that it is an efficient and reliable tool for measuring φC31 integrase activity in mammalian cells.

  14. Gated rotation mechanism of site-specific recombination by ϕC31 integrase.

    PubMed

    Olorunniji, Femi J; Buck, Dorothy E; Colloms, Sean D; McEwan, Andrew R; Smith, Margaret C M; Stark, W Marshall; Rosser, Susan J

    2012-11-27

    Integrases, such as that of the Streptomyces temperate bacteriophage ϕC31, promote site-specific recombination between DNA sequences in the bacteriophage and bacterial genomes to integrate or excise the phage DNA. ϕC31 integrase belongs to the serine recombinase family, a large group of structurally related enzymes with diverse biological functions. It has been proposed that serine integrases use a "subunit rotation" mechanism to exchange DNA strands after double-strand DNA cleavage at the two recombining att sites, and that many rounds of subunit rotation can occur before the strands are religated. We have analyzed the mechanism of ϕC31 integrase-mediated recombination in a topologically constrained experimental system using hybrid "phes" recombination sites, each of which comprises a ϕC31 att site positioned adjacent to a regulatory sequence recognized by Tn3 resolvase. The topologies of reaction products from circular substrates containing two phes sites support a right-handed subunit rotation mechanism for catalysis of both integrative and excisive recombination. Strand exchange usually terminates after a single round of 180° rotation. However, multiple processive "360° rotation" rounds of strand exchange can be observed, if the recombining sites have nonidentical base pairs at their centers. We propose that a regulatory "gating" mechanism normally blocks multiple rounds of strand exchange and triggers product release after a single round.

  15. Establishment of an improved high-efficiency thermal asymmetric interlaced PCR for identification of genomic integration sites mediated by phiC31 integrase.

    PubMed

    Zhou, Zaiwei; Ma, Haiyan; Qu, Lijuan; Xie, Fei; Ma, Qingwen; Ren, Zhaorui

    2012-03-01

    Streptomyces phage phiC31 integrase is widely used to mediate the integration of exogenous genes into host genomes for gene therapy and genomic modification, as it autonomously performs efficient, unidirectional, site-specific integration into pseudo attP sites of the host genome. Although pseudo attP sites are rarely found within exons, it is necessary to map their precise locations to avoid the risk of insertion mutagenesis. High-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) is a technique that has been developed to recover genomic sequences that flank insertion tags. We have found, however, that this technique is poorly efficient, as it amplifies many non-specific targets and frequently does not generate sufficient product for downstream analysis. Therefore, we have modified the hiTAIL-PCR procedure and re-designed the random primers. As a result, both the amount and specificity of the reaction product were enhanced for each integration site. Restriction analysis of known sequences within the integrated vector, which co-amplified with the flanking genomic sequences, validated 90% of these bands for sequencing. In contrast, only 30% of the bands produced by previous hiTAIL-PCR could be validated. Compared with the original hiTAIL-PCR, our improved hiTAIL-PCR procedure identified phiC31 integration sites more accurately and efficiently.

  16. The temperate phages RP2 and RP3 of Streptomyces rimosus.

    PubMed

    Rausch, H; Vesligaj, M; Pocta, D; Biuković, G; Pigac, J; Cullum, J; Schmieger, H; Hranueli, D

    1993-10-01

    The oxytetracycline-producing Streptomyces rimosus strains R6-65 and R7 (ATCC 10970) are lysogenic for the two narrow-host-range phages RP2 and RP3. Both phages are released at low frequency from the lysogenic strains and form plaques on 'cured' S. rimosus strains. RP2 and RP3 are of similar shape with flexible tails and contain double-stranded DNA of about 70% G+C with cohesive ends (group B1 of bacteriophage classification). The two phages also have identical, very slow, growth kinetics in S. rimosus, with a latent phase of about 6 h and a rise period of about 4 h. RP2 and RP3 are heteroimmune and they differ slightly in their size of phage particles and length of DNA (64.7 and 62.4 kb for RP2 and RP3, respectively). The restriction maps of the two phages are completely different, and hybridization experiments showed only one short region of sequence similarity (less than 430 bp); the two phages are thus essentially unrelated. Both phages lysogenize their hosts by recombination via defined attachment (att) sites. The positions of the attP sites have been localized on the restriction maps of RP2 and RP3 to restriction fragments of 800 and 300 bp, respectively. The prophages did not affect the level of oxytetracycline production or the genetic instability of this trait.

  17. The filamentous phage XacF1 causes loss of virulence in Xanthomonas axonopodis pv. citri, the causative agent of citrus canker disease

    PubMed Central

    Ahmad, Abdelmonim Ali; Askora, Ahmed; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

    2014-01-01

    In this study, filamentous phage XacF1, which can infect Xanthomonas axonopodis pv. citri (Xac) strains, was isolated and characterized. Electron microscopy showed that XacF1 is a member of the family Inoviridae and is about 600 nm long. The genome of XacF1 is 7325 nucleotides in size, containing 13 predicted open reading frames (ORFs), some of which showed significant homology to Ff-like phage proteins such as ORF1 (pII), ORF2 (pV), ORF6 (pIII), and ORF8 (pVI). XacF1 showed a relatively wide host range, infecting seven out of 11 strains tested in this study. Frequently, XacF1 was found to be integrated into the genome of Xac strains. This integration occurred at the host dif site (attB) and was mediated by the host XerC/D recombination system. The attP sequence was identical to that of Xanthomonas phage Cf1c. Interestingly, infection by XacF1 phage caused several physiological changes to the bacterial host cells, including lower levels of extracellular polysaccharide production, reduced motility, slower growth rate, and a dramatic reduction in virulence. In particular, the reduction in virulence suggested possible utilization of XacF1 as a biological control agent against citrus canker disease. PMID:25071734

  18. Differential activation of immune factors in neurons and glia contribute to individual differences in resilience/vulnerability to sleep disruption

    PubMed Central

    Dissel, Stephane; Seugnet, Laurent; Thimgan, Matthew S.; Silverman, Neal; Angadi, Veena; Thacher, Pamela V.; Burnham, Melissa M.; Shaw, Paul J.

    2014-01-01

    Individuals frequently find themselves confronted with a variety of challenges that threaten their wellbeing. While some individuals face these challenges efficiently and thrive (resilient) others are unable to cope and may suffer persistent consequences (vulnerable). Resilience/vulnerability to sleep disruption may contribute to the vulnerability to individuals exposed to challenging conditions. With that in mind we exploited individual differences in a fly’s ability to form short-term memory (STM) following 3 different types of sleep disruption to identify the underlying genes. Our analysis showed that in each category of flies examined, there are individuals that form STM in the face of sleep loss (resilient) while other individuals show dramatic declines in cognitive behavior (vulnerable). Molecular genetic studies revealed that Antimicrobial Peptides, factors important for innate immunity, were candidates for conferring resilience/vulnerability to sleep deprivation. Specifically, Metchnikowin (Mtk), drosocin (dro) and Attacin (Att) transcript levels seemed to be differentially increased by sleep deprivation in glia (Mtk), neurons (dro) or primarily in the head fat body (Att). Follow-up genetic studies confirmed that expressing Mtk in glia but not neurons, and expressing dro in neurons but not glia, disrupted memory while modulating sleep in opposite directions. These data indicate that various factors within glia or neurons can contribute to individual differences in resilience/vulnerability to sleep deprivation. PMID:25451614

  19. Staphylococcus aureus SarA is a Regulatory Protein Responsive to Redox and pH that can Support Bacteriophage Lambda Integrase-Mediated Excision/Recombination

    PubMed Central

    Fujimoto, David F.; Higginbotham, Robin H.; Maleki, Soheila J.; Segall1, Anca M.; Smeltzer, Mark S.; Hurlburt, Barry K.

    2010-01-01

    Summary Staphylococcus aureus produces a wide array of virulence factors and causes a correspondingly diverse array of infections. Production of these virulence factors is under the control of a complex network of global regulatory elements, one of which is sarA. sarA encodes a DNA-binding protein that is considered to function as a transcription factor capable of acting as either a repressor or an activator. Using competitive ELISA assays, we demonstrate that SarA is present at approximately 50,000 copies per cell, which is not characteristic of classical transcription factors. We also demonstrate that SarA is present at all stages of growth in vitro and is capable of binding DNA with high affinity but that its binding affinity and pattern of shifted complexes in electrophoretic mobility shift assays is responsive to the redox state. We also show that SarA binds to the bacteriophage lambda (λ) attachment site, attL, producing SarA-DNA complexes similar to intasomes, which consist of bacteriophage lambda integrase, E. coli integration host factor and attL DNA. In addition, SarA stimulates intramolecular excision recombination in the absence of λ excisionase, a DNA-binding accessory protein. Taken together, these data suggest that SarA may function as an architectural accessory protein. PMID:19919677

  20. Targeted Gene Knockin in Porcine Somatic Cells Using CRISPR/Cas Ribonucleoproteins

    PubMed Central

    Park, Ki-Eun; Park, Chi-Hun; Powell, Anne; Martin, Jessica; Donovan, David M.; Telugu, Bhanu P.

    2016-01-01

    The pig is an ideal large animal model for genetic engineering applications. A relatively short gestation interval and large litter size makes the pig a conducive model for generating and propagating genetic modifications. The domestic pig also shares close similarity in anatomy, physiology, size, and life expectancy, making it an ideal animal for modeling human diseases. Often, however, the technical difficulties in generating desired genetic modifications such as targeted knockin of short stretches of sequences or transgenes have impeded progress in this field. In this study, we have investigated and compared the relative efficiency of CRISPR/Cas ribonucleoproteins in engineering targeted knockin of pseudo attP sites downstream of a ubiquitously expressed COL1A gene in porcine somatic cells and generated live fetuses by somatic cell nuclear transfer (SCNT). By leveraging these knockin pseudo attP sites, we have demonstrated subsequent phiC31 integrase mediated integration of green fluorescent protein (GFP) transgene into the site. This work for the first time created an optimized protocol for CRISPR/Cas mediated knockin in porcine somatic cells, while simultaneously creating a stable platform for future transgene integration and generating transgenic animals. PMID:27240344

  1. A novel gateway-compatible binary vector series (PC-GW) for flexible cloning of multiple genes for genetic transformation of plants.

    PubMed

    Dalal, Jyoti; Yalamanchili, Roopa; La Hovary, Christophe; Ji, Mikyoung; Rodriguez-Welsh, Maria; Aslett, Denise; Ganapathy, Sowmya; Grunden, Amy; Sederoff, Heike; Qu, Rongda

    2015-09-01

    The rapidly advancing field of plant synthetic biology requires transforming plants with multiple genes. This has sparked a growing interest in flexible plant transformation vectors, which can be used for multi-gene transformations. We have developed a novel binary vector series, named the PC-GW series (GenBank: KP826769-KP826773), for Agrobacterium-mediated plant transformation. The PC-GW vectors use the pCAMBIA vector backbone, and contain NPTII, hpt, bar, mCherry or egfp genes as selectable markers for plant transformation. In a modified multiple cloning site (MCS) of the T-DNA region, we have placed the attR1, attR2 and ccdB sequences for rapid cloning of one to four genes by Gateway™-assisted recombination. In addition, we have introduced four meganuclease sites, and other restriction sites for multi-gene vector construction. Finally, we have placed a CaMV 35S promoter and a 35S terminator on the 5' and 3' ends of the MCS. The CaMV 35S promoter is flanked by PstI restriction sites that can be used to replace it with another promoter sequence if needed. The PC-GW vectors provide choices for selectable markers, cloning methods, and can accommodate up to eight gene constructs in a single T-DNA, thereby significantly reducing the number of transformations or crosses needed to generate multi-transgene expressing plants.

  2. Isolated Tubercular Osteomyelitis of Scapula – A Report of Two Cases and Review of Literature

    PubMed Central

    Balaji, Gopisankar; Arockiaraj, Justin; Roy, Alfred Cyril; Ashok, Anand

    2013-01-01

    Introduction: Isolated tuberculosis of the scapula is rare. The presentation mimics the tumors of scapula. Hence it is important to know the clinical presentation and when to suspect tubercular osteomyelitis of scapula. Few cases have been described before. We have analyzed all reports with isolated scapular tuberculosis so far and described the varied presentations. Case Report: We report two cases of isolated scapular tuberculosis. One patient underwent aspiration and was started on antituberculous drugs (ATT) and the other underwent debridement, drainage of abscess and then was started on ATT. Both had excellent outcomes at the end of one year follow up. We reviewed previous published literature of isolated scapular tuberculosis. The most common presentation of scapular tuberculosis is as follows: age less than 30 years and there is no difference with respect to gender. Swelling and pain are the commonest symptoms. Lytic areas with surrounding sclerosis is the commonest radiological finding. Body of the scapula is affected most commonly. Prognosis is excellent with adequate treatment. Conclusion: To conclude, clinicians should have high index of suspicion for diagnosing this condition as it mimics tumors. Pain and swelling is the most common presentation of scapular tuberculosis. Radiography shows lytic lesion with marginal sclerosis. The prognosis is excellent with appropriate treatment. PMID:27298922

  3. Distinct noise-controlling roles of multiple negative feedback mechanisms in a prokaryotic operon system.

    PubMed

    Nguyen, L K; Kulasiri, D

    2011-03-01

    Molecular fluctuations are known to affect dynamics of cellular systems in important ways. Studies aimed at understanding how molecular systems of certain regulatory architectures control noise therefore become essential. The interplay between feedback regulation and noise has been previously explored for cellular networks governed by a single negative feedback loop. However, similar issues within networks consisting of more complex regulatory structures remain elusive. The authors investigate how negative feedback loops manage noise within a biochemical cascade concurrently governed by multiple negative feedback loops, using the prokaryotic tryptophan (trp) operon system in Escherechia coli as the model system. To the authors knowledge, this is the first study of noise in the trp operon system. They show that the loops in the trp operon system possess distinct, even opposing, noise-controlling effects despite their seemingly analogous feedback structures. The enzyme inhibition loop, although controlling the last reaction of the cascade, was found to suppress noise not only for the tryptophan output but also for other upstream components. In contrast, the Repression (Rep) loop enhances noise for all systems components. Attenuation (Att) poses intermediate effects by attenuating noise for the upstream components but promoting noise for components downstream of its target. Regarding noise at the output tryptophan, Rep and Att can be categorised as noise-enhancing loops whereas Enzyme Inhibition as a noise-reducing loop. These findings suggest novel implications in how cellular systems with multiple feedback mechanisms control noise. [Includes supplementary material]. PMID:21405203

  4. BioSYNTHESIS: access to a knowledge network of health sciences databases.

    PubMed

    Broering, N C; Hylton, J S; Guttmann, R; Eskridge, D

    1991-04-01

    Users of the IAIMS Knowledge Network at the Georgetown University Medical Center have access to multiple in-house and external databases from a single point of entry through BioSYNTHESIS. The IAIMS project has developed a rich environment of biomedical information resources that represent a medical decision support system for campus physicians and students. The BioSYNTHESIS system is an information navigator that provides transparent access to a Knowledge Network of over a dozen databases. These multiple health sciences databases consist of bibliographic, informational, diagnostic, and research systems which reside on diverse computers such as DEC VAXs, SUN 490, AT&T 3B2s, Macintoshes, IBM PC/PS2s and the AT&T ISN and SYTEK network systems. Ethernet and TCP/IP protocols are used in the network architecture. BioSYNTHESIS also provides network links to the other campus libraries and to external institutions. As additional knowledge resources and technological advances have become available. BioSYNTHESIS has evolved from a two phase to a three phase program. Major components of the system including recent achievements and future plans are described. PMID:1661772

  5. Biphasic Effect of Rifampicin on Bilirubin- A Case Report

    PubMed Central

    Seshadri, Mandalam Subramanian

    2016-01-01

    Drug induced hepatitis is a major problem which a physician encounters in his clinical practice. In view of increasing incidence of tuberculosis in our country a large number of infected individuals are started on Antituberculous (ATT) drugs and rifampicin is invariably part of the regimen. One of the major adverse effects of ATT drugs is drug- induced hepatitis which is characterized by elevation of liver enzymes and bilirubin. Hepatotoxicity is usually idiosyncratic or dose-dependent. Rifampicin causes transient elevation of transaminases in 10-20 percent of individuals and this does not warrant dose adjustments of the drug. Rarely rifampicin can lead to severe hepatitis with hyperbilirubinaemia and marked elevations of SGOT and SGPT and in some patients this can be fatal. The exact mechanism of Rifampicin induced hepatotoxicity is not known but it is postulated to be due to idiosyncratic reaction to rifampicin metabolites which may be directly toxic or induce an immunologically mediated liver injury. Rarely rifampicin may cause hyperbilirubinaemia without enzyme elevation. Here we report a patient with bilateral pulmonary tuberculosis who developed transient severe indirect hyperbilirubinaemia on rifampicin. On review of relevant literature we find that rifampicin can have a biphasic effect on bilirubin, an initial increase in indirect bilirubin and later normalization of bilirubin. We have reported this case because of its rarity in clinical practice. PMID:27190870

  6. Spontaneous closure of an ileostomy: A rare occurrence

    PubMed Central

    Saxena, Ashish; Kumar, Lovekesh; Singh, Mahendra; Kolhe, Yuvraj; Karande, Snehal K.; Venkatesh, P.; Sahai, R.N.

    2014-01-01

    INTRODUCTION Ileostomy is an iatrogenic entero-cutaneous (EC) fistula designed for controlled evacuation of bowel contents. Once ileostomy has served its purpose, it is reverted by surgical procedure. We are reporting an interesting case of spontaneous closure of an ileostomy, obviating the need of surgical intervention. PRESENTATION OF CASE A 26 year old lady presented with perforation peritonitis. Upon exploration, a tubercular perforation of terminal ileum was found. Loop ileostomy was formed and patient was discharged on anti-tubercular treatment (ATT) after an uneventful recovery. During follow up visits, stoma was found to be retracting gradually. Retraction was not associated with any signs of peritonitis. Patient was able to pass stools per rectally. Stoma regressed completely within 8 months followed by epithelialisation of stoma site. Patient was leading an essentially normal life until her last follow up visit. DISCUSSION Considering the various factors affecting spontaneous healing of EC fistulas, all the intestinal stomas do have favourable characteristics essential for spontaneous closure. However, this is seldom seen in day to day surgical practice. The factors pertaining to this particular case that led to spontaneous closure of stoma remain poorly understood. CONCLUSION Further research is warranted to understand the mechanism behind spontaneous regression of a stoma. Relationship between this event and intestinal tuberculosis or ATT needs to be analysed. PMID:25465646

  7. A fine control of quorum-sensing communication in Agrobacterium tumefaciens

    PubMed Central

    Haudecoeur, Elise

    2010-01-01

    The bacterial pathogen Agrobacterium tumefaciens produces the quorum-sensing (QS) signal 3-oxo-octanoylhomoserine lactone (OC8HSL) for controlling horizontal transfer of its tumor inducing (Ti) plasmid that carries both the T-DNA and the virulence genes. Over-accumulation of OC8HSL also increases severity of plant symptoms (number of emerging tumors at infection site) by an unknown mechanism. A. tumefaciens strain C58 expresses two lactonases, AttM (BlcC) and AiiB, that cleave OC8HSL and are potential modulators of QS. Recent data highlight the direct contribution of lactonases AttM and AiiB in the control of OC8HSL level and QS-regulated functions such as conjugation of Ti plasmid and seriousness of plant symptoms. Expression of the two lactonases is regulated by different plant signals. A working model of QS in the course of the A. tumefaciens-plant host interaction is proposed and discussed. PMID:20585496

  8. Foldback intercoil DNA and the mechanism of DNA transposition.

    PubMed

    Kim, Byung-Dong

    2014-09-01

    Foldback intercoil (FBI) DNA is formed by the folding back at one point of a non-helical parallel track of double-stranded DNA at as sharp as 180° and the intertwining of two double helixes within each other's major groove to form an intercoil with a diameter of 2.2 nm. FBI DNA has been suggested to mediate intra-molecular homologous recombination of a deletion and inversion. Inter-molecular homologous recombination, known as site-specific insertion, on the other hand, is mediated by the direct perpendicular approach of the FBI DNA tip, as the attP site, onto the target DNA, as the attB site. Transposition of DNA transposons involves the pairing of terminal inverted repeats and 5-7-bp tandem target duplication. FBI DNA configuration effectively explains simple as well as replicative transposition, along with the involvement of an enhancer element. The majority of diverse retrotransposable elements that employ a target site duplication mechanism is also suggested to follow the FBI DNA-mediated perpendicular insertion of the paired intercoil ends by non-homologous end-joining, together with gap filling. A genome-wide perspective of transposable elements in light of FBI DNA is discussed. PMID:25317106

  9. Nanobody-based chimeric receptor gene integration in Jurkat cells mediated by PhiC31 integrase

    SciTech Connect

    Iri-Sofla, Farnoush Jafari; Rahbarizadeh, Fatemeh; Ahmadvand, Davoud; Rasaee, Mohammad J.

    2011-11-01

    The crucial role of T lymphocytes in anti-tumor immunity has led to the development of novel strategies that can target and activate T cells against tumor cells. Recombinant DNA technology has been used to generate non-MHC-restricted chimeric antigen receptors (CARs). Here, we constructed a panel of recombinant CAR that harbors the anti-MUC1 nanobody and the signaling and co-signaling moieties (CD3{zeta}/CD28) with different spacer regions derived from human IgG3 with one or two repeats of the hinge sequence or the hinge region of Fc{gamma}RII. The PhiC31 integrase system was employed to investigate if the recombination efficiency could be recruited for high and stable expression of T cell chimeric receptor genes. The effect of nuclear localization signal (NLS) and two different promoters (CMV and CAG) on efficacy of PhiC31 integrase in human T cell lines was evaluated. The presence of integrase in combination with NLS, mediated up to 7.6 and 8.5 fold increases in CAR expression in ZCHN-attB and ZCHHN-attB cassette integrated T cells, respectively. Our results showed that highly efficient and stable transduction of the Jurkat cell line by PhiC31 integrase is a feasible modality for generating anti-cancer chimeric T cells for use in cancer immunotherapy.

  10. Ultralow Density, Monolithic WS2, MoS2, and MoS2/Graphene Aerogels.

    PubMed

    Worsley, Marcus A; Shin, Swanee J; Merrill, Matthew D; Lenhardt, Jeremy; Nelson, Art J; Woo, Leta Y; Gash, Alex E; Baumann, Theodore F; Orme, Christine A

    2015-05-26

    We describe the synthesis and characterization of monolithic, ultralow density WS2 and MoS2 aerogels, as well as a high surface area MoS2/graphene hybrid aerogel. The monolithic WS2 and MoS2 aerogels are prepared via thermal decomposition of freeze-dried ammonium thio-molybdate (ATM) and ammonium thio-tungstate (ATT) solutions, respectively. The densities of the pure dichalcogenide aerogels represent 0.4% and 0.5% of full density MoS2 and WS2, respectively, and can be tailored by simply changing the initial ATM or ATT concentrations. Similar processing in the presence of the graphene aerogel results in a hybrid structure with MoS2 sheets conformally coating the graphene scaffold. This layered motif produces a ∼50 wt % MoS2 aerogel with BET surface area of ∼700 m(2)/g and an electrical conductivity of 112 S/m. The MoS2/graphene aerogel shows promising results as a hydrogen evolution reaction catalyst with low onset potential (∼100 mV) and high current density (100 mA/cm(2) at 260 mV).

  11. CO near the Pleiades: Encounter of a star cluster with a small molecular cloud

    NASA Technical Reports Server (NTRS)

    Bally, J.; White, R. E.

    1986-01-01

    Although there is a large amount of interstellar matter near the Pleiades star cluster, the observed dust and gas is not a remnant of the placental molecular cloud from which the star cluster was formed. Carbon monoxide (CO) associated with the visible reflection nebulae was discovered by Cohen (1975). Its radial velocity differs from that of the cluster by many times the cluster escape velocity, which implies that the cloud-cluster association is the result of a chance encounter. This circumstance and the proximity of the Pleiades to the sun creates an unique opportunity for study of interstellar processes at high spatial resolution. To study the molecular component of the gas, a 1.7 square degree field was mapped with the AT&T Bell Laboratories 7-meter antenna (1.7' beam) on a 1' grid in the J=1.0 C(12)O line, obtaining over 6,000 spectra with 50 kHz resolution. The cloud core was mapped in the J=1-0 line of C(13)O. Further observations include an unsuccessful search for CS (J=2-1) at AT&T BL, and some C(12)O J=2-1 spectra obtained at the Millimeter Wave Observatory of the University of Texas.

  12. Mixed random walks with a trap in scale-free networks including nearest-neighbor and next-nearest-neighbor jumps.

    PubMed

    Zhang, Zhongzhi; Dong, Yuze; Sheng, Yibin

    2015-10-01

    Random walks including non-nearest-neighbor jumps appear in many real situations such as the diffusion of adatoms and have found numerous applications including PageRank search algorithm; however, related theoretical results are much less for this dynamical process. In this paper, we present a study of mixed random walks in a family of fractal scale-free networks, where both nearest-neighbor and next-nearest-neighbor jumps are included. We focus on trapping problem in the network family, which is a particular case of random walks with a perfect trap fixed at the central high-degree node. We derive analytical expressions for the average trapping time (ATT), a quantitative indicator measuring the efficiency of the trapping process, by using two different methods, the results of which are consistent with each other. Furthermore, we analytically determine all the eigenvalues and their multiplicities for the fundamental matrix characterizing the dynamical process. Our results show that although next-nearest-neighbor jumps have no effect on the leading scaling of the trapping efficiency, they can strongly affect the prefactor of ATT, providing insight into better understanding of random-walk process in complex systems. PMID:26450286

  13. The SopEPhi phage integrates into the ssrA gene of Salmonella enterica serovar Typhimurium A36 and is closely related to the Fels-2 prophage.

    PubMed

    Pelludat, Cosima; Mirold, Susanne; Hardt, Wolf-Dietrich

    2003-09-01

    Salmonella spp. are enteropathogenic gram-negative bacteria that use a large array of virulence factors to colonize the host, manipulate host cells, and resist the host's defense mechanisms. Even closely related Salmonella strains have different repertoires of virulence factors. Bacteriophages contribute substantially to this diversity. There is increasing evidence that the reassortment of virulence factor repertoires by converting phages like the GIFSY phages and SopEPhi may represent an important mechanism in the adaptation of Salmonella spp. to specific hosts and to the emergence of new epidemic strains. Here, we have analyzed in more detail SopEPhi, a P2-like phage from Salmonella enterica serovar Typhimurium DT204 that encodes the virulence factor SopE. We have cloned and characterized the attachment site (att) of SopEPhi and found that its 47-bp core sequence overlaps the 3' terminus of the ssrA gene of serovar Typhimurium. Furthermore, we have demonstrated integration of SopEPhi into the cloned attB site of serovar Typhimurium A36. Sequence analysis of the plasmid-borne prophage revealed that SopEPhi is closely related to (60 to 100% identity over 80% of the genome) but clearly distinct from the Fels-2 prophage of serovar Typhimurium LT2 and from P2-like phages in the serovar Typhi CT18 genome. Our results demonstrate that there is considerable variation among the P2-like phages present in closely related Salmonella spp.

  14. Adverse Drug Reactions Causing Admission to Medical Wards: A Cross-Sectional Survey at 4 Hospitals in South Africa.

    PubMed

    Mouton, Johannes P; Njuguna, Christine; Kramer, Nicole; Stewart, Annemie; Mehta, Ushma; Blockman, Marc; Fortuin-De Smidt, Melony; De Waal, Reneé; Parrish, Andy G; Wilson, Douglas P K; Igumbor, Ehimario U; Aynalem, Getahun; Dheda, Mukesh; Maartens, Gary; Cohen, Karen

    2016-05-01

    Limited data exist on the burden of serious adverse drug reactions (ADRs) in sub-Saharan Africa, which has high HIV and tuberculosis prevalence. We determined the proportion of adult admissions attributable to ADRs at 4 hospitals in South Africa. We characterized drugs implicated in, risk factors for, and the preventability of ADR-related admissions.We prospectively followed patients admitted to 4 hospitals' medical wards over sequential 30-day periods in 2013 and identified suspected ADRs with the aid of a trigger tool. A multidisciplinary team performed causality, preventability, and severity assessment using published criteria. We categorized an admission as ADR-related if the ADR was the primary reason for admission.There were 1951 admissions involving 1904 patients: median age was 50 years (interquartile range 34-65), 1057 of 1904 (56%) were female, 559 of 1904 (29%) were HIV-infected, and 183 of 1904 (10%) were on antituberculosis therapy (ATT). There were 164 of 1951 (8.4%) ADR-related admissions. After adjustment for age and ATT, ADR-related admission was independently associated (P ≤ 0.02) with female sex (adjusted odds ratio [aOR] 1.51, 95% confidence interval [95% CI] 1.06-2.14), increasing drug count (aOR 1.14 per additional drug, 95% CI 1.09-1.20), increasing comorbidity score (aOR 1.23 per additional point, 95% CI 1.07-1.41), and use of antiretroviral therapy (ART) if HIV-infected (aOR 1.92 compared with HIV-negative/unknown, 95% CI 1.17-3.14). The most common ADRs were renal impairment, hypoglycemia, liver injury, and hemorrhage. Tenofovir disoproxil fumarate, insulin, rifampicin, and warfarin were most commonly implicated, respectively, in these 4 ADRs. ART, ATT, and/or co-trimoxazole were implicated in 56 of 164 (34%) ADR-related admissions. Seventy-three of 164 (45%) ADRs were assessed as preventable.In our survey, approximately 1 in 12 admissions was because of an ADR. The range of ADRs and implicated drugs reflect South Africa's high HIV

  15. Processing of two homologous precursors, pro-neuropeptide Y and pro-pancreatic polypeptide, in transfected cell lines expressing different precursor convertases.

    PubMed

    Wulff, B S; Johansen, T E; Dalbøge, H; O'Hare, M M; Schwartz, T W

    1993-06-25

    The processing of two homologous precursors, pro-neuropeptide Y (pro-NPY) and pro-pancreatic poly-peptide (pro-PP), was studied in four neuroendocrine cell lines after transfection: CA-77 medullary thyroid carcinoma cells, AtT-20 corticotrope pituitary cells, RIN2A-19 pancreatic endocrine cells, and NB1 neuroblastoma cells. Northern blot analysis indicated that the AtT-20 cells only expressed precursor convertase 3; in contrast, NB1 cells only expressed precursor convertase 2, whereas the RIN2A-19 and CA-77 cells expressed both enzymes. Despite these differences in expression pattern of precursor convertases the four cell lines were, surprisingly, indistinguishable in respect to their processing of pro-PP and pro-NPY. In all four cell lines, pro-NPY was almost completely converted to NPY, and, in all four cell lines, only around 50% of the PP precursor was converted to PP. The relatively poor processing efficiency of pro-PP was rather similar to the processing efficiency of the endogenously produced precursors in the respective cell lines, pro-calcitonin (CA-77), proopiomelanocortin (AtT-20), proinsulin (RIN2A-19), and pro-vasoactive intestinal polypeptide (NB1). At least in the CA-77 cells, NPY and PP were apparently sorted to the regulated secretory pathway, as upon stimulation with secretagogue the release of the transfected peptides increased in parallel with the endogenously expressed peptide, calcitonin gene-related peptide. Mutagenesis studies showed that on the N-terminal side of the di-basic processing site, the otherwise important difference in structure between PP and NPY, a proline for glutamine in position 34, was not responsible for the difference in processing efficiency. On the C-terminal side of the processing site, the efficient processing of pro-NPY could not be transferred to pro-PP by exchanging the whole C-terminal domains of the precursors. It is concluded that pro-NPY is processed more efficiently than pro-PP in all neuroendocrine cell lines

  16. A functional cyclic AMP response element plays a crucial role in neuroendocrine cell type-specific expression of the secretory granule protein chromogranin A.

    PubMed Central

    Wu, H; Mahata, S K; Mahata, M; Webster, N J; Parmer, R J; O'Connor, D T

    1995-01-01

    Chromogranin A, a soluble acidic protein, is a ubiquitous component of secretory vesicles throughout the neuroendocrine system. We reported previously the cloning and initial characterization of the mouse chromogranin A gene promoter, which showed that the promoter contains both positive and negative domains and that a proximal promoter spanning nucleotides -147 to +42 bp relative to the transcriptional start site is sufficient for neuroendocrine cell type-specific expression. The current study was undertaken to identify the particular elements within this proximal promoter that control tissue-specific expression. We found that deletion or point mutations in the potential cAMP response element (CRE) site at -68 bp virtually abolished promoter activity specifically in neuroendocrine (PC12 chromaffin or AtT20 corticotrope) cells, with little effect on activity in control (NIH3T3 fibroblast) cells; thus, the CRE box is necessary for neuroendocrine cell type-specific activity of the chromogranin A promoter. Furthermore, the effect of the CRE site is enhanced in the context of intact (wild-type) promoter sequences between -147 and -100 bp. DNase I footprint analysis showed that these regions (including the CRE box) bind nuclear proteins present in both neuroendocrine (AtT20) and control (NIH3T3) cells. In AtT20 cells, electrophoretic mobility shift assays and factor-specific antibody supershifts showed that an oligonucleotide containing the chromogranin A CRE site formed a single, homogeneous protein-DNA complex containing the CRE-binding protein CREB. However, in control NIH3T3 cells we found evidence for an additional immunologically unrelated protein in this complex. A single copy of this oligonucleotide was able to confer neuroendocrine-specific expression to a heterologous (thymidine kinase) promoter, albeit with less fold selectivity than the full proximal chromogranin A promoter. Hence, the CRE site was partially sufficient to explain the neuroendocrine cell type

  17. Photoaffinity labeling of the somatostatin receptor: identification of molecular subtypes.

    PubMed

    Srikant, C B; Murthy, K K; Escher, E E; Patel, Y C

    1992-05-01

    Pharmacological studies have suggested that the somatostatin (SS) receptor is heterogeneous and may exhibit subtypes selective for SS-14 and SS-28. Whether this heterogeneity can be explained by separate molecular forms of the receptor protein is unclear. In the present study, we have developed a novel photosensitive azido derivative of the octapeptide SS analog Tyr3 SMS (EE 581) and used it as a photoaffinity probe to characterize the molecular components of the SS receptor in five receptor positive tissues (normal rat brain, pituitary, pancreas, and adrenal cortex, and mouse AtT-20 pituitary tumor cells). [125I]EE-581 labeled specific high affinity binding sites in all these tissues (Kd range 1.3-1.67 nM). Photoaffinity labeled membrane SS receptors were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. Three specifically labeled SS receptor proteins of 80 kilodaltons (kDa), 58 kDa, and 32 kDa were identified and exhibited a tissue-specific distribution. The 58 kDa species was the exclusive form in pancreas, adrenal cortex, and AtT-20 cells and the dominant form in brain. The 32 kDa receptor protein was expressed as a minor form (ratio of 58 kDa:32 kDa 3:1), exclusively in brain. The 80 kDa receptor was found only in the pituitary where it occurred as the sole SS receptor species. Competition experiments showed that the 58 kDa and 32 kDa receptor proteins in brain reacted with SS-14 greater than SS-28; in contrast, the 58 kDa protein in AtT-20 cells bound SS-28 greater than SS-14 suggesting the existence of distinct subtypes of the 58 kDa receptor in these two tissues. These data represent the first systematic evaluation of the molecular forms of SS receptor proteins by photoaffinity labeling in different target tissues and provide direct evidence for molecular heterogeneity and SS-14/SS-28 selectivity; a major 58 kDa protein present in most tissues, an additional 32 kDa protein uniquely expressed in brain, and an

  18. Correlation between biological agents and levels of heavy metals in municipal sewage sludge.

    PubMed

    Bozek, Urszula; Kłapeć, Teresa

    2008-01-01

    During the period 2003-2004, 110 samples of sewage sludge were examined from 18 treatment plants located in the territory of the whole of Poland, the majority in the Lublin Province. Samples of fermented and dehydrated sewage sludge were taken from sludge drying beds. The bacteriological and parasitological determinations covered bacteria of the Salmonella genus and eggs of the helminths Ascaris spp., Trichuris spp., and Toxocara spp. Chemical examinations for the presence of the heavy metals: chromium, zinc, cadmium, copper, nickel and lead were carried out by the method of atomic absorption flame spectrophotometry. The results were subjected to statistical analysis with the use of ANOVA and Student's t-tests. In the sewage sludge samples examined, Salmonella bacteria were isolated from 9 samples (8.2 % ), and the eggs of helminths from 31 samples (28.1 % ). The eggs of Ascaris spp. were most frequently detected--in 24 samples of sewage sludge (21.8 % ), followed by Trichuris spp. eggs--in 11 samples (10.0 % ), and Toxocara spp. eggs--in 6 samples (5.4 % ). Several samples of sewage sludge were infected by eggs of all the parasites examined (ATT). A very high variation was observed with respect to Salmonella bacteria and ATT eggs according to the origin, type of sanitation and duration of deposition of sludge. Levels of chromium, zinc, copper and lead in the sewage sludge samples, in the majority of samples examined, remained below allowable values. Very high values exceeding the allowable values for the levels of heavy metals were observed only in the samples of sewage sludge from one treatment plant. This concerned a cadmium level (201.1 mg/kg d.m.) which exceeded by 20 times the most rigorous allowable contents of this metal for sewage sludge designed for agricultural use, and nickel (288.7 mg/kg d.m.)--the standard being exceeded 3 times. A significant variation of the levels of heavy metals was observed depending on treatment type. A longer period of

  19. Investigating pilot performance using mixed-modality simulated data link

    NASA Astrophysics Data System (ADS)

    Lancaster, Jeff A.

    Empirical studies of general aviation (GA) pilot performance are lacking, especially with respect to envisioned future requirements. Two research studies were conducted to evaluate human performance using new technologies. In the first study, ten participants completed the Modified Rhyme Test (MRT) in an effort to compare the intelligibility of two text-to-speech (TTS) engines (DECtalk and AT&T's Natural Voices) as presented in 85 dB(A) aircraft cockpit engine noise. Results indicated significant differences in intelligibility (p ≤ 0.05) between the two speech synthesizers across the tested speech-to-noise ratios (S/N) (i.e., -5 dB, -8 dB, and -11 dB S/N) with the AT&T engine resulting in superior intelligibility in all of the S/N. The AT&T product was therefore selected as the TTS engine for the second study. In the second study, 16 visual flight rules (VFR) rated pilots were evaluated for their data link performance using a flight simulator (ELITE i-GATE) equipped with a mixed-modality simulated data link within one of two flight conditions. Data link modalities included textual, synthesized speech, digitized speech, and synthesized speech/textual combination. Flight conditions included VFR (unlimited ceiling, visibility) or marginal VFR (MVFR) flight conditions (clouds 2800 feet above ground level [AGL], three miles visibility). Evaluation focused on the time required accessing, understanding, and executing data link commands. Additional data were gathered to evaluate workload, situation awareness, and subjective preference. Results indicated significant differences in pilot performance, mental workload, and situation awareness across the data link modalities and between flight conditions. Textual data link resulted in decreased performance while the other three data link conditions did not differ in performance. Workload evaluation indicated increased workload in the textual data link condition. Situation awareness (SA) measures indicated differences in

  20. Adverse Drug Reactions Causing Admission to Medical Wards

    PubMed Central

    Mouton, Johannes P.; Njuguna, Christine; Kramer, Nicole; Stewart, Annemie; Mehta, Ushma; Blockman, Marc; Fortuin-De Smidt, Melony; De Waal, Reneé; Parrish, Andy G.; Wilson, Douglas P.K.; Igumbor, Ehimario U.; Aynalem, Getahun; Dheda, Mukesh; Maartens, Gary; Cohen, Karen

    2016-01-01

    Abstract Limited data exist on the burden of serious adverse drug reactions (ADRs) in sub-Saharan Africa, which has high HIV and tuberculosis prevalence. We determined the proportion of adult admissions attributable to ADRs at 4 hospitals in South Africa. We characterized drugs implicated in, risk factors for, and the preventability of ADR-related admissions. We prospectively followed patients admitted to 4 hospitals’ medical wards over sequential 30-day periods in 2013 and identified suspected ADRs with the aid of a trigger tool. A multidisciplinary team performed causality, preventability, and severity assessment using published criteria. We categorized an admission as ADR-related if the ADR was the primary reason for admission. There were 1951 admissions involving 1904 patients: median age was 50 years (interquartile range 34–65), 1057 of 1904 (56%) were female, 559 of 1904 (29%) were HIV-infected, and 183 of 1904 (10%) were on antituberculosis therapy (ATT). There were 164 of 1951 (8.4%) ADR-related admissions. After adjustment for age and ATT, ADR-related admission was independently associated (P ≤ 0.02) with female sex (adjusted odds ratio [aOR] 1.51, 95% confidence interval [95% CI] 1.06–2.14), increasing drug count (aOR 1.14 per additional drug, 95% CI 1.09–1.20), increasing comorbidity score (aOR 1.23 per additional point, 95% CI 1.07–1.41), and use of antiretroviral therapy (ART) if HIV-infected (aOR 1.92 compared with HIV-negative/unknown, 95% CI 1.17–3.14). The most common ADRs were renal impairment, hypoglycemia, liver injury, and hemorrhage. Tenofovir disoproxil fumarate, insulin, rifampicin, and warfarin were most commonly implicated, respectively, in these 4 ADRs. ART, ATT, and/or co-trimoxazole were implicated in 56 of 164 (34%) ADR-related admissions. Seventy-three of 164 (45%) ADRs were assessed as preventable. In our survey, approximately 1 in 12 admissions was because of an ADR. The range of ADRs and implicated drugs reflect

  1. The fas operon of Rhodococcus fascians encodes new genes required for efficient fasciation of host plants.

    PubMed Central

    Crespi, M; Vereecke, D; Temmerman, W; Van Montagu, M; Desomer, J

    1994-01-01

    Three virulence loci (fas, att, and hyp) of Rhodococcus fascians D188 have been identified on a 200-kb conjugative linear plasmid (pFiD188). The fas locus was delimited to a 6.5-kb DNA fragment by insertion mutagenesis, single homologous disruptive recombination, and in trans complementation of different avirulent insertion mutants. The locus is arranged as a large operon containing six open reading frames whose expression is specifically induced during the interaction with host plants. One predicted protein is homologous to P-450 cytochromes from actinomycetes. The putative ferredoxin component is of a novel type containing additional domains homologous to transketolases from chemoautotrophic, photosynthetic, and methylotrophic microorganisms. Genetic analysis revealed that fas encodes, in addition to the previously identified ipt, at least two new genes that are involved in fasciation development, one of which is only required on older tobacco plants. PMID:8169198

  2. Master schedule for CY-1982 Hanford environmental surveillance routine program

    SciTech Connect

    Blumer, P.J.; Sula, M.J.; Eddy, P.A.

    1981-12-01

    This report provides the current schedule of data collection for the routine environmental surveillance program at the Hanford Site. The environmental surveillance program objectives are to evaluate and report the levels of radioactive and nonradioactive pollutants in the Hanford environs, as required in DOE Order 5484.1. The routine sampling schedule provided does not include samples which are planned to be collected during FY-1982 in support of special studies or for quality control purposes. In addition, the routine program outlined in this schedule is subject to modification during the year in response to changes in Site operations, program requirements, or unusual sample results. Sampling schedules are presented for the following: air; Columbia River; sanitary water; surface water; ground water; foodstuffs; wildlife; soil and vegetation; external radiation measurements; portable instrument surveys; and surveillance of waste disposal sites. (ATT)

  3. Effects of pre-stressing and flux on the flow of solder on PWB copper surfaces

    SciTech Connect

    Hernandez, C.L.; Hosking, F.M.

    1994-12-31

    A variety of test methods are available to evaluate the solderability of printed wiring board [PWB] surface finishes. A new test has been developed which better simulates the capillary flow physics of typical solder assembly processing, especially surface mount soldering. The work was conducted under a cooperative research and development agreement between Sandia National Laboratories, the National Center for Manufacturing Sciences, and several PWB fabricators (AT&T, IBM, Texas Instruments, and United Technologies Corporation/Hamilton Standard) to advance PWB interconnect systems technology. Particular attention has been given at Sandia to characterizing the effects of accelerated aging in a simulated indoor industrial environment on subsequent PWB solderability. The program`s baseline surface finish was copper. Solderability testing on ``as-fabricated`` and ``pre-stressed copper`` pad-strip geometries was performed with Sn-Pb eutectic solder and three different fluxes at four different reflow temperatures.

  4. Changes coming for Science Board

    NASA Astrophysics Data System (ADS)

    The role of a National Academy of Science panel that advises NASA on goals for space science and exploration will be reexamined this fall by scientists, NASA officials, and congressional staff.The NAS Space Science Board will be the subject of two conferences, the first scheduled for the week of October 24 in Virginia, according to Joyce Purcell of NAS. About 60 scientists and policymakers will discuss the future of space science and how the structure and purpose of the board might change. The sessions were proposed by Louis Lanzerotti of AT&T Bell Labs in Murray Hill, N.J., who succeeded Thomas Donahue of the University of Michigan at Ann Arbor as chairman of the SSB July 1.

  5. Genomic Changes in an Attenuated ZB Strain of Foot-and-Mouth Disease Virus Serotype Asia1 and Comparison with Its Virulent Parental Strain

    PubMed Central

    Zhu, Mingwang; Peng, Zhenqi; Hu, Qi; Shi, Chenhong; Liao, Defang; Wang, Jihua; Li, Huachun

    2014-01-01

    The molecular basis of attenuation of foot-and-mouth disease virus (FMDV) serotype Asia1 ZB strain remains unknown. To understand the genetic changes of attenuation, we compared the entire genomes of three different rabbit-passaged attenuated ZB strains (ZB/CHA/58(att), ZBRF168, and ZBRF188) and their virulent parental strains (ZBCF22 and YNBS/58). The results showed that attenuation may be brought about by 28 common amino acid substitutions in the coding region, with one nucleotide point mutation in the 5′-untranslated region (5′-UTR) and another one in the 3′-UTR. In addition, a total of 21 nucleotides silent mutations had been found after attenuation. These substitutions, alone or in combination, may be responsible for the attenuated phenotype of the ZB strain in cattle. This will contribute to elucidation of attenuating molecular basis of the FMDV ZB strain. PMID:25386556

  6. The complete mitochondrial genome of the codling moth Cydia pomonella (Lepidoptera: Tortricidae).

    PubMed

    Shi, Bao-Cai; Liu, Wei; Wei, Shu-Jun

    2013-02-01

    The complete mitochondrial genome of the codling moth Cydia pomonella (Lepidoptera: Tortricidae) was determined. The genome is 15,253 bp long with 37 typical animal mitochondrial genes and an A+T-rich region. All genes are arranged in their conserved positions compared with the pupative ancestral arrangement of insects except for trnM, which was translocated to the upstream of the transfer RNA cluster trnI-trnQ as in all previously reported lepidopteran mitochondiral genomes. Seven portein-coding genes use ATG start codon and five use ATT. However, the cox1 gene uses the CGA start codon as it is found in all previous reported mitochondrial genomes of Lepidoptera. Nine protein-coding genes stop with termination codon TAA. Four protein-coding genes use incomplete stop codons TA or T. The A+T region is located between rrnS and trnM with a length of 331 bp.

  7. Ceremonial Session

    NASA Astrophysics Data System (ADS)

    Schrieffer, R.

    1996-03-01

    1995 Edward A. Bouchet Award formerly the Visiting Minority Lectureship Award) to: Anthony M. Johnson, New Jersey Institute of Technology 1995 John H. Dillon Medal to: Julia Ann Kornfield, California Institute of Technology 1995 Maria Goeppert-Mayer Award to: Marjorie Olmstead, University of Washington 1995 Biological Physics Prize to: Sieji Ogawa, AT&T Bell Laboratories 1995 High Polymer Physics Prize to: Alan N. Gent, University of Akron 1995 Earl K. Plyler Prize to: Charles S. Parmenter, University of Indiana 1995 Frank Isakson Prize to : David E. Aspnes, North Carolina State University 1995 Oliver E. Buckley Prize to: Charles Pence Slichter, University of Illinois 1995 George E. Pake Prize to: Charles V. Shank, Lawrence Berkeley National Laboratory

  8. Démonstration expérimentale d'une distribution quantique de clé par codage temporel

    NASA Astrophysics Data System (ADS)

    Boucher, W.; Debuisschert, T.

    2006-10-01

    Nous avons démontré expérimentalement la transmission quantique de clé par codage temporel. Alice envoie des impulsions cohérentes atténuées. La clé est codée dans l'instant de détection des photons chez Bob. Une mesure de la durée de cohérence par moyen interférométrique permet d'assurer un fonctionnement en régime quantique. Nous avons mesuré un taux d'erreur quantique de 3.3% et une chute relative de contraste de l'interféromètre de 8.4%. Ces valeurs permettent d'évaluer un avantage d'information de 0.49 bit/impulsion pour Alice et Bob par rapport à un espion éventuel.

  9. Primary Amoebic Meningoencephalitis.

    PubMed

    Gupta, Ritu; Parashar, M K; Kale, Aditya

    2015-04-01

    Primary amoebic meningoencephalitis due to free living amoeba, also called 'brain eating amoeba', Naegleria fowleri, was detected in retroviral disease patient of 40 years who has history of using well water. Patient was admitted with severe headache, fever intermittent, nausea, vomiting and slurring of speech. CT scan and MRI scan findings were normal. CSF examination showed increased protein, low sugar and predominant lymphocytes. CSF was negative for cryptococcal antigen but wet mount preparation showed highly motile free living amoeba Naegleria fowleri. Patient was put on Amphotericin B, Metronidazole, Rifampicin in addition to ART and ATT and other supportive medications. His headache was relieved and patient improved and was discharged on request. Earlier eight cases have been reported from India of which four cases survived the acute episode. PMID:26591176

  10. Resistance, regulatory and production genes for the antibiotic methylenomycin are clustered.

    PubMed

    Chater, K F; Bruton, C J

    1985-07-01

    At least 17 kb of DNA from the large unisolatable Streptomyces coelicolor A3(2) plasmid SCP1 are concerned with methylenomycin biosynthesis. Mutational cloning analysis, using insert-directed integration of att site deleted phage vectors into an SCP1-containing host, provided evidence of two large transcription units, of at least 6.6 kb and 9.5 kb. At the leftmost apparent end of the larger (left-hand) transcription unit is a region apparently involved in negative regulation of methylenomycin biosynthesis: when fragments from this region were used to direct phage integration, marked overproduction of methylenomycin resulted. The methylenomycin resistance determinant is located at the rightmost end of this same transcription unit. Hybridisation analysis with 13 kb of the cloned mmy region showed that it was closely similar to a segment of pSV1, a plasmid that specifies methylenomycin biosynthesis in S. violaceus-ruber SANK 95570. PMID:2992952

  11. Equity Oil Company BX in situ oil shale project, Piceance Basin, Colorado. Environmental quarter report, March 1-May 31, 1981

    SciTech Connect

    Not Available

    1981-06-12

    This report has been organized into three sections for ease of review. Section A - Air Resources covers data collected by the 100-foot tower between September and December 1980. Section B - Water Resources covers data collected since the last Quarterly Report. Section C - Aquatic Biology covers data collected since January 1981. Laboratory analysis results of all water samples show no significant changes from previous sample have occurred. The alluvial wells did exhibit slightly higher concentrations of phenols. Presently no baseline record exists for these wells to establish the normal levels of phenols in the alluvial ground water. Laboratory analysis of all sampling locations are presented in Appendix B. Aquatic biology covers data collected during the initial two aquatic biology field sampling trips of 1981. Benthic invertebrates, periphyton algae, and a fishery survey was conducted. Stream observations indicate that the stream habitat has remained unchanged since the August 1980 field visit. (ATT)

  12. Method for exponentiating in cryptographic systems

    DOEpatents

    Brickell, Ernest F.; Gordon, Daniel M.; McCurley, Kevin S.

    1994-01-01

    An improved cryptographic method utilizing exponentiation is provided which has the advantage of reducing the number of multiplications required to determine the legitimacy of a message or user. The basic method comprises the steps of selecting a key from a preapproved group of integer keys g; exponentiating the key by an integer value e, where e represents a digital signature, to generate a value g.sup.e ; transmitting the value g.sup.e to a remote facility by a communications network; receiving the value g.sup.e at the remote facility; and verifying the digital signature as originating from the legitimate user. The exponentiating step comprises the steps of initializing a plurality of memory locations with a plurality of values g.sup.xi ; computi The United States Government has rights in this invention pursuant to Contract No. DE-AC04-76DP00789 between the Department of Energy and AT&T Company.

  13. Automatic recognition of emotions from facial expressions

    NASA Astrophysics Data System (ADS)

    Xue, Henry; Gertner, Izidor

    2014-06-01

    In the human-computer interaction (HCI) process it is desirable to have an artificial intelligent (AI) system that can identify and categorize human emotions from facial expressions. Such systems can be used in security, in entertainment industries, and also to study visual perception, social interactions and disorders (e.g. schizophrenia and autism). In this work we survey and compare the performance of different feature extraction algorithms and classification schemes. We introduce a faster feature extraction method that resizes and applies a set of filters to the data images without sacrificing the accuracy. In addition, we have enhanced SVM to multiple dimensions while retaining the high accuracy rate of SVM. The algorithms were tested using the Japanese Female Facial Expression (JAFFE) Database and the Database of Faces (AT&T Faces).

  14. A prototype 4800 bps voice terminal for the mobile satellite experiment

    NASA Technical Reports Server (NTRS)

    Chan, Wai-Yip; Davidson, Grant; Chen, Juin-Hwey; Gersho, Allen

    1987-01-01

    The authors report the final stage of a research project for the NASA mobile satellite experiment (MSAT-X). The work has resulted in two alternative coding techniques which have achieved good communications quality at 4.8 kb/s while tolerating vehicle noise, fading, and a fairly high-error-rate, bursty channel. The authors have constructed a circuit board based on three AT&T 32-bit floating-point DSP32 digital signal processors (DSP) and providing both excess processing power and considerable flexibility for handling different variations of the algorithms. A Motorola 68HC11 microcomputer chip serves as the board controller and interface handler. Mounted on a wirewrapped card, the prototype's circuit footprint amounts to only 225 cm2 and consumes about 13 W of power. A description is given of the architecture, signal processing and communication functions, circuitry, and firmware of the prototype. Issues such as synchronization, framing, error control, and modem interface are also addressed.

  15. Reduction of Carbon Monoxide. Past Research Summary

    DOE R&D Accomplishments Database

    Schrock, R. R.

    1982-01-01

    Research programs for the year on the preparation, characterization, and reactions of binuclear tantalum complexes are described. All evidence to date suggest the following of these dimeric molecules: (1) the dimer does not break into monomers under mild conditions; (2) intermolecular hydride exchange is not negligible, but it is slow; (3) intermolecular non-ionic halide exchange is fast; (4) the ends of the dimers can rotate partially with respect to one another. The binuclear tantalum hydride complexes were found to react with carbon monoxide to give a molecule which is the only example of reduction of CO by a transition metal hydride to give a complex containing a CHO ligand. Isonitrides also reacted in a similar manner with dimeric tantalum hydride. (ATT)

  16. Cerebral salt wasting syndrome due to tuberculous meningitis; a case report

    PubMed Central

    Ahmad, Syed; Majid, Zain; Mehdi, Mehwish; Mubarak, Muhammed

    2016-01-01

    A 58-year-old male presented with fever, nausea, and vomiting since 15 days along with irritability and confusion since 5 days. His laboratory reports showed low serum sodium, serum osmolality and uric acid. Computerized tomography (CT) scan of brain revealed age-related changes. While on lumbar puncture (LP) and cerebrospinal fluid (CSF) examination, CSF protein, lactate dehydrogenase (LDH) and total leukocyte count (predominant lymphocytes) were all increased. On his 14th day of admission, his serum sodium was 116 mEq/l and he had a high urine output. Fluid restriction was tried in order to rule out syndrome of inappropriate antidiuretic hormone secretion (SIADH) but the patient did not respond to it. Keeping in view the above findings, a final diagnosis of tuberculous meningitis leading to cerebral salt wasting syndrome was made. The patient was started on 3% hypertonic saline, mineralocorticoids and anti-tuberculous therapy (ATT), to which he responded favorably and was later discharged. PMID:27069970

  17. Cerebral salt wasting syndrome due to tuberculous meningitis; a case report.

    PubMed

    Ahmad, Syed; Majid, Zain; Mehdi, Mehwish; Mubarak, Muhammed

    2016-01-01

    A 58-year-old male presented with fever, nausea, and vomiting since 15 days along with irritability and confusion since 5 days. His laboratory reports showed low serum sodium, serum osmolality and uric acid. Computerized tomography (CT) scan of brain revealed age-related changes. While on lumbar puncture (LP) and cerebrospinal fluid (CSF) examination, CSF protein, lactate dehydrogenase (LDH) and total leukocyte count (predominant lymphocytes) were all increased. On his 14th day of admission, his serum sodium was 116 mEq/l and he had a high urine output. Fluid restriction was tried in order to rule out syndrome of inappropriate antidiuretic hormone secretion (SIADH) but the patient did not respond to it. Keeping in view the above findings, a final diagnosis of tuberculous meningitis leading to cerebral salt wasting syndrome was made. The patient was started on 3% hypertonic saline, mineralocorticoids and anti-tuberculous therapy (ATT), to which he responded favorably and was later discharged.

  18. Integration-dependent bacteriophage immunity provides insights into the evolution of genetic switches.

    PubMed

    Broussard, Gregory W; Oldfield, Lauren M; Villanueva, Valerie M; Lunt, Bryce L; Shine, Emilee E; Hatfull, Graham F

    2013-01-24

    Genetic switches are critical components of developmental circuits. Because temperate bacteriophages are vastly abundant and greatly diverse, they are rich resources for understanding the mechanisms and evolution of switches and the molecular control of genetic circuitry. Here, we describe a new class of small, compact, and simple switches that use site-specific recombination as the key decision point. The phage attachment site attP is located within the phage repressor gene such that chromosomal integration results in removal of a C-terminal tag that destabilizes the virally encoded form of the repressor. Integration thus not only confers prophage stability but also is a requirement for lysogenic establishment. The variety of these self-contained integration-dependent immunity systems in different genomic contexts suggests that these represent ancestral states in switch evolution from which more-complex switches have evolved. They also provide a powerful toolkit for building synthetic biological circuits. PMID:23246436

  19. Case Series: Evaluation of Behavioral Sleep Intervention for Medicated Children With ADHD.

    PubMed

    Vetrayan, Jayachandran; Othman, Suhana; Victor Paulraj, Smily Jesu Priya

    2013-03-25

    Objective: To assess the effectiveness and feasibility of behavioral sleep intervention for medicated children with ADHD. Method: Six medicated children (five boys, one girl; aged 6-12 years) with ADHD participated in a 4-week sleep intervention program. The main behavioral strategies used were Faded Bedtime With Response Cost (FBRC) and positive reinforcement. Within a case-series design, objective measure (Sleep Disturbance Scale for Children [SDSC]) and subjective measure (sleep diaries) were used to record changes in children's sleep. Results: For all six children, significant decrease was found in the severity of children's sleep problems (based on SDSC data). Bedtime resistance and mean sleep onset latency were reduced following the 4-week intervention program according to sleep diaries data. Gains were generally maintained at the follow-up. Parents perceived the intervention as being helpful. Conclusion: Based on the initial data, this intervention shows promise as an effective and feasible treatment. (J. of Att. Dis. 2013; XX(X) 1-XX).

  20. Immobilization of Thiadiazole Derivatives on Magnetite Mesoporous Silica Shell Nanoparticles in Application to Heavy Metal Removal from Biological Samples

    NASA Astrophysics Data System (ADS)

    Emadi, Masoomeh; Shams, Esmaeil

    2010-12-01

    In this report magnetite was synthesized by a coprecipitation method, then coated with a layer of silica. Another layer of mesoporous silica was added by a sol-gel method, then 5-amino-1,3,4-thiadiazole-thiol (ATT) was immobilized onto the synthesized nanoparticles with a simple procedure. This was followed by a series of characterizations, including transmission electron microscopy (TEM), FT-IR spectrum, elemental analysis and XRD. Heavy metal uptake of the modified nanoparticles was examined by atomic absorption spectroscopy. For further investigation we chose Cu2+ as the preferred heavy metal to evaluate the amount of adsorption, as well as the kinetics and mechanism of adsorption. Finally, the capacity of our nanoparticles for the heavy metal removal from blood was shown. We found that the kinetic rate of Cu2+ adsorption was 0.05 g/mg/min, and the best binding model was the Freundlich isotherm.

  1. The sequence dependence of circular dichroism spectra of DNA duplexes.

    PubMed

    Arnott, S; Arnott, S

    1975-09-01

    The three satellite DNAs of Drosophila virilis, that approximate to poly d(CAAACTA)-poly d(TAGTTTG), poly d(TAAACTA)-poly d(TAGTTTA), poly d(CAAATTA)-poly d(TAATTTG), the satellite DNA of Drosophila melanogaster that approximates to poly d(AATAT)-poly d(ATATT), the synthetic DNA duplexes, poly dG-poly dC, poly d(AT)-poly d(AT), poly d(AAT)-poly d(ATT), poly d(AAC)-poly d(GTT), poly d(TAC)-poly d(GTA) and the block copolymer d(C15A15)-d(T15G15) all have circular dichroism spectra consistent with the propositions that they have the same molecular geometry in solution and that it is the kind and frequency of nucleotide triplet sequences that determines their spectral characteristics. Poly dA-poly dT is apparently an exception.

  2. Development of a 32-bit UNIX-based ELAS workstation

    NASA Technical Reports Server (NTRS)

    Spiering, Bruce A.; Pearson, Ronnie W.; Cheng, Thomas D.

    1987-01-01

    A mini/microcomputer UNIX-based image analysis workstation has been designed and is being implemented to use the Earth Resources Laboratory Applications Software (ELAS). The hardware system includes a MASSCOMP 5600 computer, which is a 32-bit UNIX-based system (compatible with AT&T System V and Berkeley 4.2 BSD operating system), a floating point accelerator, a 474-megabyte fixed disk, a tri-density magnetic tape drive, and an 1152 by 910 by 12-plane color graphics/image interface. The software conversion includes reconfiguring the ELAs driver Master Task, recompiling and then testing the converted application modules. This hardware and software configuration is a self-sufficient image analysis workstation which can be used as a stand-alone system, or networked with other compatible workstations.

  3. The Sandia Airborne Computer (SANDAC)

    SciTech Connect

    Nava, E.J.

    1992-06-01

    The Sandia Airborne Computer (SANDAC) is a small, modular, high performance, multiprocessor computer originally designed for aerospace applications. It can use a combination of Motorola 68020 and 68040 based processor modules along with AT&T DSP32C based signal processing modules. The system is designed to use up to 15 processors in almost any combination and a complete system can include up to 20 modules. Depending on the mix of processors, total computational throughput can range from 2.5 to greater than 225 Million Instructions Per Second (MIPS). The system is designed so that processors can access all resources in the machine and the inter-processor communication details are completely transparent to the software. In addition to processors, the system includes input/output, memory, and special function modules. Because of its ease of use, small size, durability, and configuration flexibility, SANDAC has been used on applications ranging from missile navigation, guidance, and control systems to medical imaging systems.

  4. Digital control of magnetic bearings supporting a multimass flexible rotor

    NASA Technical Reports Server (NTRS)

    Keith, F. J.; Williams, R. D.; Allaire, P. E.; Schafer, R. M.

    1993-01-01

    The characteristics of magnetic bearings used to support a three mass flexible rotor operated at speeds up to 14,000 RPM are discussed. The magnetic components of the bearing are of a type reported in the literature previously, but the earlier analog controls were replaced by digital ones. Analog-to-digital and digital-to-analog converters and digital control software were installed in an AT&T PC. This PC-based digital controller was used to operate one of the magnetic bearings on the test rig. Basic proportional-derivative control was applied to the bearings, and the bearing stiffness and damping characteristics were evaluated. Particular attention is paid to the frequency dependent behavior of the stiffness and damping properties, and comparisons are made between the actual controllers and ideal proportional-derivative control.

  5. A High-Average-Power Free Electron Laser for Microfabrication and Surface Applications

    NASA Technical Reports Server (NTRS)

    Dylla, H. F.; Benson, S.; Bisognano, J.; Bohn, C. L.; Cardman, L.; Engwall, D.; Fugitt, J.; Jordan, K.; Kehne, D.; Li, Z.; Liu, H.; Merminga, L.; Neil, G. R.; Neuffer, D.; Shinn, M.; Sinclair, C.; Wiseman, M.; Brillson, L. J.; Henkel, D. P.; Helvajian, H.; Kelley, M. J.; Nair, Shanti

    1995-01-01

    CEBAF has developed a comprehensive conceptual design of an industrial user facility based on a kilowatt ultraviolet (UV) (160-1000 mm) and infrared (IR) (2-25 micron) free electron laser (FEL) driven by a recirculating, energy recovering 200 MeV superconducting radio frequency (SRF) accelerator. FEL users, CEBAF's partners in the Lase Processing Consortium, including AT&T, DuPont, IBM, Northrop Grumman, 3M, and Xerox, are developing applications such as metal, ceramic, and electronic material micro-fabrication and polymer and metal surface processing, with the overall effort leading to later scale-up to industrial systems at 50-100 kW. Representative applications are described. The proposed high-average-power FEL overcomes limitations of conventional laser sources in available power, cost-effectiveness, tunability, and pulse structure.

  6. Ulcerative lupus vulgaris of face: an uncommon presentation in India.

    PubMed

    Padmavathy, L; Rao, L Lakshmana; Ethirajan, N; Krishnaswami, B

    2007-01-01

    Tuberculosis affects the population world wide, more among those living in developing countries. The incidence of tuberculosis registered an upward trend even in developed countries, with the advent of HIV infection. Cutaneous tuberculosis accounts for about 1% of cases of extra-pulmonary tuberculosis. Cutaneous tuberculosis presents with various lesions ranging from ulcerative to proliferative or hyperkeratotic lesions. The lesions may sometimes be associated with marked destruction of the tissues resulting in marked disfigurement, especially when it involves face as seen in cases of Lupus Vulgaris. A case of Lupus Vulgaris in a young woman with extensive ulceration of face which responded to ATT resulting in scarring of the face is reported for its rarity amongst Indian population as against western population.

  7. Site-Specific Cassette Exchange Systems in the Aedes aegypti Mosquito and the Plutella xylostella Moth

    PubMed Central

    Haghighat-Khah, Roya Elaine; Scaife, Sarah; Martins, Sara; St John, Oliver; Matzen, Kelly Jean; Morrison, Neil; Alphey, Luke

    2015-01-01

    Genetically engineered insects are being evaluated as potential tools to decrease the economic and public health burden of mosquitoes and agricultural pest insects. Here we describe a new tool for the reliable and targeted genome manipulation of pest insects for research and field release using recombinase mediated cassette exchange (RMCE) mechanisms. We successfully demonstrated the established ΦC31-RMCE method in the yellow fever mosquito, Aedes aegypti, which is the first report of RMCE in mosquitoes. A new variant of this RMCE system, called iRMCE, combines the ΦC31-att integration system and Cre or FLP-mediated excision to remove extraneous sequences introduced as part of the site-specific integration process. Complete iRMCE was achieved in two important insect pests, Aedes aegypti and the diamondback moth, Plutella xylostella, demonstrating the transferability of the system across a wide phylogenetic range of insect pests. PMID:25830287

  8. Final Technical Report

    SciTech Connect

    Maxwell, Mike, J., P.E.

    2012-08-30

    The STI product is the Final Technical Report from ReliOn, Inc. for contract award DE-EE0000487: Recovery Act PEM Fuel Cell Systems Providing Emergency Reserve and Backup Power. The program covered the turnkey deployment of 431 ReliOn fuel cell systems at 189 individual sites for AT&T and PG&E with ReliOn functioning as the primary equipment supplier and the project manager. The Final Technical Report provides an executive level summary, a comparison of the actual accomplishments vs. the goals and objectives of the project, as well as a summary of the project activity from the contract award date of August 1, 2009 through the contract expiration date of December 31, 2011. Two photos are included in the body of the report which show hydrogen storage and bulk hydrogen refueling technologies developed as a result of this program.

  9. Cognitive Processes in ADHD and Asperger's Disorder: Overlaps and Differences in PASS Profiles.

    PubMed

    Taddei, Stefano; Contena, Bastianina

    2013-11-01

    Objective: Many studies report on the usefulness of the evaluation of Executive Functions (EF) in the assessment of participants with ADHD, while others underline how deficits of EF in these participants are not consistent and that the same executive deficits are present in many other disorders, particularly in Asperger's disorder. Using the Planning Attention Simultaneous Successive (PASS) theory, the present study explores the cognitive profiles of participants with ADHD or Asperger's disorder and compares the cognitive functioning of these two diagnostic groups. Method: Forty-four children, 24 with a diagnosis of ADHD and 20 with a diagnosis of Asperger's disorder, participated and their cognitive processes were evaluated with the Cognitive Assessment System. Results: Results underline specific cognitive profiles in ADHD and Asperger's disorder characterized by weaknesses in planning and attention, but with a diverse level of severity. Conclusion: Implications of the different cognitive profiles of these diagnostic groups are discussed. (J. of Att. Dis. 2013; XX(X) 1-XX).

  10. The complete mitochondrial genome of wolf spider Pirata subpiraticus Boes.et str. (Araneae: Lycosidae).

    PubMed

    Wang, Zheng-Liang; Li, Chao; Fang, Wen-Yuan; Yu, Xiao-Ping

    2016-05-01

    The complete mitochondrial genome of a wolf spider Pirata subpiraticus was determined. It is a circular molecule of 14,528 bp in length and contains 13 protein-coding genes, 2 ribosomal RNAs, 21 transfer RNAs (tRNA(Gly) is lost) and a control region. The A + T content of the overall base composition of majority strand (J-strand) is 75.6% (T: 42.0%; C: 8.6%; A:33.6%; G: 15.8%). Among protein-coding genes, one genes (COIII) start with TTG, four genes (ND1, ND2, ATP8 and Cytb) begin with ATT and other eight genes use ATA as initiation codon. ND2 and Cytb are terminated with TAG as stop codon, while all other genes end with TAA. Three isolated sequences repeat regions (212 bp) were found in the A+T-rich region (D-loop).

  11. A complicated case of altered sensorium

    PubMed Central

    Vikram, Kushaal; KN, Shivashankara; Chakraborty, Joydeep; Umakanth, Shashikiran

    2013-01-01

    Tuberculosis and malaria are two common diseases afflicting the population of the Indian subcontinent. Both are associated with fatal neurological complications. Clinically, it may be difficult to differentiate between the two, except for that the focal neurological complications are more common with central nervous system (CNS) tuberculosis than with cerebral malaria. We report the case of a 48-year-old woman who presented with a history of fever and altered sensorium of short duration. She was initially treated for falciparum malaria on the basis of positive peripheral smear and normal CT scan of the brain, but later on diagnosed to have CNS tuberculosis on the basis of persistence of symptoms, development of focal neurological symptoms and MRI of the brain showing multiple tuberculomas. She improved significantly with a course of antitubercular treatment (ATT) and systemic corticosteroids. PMID:23955980

  12. [Construction of Fat-1 eukaryotic expression vector of excision markers and the establishment of transgenic sheep cell lines].

    PubMed

    Lima, A; Zhu, Heping; Wang, Ruiyao; Yan, Tao; Su, Xiaohu; Li, Lu; Wang, Bingping; Na, Shunwendoule; Qi, Guichun; Zhou, Huanmin

    2016-02-01

    In order to establish marker-free transgenic cell lines, we cloned Fat-1 gene, attB and Loxp sequences by PCR. Then we inserted these sequences to pN1-EGFP vector and got pEGFP-N1-Fat-1 expression vector. PhiC31 integrase mRNA which was generated by in vitro transcription and a pEGFP-N1-Fat-1 expression vector co-electroporated into sheep fetal fibroblasts, and then we got transgenic cell lines expressing green fluorescence. Prokaryotic expression and purification of Cre recombinant protein was performed. Cre recombinant protein was transducted into stably-transfected cell colonies. We identified cell colonies by sequencing and established marker-free transgenic cell lines and eventually- established marker-free transgenic cell lines which were building more safely basic for producing Fat-1 transgenic animals. PMID:27382771

  13. A "Sweet 16" of Rules About Teamwork

    NASA Technical Reports Server (NTRS)

    Laufer, Alexander (Editor)

    2002-01-01

    The following "Sweet 16" rules included in this paper derive from a longer paper by APPL Director Dr. Edward Hoffman and myself entitled " 99 Rules for Managing Faster, Better, Cheaper Projects." Our sources consisted mainly of "war stories" told by master project managers in my book Simultaneous Management: Managing Projects in a Dynamic Environment (AMACOM, The American Management Association, 1996). The Simultaneous Management model was a result of 10 years of intensive research and testing conducted with the active participation of master project managers from leading private organizations such as AT&T, DuPont, Exxon, General Motors, IBM, Motorola and Procter & Gamble. In a more recent study, led by Dr. Hoffman, we learned that master project managers in leading public organizations employ most of these rules as well. Both studies, in private and public organizations, found that a dynamic environment calls for dynamic management, and that is especially clear in how successful project managers think about their teams.

  14. Insights into transcription enhancer factor 1 (TEF-1) activity from the solution structure of the TEA domain

    PubMed Central

    Anbanandam, Asokan; Albarado, Diana C.; Nguyen, Catherine T.; Halder, Georg; Gao, Xiaolian; Veeraraghavan, Sudha

    2006-01-01

    Transcription enhancer factor 1 is essential for cardiac, skeletal, and smooth muscle development and uses its N-terminal TEA domain (TEAD) to bind M-CAT elements. Here, we present the first structure of TEAD and show that it is a three-helix bundle with a homeodomain fold. Structural data reveal how TEAD binds DNA. Using structure-function correlations, we find that the L1 loop is essential for cooperative loading of TEAD molecules on to tandemly duplicated M-CAT sites. Furthermore, using a microarray chip-based assay, we establish that known binding sites of the full-length protein are only a subset of DNA elements recognized by TEAD. Our results provide a model for understanding the regulation of genome-wide gene expression during development by TEA/ATTS family of transcription factors. PMID:17085591

  15. Dynamic evaluation of growth hormone (GH) and prolactin (hPRL) secretion in active acromegaly with high and low GH output.

    PubMed

    Tolis, G; Kovacs, L; Friesen, H; Martin, J B

    1975-02-01

    Ten patients with active acromegaly were studied. In 9 plasma GH levels failed to suppress after glucose (OGTT), in 8 an increase in serum GH occurred after thyrotropin releasing hormone (TRH). After L-Dopa, 4 patients showed no change in serum GH, 3 exhibited a decrease and in 3 an increase in serum hGH occurred. With a combined insulin (ITT) and arginine (ATT) test, 2 patients exhibited an increase in hGH, and in 6 no change occurred. Fasting serum GH concentration was less than 11 ng/ml in 5 patients. Basal prolactin (hPRL) levels were normal in all patients including two with galactorrhea. L-Dopa suppressed and TRH stimulated hPRL secretion in all, but the responses which were seen were subnormal. Hydrocortisone infusion in two acromegalics did not affect the prolactin induced increase after TRH but blunted the GH increase after TRH.

  16. Symposium on the 50th anniversary of the discovery of deuterium: divisions of history of chemistry, geochemistry, nuclear and physical chemistry

    SciTech Connect

    Bigeleisen, J.

    1981-01-01

    In tribute to Harold Urey, the author cites Urey's accomplishments as a scientist and public servant and gives insight of the man. Some of his accomplishments in science are: discovered deuterium for which he received the Nobel prize in 1934; worked with Rittenberg and Greiff on the theory and calculation of the differences in the thermodynamic properties of isotopic compounds; with Thode, Hutchison and others, separated the isotopes of the light elements by chemical methods on a laboratory scale; during World War II headed the Substitute Alloy Materials Laboratories of Columbia University where his group developed industrial scale processes for the separation of /sup 2/H, /sup 10/B, and /sup 235/U; with his associates developed the /sup 18/O paleotemperature scale; made fundamental contributions to a widely accepted theory of the origin of the earth. (ATT)

  17. The complete mitochondrial genome of Celastrina hersilia (Lepidoptera: Lycaenidae).

    PubMed

    Lei, Ying; Xu, Chang; Xu, Chongren; Wang, Rongjiang

    2016-01-01

    The complete mitochondrial genome of Celastrina hersilia (Lepidoptera: Lycaenidae) is determined in this work. The mitochondrial genome is 15,304 bp in length, which contains typical 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes and 1 non-coding A + T-rich region. All PCGs are initiated by ATA or ATT codons, except for COI, which uses CGA as a start codon. Four PCGs (COI, COII, ND5, and ND4) terminate with incomplete termination codons TA or T, while the others use TAA as stop codons. Most of the tRNA genes can be folded into a typical cloverleaf structure. The A + T-rich region is 370 bp in length, which contains several features common to the other lepidopteran species. PMID:24495135

  18. Properties of the streptomycete temperate bacteriophage FP43.

    PubMed

    Hahn, D R; McHenney, M A; Baltz, R H

    1991-06-01

    FP43 is a temperate bacteriophage for Streptomyces griseofuscus that forms plaques on many Streptomyces species. FP43 virions contain 56 kb of double-strand DNA that is circularly permuted and terminally redundant, and contains 65% G + C. A physical map of the FP43 genome was constructed, and the origin for headful packaging (pac) was localized to an 8.8-kb region of the genome (hft) that mediates high-frequency transduction by FP43 of plasmid pRHB101. The phage attachment site (attP), a replication origin (rep), a region that inhibits plaque formation (pin), and a 3-kb deletion (rpt) that caused a 100-fold reduction in plasmid transduction were mapped.

  19. Using CLIPS in a distributed system: The Network Control Center (NCC) expert system

    NASA Technical Reports Server (NTRS)

    Wannemacher, Tom

    1990-01-01

    This paper describes an intelligent troubleshooting system for the Help Desk domain. It was developed on an IBM-compatible 80286 PC using Microsoft C and CLIPS and an AT&T 3B2 minicomputer using the UNIFY database and a combination of shell script, C programs and SQL queries. The two computers are linked by a lan. The functions of this system are to help non-technical NCC personnel handle trouble calls, to keep a log of problem calls with complete, concise information, and to keep a historical database of problems. The database helps identify hardware and software problem areas and provides a source of new rules for the troubleshooting knowledge base.

  20. Immobilization of Thiadiazole Derivatives on Magnetite Mesoporous Silica Shell Nanoparticles in Application to Heavy Metal Removal from Biological Samples

    SciTech Connect

    Emadi, Masoomeh; Shams, Esmaeil

    2010-12-02

    In this report magnetite was synthesized by a coprecipitation method, then coated with a layer of silica. Another layer of mesoporous silica was added by a sol-gel method, then 5-amino-1,3,4-thiadiazole-thiol (ATT) was immobilized onto the synthesized nanoparticles with a simple procedure. This was followed by a series of characterizations, including transmission electron microscopy (TEM), FT-IR spectrum, elemental analysis and XRD. Heavy metal uptake of the modified nanoparticles was examined by atomic absorption spectroscopy. For further investigation we chose Cu{sup 2+} as the preferred heavy metal to evaluate the amount of adsorption, as well as the kinetics and mechanism of adsorption. Finally, the capacity of our nanoparticles for the heavy metal removal from blood was shown. We found that the kinetic rate of Cu{sup 2+} adsorption was 0.05 g/mg/min, and the best binding model was the Freundlich isotherm.

  1. The complete mitochondrial genome and phylogenetic analysis of the giant panda (Ailuropoda melanoleuca).

    PubMed

    Peng, Rui; Zeng, Bo; Meng, Xiuxiang; Yue, Bisong; Zhang, Zhihe; Zou, Fangdong

    2007-08-01

    The complete mitochondrial genome sequence of the giant panda, Ailuropoda melanoleuca, was determined by the long and accurate polymerase chain reaction (LA-PCR) with conserved primers and primer walking sequence methods. The complete mitochondrial DNA is 16,805 nucleotides in length and contains two ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes and one control region. The total length of the 13 protein-coding genes is longer than the American black bear, brown bear and polar bear by 3 amino acids at the end of ND5 gene. The codon usage also followed the typical vertebrate pattern except for an unusual ATT start codon, which initiates the NADH dehydrogenase subunit 5 (ND5) gene. The molecular phylogenetic analysis was performed on the sequences of 12 concatenated heavy-strand encoded protein-coding genes, and suggested that the giant panda is most closely related to bears. PMID:17499457

  2. Characterization of the mitochondrial genome of Bufo gargarizans minshanicus (Anura: Bufonidae).

    PubMed

    Yang, Jiandong; Liu, Jiabin; Xue, Rui; Chen, Limin

    2016-09-01

    The complete mitogenome of Bufo gargarizans minshanicus is 17,719 bp in size, and shares a conserved gene arrangement, gene content and base composition with other Bufonidae species. The base composition of the H-strand is biased toward A + T content at 56.97% (T 28.14%, C 27.67%, A 28.83% and G 15.36%). Most protein-coding genes (PCGs) begin with ATG, while COX1 and ND2 initiate with ATA and ND1 starts at ATT. Seven PCGs have the complete stop codons AGA and TAA, whereas the remaining four PCGs end with TA or T. The putative control region possesses two repeat motifs, and four termination-associated sequences and six conserved sequence blocks at the 5' and 3' ends.

  3. Plasmids for increased efficiency of vector construction and genetic engineering in filamentous fungi.

    PubMed

    Schoberle, Taylor J; Nguyen-Coleman, C Kim; May, Gregory S

    2013-01-01

    Fungal species are continuously being studied to not only understand disease in humans and plants but also to identify novel antibiotics and other metabolites of industrial importance. Genetic manipulations, such as gene deletion, gene complementation, and gene over-expression, are common techniques to investigate fungal gene functions. Although advances in transformation efficiency and promoter usage have improved genetic studies, some basic steps in vector construction are still laborious and time-consuming. Gateway cloning technology solves this problem by increasing the efficiency of vector construction through the use of λ phage integrase proteins and att recombination sites. We developed a series of Gateway-compatible vectors for use in genetic studies in a range of fungal species. They contain nutritional and drug-resistance markers and can be utilized to manipulate different filamentous fungal genomes.

  4. Sandia`s network for supercomputing `95: Validating the progress of Asynchronous Transfer Mode (ATM) switching

    SciTech Connect

    Pratt, T.J.; Vahle, O.; Gossage, S.A.

    1996-04-01

    The Advanced Networking Integration Department at Sandia National Laboratories has used the annual Supercomputing conference sponsored by the IEEE and ACM for the past three years as a forum to demonstrate and focus communication and networking developments. For Supercomputing `95, Sandia elected: to demonstrate the functionality and capability of an AT&T Globeview 20Gbps Asynchronous Transfer Mode (ATM) switch, which represents the core of Sandia`s corporate network, to build and utilize a three node 622 megabit per second Paragon network, and to extend the DOD`s ACTS ATM Internet from Sandia, New Mexico to the conference`s show floor in San Diego, California, for video demonstrations. This paper documents those accomplishments, discusses the details of their implementation, and describes how these demonstrations supports Sandia`s overall strategies in ATM networking.

  5. Approaches to eliminating chlorofluorocarbon use in manufacturing.

    PubMed

    Boyhan, W S

    1992-02-01

    Until quite recently, chlorofluorocarbons (CFCs) had been considered the safest and most benign of industrial chemicals. Their physical and chemical properties made them an integral part of manufacturing processes for electronics products. The recognition that CFCs destroy the stratospheric ozone layer, with consequent enormous consequences to all forms of life on earth, has led to international agreements which will end virtually all possibly before. This impending phaseout of CFCs has caused electronics manufacturers to examine alternative chemicals and processing methods. This manuscript documents the steps AT&T has taken to reach its goal of 100% phaseout of CFCs by years-end 1994. These actions include top-down management support with combined bottom-up thrusts, an internal information gathering and dissemination center, internal technology transfer, and external corporate activism.

  6. A prototype 4800 bps voice terminal for the mobile satellite experiment

    NASA Astrophysics Data System (ADS)

    Chan, Wai-Yip; Davidson, Grant; Chen, Juin-Hwey; Gersho, Allen

    The authors report the final stage of a research project for the NASA mobile satellite experiment (MSAT-X). The work has resulted in two alternative coding techniques which have achieved good communications quality at 4.8 kb/s while tolerating vehicle noise, fading, and a fairly high-error-rate, bursty channel. The authors have constructed a circuit board based on three AT&T 32-bit floating-point DSP32 digital signal processors (DSP) and providing both excess processing power and considerable flexibility for handling different variations of the algorithms. A Motorola 68HC11 microcomputer chip serves as the board controller and interface handler. Mounted on a wirewrapped card, the prototype's circuit footprint amounts to only 225 cm2 and consumes about 13 W of power. A description is given of the architecture, signal processing and communication functions, circuitry, and firmware of the prototype. Issues such as synchronization, framing, error control, and modem interface are also addressed.

  7. A high-average-power FEL for industrial applications

    SciTech Connect

    Dylla, H.F.; Benson, S.; Bisognano, J.

    1995-12-31

    CEBAF has developed a comprehensive conceptual design of an industrial user facility based on a kilowatt UV (150-1000 nm) and IR (2-25 micron) FEL driven by a recirculating, energy-recovering 200 MeV superconducting radio-frequency (SRF) accelerator. FEL users{endash}CEBAF`s partners in the Laser Processing Consortium, including AT&T, DuPont, IBM, Northrop-Grumman, 3M, and Xerox{endash}plan to develop applications such as polymer surface processing, metals and ceramics micromachining, and metal surface processing, with the overall effort leading to later scale-up to industrial systems at 50-100 kW. Representative applications are described. The proposed high-average-power FEL overcomes limitations of conventional laser sources in available power, cost-effectiveness, tunability and pulse structure. 4 refs., 3 figs., 2 tabs.

  8. A Multiprocessor Operating System Simulator

    NASA Technical Reports Server (NTRS)

    Johnston, Gary M.; Campbell, Roy H.

    1988-01-01

    This paper describes a multiprocessor operating system simulator that was developed by the authors in the Fall semester of 1987. The simulator was built in response to the need to provide students with an environment in which to build and test operating system concepts as part of the coursework of a third-year undergraduate operating systems course. Written in C++, the simulator uses the co-routine style task package that is distributed with the AT&T C++ Translator to provide a hierarchy of classes that represents a broad range of operating system software and hardware components. The class hierarchy closely follows that of the 'Choices' family of operating systems for loosely- and tightly-coupled multiprocessors. During an operating system course, these classes are refined and specialized by students in homework assignments to facilitate experimentation with different aspects of operating system design and policy decisions. The current implementation runs on the IBM RT PC under 4.3bsd UNIX.

  9. Next-generation avionics packaging and cooling 'test results from a prototype system'

    NASA Astrophysics Data System (ADS)

    Seals, J. D.

    The author reports on the design, material characteristics, and test results obtained under the US Air Force's advanced aircraft avionics packaging technologies (AAAPT) program, whose charter is to investigate new designs and technologies for reliable packaging, interconnection, and thermal management. Under this program, AT&T Bell Laboratories has completed the preliminary testing of and is evaluating a number of promising materials and technologies, including conformal encapsulation, liquid flow-through cooling, and a cyanate ester backplane. A fifty-two module system incorporating these and and other technologies has undergone preliminary cooling efficiency, shock, sine and random vibration, and maintenance testing. One of the primary objectives was to evaluate the interaction compatibility of new materials and designs with other components in the system.

  10. The complete mitochondrial genome of Lithobates catesbeianus (Anura: Ranidae).

    PubMed

    Lin, Yubo; Tao, Bofang; Fang, Xindong; Wang, Tingting; Zhang, Jiayong

    2014-12-01

    The complete mitochondrial genome of Lithobates catesbeianus (Anura: Ranidae) is sequenced to analyze the gene arrangement. It is a circular molecule of 18,241 bp in length including 37 genes typically found in other frogs. The AT content of the overall base composition of L. catesbeianus is 59.9%. The length of control region is 2783 bp with 66.0% AT content. Protein-coding genes begin with ATG as start codon except except ND1 and ATP6 began with ATA, COI and ND4L with GTG, and ND2 with ATT. COI end with AGG as stop codon, COII and ND6 end with AGA, ND2 end with TAG, ATP8. ND4L. ND5 and Cytb end with TAA, and the other five PCGs end with a incomplete stop codon (a single stop nucleotide T).

  11. Position, rotation, and intensity invariant recognizing method

    DOEpatents

    Ochoa, Ellen; Schils, George F.; Sweeney, Donald W.

    1989-01-01

    A method for recognizing the presence of a particular target in a field of view which is target position, rotation, and intensity invariant includes the preparing of a target-specific invariant filter from a combination of all eigen-modes of a pattern of the particular target. Coherent radiation from the field of view is then imaged into an optical correlator in which the invariant filter is located. The invariant filter is rotated in the frequency plane of the optical correlator in order to produce a constant-amplitude rotational response in a correlation output plane when the particular target is present in the field of view. Any constant response is thus detected in the output The U.S. Government has rights in this invention pursuant to Contract No. DE-AC04-76DP00789 between the U.S. Department of Energy and AT&T Technologies, Inc.

  12. Geologic analysis of Devonian Shale cores

    SciTech Connect

    1982-02-01

    Cleveland Cliffs Iron Company was awarded a DOE contract in December 1977 for field retrieval and laboratory analysis of cores from the Devonian shales of the following eleven states: Michigan, Illinois, Indiana, Ohio, New York, Pennsylvania, West Virginia, Maryland, Kentucky, Tennessee and Virginia. The purpose of this project is to explore these areas to determine the amount of natural gas being produced from the Devonian shales. The physical properties testing of the rock specimens were performed under subcontract at Michigan Technological University (MTU). The study also included LANDSAT information, geochemical research, structural sedimentary and tectonic data. Following the introduction, and background of the project this report covers the following: field retrieval procedures; laboratory procedures; geologic analysis (by state); references and appendices. (ATT)

  13. Drosophila topoisomerase II double-strand DNA cleavage: analysis of DNA sequence homology at the cleavage site.

    PubMed Central

    Sander, M; Hsieh, T S

    1985-01-01

    In order to study the sequence specificity of double-strand DNA cleavage by Drosophila topoisomerase II, we have mapped and sequenced 16 strong and 47 weak cleavage sites in the recombinant plasmid p pi 25.1. Analysis of the nucleotide and dinucleotide frequencies in the region near the site of phosphodiester bond breakage revealed a nonrandom distribution. The nucleotide frequencies observed would occur by chance with a probability less than 0.05. The consensus sequence we derived is 5'GT.A/TAY decrease ATT.AT..G 3', where a dot means no preferred nucleotide, Y is for pyrimidine, and the arrow shows the point of bond cleavage. On average, strong sites match the consensus better than weak sites. Images PMID:2987816

  14. Averaging to minimize or eliminate regression toward the mean to measure pure experimental effects.

    PubMed

    Toh, Rex S; Hu, Michael Y

    2008-06-01

    This paper explains how regression toward the mean can contaminate diary data, making it difficult to measure the pure effects of an experimental variable over time. Using a large scale real-life database collected by AT&T, a method of measuring this mathematical artifact is advanced. It is shown to manifest very quickly as a result of a spontaneous reaction toward happenstance, with the most extreme initial values gravitating most toward the mean. Then averaging over longer and longer periods of time to define use categories is shown to dilute happenstance increasingly, and therefore progressively minimizes or eliminates regression toward the mean. Finally, regression toward the mean is very pervasive and very persistent.

  15. Adolescents With ADHD: Experiences of Having an ADHD Diagnosis and Negotiations of Self-Image and Identity.

    PubMed

    Jones, Sheila; Hesse, Morten

    2014-02-18

    Objective: To develop an understanding of the meanings and consequences of a diagnosis of ADHD for young people during the transition from childhood to adolescence. Method: Nine youth with an ADHD diagnosis were interviewed using a semi-structured interview guide on topics of management of identity and their own experiences of their difficulties relating to ADHD. Results: The meanings and consequences of the ADHD diagnosis is best understood in terms of overall identity development and social connectedness. In addition, cultural understandings of what ADHD is play a major role in negotiating the role of ADHD in identity. Conclusion: Youth with ADHD moving from childhood to adulthood need support for identity development. Health care professionals should be aware of young people's experiences of ADHD symptoms in the context of their life situations and the cultural understanding of ADHD during treatment and psychoeducation. (J. of Att. Dis. 2014; XX(X) 1-XX).

  16. Primary Amoebic Meningoencephalitis.

    PubMed

    Gupta, Ritu; Parashar, M K; Kale, Aditya

    2015-04-01

    Primary amoebic meningoencephalitis due to free living amoeba, also called 'brain eating amoeba', Naegleria fowleri, was detected in retroviral disease patient of 40 years who has history of using well water. Patient was admitted with severe headache, fever intermittent, nausea, vomiting and slurring of speech. CT scan and MRI scan findings were normal. CSF examination showed increased protein, low sugar and predominant lymphocytes. CSF was negative for cryptococcal antigen but wet mount preparation showed highly motile free living amoeba Naegleria fowleri. Patient was put on Amphotericin B, Metronidazole, Rifampicin in addition to ART and ATT and other supportive medications. His headache was relieved and patient improved and was discharged on request. Earlier eight cases have been reported from India of which four cases survived the acute episode.

  17. Understanding radioactive waste

    SciTech Connect

    Murray, R.L.

    1981-12-01

    This document contains information on all aspects of radioactive wastes. Facts are presented about radioactive wastes simply, clearly and in an unbiased manner which makes the information readily accessible to the interested public. The contents are as follows: questions and concerns about wastes; atoms and chemistry; radioactivity; kinds of radiation; biological effects of radiation; radiation standards and protection; fission and fission products; the Manhattan Project; defense and development; uses of isotopes and radiation; classification of wastes; spent fuels from nuclear reactors; storage of spent fuel; reprocessing, recycling, and resources; uranium mill tailings; low-level wastes; transportation; methods of handling high-level nuclear wastes; project salt vault; multiple barrier approach; research on waste isolation; legal requiremnts; the national waste management program; societal aspects of radioactive wastes; perspectives; glossary; appendix A (scientific American articles); appendix B (reference material on wastes). (ATT)

  18. Psychiatric morbidity in vitiligo and psoriasis: a comparative study from India.

    PubMed

    Mattoo, S K; Handa, S; Kaur, I; Gupta, N; Malhotra, R

    2001-08-01

    In a tertiary-care teaching hospital in India, dermatology outpatients with vitiligo (N=113) and psoriasis (N=103) were studied for psychiatric morbidity. The two groups were similar with regard to education, locality, religion, and attitude to appearance (ATT). Psoriasis cases were older, more often male, and more often married. The General Health Questionnaire (GHQ) assessed psychiatric morbidity rates at 33.63% and 24.7% for vitiligo and psoriasis, respectively. The ICD-10 psychiatric diagnoses in GHQ positive cases were: adjustment disorder (56% vs 62%), depressive episode (22% vs 29%) and dysthymia (9% vs 4%) in vitiligo and psoriasis, respectively. The Comprehensive Psychopathological Rating Scale (CPRS) assessed that depression, anxiety, and total psychopathology levels were similar in the two GHQ positive subgroups. Significant correlations were noted between psychopathology (GHQ CRPS), dysfunction as per Dysfunction Analysis Questionnaire (DAQ), and behavior change as per Impact of Skin Disease Scale (IMPACT), and all were more prominent in vitiligo.

  19. A new synthetic route to a family of non-classical addition-type thermoplastics. I - Concept and demonstration

    NASA Technical Reports Server (NTRS)

    Pater, Ruth H.

    1988-01-01

    A synthesis scheme was developed for a host of nonclassical addition-type thermoplastics (ATT) that can be cured by an addition reaction, leading to a linear polymer structure. The synthesis involves the reaction of an acetylene-terminated prepolymer with either a bismaleimide (BMI) or a biscitraconimide. A new polymer, designated LaRC-RP80, synthesized using this scheme, was found to exhibit several significantly improved properties over the commercial BMI, Kerimid 601, including an eight-fold increase in toughness while maintaining a high Tg, a higher (by 167 C) heat stability, and a 50-percent increase in moisture resistance. In addition, LaRC-RP80 has good hot/wet lap shear strength and processes easily at 288 C without voids in the finished product.

  20. Operation and performance of a longitudinal damping system using parallel digital signal processing

    SciTech Connect

    Fox, J.D.; Hindi, H.; Linscott, I.

    1994-06-01

    A programmable longitudinal feedback system based on four AT&T 1610 digital signal processors has been developed as a component of the PEP-II R&D program. This Longitudinal Quick Prototype is a proof of concept for the PEP-II system and implements full speed bunch-by-bunch signal processing for storage rings with bunch spacings of 4 ns. The design implements, via software, a general purpose feedback controller which allows the system to be operated at several accelerator facilities. The system configuration used for tests at the LBL Advanced Light Source is described. Open and closed loop results showing the detection and calculation of feedback signals from bunch motion are presented, and the system is shown to damp coupled-bunch instabilities in the ALS. Use of the system for accelerator diagnostics is illustrated via measurement of injection transients and analysis of open loop bunch motion.

  1. [Construction of Fat-1 eukaryotic expression vector of excision markers and the establishment of transgenic sheep cell lines].

    PubMed

    Lima, A; Zhu, Heping; Wang, Ruiyao; Yan, Tao; Su, Xiaohu; Li, Lu; Wang, Bingping; Na, Shunwendoule; Qi, Guichun; Zhou, Huanmin

    2016-02-01

    In order to establish marker-free transgenic cell lines, we cloned Fat-1 gene, attB and Loxp sequences by PCR. Then we inserted these sequences to pN1-EGFP vector and got pEGFP-N1-Fat-1 expression vector. PhiC31 integrase mRNA which was generated by in vitro transcription and a pEGFP-N1-Fat-1 expression vector co-electroporated into sheep fetal fibroblasts, and then we got transgenic cell lines expressing green fluorescence. Prokaryotic expression and purification of Cre recombinant protein was performed. Cre recombinant protein was transducted into stably-transfected cell colonies. We identified cell colonies by sequencing and established marker-free transgenic cell lines and eventually- established marker-free transgenic cell lines which were building more safely basic for producing Fat-1 transgenic animals.

  2. An avian live attenuated master backbone for potential use in epidemic and pandemic influenza vaccines.

    PubMed

    Hickman, Danielle; Hossain, Md Jaber; Song, Haichen; Araya, Yonas; Solórzano, Alicia; Perez, Daniel R

    2008-11-01

    The unprecedented emergence in Asia of multiple avian influenza virus (AIV) subtypes with a broad host range poses a major challenge in the design of vaccination strategies that are both effective and available in a timely manner. The present study focused on the protective effects of a genetically modified AIV as a source for the preparation of vaccines for epidemic and pandemic influenza. It has previously been demonstrated that a live attenuated AIV based on the internal backbone of influenza A/Guinea fowl/Hong Kong/WF10/99 (H9N2), called WF10att, is effective at protecting poultry species against low- and high-pathogenicity influenza strains. More importantly, this live attenuated virus provided effective protection when administered in ovo. In order to characterize the WF10att backbone further for use in epidemic and pandemic influenza vaccines, this study evaluated its protective effects in mice. Intranasal inoculation of modified attenuated viruses in mice provided adequate protective immunity against homologous lethal challenges with both the wild-type influenza A/WSN/33 (H1N1) and A/Vietnam/1203/04 (H5N1) viruses. Adequate heterotypic immunity was also observed in mice vaccinated with modified attenuated viruses carrying H7N2 surface proteins. The results presented in this report suggest that the internal genes of a genetically modified AIV confer similar protection in a mouse model and thus could be used as a master donor strain for the generation of live attenuated vaccines for epidemic and pandemic influenza.

  3. Hydrogel Microencapsulated Insulin-Secreting Cells Increase Keratinocyte Migration, Epidermal Thickness, Collagen Fiber Density, and Wound Closure in a Diabetic Mouse Model of Wound Healing

    PubMed Central

    Faulknor, Renea; Berthiaume, François; Olabisi, Ronke M.

    2015-01-01

    Wound healing is a hierarchical process of intracellular and intercellular signaling. Insulin is a potent chemoattractant and mitogen for cells involved in wound healing. Insulin's potential to promote keratinocyte growth and stimulate collagen synthesis in fibroblasts is well described. However, there currently lacks an appropriate delivery mechanism capable of consistently supplying a wound environment with insulin; current approaches require repeated applications of insulin, which increase the chances of infecting the wound. In this study, we present a novel cell-based therapy that delivers insulin to the wound area in a constant or glucose-dependent manner by encapsulating insulin-secreting cells in nonimmunogenic poly(ethylene glycol) diacrylate (PEGDA) hydrogel microspheres. We evaluated cell viability and insulin secretory characteristics of microencapsulated cells. Glucose stimulation studies verified free diffusion of glucose and insulin through the microspheres, while no statistical difference in insulin secretion was observed between cells in microspheres and cells in monolayers. Scratch assays demonstrated accelerated keratinocyte migration in vitro when treated with microencapsulated cells. In excisional wounds on the dorsa of diabetic mice, microencapsulated RIN-m cells accelerated wound closure by postoperative day 7; a statistically significant increase over AtT-20ins-treated and control groups. Histological results indicated significantly greater epidermal thickness in both microencapsulated RIN-m and AtT-20ins-treated wounds. The results suggest that microencapsulation enables insulin-secreting cells to persist long enough at the wound site for a therapeutic effect and thereby functions as an effective delivery vehicle to accelerate wound healing. PMID:26239745

  4. Morphological and Morphometrical Characterization of Meloidogyne incognita from Different Host Plants in Four Districts of Punjab, India.

    PubMed

    Kaur, Harpreet; Attri, Rajni

    2013-06-01

    The population of M. incognita, the root knot nematode (RKN) was found infesting five different host plants (okra, banana, sunflower, bottle gourd, and brinjal) out of 24 examined from four districts of Punjab, India (Gurdaspur, Ludhiana, Patiala, and Hoshiarpur). Morphological and morphometrical characterization indicated that in the case of mature female, the characters of body length and width, neck length, ratio 'a', anus to tail terminus (ATT), interphasmid distance (IPD), and perineal pattern were recorded as stable characters. These taxonomic characters can be reliable for identification. All characters of second-stage juvenile (J2) such as body length, stylet length, head to median bulb length (H-MB), distance from median bulb to excretory pore (MB-EP), tail length, anal body width (ABW), and ratios C and C' were highly variable. Analysis of interpopulation morphometric characters of mature female of M. incognita, namely, body length, width, and ratio 'a' were moderately variable characters (CV 0.26% to 20%) and stylet length, neck length, length of median bulb (LMB), and width of median bulb (WMB) were highly variable (CV 1.0% to 36.1%). In the perineal pattern, the two characters ATT and IPD were moderately variable (CV 8.8% to 17.6%) and two characters, anus to vulval slit (AVS) and length of vulval slit (LVS), were highly variable (CV 2.1% to 40.5%). In J2, body length, stylet length, H-MB, MB-EP, ABW, tail length, ratios C, and C' were highly variable characters (CV > 12%). PMID:23833327

  5. Doppler ultrasound and tibial tuberosity maturation status predicts pain in adolescent male athletes with Osgood-Schlatter's disease: a case series with comparison group and clinical interpretation

    PubMed Central

    Sailly, Matthieu; Whiteley, Rod; Johnson, Amanda

    2013-01-01

    Background The pathogenesis of the Osgood-Schlatter's disease (OSD) is still debated. The fragmentation of the ossification centre has been questioned as a definitive sign of OSD and has been seen as a normal development of the anterior tibial tubercle (ATT). Objectives It is unknown if such changes are present in the presumed pathological tendon insertion seen in OSD, nor the relation of Doppler-positive changes to pain on clinical examination. Methods A prospective analysis was carried out on 20 consecutive symptomatic male athletes (13.9 years±1.3) and a comparison group of asymptomatic subjects. All underwent a comparative clinical assessment and ultrasound with colour Doppler scan on both knees. Subjective pain was recorded with a visual analogue scale (VAS) during provocative manoeuvres: palpation, resisted contraction and single leg squat. Results Positive Doppler US (within the distal end of the patellar tendon) was associated with higher pain on palpation (47±24.5 vs 18±11.4, p<0.01) and resisted static contraction (59±20.2 vs 27±12.5, p<0.001) compared with Doppler-negative subjects. No Doppler activity was found in the comparison group. VAS for palpation and resisted contraction of the athletes graded as stage 2 (51.1±22.0 and 60.0±21.2) were significantly higher than stage 3 (17.8±12.0 and 18.9±16.9) and stage 4 (15.0±7.1 and 25.0±7.1; p<0.01). Conclusions More painful OSD is associated with the presence of neo-vessels. This may be linked with a particular stage of ATT maturation and applied compressive forces. A Doppler ultrasound scan adds practical information to develop the care plan of the patient. PMID:22952406

  6. Age-related differences when walking downhill on different sloped terrains.

    PubMed

    Scaglioni-Solano, Pietro; Aragón-Vargas, Luis Fernando

    2015-01-01

    Despite the common situation of walking on different sloped terrains, previous work on gait has focused on level terrain. This study aims to assess whether any age-related differences exist in spatiotemporal and stability parameters when walking downhill on three different sloped walkways. Two tri-axial accelerometers were used at the levels of head and pelvis to investigate spatiotemporal parameters, magnitude (root mean square, RMS), harmonic content of accelerations (harmonic ratios, HR) and attenuation between body levels (ATT) in 35 older adults (OA, 69 ± 4.5 y.o.) and 22 young adults (YA, 22.1 ± 1.9 y.o.). Older adults walked at the same speed and cadence as young adults in flat terrain (FL, 0%) and moderate hill (MH, 8%). In the highest slope (PH, 20%), older adults reduced speed and step length and both groups increased cadence. Age had no effect on attenuation and RMS profiles. RMS increased with slope in all directions at both head and pelvis, except, for medio-lateral direction (ML), with similar head RMS in all slopes. There is an important shift in ATT from anteroposterior direction (AP) to ML at the highest slope, resulting in smaller antero-posterior attenuation and greater medio-lateral attenuation. Age differences appeared in the smoothness (HR) at the flat terrain, with increased vertical and antero-posterior values for young adults. As slope increased, group differences disappeared and HR decreased for all directions of motion. In general, spatiotemporal adaptations to increased slope seem to be part of a mechanism to improve ML attenuation, in both young and old adults. PMID:25455702

  7. State Soil Geographic (STATSGO) Data Base for the Conterminous United States

    USGS Publications Warehouse

    Schwarz, Gregory E.; Alexander, R.B.

    1995-01-01

    USSOILS is an Arc 7.0 coverage containing hydrology-relevant information for 10,498 map units covering the entire conterminous United States. The coverage was compiled from individual State coverages contained in the October 1994 State Soil Geographic (STATSGO) Data Base produced on CD-ROM. The geo-dataset USSOILS.PAT relates (on the basis of a map unit identifier) the 10,498 map units to 78,518 polygons. The scale of the geo-dataset is 1:250,000. The INFO attribute table USSOILS.MUID_ATTS contains selected variables from the STATSGO data set for 10,501 map units (an extra 3 map units are contained in the attribute table that are not in the geo-dataset - see the 'Procedures' section below), including: the map unit identifier, a 2-character state abbreviation, available water capacity of the soil, percent clay in the soil, the actual k-factor used in the water erosion component of the universal soil loss equation, the organic material in soil, soil permeability, cumulative thickness of all soil layers, hydrologic characteristics of the soil, quality of drainage, surface slope, liquid limit of the soil, share of a map unit having hydric soils, and the annual frequency of flooding. To facilitate mapping the attribute data, the narrative section below contains instructions for transferring the information contained in the attribute table USSOILS.MUID_ATTS to the polygon attribute table USSOILS.PAT. STATSGO United States Soil Water Capacity Clay Organic material Permeability Infiltration Drainage Hydric Flood frequency Slope

  8. Assessment of treatment interruption among pulmonary tuberculosis patients: A cross-sectional study

    PubMed Central

    Gorityala, Satya Bhgath; Mateti, Uday Venkat; Konuru, Venkateswarlu; Martha, Srinivas

    2015-01-01

    Background: Multi-drug resistant tuberculosis (TB) is a rising peril of the TB control in India caused mostly by incomplete treatment. Aim: The aim was to assess the treatment interruption among pulmonary TB (PTB) patients. Materials and Methods: A cross-sectional study was carried out for a period of 9 months among PTB patients. Patients admitted with active pulmonary Koch's and history of anti-TB treatment (ATT) for 1-month or more from any source and who returns to treatment after not taking ATT consecutively for 2 months or more were included in the study. The data were collected from the patients or their caretakers to obtain the source of treatment given previously before default, number of treatment interruptions, phase and reasons for treatment interruption treatment. Results: A total of 107 defaulters were identified during the study period. In the present study, 62.6% of the patients interrupted treatment only once, 55.34% of the patient's early continuation (3–4 months) treatment, and 47.66% of the patient's only one reason for the treatment interruptions during the course of the treatment. The most common reason for the treatment interruptions were felt well with TB treatment (29.53%) followed by side effects (16.06%), lack of money (8.29%), and other reasons. Conclusion: The study revealed that most of the defaulters were in the age group between 35 and 60 years, male gender, illiterates, daily wage labor, and married. The treatment interruptions were minimized by putting the efforts to improve direct supervision; pretreatment counseling and retrieve treatment interrupters were recommended. PMID:26229358

  9. A novel, broad-range, CTXΦ-derived stable integrative expression vector for functional studies.

    PubMed

    Das, Bhabatosh; Kumari, Reena; Pant, Archana; Sen Gupta, Sourav; Saxena, Shruti; Mehta, Ojasvi; Nair, Gopinath Balakrish

    2014-12-01

    CTXΦ, a filamentous vibriophage encoding cholera toxin, uses a unique strategy for its lysogeny. The single-stranded phage genome forms intramolecular base-pairing interactions between two inversely oriented XerC and XerD binding sites (XBS) and generates a functional phage attachment site, attP(+), for integration. The attP(+) structure is recognized by the host-encoded tyrosine recombinases XerC and XerD (XerCD), which enables irreversible integration of CTXΦ into the chromosome dimer resolution site (dif) of Vibrio cholerae. The dif site and the XerCD recombinases are widely conserved in bacteria. We took advantage of these conserved attributes to develop a broad-host-range integrative expression vector that could irreversibly integrate into the host chromosome using XerCD recombinases without altering the function of any known open reading frame (ORF). In this study, we engineered two different arabinose-inducible expression vectors, pBD62 and pBD66, using XBS of CTXΦ. pBD62 replicates conditionally and integrates efficiently into the dif of the bacterial chromosome by site-specific recombination using host-encoded XerCD recombinases. The expression level of the gene of interest could be controlled through the PBAD promoter by modulating the functions of the vector-encoded transcriptional factor AraC. We validated the irreversible integration of pBD62 into a wide range of pathogenic and nonpathogenic bacteria, such as V. cholerae, Vibrio fluvialis, Vibrio parahaemolyticus, Escherichia coli, Salmonella enterica, and Klebsiella pneumoniae. Gene expression from the PBAD promoter of integrated vectors was confirmed in V. cholerae using the well-studied reporter genes mCherry, eGFP, and lacZ.

  10. Unified Model of Multiple Wind Turbines

    NASA Astrophysics Data System (ADS)

    Mutule, A.; Kochukov, O.

    2014-08-01

    An approach is proposed to the modelling of wind farms in the electric power system long-term planning. It allows a specialist to perform calculations based on scanty information and offers a set of ready-to-use data for easy, fast, and precise modelling. The authors exemplify the calculations of wind speed probability density and power curves and give an idea for relevant corrections. They also show how to pass from a single wind turbine model to the unified model of multiple wind turbines which would meet the requirements of long-term planning tasks. The paper presents the data on wind farms that are operating in UK and Oceania Rakstā ir apskatīta vēja elektrostaciju modelēšana ilgtermiņa attīstības plānošanas uzdevumos. Modelēšana tika veikta, izmantojot ierobežotu datu apjomu, kuri bija piejami lietotājam. Gatavie dati deva iespēju veikt ātru un precīzu modelēšanu. Raksts piedāva metodi kā pāriet no viena vēja ģeneratora modeli uz vēja elektrostaciju (vairāki vēja ģeneratori) modeli, kas atbilst ilgtermiņa attīstības plānošanas prasībām. Rakstā atspoguļoti dati no Okeānijas un Lielbritānijas eksistējošām vēja elektrostacijām

  11. Effects of RXR Agonists on Cell Proliferation/Apoptosis and ACTH Secretion/Pomc Expression

    PubMed Central

    Saito-Hakoda, Akiko; Uruno, Akira; Yokoyama, Atsushi; Shimizu, Kyoko; Parvin, Rehana; Kudo, Masataka; Saito-Ito, Takako; Sato, Ikuko; Kogure, Naotaka; Suzuki, Dai; Shimada, Hiroki; Yoshikawa, Takeo; Fujiwara, Ikuma; Kagechika, Hiroyuki; Iwasaki, Yasumasa; Kure, Shigeo; Ito, Sadayoshi; Sugawara, Akira

    2015-01-01

    Various retinoid X receptor (RXR) agonists have recently been developed, and some of them have shown anti-tumor effects both in vivo and in vitro. However, there has been no report showing the effects of RXR agonists on Cushing’s disease, which is caused by excessive ACTH secretion in a corticotroph tumor of the pituitary gland. Therefore, we examined the effects of synthetic RXR pan-agonists HX630 and PA024 on the proliferation, apoptosis, ACTH secretion, and pro-opiomelanocortin (Pomc) gene expression of murine pituitary corticotroph tumor AtT20 cells. We demonstrated that both RXR agonists induced apoptosis dose-dependently in AtT20 cells, and inhibited their proliferation at their higher doses. Microarray analysis identified a significant gene network associated with caspase 3 induced by high dose HX630. On the other hand, HX630, but not PA024, inhibited Pomc transcription, Pomc mRNA expression, and ACTH secretion dose-dependently. Furthermore, we provide new evidence that HX630 negatively regulates the Pomc promoter activity at the transcriptional level due to the suppression of the transcription factor Nur77 and Nurr1 mRNA expression and the reduction of Nur77/Nurr1 heterodimer recruiting to the Pomc promoter region. We also demonstrated that the HX630-mediated suppression of the Pomc gene expression was exerted via RXRα. Furthermore, HX630 inhibited tumor growth and decreased Pomc mRNA expression in corticotroph tumor cells in female nude mice in vivo. Thus, these results indicate that RXR agonists, especially HX630, could be a new therapeutic candidate for Cushing’s disease. PMID:26714014

  12. Online Community Use Predicts Abstinence in Combined Internet/Phone Intervention for Smoking Cessation

    PubMed Central

    Papandonatos, George D.; Erar, Bahar; Stanton, Cassandra A.; Graham, Amanda L.

    2016-01-01

    Objective To estimate the causal effects of online community use on 30-day point prevalence abstinence at 3 months among smokers randomized to combined Internet+Phone intervention for smoking cessation. Method Participants were N=399 adult smokers in the Internet+Phone arm of The iQUITT Study, a randomized trial of Internet and proactive telephone counseling for smoking cessation. All participants accessed a web-based smoking-cessation program with an established online community and received telephone counseling. Automated tracking metrics of passive (e.g., reading posts, viewing profiles) and active (e.g., writing posts, sending messages) community use were extracted at 3 months. Self-selected community use defines the groups of interest: None, Passive, and Both (passive+active). Inverse probability of treatment weighting corrected for baseline imbalances on demographic, smoking, and psychosocial variables. Propensity weights estimated via generalized boosted models were used to calculate Average Treatment Effects (ATE) and Average Treatment effects on the Treated (ATT). Results Patterns of community use were: None=145 (36.3%), Passive=82 (20.6%), and Both=172 (43.1%). ATE-weighted abstinence rates were: None=12.2% (95% CI=6.7–17.7); Passive=25.2% (95% CI=15.1–35.2); Both=35.5% (95% CI=28.1–42.9). ATT-weighted abstinence rates indicated even greater benefits of passive community use by non-users. Conclusions More than one third of participants who received telephone counseling and used the community both passively and actively achieved abstinence. Participation in an established online community as part of a combined Internet+phone intervention has the potential to promote short-term abstinence. Results also demonstrated that information and support that originate in the community can serve as a resource for all users. PMID:27100127

  13. Mycobacterium tuberculosis modulates the gene interactions to activate the HIV replication and faster disease progression in a co-infected host.

    PubMed

    Toor, Jaideep S; Singh, Sukhvinder; Sharma, Aman; Arora, Sunil K

    2014-01-01

    Understanding of the chronic immune activation, breakdown of immune defense and synergistic effect between HIV and Mycobacterium tuberculosis (Mtb) may provide essential information regarding key factors involved in the pathogenesis of HIV disease. In this study, we aimed to highlight a few of the immunological events that may influence and accelerate the progression of HIV disease in the presence of co-infecting Mtb. A cross-sectional study was performed on cohorts, including anti-tubercular therapy (ATT) naïve active pulmonary tuberculosis (PTB) patients, antiretroviral therapy (ART) naïve HIV-1 infected individuals at different stages of disease, ATT and ART naïve HIV-PTB co-infected individuals and healthy controls. A significantly higher T-regulatory cell (Treg) frequency coupled with the high FoxP3 expression in the CD4 T-cells indicated an immunosuppressive environment in the advance stage of HIV-1 infection. This is further substantiated by high HO-1 expression favoring TB co-infection. Functionally, this change in Treg frequency in HIV-1 infected individuals correlated well with suppression of T-cell proliferation. Mtb infection seems to facilitate the expansion of the Treg pool along with increased expression of FoxP3, specifically the variant-1, as evident from the data in HIV-1 co-infected as well as in patients with only PTB. A significantly lower expression of HO-1 in co-infected individuals compared to patients with only HIV-infection having comparable CD4 count correlated well with increased expression of CCR5 and CxCR4 as well as NF-κB and inflammatory cytokines IL-6 and TNF-α, which collectively may contribute to enhanced viral replication and increased cell death, hence faster disease progression in co-infected individuals.

  14. Clinical Correlates and Drug Resistance in HIV-Infected and -Uninfected Pulmonary Tuberculosis Patients in South India

    PubMed Central

    Sara, Chandy; Elsa, Heylen; Baijayanti, Mishra; Lennartsdotter, Ekstrand Maria

    2016-01-01

    Objectives To examine demographics, clinical correlates, sputum AFB (acid fast bacilli) smear grading DOTS (Directly Observed Therapy Short Course) uptake, and drug resistance in a cohort of newly-diagnosed, smear positive pulmonary tuberculosis (TB) patients with respect to HIV status at baseline, and compare smear conversion rates, side effects and mortality after two months. Design A prospective study among 54 HIV positive and 41 HIV negative pulmonary TB patients. Data were collected via face-to-face interviews, review of medical records, and lab tests. Results HIVTB co-infected patients, though more symptomatic at baseline, showed more improvement in their symptoms compared to HIV-uninfected TB patients at follow-up. The HIV co-infected group had more prevalent perceived side effects, and sputum smear positivity was marginally higher compared to the HIV negative group at follow-up. Mortality was higher among the HIV-infected group. Both groups had high rates of resistance to first-line anti-tubercular drugs, particularly isoniazid. There was no significant difference in the drug resistance patterns between the groups. Conclusions Prompt initiation and provision of daily regimens of ATT (Anti-Tubercular treatment) along with ART (Anti-Retroviral treatment) via ART centers is urgently needed in India. As resistance to ART and/or ATT is directly linked to medication non-adherence, the use of counseling, regular reinforcement, early detection and appropriate intervention strategies to tackle this complex issue could help prevent premature mortality and development of resistance in HIV-TB co-infected patients. The high rate of isoniazid resistance might preclude its use in India as prophylaxis for latent TB in HIV infected persons as per the World Health Organization (WHO) guideline.

  15. Attrition of TCR Vα7.2+ CD161++ MAIT Cells in HIV-Tuberculosis Co-Infection Is Associated with Elevated Levels of PD-1 Expression

    PubMed Central

    Saeidi, Alireza; Tien Tien, Vicky L.; Al-Batran, Rami; Al-Darraji, Haider A.; Tan, Hong Y.; Yong, Yean K.; Ponnampalavanar, Sasheela; Barathan, Muttiah; Rukumani, Devi V.; Ansari, Abdul W.; Velu, Vijayakumar; Kamarulzaman, Adeeba; Larsson, Marie; Shankar, Esaki M.

    2015-01-01

    Mucosal-associated invariant T (MAIT) cells are evolutionarily conserved antimicrobial MR1-restricted CD8+ T cells co-expressing the semi-invariant TCR Vα7.2, and are numerous in the blood and mucosal tissues of humans. MAIT cells appear to undergo exhaustion in chronic viral infections. However, their role in human immunodeficiency virus type 1 (HIV-1) mono-infection and HIV/tuberculosis (TB) co-infection have seldom been elaborately investigated. We conducted a cross-sectional study to investigate the frequencies and phenotypes of CD161++CD8+ T cells among anti-retroviral therapy (ART)/anti-TB therapy (ATT) treatment-naïve HIV/TB co-infected, ART/TB treated HIV/TB co-infected, ART naïve HIV-infected, ART-treated HIV-infected patients, and HIV negative healthy controls (HCs) by flow cytometry. Our data revealed that the frequency of MAIT cells was severely depleted in HIV mono- and HIV/TB co-infections. Further, PD-1 expression on MAIT cells was significantly increased in HIV mono- and HIV-TB co-infected patients. The frequency of MAIT cells did not show any significant increase despite the initiation of ART and/or ATT. Majority of the MAIT cells in HCs showed a significant increase in CCR6 expression as compared to HIV/TB co-infections. No marked difference was seen with expressions of chemokine co-receptor CCR5 and CD103 among the study groups. Decrease of CCR6 expression appears to explain why HIV-infected patients display weakened mucosal immune responses. PMID:25894562

  16. Telemedicine: the slow revolution.

    PubMed

    Moncrief, Jack W

    2014-01-01

    The use of interactive video has been recognized as a means of delivering medical support to isolated areas since the 1950s. The Department of Defense recognized early the capacity of telemedicine to deliver medical care and support to front-line military personnel. In 1989, the Texas Telemedicine Project received grants and support from the then American Telephone and Telegraph Company (now AT&T) and the Meadows Foundation of Dallas, Texas, to establish and evaluate telemedicine delivery in central Texas. That project had 6 connected telemedicine sites: 3 in Austin, Texas, and 3 in Giddings, Texas (a small community 55 miles to the southeast of Austin). The sites in Giddings included a chronic outpatient dialysis facility, an inpatient psychiatric hospital, and the emergency department at Giddings Hospital. Patient contact began in April 1991 and continued through March 1993. During that period, data on the 1500 patient contacts made were recorded. After termination of the Texas Telemedicine Project, AT&T continued to provide the transmission lines, and between 1993 and 1996, another 12,000 patient contacts were made. Approximately 80% were dialysis evaluations and 20% were non-dialysis primary care contacts. The original cost of materials and equipment in the Texas Telemedicine Project exceeded $50,000 per site. Today, a secure Internet connection with full-motion video and wireless data transfer to almost any location in the world is achievable with an iPad. Multiple inexpensive applications with connections for electrocardiogram, otoscope, and stethoscope, among others, make this technology extremely inexpensive and user-friendly. The revolution now is rapidly moving forward, with Medicare reimbursing telemedicine contacts in medically underserved areas. Multiple bills are before Congress to expand Medicare and therefore private insurance payment for this service.

  17. phiC31 integrase-mediated site-specific recombination in barley.

    PubMed

    Kapusi, Eszter; Kempe, Katja; Rubtsova, Myroslava; Kumlehn, Jochen; Gils, Mario

    2012-01-01

    The Streptomyces phage phiC31 integrase was tested for its feasibility in excising transgenes from the barley genome through site-specific recombination. We produced transgenic barley plants expressing an active phiC31 integrase and crossed them with transgenic barley plants carrying a target locus for recombination. The target sequence involves a reporter gene encoding green fluorescent protein (GFP), which is flanked by the attB and attP recognition sites for the phiC31 integrase. This sequence disruptively separates a gusA coding sequence from an upstream rice actin promoter. We succeeded in producing site-specific recombination events in the hybrid progeny of 11 independent barley plants carrying the above target sequence after crossing with plants carrying a phiC31 expression cassette. Some of the hybrids displayed fully executed recombination. Excision of the GFP gene fostered activation of the gusA gene, as visualized in tissue of hybrid plants by histochemical staining. The recombinant loci were detected in progeny of selfed F(1), even in individuals lacking the phiC31 transgene, which provides evidence of stability and generative transmission of the recombination events. In several plants that displayed incomplete recombination, extrachromosomal excision circles were identified. Besides the technical advance achieved in this study, the generated phiC31 integrase-expressing barley plants provide foundational stock material for use in future approaches to barley genetic improvement, such as the production of marker-free transgenic plants or switching transgene activity.

  18. Delivering dispersion-managed soliton and Q-switched pulse in fiber laser based on graphene and nonlinear optical loop mirror

    NASA Astrophysics Data System (ADS)

    Wang, W. B.; Wang, F.; Yu, Q.; Zhang, X.; Lu, Y. X.; Gu, J.

    2016-11-01

    We propose and experimentally demonstrate a bidirectional erbium-doped fiber laser delivering dispersion-managed soliton (DMS) and Q-switched pulse based on a graphene-polyvinyl alcohol (PVA) and nonlinear optical loop mirror (NOLM) saturable absorbers (SAs). In proposed structure, the DMS is achieved in clockwise (CW) direction and Q-switched pulse is obtained in counter-clockwise (CCW) direction. By properly adjusting the intracavity attenuators (ATT) and polarizer controllers (PCs), DMS in the CW direction and Q-switched pulse in the CCW direction can be obtained, respectively or simultaneously. The DMS with full width at half maximum (FWHM) of ~480 fs, signal to noise ratio (SNR) of ~60 dB and repetition frequency about 3.907 MHz is obtained. The Q-switched pulse is established at a pump power of 180 mW with a repetition rate of ~43.5 kHz and FWHM of ~8.18 μs. When the pump power is increased to 700 mW, Q-switched pulse with a repetition rate of ~107.1 kHz and FWHM of ~2.15 μs is generated. When the two type pulses are formed simultaneously, the maximum repetition rate of Q-switched pulse is 55.8 kHz and minimum FWHM is 2.81 μs, the DMS can be formed by properly adjusting PC and ATT in this case. To the best of our knowledge, it is the first time that Q-switched pulse and DMS have been acquired respectively or simultaneously in a fiber laser.

  19. An on-line high-performance liquid chromatography-diode-array detector-multi-stage mass spectrometry-deoxyribonucleic acid-4',6-diamidino-2-phenylindole-fluorescence detector system for screening the DNA-binding active compounds in Fufang Banbianlian Injection.

    PubMed

    Li, Sensen; Jiang, Haixiu; Lin, Zongtao; Deng, Shanshan; Guan, Yanqing; Wang, Hong; Chen, Shizhong

    2015-12-11

    Fufang Banbianlian Injection (FBI), a well-known traditional Chinese medicine formula, has been recently approved and extensively used as a newly anti-inflammatory and anti-tumor drug. This prescription comprises an equal ratio of three traditional Chinese herbs, Lobelia chinensis Lour, Scutellaria barbata D. Don and Hedyotis diffusa Willd. The relationships between its chemical compositions and activities have not been understood well yet. To investigate the ingredients and their DNA-binding activities in FBI, an on-line high-performance liquid chromatography-diode-array detector-multi-stage mass spectrometry-deoxyribonucleic acid-4',6-diamidino-2-phenylindole-fluorescence detector (HPLC-DAD-MS(n)-DNA-DAPI-FLD) system was developed using a combination of chromatographic, mass spectrometric and fluorescent detection techniques. 4',6-Diamidino-2-phenylindole (DAPI) specifically binds to three ATT base pairs on the DNA minor groove, and thus can be used as a fluorescent probe for screening active compounds that compete ATT sequences with DAPI. Using this system, 21 of 58 identified or tentatively characterized compounds in FBI showed DNA-binding activities, with most of the active compounds being flavone glycosides. In addition, the structure-activity relationships of these active compounds suggested that conjugated planar structures are favorable for DNA-binding activities, and adjacent hydroxyl groups in flavonoids can significantly improve their activities. This is, to the best of our knowledge, the first application of DAPI as a fluorescent probe for the screening of DNA-binding active compounds in complex samples. PMID:26592560

  20. Prognostic impact of coronary microcirculation abnormalities in systemic sclerosis: a prospective study to evaluate the role of non-invasive tests

    PubMed Central

    2013-01-01

    Introduction Microcirculation dysfunction is a typical feature of systemic sclerosis (SSc) and represents the earliest abnormality of primary myocardial involvement. We assessed coronary microcirculation status by combining two functional tests in SSc patients and estimating its impact on disease outcome. Methods Forty-one SSc patients, asymptomatic for coronary artery disease, were tested for coronary flow velocity reserve (CFR) by transthoracic-echo-Doppler with adenosine infusion (A-TTE) and for left ventricular wall motion abnormalities (WMA) by dobutamine stress echocardiography (DSE). Myocardial multi-detector computed tomography (MDCT) enabled the presence of epicardial stenosis, which could interfere with the accuracy of the tests, to be excluded. Patient survival rate was assessed over a 6.7- ± 3.5-year follow-up. Results Nineteen out of 41 (46%) SSc patients had a reduced CFR (≤2.5) and in 16/41 (39%) a WMA was observed during DSE. Furthermore, 13/41 (32%) patients showed pathological CFR and WMA. An inverse correlation between wall motion score index (WMSI) during DSE and CFR value (r = -0.57, P <0.0001) was observed; in addition, CFR was significantly reduced (2.21 ± 0.38) in patients with WMA as compared to those without (2.94 ± 0.60) (P <0.0001). In 12 patients with abnormal DSE, MDCT was used to exclude macrovasculopathy. During a 6.7- ± 3.5-year follow-up seven patients with abnormal coronary functional tests died of disease-related causes, compared to only one patient with normal tests. Conclusions A-TTE and DSE tests are useful tools to detect non-invasively pre-clinical microcirculation abnormalities in SSc patients; moreover, abnormal CFR and WMA might be related to a worse disease outcome suggesting a prognostic value of these tests, similar to other myocardial diseases. PMID:23302110

  1. Intergeneric conjugation in Streptomyces peucetius and Streptomyces sp. strain C5: chromosomal integration and expression of recombinant plasmids carrying the chiC gene.

    PubMed

    Paranthaman, Senthamaraikannan; Dharmalingam, Kuppamuthu

    2003-01-01

    Intergeneric conjugal transfer of plasmid DNA from Escherichia coli to Streptomyces circumvents problems such as host-controlled restriction and instability of foreign DNA during the transformation of Streptomyces protoplasts. The anthracycline antibiotic-producing strains Streptomyces peucetius and Streptomyces sp. strain C5 were transformed using E. coli ET12567(pUZ8002) as a conjugal donor. When this donor species, carrying pSET152, was mated with Streptomyces strains, the resident plasmid was mobilized to the recipient and the transferred DNA was also integrated into the recipient chromosome. Analysis of the exconjugants showed stable integration of the plasmid at a single chromosomal site (attB) of the Streptomyces genome. The DNA sequence of the chromosomal integration site was determined and shown to be conserved. However, the core sequence, where the crossover presumably occurred in C5 and S. peucetius, is TTC. These results also showed that the phiC31 integrative recombination is active and the phage attP site is functional in S. peucetius as well as in C5. The efficiency and specificity of phiC31-mediated site-specific integration of the plasmid in the presence of a 3.7-kb homologous DNA sequence indicates that integrative recombination is preferred under these conditions. The integration of plasmid DNA did not affect antibiotic biosynthesis or biosynthesis of essential amino acids. Integration of a single copy of a mutant chiC into the wild-type S. peucetius chromosome led to the production of 30-fold more chitinase.

  2. Characterization of the integrase of NBU1, a Bacteroides mobilizable transposon.

    PubMed

    Rajeev, Lara; Salyers, Abigail A; Gardner, Jeffrey F

    2006-08-01

    NBU1 is a Bacteroides mobilizable transposon (MTn) that is integrated within the host chromosome and requires CTnDOT functions for its excision and transfer into a new host. The NBU1 integrase IntN1 has been classified as a tyrosine recombinase based on the presence of conserved residues. We created alanine mutants of the residues R291, K314, H393, R396, H419 and the conserved substitution Y429F and tested them for integration efficiency. The results suggest that these residues in IntN1 are important for integration, and Y429 could be the catalytic nucleophile. We employed suicide substrates and partially purified IntN1 to determine the positions of IntN1 cleavage within the 14 bp common core region that is identical in both NBU1 att sites. We show that IntN1 makes 7 bp staggered cuts on the top and bottom strands. From previous mutational analysis of the att sites, we show that two specific mutations near the site of bottom strand cleavage within this 7 bp region increased integration, and mutations of the two bases near top strand cleavage site had no effect on integration. These results indicate that IntN1 lacks the strict requirement for homology between the recombining sites seen with other tyrosine recombinases. We also show that phosphorothioate substitutions at the cleavage site and 1 bp downstream inhibited cleavage by IntN1. This differs from other studied tyrosine recombinases where inhibition occurs by substitutions at the cleavage site only.

  3. Rapid Optimization of Engineered Metabolic Pathways with Serine Integrase Recombinational Assembly (SIRA).

    PubMed

    Merrick, C A; Wardrope, C; Paget, J E; Colloms, S D; Rosser, S J

    2016-01-01

    Metabolic pathway engineering in microbial hosts for heterologous biosynthesis of commodity compounds and fine chemicals offers a cheaper, greener, and more reliable method of production than does chemical synthesis. However, engineering metabolic pathways within a microbe is a complicated process: levels of gene expression, protein stability, enzyme activity, and metabolic flux must be balanced for high productivity without compromising host cell viability. A major rate-limiting step in engineering microbes for optimum biosynthesis of a target compound is DNA assembly, as current methods can be cumbersome and costly. Serine integrase recombinational assembly (SIRA) is a rapid DNA assembly method that utilizes serine integrases, and is particularly applicable to rapid optimization of engineered metabolic pathways. Using six pairs of orthogonal attP and attB sites with different central dinucleotide sequences that follow SIRA design principles, we have demonstrated that ΦC31 integrase can be used to (1) insert a single piece of DNA into a substrate plasmid; (2) assemble three, four, and five DNA parts encoding the enzymes for functional metabolic pathways in a one-pot reaction; (3) generate combinatorial libraries of metabolic pathway constructs with varied ribosome binding site strengths or gene orders in a one-pot reaction; and (4) replace and add DNA parts within a construct through targeted postassembly modification. We explain the mechanism of SIRA and the principles behind designing a SIRA reaction. We also provide protocols for making SIRA reaction components and practical methods for applying SIRA to rapid optimization of metabolic pathways. PMID:27417934

  4. Rifampicin and anti-hypertensive drugs in chronic kidney disease: Pharmacokinetic interactions and their clinical impact

    PubMed Central

    Agrawal, A.; Agarwal, S. K.; Kaleekal, T.; Gupta, Y. K.

    2016-01-01

    Patients on dialysis have an increased incidence of tuberculosis (TB). Rifampicin, a first-line antitubercular therapy (ATT) drug, is a potent inducer of hepatic cytochrome P450 (CYP). There is potential for pharmacokinetic interaction between rifampicin and anti-hypertensives that are CYP substrates: amlodipine and metoprolol. Therefore, hypertensive patients receiving rifampicin-based ATT are at risk for worsening of hypertension. However, this hypothesis has not yet been systematically studied. In this prospective study, hypertensive CKD 5D patients with TB were followed after rifampicin initiation. Blood pressure (BP) was ≤140/90 mmHg with stable anti-HT requirement at inclusion. Serum amlodipine, metoprolol, and prazosin levels were estimated by high-performance liquid chromatography at baseline and 3, 7, 10, and 14 days after rifampicin initiation. BP and anti-HT requirement were monitored for 2 weeks or until stabilization. All 24 patients in the study had worsening of hypertension after rifampicin and 83.3% required increase in drugs to maintain BP <140/90 mmHg. Serial amlodipine levels were estimated in 16 patients; metoprolol and prazosin in four patients each. Drug levels declined by >50% in all patients and became undetectable in 50-75%. Drug requirement increased from 4.5 ± 3.6 to 8.5 ± 6.4 units (P < 0.0001). Mean time to first increase in dose was 6.5 ± 3.6 days. Eleven (46%) patients experienced a hypertensive crisis at 9.1 ± 3.8 days. Three of them had a hypertensive emergency with acute pulmonary edema. In two patients, rifampicin had to be discontinued to achieve BP control. In conclusion, rifampicin caused a significant decrease in blood levels of commonly used anti hypertensives. This decrease in levels correlated well with worsening of hypertension. Thus, we suggest very close BP monitoring in CKD patients after rifampicin initiation. PMID:27795624

  5. Clinical Correlates and Drug Resistance in HIV-Infected and -Uninfected Pulmonary Tuberculosis Patients in South India

    PubMed Central

    Sara, Chandy; Elsa, Heylen; Baijayanti, Mishra; Lennartsdotter, Ekstrand Maria

    2016-01-01

    Objectives To examine demographics, clinical correlates, sputum AFB (acid fast bacilli) smear grading DOTS (Directly Observed Therapy Short Course) uptake, and drug resistance in a cohort of newly-diagnosed, smear positive pulmonary tuberculosis (TB) patients with respect to HIV status at baseline, and compare smear conversion rates, side effects and mortality after two months. Design A prospective study among 54 HIV positive and 41 HIV negative pulmonary TB patients. Data were collected via face-to-face interviews, review of medical records, and lab tests. Results HIVTB co-infected patients, though more symptomatic at baseline, showed more improvement in their symptoms compared to HIV-uninfected TB patients at follow-up. The HIV co-infected group had more prevalent perceived side effects, and sputum smear positivity was marginally higher compared to the HIV negative group at follow-up. Mortality was higher among the HIV-infected group. Both groups had high rates of resistance to first-line anti-tubercular drugs, particularly isoniazid. There was no significant difference in the drug resistance patterns between the groups. Conclusions Prompt initiation and provision of daily regimens of ATT (Anti-Tubercular treatment) along with ART (Anti-Retroviral treatment) via ART centers is urgently needed in India. As resistance to ART and/or ATT is directly linked to medication non-adherence, the use of counseling, regular reinforcement, early detection and appropriate intervention strategies to tackle this complex issue could help prevent premature mortality and development of resistance in HIV-TB co-infected patients. The high rate of isoniazid resistance might preclude its use in India as prophylaxis for latent TB in HIV infected persons as per the World Health Organization (WHO) guideline. PMID:27708985

  6. Bolus-tracking arterial spin labelling: theoretical and experimental results

    NASA Astrophysics Data System (ADS)

    Kelly, M. E.; Blau, C. W.; Kerskens, C. M.

    2009-03-01

    Arterial spin labelling (ASL) is a magnetic resonance imaging (MRI) technique that can be used to provide a quantitative assessment of cerebral perfusion. Despite the development of a number of theoretical models to facilitate quantitative ASL, some key challenges still remain. The purpose of this study is to develop a novel quantitative ASL method based on a macroscopic model that reduces the number of variables required to describe the physiological processes involved. To this end, a novel Fokker-Planck equation consisting of stochastically varying macroscopic variables was derived from a general Langevin equation. ASL data from the rat brain was acquired using a bolus-tracking ASL protocol where a bolus of labelled spins flowing from an inversion plane in the neck into an imaging plane in the brain can be observed. Bolus durations of 1.5 s, 2.0 s and 3.0 s were used and the solution to the Fokker-Planck equation for the boundary conditions of bolus-tracking ASL was fitted to the experimental data using a least-squares fit. The mean transit time (MTT) and capillary transit time (CTT) were calculated from the first and second moments of the resultant curve respectively and the arterial transit time (ATT) was calculated by subtracting the CTT from the MTT. The average MTT, CTT and ATT values were 1.75 ± 0.22 s, 1.43 ± 0.12 s and 0.32 ± 0.04 s respectively. In conclusion, a new ASL protocol has been developed by combining the theoretical model with ASL experiments. The technique has the unique ability to provide solutions for varying bolus volumes and the generality of the new model is demonstrated by the derivation of additional solutions for the continuous and pulsed ASL (CASL and PASL) techniques.

  7. Advanced Technology Lunar Telescopes I. Overview and Progress Report On Ultra-Lightweight Optics

    NASA Astrophysics Data System (ADS)

    Chen, P. C.; Pitts, R. E.; Oliversen, R. J.; Stolarik, J. D.; Segal, K.; Wilson, T. L.; Lin, E. I.; Hull, J. R.; Romeo, R.; Hojaji, H.; Ma, K. B.; Chen, Q. Y.; Chu, W. K.; Chu, C. W.

    1993-12-01

    The materials and technology already exist to build fully functional steerable telescopes for use on the moon, telescopes that are cost effective, that can be deployed using existing launchers, and that can function for extended periods without human maintenance. We describe our concept of advanced technology telescopes (ATT) which combines the elements of i) ultra-lightweight precision optics and structures, ii) non-contact, electronically controlled superconductor bearings and drive mechanisms, and iii) high dynamic range radiation resistant sensors. Unlike previous transit telescope designs, the ATTs can point and track objects anywhere in the sky over the entire lunar night (or day), can be deployed in multiple unit arrays, and can be equipped with standard astronomical instruments including spectrographs, imagers, or even interferometers. We first describe the optics. Lightweight optics are crucial because they minimize the mass of the telescope assembly and its support structure and ultimately the entire payload. By using materials and fabrication technology similar to that already refined by ESA and proven for space applications, we show that it is possible to produce precision optical elements of very low areal density (< 2 kg per sq. m). The process also has much lower per unit cost compared to traditional mirror fabrication techniques. By supporting the optical elements with a class of very lightweight but stiff material already developed by NASA, a telescope assembly can be made that has essentially the minimum possible mass. Such ultra-lightweight construction makes possible astronomical payloads that can be sent to the moon using existing small and medium size rockets. The very low per unit cost permits the production and deployment of multiple units, thereby increasing the versatility and productivity of a lunar observatory while providing good redundancy. We demonstrate a proof-of-concept optical telescope assembly that has a 31 cm diameter primary

  8. The effects of mountain bike suspension systems on energy expenditure, physical exertion, and time trial performance during mountain bicycling.

    PubMed

    Seifert, J G; Luetkemeier, M J; Spencer, M K; Miller, D; Burke, E R

    1997-04-01

    The purpose of this 3-Phase study was to investigate the effects of suspension systems on muscular stress, energy expenditure, and time trial performance during mountain biking. Three suspension systems were tested, a rigid frame bike (RIG), a suspension fork bike (FS), and a front and rear suspension bike (FSR). Phase I and II consisted of cycling at 16.1 km.hr-1 over a flat, bumpy course for 63 min. Phase III consisted of ascending (ATT), descending (DTT), and cross country (XTT) time trials. Phase I assessed muscular stress by 24 h change in CK, Phase II assessed HR, VO2, VE, and Phase III assessed performance responses to the suspension systems. The 24 hr change in CK was greater for RIG than FS and FSR (+91.9 +/- 79.5 IU vs +8.6 +/- 17.5 IU and +9.7 +/- 21.8 IU). Mean HR was greater for RIG than FS and FSR (153.7 +/- 15.6 bpm vs 146.7 +/- 15.4 bpm, 146.3 +/- 16.2 bpm). Subjects rode significantly faster on FS than FSR and RIG during the XTT (30.9 +/- 2.0 min vs 32.3 +/- 3.6 min, 32.3 +/- 3.2 min). Subjects RPE was lower for FSR than FS and RIG, however, no differences were observed for VO2, VE, ATT, or DTT. Cyclists incurred less muscular stress, indicated by CK and HR, when riding the FS and FSR. Although the FS and FSR weigh from 0.7 to 2.2 kg more than RIG, no differences were observed for energy expenditure and that riding the FS in a XTT resulted in a faster finishing time than FSR or RIG.

  9. Telemedicine: the slow revolution.

    PubMed

    Moncrief, Jack W

    2014-01-01

    The use of interactive video has been recognized as a means of delivering medical support to isolated areas since the 1950s. The Department of Defense recognized early the capacity of telemedicine to deliver medical care and support to front-line military personnel. In 1989, the Texas Telemedicine Project received grants and support from the then American Telephone and Telegraph Company (now AT&T) and the Meadows Foundation of Dallas, Texas, to establish and evaluate telemedicine delivery in central Texas. That project had 6 connected telemedicine sites: 3 in Austin, Texas, and 3 in Giddings, Texas (a small community 55 miles to the southeast of Austin). The sites in Giddings included a chronic outpatient dialysis facility, an inpatient psychiatric hospital, and the emergency department at Giddings Hospital. Patient contact began in April 1991 and continued through March 1993. During that period, data on the 1500 patient contacts made were recorded. After termination of the Texas Telemedicine Project, AT&T continued to provide the transmission lines, and between 1993 and 1996, another 12,000 patient contacts were made. Approximately 80% were dialysis evaluations and 20% were non-dialysis primary care contacts. The original cost of materials and equipment in the Texas Telemedicine Project exceeded $50,000 per site. Today, a secure Internet connection with full-motion video and wireless data transfer to almost any location in the world is achievable with an iPad. Multiple inexpensive applications with connections for electrocardiogram, otoscope, and stethoscope, among others, make this technology extremely inexpensive and user-friendly. The revolution now is rapidly moving forward, with Medicare reimbursing telemedicine contacts in medically underserved areas. Multiple bills are before Congress to expand Medicare and therefore private insurance payment for this service. PMID:25338434

  10. Effect of a Primary Care based Brief Intervention Trial among Risky Drug Users on Health-related Quality of Life

    PubMed Central

    Baumeister, Sebastian E.; Gelberg, Lillian; Leake, Barbara D.; Yacenda-Murphy, Julia; Vahidi, Mani; Andersen, Ronald M.

    2014-01-01

    Background Improvement in quality of life (QOL) is a long term goal of drug treatment. Although some brief interventions have been found to reduce illicit drug use, no trial among adult risky (moderate non-dependent) drug users has tested effects on health-related quality of life. Methods A single-blind randomized controlled trial of patients enrolled from February 2011 to November 2012 was conducted in waiting rooms of five federally qualified health centers. 413 adult primary care patients were identified as risky drug users using the WHO-ASSIST and 334 (81% response; 171 intervention, 163 control) consented to participate in the trial. Three-month follow-ups were completed by 261 patients (78%). Intervention patients received the QUIT intervention of brief clinician advice and up to two drug-use health telephone sessions. The control group received usual care and information on cancer screening. Outcomes were three-month changes in the Short Form Health Survey (SF-12) mental health component summary score (MCS) and physical health component summary score (PCS). Results The average treatment effect (ATE) was non-significant for MCS (0.2 points, p-value=0.87) and marginally significant for PCS (1.7 points, p-value=0.08). The average treatment effect on the treated (ATT) was 0.1 (p-value=0.93) for MCS and 1.9 (p-value=0.056) for PCS. The effect on PCS was stronger at higher (above median) baseline number of drug use days: ATE=2.7, p-value=0.04; ATT=3.21, p-value=0.02. Conclusions The trial found a marginally significant effect on improvement in PCS, and significant and stronger effect on the SF-12 physical component among patients with greater frequency of initial drug use. PMID:25042213

  11. Use of an Online Smoking Cessation Community Promotes Abstinence: Results of Propensity Score Weighting

    PubMed Central

    Graham, Amanda L.; Papandonatos, George D.; Erar, Bahar; Stanton, Cassandra A.

    2015-01-01

    Objective To estimate the causal effects of use of an online smoking cessation community on 30-day point prevalence abstinence at 3 months. Methods Participants were N=492 adult current smokers in the enhanced Internet arm of The iQUITT Study, a randomized trial of Internet and telephone treatment for smoking cessation. All participants accessed a web-based smoking-cessation program that included a large, established online community. Automated tracking metrics of passive (e.g., reading forum posts, viewing member profiles) and active (e.g., writing forum posts, sending private messages) community use were extracted from the site at 3 months. Self-selected community use defines the groups of interest: “None”, “Passive”, and “Both” (passive+active). Inverse probability of treatment weighting corrected for baseline imbalances on demographic, smoking, psychosocial, and medical history variables. Propensity weights estimated via generalized boosted models were used to calculate Average Treatment Effects (ATE) and Average Treatment effects on the Treated (ATT). Results Patterns of community use were: None=198 (40.2%), Passive=110 (22.4%), and Both=184 (37.4%). ATE-weighted abstinence rates were: None=4.2% (95% CI=1.5–6.9); Passive=15.1% (95% CI=8.4–21.9); Both=20.4% (95% CI=13.9–26.8). ATT-weighted abstinence rates indicated even greater benefits of community use. Conclusions Community users were more likely to quit smoking at 3 months than nonusers. The estimated benefit from use of online community resources was even larger among subjects with high propensity to use them. No differences in abstinence emerged between passive and passive/active users. Results suggest that lurking in online communities confers specific abstinence benefits. Implications of these findings for online cessation communities are discussed. PMID:26651470

  12. Genomic characterization of Ralstonia solanacearum phage phiRSA1 and its related prophage (phiRSX) in strain GMI1000.

    PubMed

    Fujiwara, Akiko; Kawasaki, Takeru; Usami, Shoji; Fujie, Makoto; Yamada, Takashi

    2008-01-01

    PhiRSA1 is a wide-host-range bacteriophage isolated from Ralstonia solanacearum. In this study, the complete nucleotide sequence of the phiRSA1 genomic DNA was determined. The genome was 38,760 bp of double-stranded DNA (65.3% G+C) with 19-bp 5'-extruding cohesive ends (cos) and contained 51 open reading frames (ORFs). Two-thirds of the phiRSA1 genomic region encodes the phage structural modules, and they are very similar to those reported for coliphage P2 and P2-like phages. A phiRSA1 minireplicon with an 8.2-kbp early-expressing region was constructed. A late-expression promoter sequence motif was predicted for these phiRSA1 genes as 5' TGTTGT-(X)13-ACAACA. The genomic sequence similarity between phiRSA1 and related phages phi52237 and phiCTX was interrupted by three AT islands, one of which contained an insertion sequence element, suggesting that they were recombinational hot spots. phiRSA1 was found to be integrated into at least three different strains of R. solanacearum, and the chromosomal integration site (attB) was identified as the 3' portion of the arginine tRNA(CCG) gene. In the light of the phiRSA1 gene arrangement, one possible prophage sequence previously detected on the chromosome of R. solanacearum strain GMI1000 was characterized as a phiRSA1-related prophage (designated phiRSX). phiRSX was found to be integrated at the serine tRNA (GGA) gene as an att site, and its size was determined to be 40,713 bp. phiRSX ORFs shared very high amino acid identity with their phiRSA1 counterparts. The relationships and evolution of these P2-like phages are discussed.

  13. Tight Regulation of the intS Gene of the KplE1 Prophage: A New Paradigm for Integrase Gene Regulation

    PubMed Central

    Panis, Gaël; Duverger, Yohann; Courvoisier-Dezord, Elise; Champ, Stéphanie; Talla, Emmanuel; Ansaldi, Mireille

    2010-01-01

    Temperate phages have the ability to maintain their genome in their host, a process called lysogeny. For most, passive replication of the phage genome relies on integration into the host's chromosome and becoming a prophage. Prophages remain silent in the absence of stress and replicate passively within their host genome. However, when stressful conditions occur, a prophage excises itself and resumes the viral cycle. Integration and excision of phage genomes are mediated by regulated site-specific recombination catalyzed by tyrosine and serine recombinases. In the KplE1 prophage, site-specific recombination is mediated by the IntS integrase and the TorI recombination directionality factor (RDF). We previously described a sub-family of temperate phages that is characterized by an unusual organization of the recombination module. Consequently, the attL recombination region overlaps with the integrase promoter, and the integrase and RDF genes do not share a common activated promoter upon lytic induction as in the lambda prophage. In this study, we show that the intS gene is tightly regulated by its own product as well as by the TorI RDF protein. In silico analysis revealed that overlap of the attL region with the integrase promoter is widely encountered in prophages present in prokaryotic genomes, suggesting a general occurrence of negatively autoregulated integrase genes. The prediction that these integrase genes are negatively autoregulated was biologically assessed by studying the regulation of several integrase genes from two different Escherichia coli strains. Our results suggest that the majority of tRNA-associated integrase genes in prokaryotic genomes could be autoregulated and that this might be correlated with the recombination efficiency as in KplE1. The consequences of this unprecedented regulation for excisive recombination are discussed. PMID:20949106

  14. Further Studies on the Immunogenicity of Haemophilus influenzae Type b and Pneumococcal Type 6A Polysaccharide-Protein Conjugates

    PubMed Central

    Chu, Chiayung; Schneerson, Rachel; Robbins, John B.; Rastogi, Suresh C.

    1983-01-01

    Conjugates were prepared by carbodiimide-mediated coupling of adipic acid hydrazide derivatives of Haemophilus influenzae type b (Hib), Escherichia coli K100, and pneumococcal 6A (Pn6A) polysaccharides with tetanus toxoid (TT), as an example of a “useful” carrier, and horseshoe crab hemocyanin (HCH), as an example of a “nonsense” carrier. These conjugates were injected into NIH mice, and their serum antibody responses to the polysaccharides and proteins were characterized. As originally reported, Hib conjugates increased the immunogenicity of the capsular polysaccharide and elicited greater than the estimated protective levels of anti-Hib antibodies in most recipients after one injection and in all after the third injection (Schneerson et al., J. Exp. Med. 152:361-376, 1980). Both Hib conjugates induced similar anti-Hib responses. The K100-HCH conjugate was more immunogenic than the K100-TT conjugate and elicited anti-Hib responses similar to the Hib conjugates after the third injection. Simultaneous injection of the K100 and the Hib conjugates did not enhance the anti-Hib response. The Pn6A-TT conjugate induced low levels of anti-Hib antibodies; when injected simultaneously with the Hib conjugates, the anti-Hib response was enhanced, as all mice responded after the first injection and with higher levels of anti-Hib than observed with the Hib conjugates alone (P < 0.05). The Pn6A conjugates were not as immunogenic as the Hib conjugates. Pn6A-TT was more effective than was Pn6A-HCH; it elicited anti-Pn6A (>100 ng of antibody nitrogen per ml) in 6 of 10 mice after the third injection. The addition of the Hib-HCH conjugate to the Pn6A-TT conjugate increased the anti-Pn6A response with a higher geometric mean antibody titer, and 9 of 10 mice responded after the third injection. A preparation of diphtheria toxoid, TT, and pertussis vaccine increased the anti-Hib antibody levels after the first injection only in mice receiving Hib-TT, but not in mice receiving

  15. Typing of Panton-Valentine Leukocidin-Encoding Phages and lukSF-PV Gene Sequence Variation in Staphylococcus aureus from China

    PubMed Central

    Zhao, Huanqiang; Hu, Fupin; Jin, Shu; Xu, Xiaogang; Zou, Yuhan; Ding, Baixing; He, Chunyan; Gong, Fang; Liu, Qingzhong

    2016-01-01

    Panton-Valentine leukocidin (PVL, encoded by lukSF-PV genes), a bi-component and pore-forming toxin, is carried by different staphylococcal bacteriophages. The prevalence of PVL in Staphylococcus aureus has been reported around the globe. However, the data on PVL-encoding phage types, lukSF-PV gene variation and chromosomal phage insertion sites for PVL-positive S. aureus are limited, especially in China. In order to obtain a more complete understanding of the molecular epidemiology of PVL-positive S. aureus, an integrated and modified PCR-based scheme was applied to detect the PVL-encoding phage types. Phage insertion locus and the lukSF-PV variant were determined by PCR and sequencing. Meanwhile, the genetic background was characterized by staphylococcal cassette chromosome mec (SCCmec) typing, staphylococcal protein A (spa) gene polymorphisms typing, pulsed-field gel electrophoresis (PFGE) typing, accessory gene regulator (agr) locus typing and multilocus sequence typing (MLST). Seventy eight (78/1175, 6.6%) isolates possessed the lukSF-PV genes and 59.0% (46/78) of PVL-positive strains belonged to CC59 lineage. Eight known different PVL-encoding phage types were detected, and Φ7247PVL/ΦST5967PVL (n = 13) and ΦPVL (n = 12) were the most prevalent among them. While 25 (25/78, 32.1%) isolates, belonging to ST30, and ST59 clones, were unable to be typed by the modified PCR-based scheme. Single nucleotide polymorphisms (SNPs) were identified at five locations in the lukSF-PV genes, two of which were non-synonymous. Maximum-likelihood tree analysis of attachment sites sequences detected six SNP profiles for attR and eight for attL, respectively. In conclusion, the PVL-positive S. aureus mainly harbored Φ7247PVL/ΦST5967PVL and ΦPVL in the regions studied. lukSF-PV gene sequences, PVL-encoding phages, and phage insertion locus generally varied with lineages. Moreover, PVL-positive clones that have emerged worldwide likely carry distinct phages. PMID:27536288

  16. Quantitative and qualitative analyses of serum antibodies elicited in adults by Haemophilus influenzae type b and pneumococcus type 6A capsular polysaccharide-tetanus toxoid conjugates.

    PubMed Central

    Schneerson, R; Robbins, J B; Parke, J C; Bell, C; Schlesselman, J J; Sutton, A; Wang, Z; Schiffman, G; Karpas, A; Shiloach, J

    1986-01-01

    Covalent binding to immunogenic proteins increases the immunogenicity of the capsular polysaccharides of Haemophilus influenzae type b (Hib) and pneumococcus type 6A (Pn6A). Conjugates composed of Hib, Pn6A, or the cross-reacting Escherichia coli K100 covalently bound to tetanus toxoid (TT) were injected into young adult volunteers. Local reactions were common and were probably due to Arthus reactivity mediated by the preexisting antibodies reacting with the TT component of the conjugates. Fever occurred in about 10% of the volunteers after the first injection; no volunteers had fever after the second injection. Similar levels of Hib or Pn6A antibodies were elicited by either 50- or 100-micrograms doses or by concurrent injection of two different conjugates (Hib-TT and Pn6A-TT or Hib-TT and K100-TT). The Hib-TT elicited about a 180-fold increase in Hib antibodies, and the Pn6A-TT conjugate elicited about an 8-fold increase in Pn6A antibodies after one injection. Booster reactions were not elicited in adults; similar levels of antibodies in the five experimental groups suggested that the responses elicited by the conjugates were maximal. A one-way cross-reaction was noted as Pn6A conjugates elicited rises of Hib antibodies in 13 of 20 volunteers; only 4 of 59 volunteers immunized with Hib-TT had increases in Pn6A antibodies. The preimmunization Hib antibodies were composed of immunoglobulin M (IgM), IgA, and IgG. The postimmunization sera showed an increase in all three isotypes; the elevation of the IgG was the highest of the three isotypes. Conjugate-induced antibodies to both the polysaccharide and TT exerted biological activities that have been correlated with immunity. Adsorption of the Hib-TT onto aluminium hydroxide resulted in higher levels and an earlier Hib antibody response in infant rhesus. These results encourage the evaluation of Hib and Pn6A conjugates in human children and infants. PMID:3516876

  17. Identification of aromatase activity in rodent pituitary cell strains.

    PubMed

    Callard, G V; Petro, Z; Tashjian, A H

    1983-07-01

    To date, biochemical evidence has been presented for hypophysial aromatization in only one species, a teleost fish, although the pituitary glands of several mammals have been reported to be aromatase negative. To reinvestigate this problem, established clonal strains of rodent pituitary cells (GH3, GH4C1, and AtT20/D16) were incubated at 37 C for 6-48 h in serum-less medium containing [7-3H]androstenedione. Radiolabeled metabolites were isolated by solvent extraction, thin layer chromatography, and phenolic partition. The authenticity of the estrogenic products in both cells and incubation medium was verified by methylation and recrystallization to constant specific activity. Measurement of androgen metabolites was also validated by recrystallization of selected samples. Authentic estrone and 17 beta-estradiol were identified in cultures of the two PRL- and GH-secreting clones, and there were strain differences in the quantity of estrogen produced (GH3 greater than GH4C1). Under the same conditions, aromatization was not detectable in the ACTH-secreting line (AtT20/D16). A time-yield analysis of androgen metabolism in GH4C1 cells showed that aromatization was linear for 12 h after labeling, but that substrate was diverted mainly to 5 alpha-reducing pathways. Large amounts of highly polar metabolites accumulated 24 and 48 h after the addition of [3H]androgen, and subsequent hydrolysis revealed that these were sulfo- and glucuronoconjugates. The metabolic fate of estrogen in GH4C1 cultures was investigated indirectly by adding a radioinert estrone trap together with the radiolabeled androgen substrate and was also tested in separate cultures by adding [3H]estrone and [3H]estradiol directly. Although the two estrogens were interconverted, there was no evidence that formed or added estrogen was extensively metabolized or conjugated. We conclude that the expression of aromatase activity in hypophysial cells is not a property of all transformed lines but may be dictated

  18. TB-HIV co-infection: a catastrophic comradeship.

    PubMed

    Narendran, G; Swaminathan, S

    2016-04-01

    The symbiotic association of tuberculosis (TB) and HIV poses a challenge to human survival. HIV complicates every aspect of TB including presentation, diagnosis and treatment. HIV-TB patients encounter unique problems like drug-drug interactions, cumulative toxicity, immune reconstitution inflammatory syndrome (IRIS), lower plasma drug levels and emergence of drug resistance during treatment despite adherence. TB may also be overdiagnosed in HIV due to a number of diseases that closely resemble TB. Notable among them are non-tuberculous mycobacteria, Pneumocystis Jirovecii and Nocardia. Even though diagnostic procedures have improved over the years, patients in developing countries usually seek health care at later stage of the disease. Research data ascertains the duration of therapy for TB to be 6 months with rifampicin and isoniazid, reinforced with ethambutol and pyrazinamide in the first 2 months. The schedule of therapy is still debatable with daily regimens being preferred in the context of HIV. Many reasons exist for persistence of Mycobacterium Tuberculosis (M.TB) in sputum, or delayed-clearance of TB from sputum smears in HIV, apart from emergence of drug resistance and non-compliance. Acquired rifampicin resistance (ARR) is a unique phenomenon complicating HIV-associated TB when an intermittent regimen of antituberculosis therapy (ATT) is used without timely initiation of highly active antiretroviral therapy (HAART), especially in patients harbouring isoniazid-resistant strains Immune restoration is often incomplete ('swiss cheese' pattern) even with effective HAART if not started early. Immune reconstitution inflammatory syndrome (IRIS) is the paradoxical worsening of the patient's condition often with radiological deterioration, due to an enhanced immune response with HAART. IRIS occurs despite an effective virological suppression and a favourable response to ATT. The incidence of IRIS in HIV has reached up to 54%, requiring utilization of experts

  19. [Construction of the new Escherichia coli K-12 wild-type strain with improved growth characteristics for application in metabolic engineering].

    PubMed

    Biriukova, I V; Krylov, A A; Kiseleva, E M; Minaeva, N I; Mashko, S V

    2010-03-01

    MG1655 of Escherichia coli K-12 is frequently used in metabolic engineering as the wild-type strain. However, its two mutations, ilvG and rph-1 provide a negative effect on culture growth. The "polar effect" of rph-1 decreases the level of pyrE expression, causing partial auxotrophy for pyrimidines. Mutation ilvG leading to the appearance of Val(S) phenotype causes retardation of cell growth rate on media containing amino acids. In this work, the substitution of two loci in the genome of MG1655 with the recovery of the wild-type phenotype was accomplished. Gene rph(wt) from the chromosome of E. coli TG1 was marked via Red-dependent integration of DNA fragment carrying lambda attL-Cm(R)-lambda attR and transduced with phage P1 into MG1655; later, the Cm(R) marker was removed with the use of lambda Xis/Int recombinase. Parallel to this procedure, a spontaneous Val(R) mutant of E. coli MG1655 yielding colonies of maximal size on M9 medium with glucose in the presence of Val (50 microg/ml) was isolated. It was shown that a nucleotide deletion in the isolated Val(R) strain had been generated in the region of the identified E. coli K-12 ilvG mutation, which led to the recovery of the reading frame and active protein synthesis. This mutation named ilvG-15, which is the only reason for the Val(R) phenotype in the obtained strain, was transferred to MG1655-rph(wt) using cotransduction, by analogy to the transfer of rph(wt). Evaluation of rates of aerobically growing cells (microm, hour(-1)) on M9 medium with glucose produced the following values: 0.56, 0.69, and 0.73 for strains MG1655, MG1655-rph(wt), and MG1655-(rph(wt), ilvG-15), respectively.

  20. Identification of aromatase activity in rodent pituitary cell strains.

    PubMed

    Callard, G V; Petro, Z; Tashjian, A H

    1983-07-01

    To date, biochemical evidence has been presented for hypophysial aromatization in only one species, a teleost fish, although the pituitary glands of several mammals have been reported to be aromatase negative. To reinvestigate this problem, established clonal strains of rodent pituitary cells (GH3, GH4C1, and AtT20/D16) were incubated at 37 C for 6-48 h in serum-less medium containing [7-3H]androstenedione. Radiolabeled metabolites were isolated by solvent extraction, thin layer chromatography, and phenolic partition. The authenticity of the estrogenic products in both cells and incubation medium was verified by methylation and recrystallization to constant specific activity. Measurement of androgen metabolites was also validated by recrystallization of selected samples. Authentic estrone and 17 beta-estradiol were identified in cultures of the two PRL- and GH-secreting clones, and there were strain differences in the quantity of estrogen produced (GH3 greater than GH4C1). Under the same conditions, aromatization was not detectable in the ACTH-secreting line (AtT20/D16). A time-yield analysis of androgen metabolism in GH4C1 cells showed that aromatization was linear for 12 h after labeling, but that substrate was diverted mainly to 5 alpha-reducing pathways. Large amounts of highly polar metabolites accumulated 24 and 48 h after the addition of [3H]androgen, and subsequent hydrolysis revealed that these were sulfo- and glucuronoconjugates. The metabolic fate of estrogen in GH4C1 cultures was investigated indirectly by adding a radioinert estrone trap together with the radiolabeled androgen substrate and was also tested in separate cultures by adding [3H]estrone and [3H]estradiol directly. Although the two estrogens were interconverted, there was no evidence that formed or added estrogen was extensively metabolized or conjugated. We conclude that the expression of aromatase activity in hypophysial cells is not a property of all transformed lines but may be dictated