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Sample records for 35s core promoter

  1. Characteristics of a strong promoter from figwort mosaic virus: comparison with the analogous 35S promoter from cauliflower mosaic virus and the regulated mannopine synthase promoter.

    PubMed

    Sanger, M; Daubert, S; Goodman, R M

    1990-03-01

    A segment of DNA from the genome of figwort mosaic virus (FMV) strain M3 possesses promoter activity when tested in electroporated protoplasts from, and transgenic plants of, Nicotiana tabacum cv. Xanthi nc. The 1.1 kb DNA segment, designated the '34S' promoter, is derived from a position on the FMV genome comparable to the position on the cauliflower mosaic virus (CaMV) genome containing the 35S promoter. The 34S and 35S promoters show approximately 63% nucleotide homology in the TATA, CCACT, and -18 to +1 domains, but in sequences further upstream the homology drops below 50%. Promoter activities were estimated using beta-glucuronidase and neomycin phosphotransferase II reporter gene systems. The activity of the 34S promoter segment approximates that of the 35S promoter in both protoplast transient expression assays and in stably transformed tobacco plants. Truncation of 5' sequences from the 34S promoter indicates that promoter strength depends upon DNA sequences located several hundred nucleotides upstream from the TATA box. In leaf tissue the 34S promoter is 20-fold more active than the mannopine synthase (MAS) promoter from Agrobacterium tumefaciens T-DNA. The 34S promoter lacks the root-specific and wound-stimulated expression of the MAS promoter, showing relatively uniform root, stem, leaf, and floral activities.

  2. Development of a general method for detection and quantification of the P35S promoter based on assessment of existing methods

    PubMed Central

    Wu, Yuhua; Wang, Yulei; Li, Jun; Li, Wei; Zhang, Li; Li, Yunjing; Li, Xiaofei; Li, Jun; Zhu, Li; Wu, Gang

    2014-01-01

    The Cauliflower mosaic virus (CaMV) 35S promoter (P35S) is a commonly used target for detection of genetically modified organisms (GMOs). There are currently 24 reported detection methods, targeting different regions of the P35S promoter. Initial assessment revealed that due to the absence of primer binding sites in the P35S sequence, 19 of the 24 reported methods failed to detect P35S in MON88913 cotton, and the other two methods could only be applied to certain GMOs. The rest three reported methods were not suitable for measurement of P35S in some testing events, because SNPs in binding sites of the primer/probe would result in abnormal amplification plots and poor linear regression parameters. In this study, we discovered a conserved region in the P35S sequence through sequencing of P35S promoters from multiple transgenic events, and developed new qualitative and quantitative detection systems targeting this conserved region. The qualitative PCR could detect the P35S promoter in 23 unique GMO events with high specificity and sensitivity. The quantitative method was suitable for measurement of P35S promoter, exhibiting good agreement between the amount of template and Ct values for each testing event. This study provides a general P35S screening method, with greater coverage than existing methods. PMID:25483893

  3. Development of a general method for detection and quantification of the P35S promoter based on assessment of existing methods.

    PubMed

    Wu, Yuhua; Wang, Yulei; Li, Jun; Li, Wei; Zhang, Li; Li, Yunjing; Li, Xiaofei; Li, Jun; Zhu, Li; Wu, Gang

    2014-12-08

    The Cauliflower mosaic virus (CaMV) 35S promoter (P35S) is a commonly used target for detection of genetically modified organisms (GMOs). There are currently 24 reported detection methods, targeting different regions of the P35S promoter. Initial assessment revealed that due to the absence of primer binding sites in the P35S sequence, 19 of the 24 reported methods failed to detect P35S in MON88913 cotton, and the other two methods could only be applied to certain GMOs. The rest three reported methods were not suitable for measurement of P35S in some testing events, because SNPs in binding sites of the primer/probe would result in abnormal amplification plots and poor linear regression parameters. In this study, we discovered a conserved region in the P35S sequence through sequencing of P35S promoters from multiple transgenic events, and developed new qualitative and quantitative detection systems targeting this conserved region. The qualitative PCR could detect the P35S promoter in 23 unique GMO events with high specificity and sensitivity. The quantitative method was suitable for measurement of P35S promoter, exhibiting good agreement between the amount of template and Ct values for each testing event. This study provides a general P35S screening method, with greater coverage than existing methods.

  4. 35S Promoter Methylation in Kanamycin-Resistant Kalanchoe (Kalanchoe pinnata L.) Plants Expressing the Antimicrobial Peptide Cecropin P1 Transgene.

    PubMed

    Shevchuk, T V; Zakharchenko, N S; Tarlachkov, S V; Furs, O V; Dyachenko, O V; Buryanov, Y I

    2016-09-01

    Transgenic kalanchoe plants (Kalanchoe pinnata L.) expressing the antimicrobial peptide cecropin P1 gene (cecP1) under the control of the 35S cauliflower mosaic virus 35S RNA promoter and the selective neomycin phosphotransferase II (nptII) gene under the control of the nopaline synthase gene promoter were studied. The 35S promoter methylation and the cecropin P1 biosynthesis levels were compared in plants growing on media with and without kanamycin. The low level of active 35S promoter methylation further decreases upon cultivation on kanamycin-containing medium, while cecropin P1 synthesis increases.

  5. A DNA probe based on phosphorescent resonance energy transfer for detection of transgenic 35S promoter DNA.

    PubMed

    Lv, Jinzhi; Miao, Yanming; Yang, Jiajia; Qin, Jin; Li, Dongxia; Yan, Guiqin

    2017-05-15

    A QDs-DNA nano-probe was made by combining Mn-doped ZnS room-temperature phosphorescence (RTP) quantum dots (QDs) and DNA. Then an RTP sensor for quantitative detection of genetically-modified mark sequence cauliflower mosaic virus 35S promoter (Ca MV 35S) DNA was built on basis of phosphorescent resonance energy transfer (PRET). The underlying principles were that a QDs-DNA water-soluble nano-probe was built by connecting single-strand DNA to the surfaces of QDs via a ligand exchange method. This probe had good RTP performance and could well identify Ca MV 35S. Thereby, the simple, rapid and efficient detection of genetically-modified organisms was realized. With the increase of target DNA sequence, the phosphorescent intensity of QDs was gradually reduced due to the energy transfer between QDs and the organic quencher BHQ2. This sensor had a detection limit of 4.03nM and a detection range of 12-300nM. Moreover, this sensor had high selectivity. This sensor could effectively detect the target DNA compared with mismatched and random sequences. Thus, this method is very promising for biological analysis.

  6. Detection of the 35S promoter in transgenic maize via various isothermal amplification techniques: a practical approach.

    PubMed

    Zahradnik, Celine; Kolm, Claudia; Martzy, Roland; Mach, Robert L; Krska, Rudolf; Farnleitner, Andreas H; Brunner, Kurt

    2014-11-01

    In 2003 the European Commission introduced a 0.9% threshold for food and feed products containing genetically modified organism (GMO)-derived components. For commodities containing GMO contents higher than this threshold, labelling is mandatory. To provide a DNA-based rapid and simple detection method suitable for high-throughput screening of GMOs, several isothermal amplification approaches for the 35S promoter were tested: strand displacement amplification, nicking-enzyme amplification reaction, rolling circle amplification, loop-mediated isothermal amplification (LAMP) and helicase-dependent amplification (HDA). The assays developed were tested for specificity in order to distinguish between samples containing genetically modified (GM) maize and non-GM maize. For those assays capable of this discrimination, tests were performed to determine the lower limit of detection. A false-negative rate was determined to rule out whether GMO-positive samples were incorrectly classified as GMO-negative. A robustness test was performed to show reliable detection independent from the instrument used for amplification. The analysis of three GM maize lines showed that only LAMP and HDA were able to differentiate between the GMOs MON810, NK603, and Bt11 and non-GM maize. Furthermore, with the HDA assay it was possible to realize a detection limit as low as 0.5%. A false-negative rate of only 5% for 1% GM maize for all three maize lines shows that HDA has the potential to be used as an alternative strategy for the detection of transgenic maize. All results obtained with the LAMP and HDA assays were compared with the results obtained with a previously reported real-time PCR assay for the 35S promoter in transgenic maize. This study presents two new screening assays for detection of the 35S promoter in transgenic maize by applying the isothermal amplification approaches HDA and LAMP.

  7. Use of a novel metal indicator to judge loop-mediated isothermal amplification for detecting the 35S promoter.

    PubMed

    Wang, Xiaofu; Fu, Zhenfang; Chen, Xiaoyun; Peng, Cheng; Xu, Xiaoli; Wei, Wei; Li, Feiwu; Xu, Junfeng

    2017-02-01

    Loop-mediated isothermal amplification (LAMP) is a widely used isothermal nucleic acid amplification method. Here we developed a new closed-tube colorimetric method for judging LAMP with a novel metal indicator. First, the metal indicator, acid chrome blue K (ACBK), was evaluated in the LAMP reaction with various combinations of reaction reagents, such as reaction buffer, dNTP mixtures, primer mixtures, or Mg(2+). We found that the solution color of the LAMP reaction with ACBK changed from red to blue based on a decrease in the Mg(2+) concentration in the reaction solution. We then optimized the LAMP with ACBK method for detecting the Cauliflower Mosaic Virus 35S promoter. Further, the specificity of the new colorimetric assay using ACBK in the LAMP reaction for detecting the 35S promoter was tested with diverse transgenic events in different crops, and the sensitivity threshold of the assay was ∼50 copies for transgenic rice genomic DNA and 100 ng of 0.1 % DNA from rice, soybean, rapeseed, and maize. Finally, the applicability of the LAMP assay was successfully validated using practical maize samples. All the detection results could be easily discerned either by UV-vis spectroscopy or the naked eye. Graphical Abstract The visual detect LAMP amplification by the addition of ACBK as a signal indicator. The color of the LAMP-ACBK solution turned from red to blue as the concentration of free Mg(2+) decreases. The detection results could be easily discerned either by UV-vis spectroscopy or the naked eye.

  8. Expression of. beta. -conglycinin gene driven by CaMV /sup 35/S promoter in transgenic plants

    SciTech Connect

    Nakamura, I.; Dube, P.H.; Beachy, R.N.

    1987-04-01

    ..beta..-conglycinin is a abundant protein stored in protein bodies of soybean seeds. This protein consists of three major subunits, ..cap alpha..' (76 kDa), ..cap alpha.. (72 kDa) and ..beta.. (53 kDa), and accumulates in developing soybean embryos during the mid- to late-maturation stages of seed development. Coding sequence of an ..cap alpha..'-subunit gene was expressed in transgenic petunia plants under control of the promoter from the CaMV (cauliflower mosaic virus) /sup 35/S transcript. Two different types of ..cap alpha..'-protein accumulated in tissues of the transgenic plant; seed-type ..cap alpha..'-protein accumulated only in seeds during mid- to late-maturation stages, while non-seed-type ..cap alpha..'-protein was found in non-seed tissues and in early stages of seed maturation. Seed-type ..cap alpha..'-protein was the same size as soybean ..cap alpha..'-subunit, while non-seed-type ..cap alpha..'-protein was larger by about 4 kDa. Seeds contained approximately 30-fold greater levels of ..cap alpha..'-protein than did non-seed tissues. This is presumably due to differences in protein stability because the amount of ..cap alpha..'-mRNA was equivalent in each of the tissues examined. The ..cap alpha..'-protein in leaves was localized in microsomal membrane fractions. Proteins solubilized from the membranes were sedimented by sucrose gradient centrifugation and analyzed by immuno blot technique. The results suggest that the protein assembles into multimeric forms in leaf membranes, as it does in seed protein bodies.

  9. An oleosin-fusion protein driven by the CaMV35S promoter is accumulated in Arabidopsis (Brassicaceae) seeds and correctly targeted to oil bodies.

    PubMed

    Li, W; Li, L G; Sun, X F; Tang, K X

    2012-08-13

    Oleosin-fusion technology is used to express desired proteins. It was developed based on the properties of oleosin; the heterologous protein gene is fused to the oleosin gene and the fusion gene is driven by a seed-specific promoter. We replaced the seed specific promoter with the CaMV35S promoter to dive a gfp-oleosin fusion gene in transformed Arabidopsis. The heterologous oleosin-fusion protein was mainly accumulated in the transgenic Arabidopsis seeds and correctly targeted to oil bodies. This provides an alternate choice of promoter in oleosin-fusion technology.

  10. U2AF35(S34F) Promotes Transformation by Directing Aberrant ATG7 Pre-mRNA 3' End Formation.

    PubMed

    Park, Sung Mi; Ou, Jianhong; Chamberlain, Lynn; Simone, Tessa M; Yang, Huan; Virbasius, Ching-Man; Ali, Abdullah M; Zhu, Lihua Julie; Mukherjee, Siddhartha; Raza, Azra; Green, Michael R

    2016-05-19

    Recurrent mutations in the splicing factor U2AF35 are found in several cancers and myelodysplastic syndrome (MDS). How oncogenic U2AF35 mutants promote transformation remains to be determined. Here we derive cell lines transformed by the oncogenic U2AF35(S34F) mutant and identify aberrantly processed pre-mRNAs by deep sequencing. We find that in U2AF35(S34F)-transformed cells the autophagy-related factor 7 (Atg7) pre-mRNA is abnormally processed, which unexpectedly is not due to altered splicing but rather selection of a distal cleavage and polyadenylation (CP) site. This longer Atg7 mRNA is translated inefficiently, leading to decreased ATG7 levels and an autophagy defect that predisposes cells to secondary mutations, resulting in transformation. MDS and acute myeloid leukemia patient samples harboring U2AF35(S34F) have a similar increased use of the ATG7 distal CP site, and previous studies have shown that mice with hematopoietic cells lacking Atg7 develop an MDS-like syndrome. Collectively, our results reveal a basis for U2AF35(S34F) oncogenic activity.

  11. Recombinase Polymerase Amplification (RPA) of CaMV-35S Promoter and nos Terminator for Rapid Detection of Genetically Modified Crops

    PubMed Central

    Xu, Chao; Li, Liang; Jin, Wujun; Wan, Yusong

    2014-01-01

    Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37–42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence of the cauliflower mosaic virus 35S (CaMV-35S) promoter and the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator, which are widely incorporated in genetically modified (GM) crops, we designed two sets of RPA primers and established a real-time RPA detection method for GM crop screening and detection. This method could reliably detect as few as 100 copies of the target molecule in a sample within 15–25 min. Furthermore, the real-time RPA detection method was successfully used to amplify and detect DNA from samples of four major GM crops (maize, rice, cotton, and soybean). With this novel amplification method, the test time was significantly shortened and the reaction process was simplified; thus, this method represents an effective approach to the rapid detection of GM crops. PMID:25310647

  12. Recombinase polymerase amplification (RPA) of CaMV-35S promoter and nos terminator for rapid detection of genetically modified crops.

    PubMed

    Xu, Chao; Li, Liang; Jin, Wujun; Wan, Yusong

    2014-10-10

    Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37-42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence of the cauliflower mosaic virus 35S (CaMV-35S) promoter and the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator, which are widely incorporated in genetically modified (GM) crops, we designed two sets of RPA primers and established a real-time RPA detection method for GM crop screening and detection. This method could reliably detect as few as 100 copies of the target molecule in a sample within 15-25 min. Furthermore, the real-time RPA detection method was successfully used to amplify and detect DNA from samples of four major GM crops (maize, rice, cotton, and soybean). With this novel amplification method, the test time was significantly shortened and the reaction process was simplified; thus, this method represents an effective approach to the rapid detection of GM crops.

  13. Synthetic Core Promoters for Pichia pastoris

    PubMed Central

    2013-01-01

    Synthetic promoters are commonly used tools for circuit design or high level protein production. Promoter engineering efforts in yeasts, such as Saccharomyces cerevisiae and Pichia pastoris have mostly been focused on altering upstream regulatory sequences such as transcription factor binding sites. In higher eukaryotes synthetic core promoters, directly needed for transcription initiation by RNA Polymerase II, have been successfully designed. Here we report the first synthetic yeast core promoter for P. pastoris, based on natural yeast core promoters. Furthermore we used this synthetic core promoter sequence to engineer the core promoter of the natural AOX1 promoter, thereby creating a set of core promoters providing a range of different expression levels. As opposed to engineering strategies of the significantly longer entire promoter, such short core promoters can directly be added on a PCR primer facilitating library generation and are sufficient to obtain variable expression yields. PMID:24187969

  14. The ABA effect on the accumulation of an invertase inhibitor transcript that is driven by the CAMV35S promoter in ARABIDOPSIS.

    PubMed

    Koh, Eun-Ji; Lee, Sung June; Hong, Suk-Whan; Lee, Hoi Seon; Lee, Hojoung

    2008-09-30

    Invertase (beta-D-fructofuranosidase; EC 3.2.1.26) catalyzes the conversion of sucrose into glucose and fructose and is involved in an array of important processes, including phloem unloading, carbon partitioning, the response to pathogens, and the control of cell differentiation and development. Its importance may have caused the invertases to evolve into a multigene family whose members are regulated by a variety of different mechanisms, such as pH, sucrose levels, and inhibitor proteins. Although putative invertase inhibitors in the Arabidopsis genome are easy to locate, few studies have been conducted to elucidate their individual functions in vivo in plant growth and development because of their high redundancy. In this study we assessed the functional role of the putative invertase inhibitors in Arabidopsis by generating transgenic plants harboring a putative invertase inhibitor gene under the control of the CaMV35S promoter. A transgenic plant that expressed high levels of the putative invertase inhibitor transcript when grown under normal conditions was chosen for the current study. To our surprise, the stability of the invertase inhibitor transcripts was shown to be down-regulated by the phytohormone ABA (abscisic acid). It is well established that ABA enhances invertase activity in vivo but the underlying mechanisms are still poorly understood. Our results thus suggest that one way ABA regulates invertase activity is by down-regulating its inhibitor.

  15. Native Phytoremediation Potential of Urtica dioica for Removal of PCBs and Heavy Metals Can Be Improved by Genetic Manipulations Using Constitutive CaMV 35S Promoter.

    PubMed

    Viktorova, Jitka; Jandova, Zuzana; Madlenakova, Michaela; Prouzova, Petra; Bartunek, Vilem; Vrchotova, Blanka; Lovecka, Petra; Musilova, Lucie; Macek, Tomas

    2016-01-01

    Although stinging nettle (Urtica dioica) has been shown to reduce HM (heavy metal) content in soil, its wider phytoremediation potential has been neglected. Urtica dioica was cultivated in soils contaminated with HMs or polychlorinated biphenyls (PCBs). After four months, up to 33% of the less chlorinated biphenyls and 8% of HMs (Zn, Pb, Cd) had been removed. Bacteria were isolated from the plant tissue, with the endophytic bacteria Bacillus shackletonii and Streptomyces badius shown to have the most significant effect. These bacteria demonstrated not only benefits for plant growth, but also extreme tolerance to As, Zn and Pb. Despite these results, the native phytoremediation potential of nettles could be improved by biotechnologies. Transient expression was used to investigate the functionality of the most common constitutive promoter, CaMV 35S in Urtica dioica. This showed the expression of the CUP and bphC transgenes. Collectively, our findings suggest that remediation by stinging nettle could have a much wider range of applications than previously thought.

  16. Native Phytoremediation Potential of Urtica dioica for Removal of PCBs and Heavy Metals Can Be Improved by Genetic Manipulations Using Constitutive CaMV 35S Promoter

    PubMed Central

    Viktorova, Jitka; Jandova, Zuzana; Madlenakova, Michaela; Prouzova, Petra; Bartunek, Vilem; Vrchotova, Blanka; Lovecka, Petra; Musilova, Lucie; Macek, Tomas

    2016-01-01

    Although stinging nettle (Urtica dioica) has been shown to reduce HM (heavy metal) content in soil, its wider phytoremediation potential has been neglected. Urtica dioica was cultivated in soils contaminated with HMs or polychlorinated biphenyls (PCBs). After four months, up to 33% of the less chlorinated biphenyls and 8% of HMs (Zn, Pb, Cd) had been removed. Bacteria were isolated from the plant tissue, with the endophytic bacteria Bacillus shackletonii and Streptomyces badius shown to have the most significant effect. These bacteria demonstrated not only benefits for plant growth, but also extreme tolerance to As, Zn and Pb. Despite these results, the native phytoremediation potential of nettles could be improved by biotechnologies. Transient expression was used to investigate the functionality of the most common constitutive promoter, CaMV 35S in Urtica dioica. This showed the expression of the CUP and bphC transgenes. Collectively, our findings suggest that remediation by stinging nettle could have a much wider range of applications than previously thought. PMID:27930707

  17. Over-Expression of the Pikh Gene with a CaMV 35S Promoter Leads to Improved Blast Disease (Magnaporthe oryzae) Tolerance in Rice

    PubMed Central

    Azizi, Parisa; Rafii, Mohd Y.; Abdullah, Siti N. A.; Hanafi, Mohamed M.; Maziah, M.; Sahebi, Mahbod; Ashkani, Sadegh; Taheri, Sima; Jahromi, Mohammad F.

    2016-01-01

    Magnaporthe oryzae is a rice blast fungus and plant pathogen that causes a serious rice disease and, therefore, poses a threat to the world's second most important food security crop. Plant transformation technology has become an adaptable system for cultivar improvement and to functionally analyze genes in plants. The objective of this study was to determine the effects (through over-expressing and using the CaMV 35S promoter) of Pikh on MR219 resistance because it is a rice variety that is susceptible to the blast fungus pathotype P7.2. Thus, a full DNA and coding DNA sequence (CDS) of the Pikh gene, 3172 bp, and 1206 bp in length, were obtained through amplifying the gDNA and cDNA template from a PH9-resistant rice variety using a specific primer. Agrobacterium-mediated transformation technology was also used to introduce the Pikh gene into the MR219 callus. Subsequently, transgenic plants were evaluated from the DNA to protein stages using polymerase chain reaction (PCR), semi-quantitative RT-PCR, real-time quantitative PCR and high performance liquid chromatography (HPLC). Transgenic plants were also compared with a control using a real-time quantification technique (to quantify the pathogen population), and transgenic and control plants were challenged with the local most virulent M. oryzae pathotype, P7.2. Based on the results, the Pikh gene encodes a hydrophilic protein with 18 sheets, 4 helixes, and 21 coils. This protein contains 401 amino acids, among which the amino acid sequence from 1 to 376 is a non-cytoplasmic region, that from 377 to 397 is a transmembrane region, and that from 398 to 401 is a cytoplasmic region with no identified disordered regions. The Pikh gene was up-regulated in the transgenic plants compared with the control plants. The quantity of the amino acid leucine in the transgenic rice plants increased significantly from 17.131 in the wild-type to 47.865 mg g−1 in transgenic plants. The M. oryzae population was constant at 31, 48

  18. Over-Expression of the Pikh Gene with a CaMV 35S Promoter Leads to Improved Blast Disease (Magnaporthe oryzae) Tolerance in Rice.

    PubMed

    Azizi, Parisa; Rafii, Mohd Y; Abdullah, Siti N A; Hanafi, Mohamed M; Maziah, M; Sahebi, Mahbod; Ashkani, Sadegh; Taheri, Sima; Jahromi, Mohammad F

    2016-01-01

    Magnaporthe oryzae is a rice blast fungus and plant pathogen that causes a serious rice disease and, therefore, poses a threat to the world's second most important food security crop. Plant transformation technology has become an adaptable system for cultivar improvement and to functionally analyze genes in plants. The objective of this study was to determine the effects (through over-expressing and using the CaMV 35S promoter) of Pikh on MR219 resistance because it is a rice variety that is susceptible to the blast fungus pathotype P7.2. Thus, a full DNA and coding DNA sequence (CDS) of the Pikh gene, 3172 bp, and 1206 bp in length, were obtained through amplifying the gDNA and cDNA template from a PH9-resistant rice variety using a specific primer. Agrobacterium-mediated transformation technology was also used to introduce the Pikh gene into the MR219 callus. Subsequently, transgenic plants were evaluated from the DNA to protein stages using polymerase chain reaction (PCR), semi-quantitative RT-PCR, real-time quantitative PCR and high performance liquid chromatography (HPLC). Transgenic plants were also compared with a control using a real-time quantification technique (to quantify the pathogen population), and transgenic and control plants were challenged with the local most virulent M. oryzae pathotype, P7.2. Based on the results, the Pikh gene encodes a hydrophilic protein with 18 sheets, 4 helixes, and 21 coils. This protein contains 401 amino acids, among which the amino acid sequence from 1 to 376 is a non-cytoplasmic region, that from 377 to 397 is a transmembrane region, and that from 398 to 401 is a cytoplasmic region with no identified disordered regions. The Pikh gene was up-regulated in the transgenic plants compared with the control plants. The quantity of the amino acid leucine in the transgenic rice plants increased significantly from 17.131 in the wild-type to 47.865 mg g(-1) in transgenic plants. The M. oryzae population was constant at 31, 48

  19. Linking Core Promoter Classes to Circadian Transcription

    PubMed Central

    Westermark, Pål O.

    2016-01-01

    Circadian rhythms in transcription are generated by rhythmic abundances and DNA binding activities of transcription factors. Propagation of rhythms to transcriptional initiation involves the core promoter, its chromatin state, and the basal transcription machinery. Here, I characterize core promoters and chromatin states of genes transcribed in a circadian manner in mouse liver and in Drosophila. It is shown that the core promoter is a critical determinant of circadian mRNA expression in both species. A distinct core promoter class, strong circadian promoters (SCPs), is identified in mouse liver but not Drosophila. SCPs are defined by specific core promoter features, and are shown to drive circadian transcriptional activities with both high averages and high amplitudes. Data analysis and mathematical modeling further provided evidence for rhythmic regulation of both polymerase II recruitment and pause release at SCPs. The analysis provides a comprehensive and systematic view of core promoters and their link to circadian mRNA expression in mouse and Drosophila, and thus reveals a crucial role for the core promoter in regulated, dynamic transcription. PMID:27504829

  20. Strict de novo methylation of the 35S enhancer sequence in gentian.

    PubMed

    Mishiba, Kei-ichiro; Yamasaki, Satoshi; Nakatsuka, Takashi; Abe, Yoshiko; Daimon, Hiroyuki; Oda, Masayuki; Nishihara, Masahiro

    2010-03-23

    A novel transgene silencing phenomenon was found in the ornamental plant, gentian (Gentiana triflora x G. scabra), in which the introduced Cauliflower mosaic virus (CaMV) 35S promoter region was strictly methylated, irrespective of the transgene copy number and integrated loci. Transgenic tobacco having the same vector did not show the silencing behavior. Not only unmodified, but also modified 35S promoters containing a 35S enhancer sequence were found to be highly methylated in the single copy transgenic gentian lines. The 35S core promoter (-90)-introduced transgenic lines showed a small degree of methylation, implying that the 35S enhancer sequence was involved in the methylation machinery. The rigorous silencing phenomenon enabled us to analyze methylation in a number of the transgenic lines in parallel, which led to the discovery of a consensus target region for de novo methylation, which comprised an asymmetric cytosine (CpHpH; H is A, C or T) sequence. Consequently, distinct footprints of de novo methylation were detected in each (modified) 35S promoter sequence, and the enhancer region (-148 to -85) was identified as a crucial target for de novo methylation. Electrophoretic mobility shift assay (EMSA) showed that complexes formed in gentian nuclear extract with the -149 to -124 and -107 to -83 region probes were distinct from those of tobacco nuclear extracts, suggesting that the complexes might contribute to de novo methylation. Our results provide insights into the phenomenon of sequence- and species- specific gene silencing in higher plants.

  1. Quantification of the 35S promoter in DNA extracts from genetically modified organisms using real-time polymerase chain reaction and specificity assessment on various genetically modified organisms, part I: operating procedure.

    PubMed

    Fernandez, Sophie; Charles-Delobel, Chrystèle; Geldreich, Angèle; Berthier, Georges; Boyer, Francine; Collonnier, Cécile; Coué-Philippe, Géraldine; Diolez, Annick; Duplan, Marie-Noëlle; Kebdani, Naïma; Romaniuk, Marcel; Feinberg, Max; Bertheau, Yves

    2005-01-01

    A highly sensitive quantitative real-time assay targeted on the 35S promoter of a commercial genetically modified organism (GMO) was characterized (sF/sR primers) and developed for an ABI Prism 7700 Sequence Detection System and TaqMan chemistry. The specificity assessment and performance criteria of sF/sR assay were compared to other P35S-targeted published assays. sF/sR primers amplified a 79 base pair DNA sequence located in a part of P35S that is highly conserved among many caulimovirus strains, i.e., this consensus part of CaMV P35S is likely to be present in many GM events. According to the experimental conditions, the absolute limit of detection for Bt176 corn was estimated between 0.2 and 2 copies of equivalent genome (CEG). The limit of quantification was reached below 0.1% Bt176 content. A Cauliflower Mosaic Virus control (CaMV) qualitative assay targeted on the ORF III of the viral genome was also used as a control (primers 3F/3R) to assess the presence of CaMV in plant-derived products. The specificity of this test was assessed on various CaMV strains, including the Figwort Mosaic Virus (FMV) and solanaceous CaMV strains. Considering the performance of sF/sR quantification test, the highly conserved sequence, and the small size of the amplicon, this assay was tested in a collaborative study in order to be proposed as an international standard.

  2. The VviMYB80 Gene is Abnormally Expressed in Vitis vinifera L. cv. 'Zhong Shan Hong' and its Expression in Tobacco Driven by the 35S Promoter Causes Male Sterility.

    PubMed

    Zheng, Huan; Yu, Xiaojuan; Yuan, Yue; Zhang, Yaguang; Zhang, Zhen; Zhang, Jiyu; Zhang, Meng; Ji, Chenfei; Liu, Qian; Tao, Jianmin

    2016-03-01

    Anther development is a very precise and complicated process. In Arabidopsis, the AtMYB80 transcription factor regulates genes involved in pollen development and controls the timing of tapetal programmed cell death (PCD). In this study, we isolated and characterized cDNA for VviMYB80 expressed in flower buds of male-sterile Vitis vinifera L. cv. 'Zhong Shan Hong', a late-maturing cultivar derived from self-progeny of cv. 'Wink'. VviMYB80 belongs to the MYB80 subfamily and clusters with AtMYB35/TDF1 in a distinct clade. We found that in flower buds, expression of the VviMYB80 gene in cv. 'Zhong Shan Hong' sharply increased at the tetrad stage, resulting in a higher and earlier transcript level than that found in cv. 'Wink'. Expression of the VviMYB80 gene, driven by the 35S promoter, caused pleiotropic effects on the stamens, including smaller and shriveled anthers, delayed dehiscence, fewer seeds, shorter anther filaments, distorted pollen shape and a lack of cytoplasm, with the tapetum exhibiting hypertrophy in transformed tobacco. These results suggest that VviMYB80 may play an important role in stamen development and that expression of VviMYB80 driven by the 35S promoter in tobacco induces male sterility.

  3. A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles.

    PubMed

    Li, Yaqi; Sun, Li; Qian, Jing; Wang, Chengke; Liu, Qian; Han, En; Hao, Nan; Zhang, Liuping; Cai, Jianrong; Wang, Kun

    2016-12-15

    In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1-500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection.

  4. A screening method for the detection of the 35S promoter and the nopaline synthase terminator in genetically modified organisms in a real-time multiplex polymerase chain reaction using high-resolution melting-curve analysis.

    PubMed

    Akiyama, Hiroshi; Nakamura, Fumi; Yamada, Chihiro; Nakamura, Kosuke; Nakajima, Osamu; Kawakami, Hiroshi; Harikai, Naoki; Furui, Satoshi; Kitta, Kazumi; Teshima, Reiko

    2009-11-01

    To screen for unauthorized genetically modified organisms (GMO) in the various crops, we developed a multiplex real-time polymerase chain reaction high-resolution melting-curve analysis method for the simultaneous qualitative detection of 35S promoter sequence of cauliflower mosaic virus (35SP) and the nopaline synthase terminator (NOST) in several crops. We selected suitable primer sets for the simultaneous detection of 35SP and NOST and designed the primer set for the detection of spiked ColE1 plasmid to evaluate the validity of the polymerase chain reaction (PCR) analyses. In addition, we optimized the multiplex PCR conditions using the designed primer sets and EvaGreen as an intercalating dye. The contamination of unauthorized GMO with single copy similar to NK603 maize can be detected as low as 0.1% in a maize sample. Furthermore, we showed that the present method would be applicable in identifying GMO in various crops and foods like authorized GM soybean, authorized GM potato, the biscuit which is contaminated with GM soybeans and the rice which is contaminated with unauthorized GM rice. We consider this method to be a simple and reliable assay for screening for unauthorized GMO in crops and the processing food products.

  5. The core promoter: At the heart of gene expression.

    PubMed

    Danino, Yehuda M; Even, Dan; Ideses, Diana; Juven-Gershon, Tamar

    2015-08-01

    The identities of different cells and tissues in multicellular organisms are determined by tightly controlled transcriptional programs that enable accurate gene expression. The mechanisms that regulate gene expression comprise diverse multiplayer molecular circuits of multiple dedicated components. The RNA polymerase II (Pol II) core promoter establishes the center of this spatiotemporally orchestrated molecular machine. Here, we discuss transcription initiation, diversity in core promoter composition, interactions of the basal transcription machinery with the core promoter, enhancer-promoter specificity, core promoter-preferential activation, enhancer RNAs, Pol II pausing, transcription termination, Pol II recycling and translation. We further discuss recent findings indicating that promoters and enhancers share similar features and may not substantially differ from each other, as previously assumed. Taken together, we review a broad spectrum of studies that highlight the importance of the core promoter and its pivotal role in the regulation of metazoan gene expression and suggest future research directions and challenges.

  6. Complete nucleotide sequences and construction of full-length infectious cDNA clones of cucumber green mottle mosaic virus (CGMMV) in a versatile newly developed binary vector including both 35S and T7 promoters.

    PubMed

    Park, Chan-Hwan; Ju, Hye-Kyoung; Han, Jae-Yeong; Park, Jong-Seo; Kim, Ik-Hyun; Seo, Eun-Young; Kim, Jung-Kyu; Hammond, John; Lim, Hyoun-Sub

    2017-04-01

    Seed-transmitted viruses have caused significant damage to watermelon crops in Korea in recent years, with cucumber green mottle mosaic virus (CGMMV) infection widespread as a result of infected seed lots. To determine the likely origin of CGMMV infection, we collected CGMMV isolates from watermelon and melon fields and generated full-length infectious cDNA clones. The full-length cDNAs were cloned into newly constructed binary vector pJY, which includes both the 35S and T7 promoters for versatile usage (agroinfiltration and in vitro RNA transcription) and a modified hepatitis delta virus ribozyme sequence to precisely cleave RNA transcripts at the 3' end of the tobamovirus genome. Three CGMMV isolates (OMpj, Wpj, and Mpj) were separately evaluated for infectivity in Nicotiana benthamiana, demonstrated by either Agroinfiltration or inoculation with in vitro RNA transcripts. CGMMV nucleotide identities to other tobamoviruses were calculated from pairwise alignments using DNAMAN. CGMMV identities were 49.89% to tobacco mosaic virus; 49.85% to pepper mild mottle virus; 50.47% to tomato mosaic virus; 60.9% to zucchini green mottle mosaic virus; and 60.96% to kyuri green mottle mosaic virus, confirming that CGMMV is a distinct species most similar to other cucurbit-infecting tobamoviruses. We further performed phylogenetic analysis to determine relationships of our new Korean CGMMV isolates to previously characterized isolates from Canada, China, India, Israel, Japan, Korea, Russia, Spain, and Taiwan available from NCBI. Analysis of CGMMV amino acid sequences showed three major clades, broadly typified as 'Russian,' 'Israeli,' and 'Asian' groups. All of our new Korean isolates fell within the 'Asian' clade. Neither the 128 nor 186 kDa RdRps of the three new isolates showed any detectable gene silencing suppressor function.

  7. Drosophila TRF2 is a preferential core promoter regulator

    PubMed Central

    Kedmi, Adi; Zehavi, Yonathan; Glick, Yair; Orenstein, Yaron; Ideses, Diana; Wachtel, Chaim; Doniger, Tirza; Waldman Ben-Asher, Hiba; Muster, Nemone; Thompson, James; Anderson, Scott; Avrahami, Dorit; Yates, John R.; Shamir, Ron; Gerber, Doron

    2014-01-01

    Transcription of protein-coding genes is highly dependent on the RNA polymerase II core promoter. Core promoters, generally defined as the regions that direct transcription initiation, consist of functional core promoter motifs (such as the TATA-box, initiator [Inr], and downstream core promoter element [DPE]) that confer specific properties to the core promoter. The known basal transcription factors that support TATA-dependent transcription are insufficient for in vitro transcription of DPE-dependent promoters. In search of a transcription factor that supports DPE-dependent transcription, we used a biochemical complementation approach and identified the Drosophila TBP (TATA-box-binding protein)-related factor 2 (TRF2) as an enriched factor in the fractions that support DPE-dependent transcription. We demonstrate that the short TRF2 isoform preferentially activates DPE-dependent promoters. DNA microarray analysis reveals the enrichment of DPE promoters among short TRF2 up-regulated genes. Using primer extension analysis and reporter assays, we show the importance of the DPE in transcriptional regulation of TRF2 target genes. It was previously shown that, unlike TBP, TRF2 fails to bind DNA containing TATA-boxes. Using microfluidic affinity analysis, we discovered that short TRF2-bound DNA oligos are enriched for Inr and DPE motifs. Taken together, our findings highlight the role of short TRF2 as a preferential core promoter regulator. PMID:25223897

  8. Computational analysis of core promoters in the Drosophila genome

    PubMed Central

    Ohler, Uwe; Liao, Guo-chun; Niemann, Heinrich; Rubin, Gerald M

    2002-01-01

    Background The core promoter, a region of about 100 base-pairs flanking the transcription start site (TSS), serves as the recognition site for the basal transcription apparatus. Drosophila TSSs have generally been mapped by individual experiments; the low number of accurately mapped TSSs has limited analysis of promoter sequence motifs and the training of computational prediction tools. Results We identified TSS candidates for about 2,000 Drosophila genes by aligning 5' expressed sequence tags (ESTs) from cap-trapped cDNA libraries to the genome, while applying stringent criteria concerning coverage and 5'-end distribution. Examination of the sequences flanking these TSSs revealed the presence of well-known core promoter motifs such as the TATA box, the initiator and the downstream promoter element (DPE). We also define, and assess the distribution of, several new motifs prevalent in core promoters, including what appears to be a variant DPE motif. Among the prevalent motifs is the DNA-replication-related element DRE, recently shown to be part of the recognition site for the TBP-related factor TRF2. Our TSS set was then used to retrain the computational promoter predictor McPromoter, allowing us to improve the recognition performance to over 50% sensitivity and 40% specificity. We compare these computational results to promoter prediction in vertebrates. Conclusions There are relatively few recognizable binding sites for previously known general transcription factors in Drosophila core promoters. However, we identified several new motifs enriched in promoter regions. We were also able to significantly improve the performance of computational TSS prediction in Drosophila. PMID:12537576

  9. Characterization of nonconventional hepatitis B viruses lacking the core promoter.

    PubMed

    Chang, Shau-Feng; Chang, Shih-Hsuan; Li, Bi-Chen; Will, Hans; Netter, Hans Jürgen

    2004-12-20

    The core gene (C-gene) promoter and regulatory sequences play a central role in the hepatitis B virus (HBV) life cycle. They are essential for the synthesis of the pregenomic and precore mRNA. The pregenomic RNA is the template required for replication and also the template for the synthesis of the core protein and polymerase. Here, we report the in vivo existence and functional characterization of HBV variants that lack the C-gene promoter region and the regulatory sequences located therein. HBV promoter fragments were isolated by PCR from sera of chronic carriers and characterized. Truncated promoter elements were identified, and then tested in the context of wild-type genomes in the HuH-7 cell line. The expression of the recombinant HBV genome resulted in the synthesis of surface proteins, and low level of core protein as well as a transcript pattern similar to, but smaller in size to wild-type virus. The recombinant HBV genome with the truncated promoter region produced pregenomic RNA-like transcripts. These transcripts were encapsidated and reverse transcribed when complemented by sufficient core and polymerase protein. These date provide an explanation as to why such deletion mutants of HBV can be produced at all, they highlight the functional potentials of viral sequences activated by mutations and may be of relevance for viral evolution and persistence.

  10. Dynamic usage of transcription start sites within core promoters

    PubMed Central

    Kawaji, Hideya; Frith, Martin C; Katayama, Shintaro; Sandelin, Albin; Kai, Chikatoshi; Kawai, Jun; Carninci, Piero; Hayashizaki, Yoshihide

    2006-01-01

    Background Mammalian promoters do not initiate transcription at single, well defined base pairs, but rather at multiple, alternative start sites spread across a region. We previously characterized the static structures of transcription start site usage within promoters at the base pair level, based on large-scale sequencing of transcript 5' ends. Results In the present study we begin to explore the internal dynamics of mammalian promoters, and demonstrate that start site selection within many mouse core promoters varies among tissues. We also show that this dynamic usage of start sites is associated with CpG islands, broad and multimodal promoter structures, and imprinting. Conclusion Our results reveal a new level of biologic complexity within promoters - fine-scale regulation of transcription starting events at the base pair level. These events are likely to be related to epigenetic transcriptional regulation. PMID:17156492

  11. Two independent transcription initiation codes overlap on vertebrate core promoters.

    PubMed

    Haberle, Vanja; Li, Nan; Hadzhiev, Yavor; Plessy, Charles; Previti, Christopher; Nepal, Chirag; Gehrig, Jochen; Dong, Xianjun; Akalin, Altuna; Suzuki, Ana Maria; van IJcken, Wilfred F J; Armant, Olivier; Ferg, Marco; Strähle, Uwe; Carninci, Piero; Müller, Ferenc; Lenhard, Boris

    2014-03-20

    A core promoter is a stretch of DNA surrounding the transcription start site (TSS) that integrates regulatory inputs and recruits general transcription factors to initiate transcription. The nature and causative relationship of the DNA sequence and chromatin signals that govern the selection of most TSSs by RNA polymerase II remain unresolved. Maternal to zygotic transition represents the most marked change of the transcriptome repertoire in the vertebrate life cycle. Early embryonic development in zebrafish is characterized by a series of transcriptionally silent cell cycles regulated by inherited maternal gene products: zygotic genome activation commences at the tenth cell cycle, marking the mid-blastula transition. This transition provides a unique opportunity to study the rules of TSS selection and the hierarchy of events linking transcription initiation with key chromatin modifications. We analysed TSS usage during zebrafish early embryonic development at high resolution using cap analysis of gene expression, and determined the positions of H3K4me3-marked promoter-associated nucleosomes. Here we show that the transition from the maternal to zygotic transcriptome is characterized by a switch between two fundamentally different modes of defining transcription initiation, which drive the dynamic change of TSS usage and promoter shape. A maternal-specific TSS selection, which requires an A/T-rich (W-box) motif, is replaced with a zygotic TSS selection grammar characterized by broader patterns of dinucleotide enrichments, precisely aligned with the first downstream (+1) nucleosome. The developmental dynamics of the H3K4me3-marked nucleosomes reveal their DNA-sequence-associated positioning at promoters before zygotic transcription and subsequent transcription-independent adjustment to the final position downstream of the zygotic TSS. The two TSS-defining grammars coexist, often physically overlapping, in core promoters of constitutively expressed genes to enable

  12. Two independent transcription initiation codes overlap on vertebrate core promoters

    NASA Astrophysics Data System (ADS)

    Haberle, Vanja; Li, Nan; Hadzhiev, Yavor; Plessy, Charles; Previti, Christopher; Nepal, Chirag; Gehrig, Jochen; Dong, Xianjun; Akalin, Altuna; Suzuki, Ana Maria; van Ijcken, Wilfred F. J.; Armant, Olivier; Ferg, Marco; Strähle, Uwe; Carninci, Piero; Müller, Ferenc; Lenhard, Boris

    2014-03-01

    A core promoter is a stretch of DNA surrounding the transcription start site (TSS) that integrates regulatory inputs and recruits general transcription factors to initiate transcription. The nature and causative relationship of the DNA sequence and chromatin signals that govern the selection of most TSSs by RNA polymerase II remain unresolved. Maternal to zygotic transition represents the most marked change of the transcriptome repertoire in the vertebrate life cycle. Early embryonic development in zebrafish is characterized by a series of transcriptionally silent cell cycles regulated by inherited maternal gene products: zygotic genome activation commences at the tenth cell cycle, marking the mid-blastula transition. This transition provides a unique opportunity to study the rules of TSS selection and the hierarchy of events linking transcription initiation with key chromatin modifications. We analysed TSS usage during zebrafish early embryonic development at high resolution using cap analysis of gene expression, and determined the positions of H3K4me3-marked promoter-associated nucleosomes. Here we show that the transition from the maternal to zygotic transcriptome is characterized by a switch between two fundamentally different modes of defining transcription initiation, which drive the dynamic change of TSS usage and promoter shape. A maternal-specific TSS selection, which requires an A/T-rich (W-box) motif, is replaced with a zygotic TSS selection grammar characterized by broader patterns of dinucleotide enrichments, precisely aligned with the first downstream (+1) nucleosome. The developmental dynamics of the H3K4me3-marked nucleosomes reveal their DNA-sequence-associated positioning at promoters before zygotic transcription and subsequent transcription-independent adjustment to the final position downstream of the zygotic TSS. The two TSS-defining grammars coexist, often physically overlapping, in core promoters of constitutively expressed genes to enable

  13. Nuclear factor Y regulates ancient budgerigar hepadnavirus core promoter activity.

    PubMed

    Shen, Zhongliang; Liu, Yanfeng; Luo, Mengjun; Wang, Wei; Liu, Jing; Liu, Wei; Pan, Shaokun; Xie, Youhua

    2016-09-16

    Endogenous viral elements (EVE) in animal genomes are the fossil records of ancient viruses and provide invaluable information on the origin and evolution of extant viruses. Extant hepadnaviruses include avihepadnaviruses of birds and orthohepadnaviruses of mammals. The core promoter (Cp) of hepadnaviruses is vital for viral gene expression and replication. We previously identified in the budgerigar genome two EVEs that contain the full-length genome of an ancient budgerigar hepadnavirus (eBHBV1 and eBHBV2). Here, we found eBHBV1 Cp and eBHBV2 Cp were active in several human and chicken cell lines. A region from nt -85 to -11 in eBHBV1 Cp was critical for the promoter activity. Bioinformatic analysis revealed a putative binding site of nuclear factor Y (NF-Y), a ubiquitous transcription factor, at nt -64 to -50 in eBHBV1 Cp. The NF-Y core binding site (ATTGG, nt -58 to -54) was essential for eBHBV1 Cp activity. The same results were obtained with eBHBV2 Cp and duck hepatitis B virus Cp. The subunit A of NF-Y (NF-YA) was recruited via the NF-Y core binding site to eBHBV1 Cp and upregulated the promoter activity. Finally, the NF-Y core binding site is conserved in the Cps of all the extant avihepadnaviruses but not of orthohepadnaviruses. Interestingly, a putative and functionally important NF-Y core binding site is located at nt -21 to -17 in the Cp of human hepatitis B virus. In conclusion, our findings have pinpointed an evolutionary conserved and functionally critical NF-Y binding element in the Cps of avihepadnaviruses.

  14. NC2 mobilizes TBP on core promoter TATA boxes.

    PubMed

    Schluesche, Peter; Stelzer, Gertraud; Piaia, Elisa; Lamb, Don C; Meisterernst, Michael

    2007-12-01

    The general transcription factors (GTFs) of eukaryotic RNA polymerase II, in a process facilitated by regulatory and accessory factors, target promoters through synergistic interactions with core elements. The specific binding of the TATA box-binding protein (TBP) to the TATA box has led to the assumption that GTFs recognize promoters directly, producing a preinitiation complex at a defined position. Using biochemical analysis as well as biophysical single-pair Förster resonance energy transfer, we now provide evidence that negative cofactor-2 (NC2) induces dynamic conformational changes in the TBP-DNA complex that allow it to escape and return to TATA-binding mode. This can lead to movement of TBP along the DNA away from TATA.

  15. Regulatory Enhancer-Core-Promoter Communication via Transcription Factors and Cofactors.

    PubMed

    Zabidi, Muhammad A; Stark, Alexander

    2016-12-01

    Gene expression is regulated by genomic enhancers that recruit transcription factors and cofactors to activate transcription from target core promoters. Over the past years, thousands of enhancers and core promoters in animal genomes have been annotated, and we have learned much about the domain structure in which regulatory genomes are organized in animals. Enhancer-core-promoter targeting occurs at several levels, including regulatory domains, DNA accessibility, and sequence-encoded core-promoter specificities that are likely mediated by different regulatory proteins. We review here current knowledge about enhancer-core-promoter targeting, regulatory communication between enhancers and core promoters, and the protein factors involved. We conclude with an outlook on open questions that we find particularly interesting and that will likely lead to additional insights in the upcoming years.

  16. Core promoter functions in the regulation of gene expression of Drosophila dorsal target genes.

    PubMed

    Zehavi, Yonathan; Kuznetsov, Olga; Ovadia-Shochat, Avital; Juven-Gershon, Tamar

    2014-04-25

    Developmental processes are highly dependent on transcriptional regulation by RNA polymerase II. The RNA polymerase II core promoter is the ultimate target of a multitude of transcription factors that control transcription initiation. Core promoters consist of core promoter motifs, e.g. the initiator, TATA box, and the downstream core promoter element (DPE), which confer specific properties to the core promoter. Here, we explored the importance of core promoter functions in the dorsal-ventral developmental gene regulatory network. This network includes multiple genes that are activated by different nuclear concentrations of Dorsal, an NFκB homolog transcription factor, along the dorsal-ventral axis. We show that over two-thirds of Dorsal target genes contain DPE sequence motifs, which is significantly higher than the proportion of DPE-containing promoters in Drosophila genes. We demonstrate that multiple Dorsal target genes are evolutionarily conserved and functionally dependent on the DPE. Furthermore, we have analyzed the activation of key Dorsal target genes by Dorsal, as well as by another Rel family transcription factor, Relish, and the dependence of their activation on the DPE motif. Using hybrid enhancer-promoter constructs in Drosophila cells and embryo extracts, we have demonstrated that the core promoter composition is an important determinant of transcriptional activity of Dorsal target genes. Taken together, our results provide evidence for the importance of core promoter composition in the regulation of Dorsal target genes.

  17. Core promoter specificities of the Sp1 and VP16 transcriptional activation domains.

    PubMed Central

    Emami, K H; Navarre, W W; Smale, S T

    1995-01-01

    The core promoter compositions of mammalian protein-coding genes are highly variable; some contain TATA boxes, some contain initiator (Inr) elements, and others contain both or neither of these basal elements. The underlying reason for this heterogeneity remains a mystery, as recent studies have suggested that TATA-containing and Inr-containing core promoters direct transcription initiation by similar mechanisms and respond similarly to a wide variety of upstream activators. To analyze in greater detail the influence of core promoter structure on transcriptional activation, we compared activation by GAL4-VP16 and Sp1 through synthetic core promoters containing a TATA box, an Inr, or both TATA and Inr. Striking differences were found between the two activators, most notably in the relative strengths of the TATA/Inr and Inr core promoters: the TATA/Inr promoter was much stronger than the Inr promoter when transcription was activated by GAL4-VP16, but the strengths of the two promoters were more comparable when transcription was activated by Sp1. To define the domains of Sp1 responsible for efficient activation through an Inr, several Sp1 deletion mutants were tested as GAL4 fusion proteins. The results reveal that the glutamine-rich activation domains, which previously were found to interact with Drosophila TAF110, preferentially stimulate Inr-containing core promoters. In contrast, efficient activation through TATA appears to require additional domains of Sp1. These results demonstrate that activation domains differ in their abilities to function with specific core promoters, suggesting that the core promoter structure found in a given gene may reflect a preference of the regulators of that gene. Furthermore, the core promoter preference of an activation domain may be related to a specific mechanism of action, which may provide a functional criterion for grouping activation domains into distinct classes. PMID:7565743

  18. The CompHP Core Competencies Framework for Health Promotion in Europe

    ERIC Educational Resources Information Center

    Barry, Margaret M.; Battel-Kirk, Barbara; Dempsey, Colette

    2012-01-01

    Background: The CompHP Project on Developing Competencies and Professional Standards for Health Promotion in Europe was developed in response to the need for new and changing health promotion competencies to address health challenges. This article presents the process of developing the CompHP Core Competencies Framework for Health Promotion across…

  19. Synthetic Core Promoters as Universal Parts for Fine-Tuning Expression in Different Yeast Species

    PubMed Central

    2016-01-01

    Synthetic biology and metabolic engineering experiments frequently require the fine-tuning of gene expression to balance and optimize protein levels of regulators or metabolic enzymes. A key concept of synthetic biology is the development of modular parts that can be used in different contexts. Here, we have applied a computational multifactor design approach to generate de novo synthetic core promoters and 5′ untranslated regions (UTRs) for yeast cells. In contrast to upstream cis-regulatory modules (CRMs), core promoters are typically not subject to specific regulation, making them ideal engineering targets for gene expression fine-tuning. 112 synthetic core promoter sequences were designed on the basis of the sequence/function relationship of natural core promoters, nucleosome occupancy and the presence of short motifs. The synthetic core promoters were fused to the Pichia pastoris AOX1 CRM, and the resulting activity spanned more than a 200-fold range (0.3% to 70.6% of the wild type AOX1 level). The top-ten synthetic core promoters with highest activity were fused to six additional CRMs (three in P. pastoris and three in Saccharomyces cerevisiae). Inducible CRM constructs showed significantly higher activity than constitutive CRMs, reaching up to 176% of natural core promoters. Comparing the activity of the same synthetic core promoters fused to different CRMs revealed high correlations only for CRMs within the same organism. These data suggest that modularity is maintained to some extent but only within the same organism. Due to the conserved role of eukaryotic core promoters, this rational design concept may be transferred to other organisms as a generic engineering tool. PMID:27973777

  20. Synthetic Core Promoters as Universal Parts for Fine-Tuning Expression in Different Yeast Species.

    PubMed

    Portela, Rui M C; Vogl, Thomas; Kniely, Claudia; Fischer, Jasmin E; Oliveira, Rui; Glieder, Anton

    2017-03-17

    Synthetic biology and metabolic engineering experiments frequently require the fine-tuning of gene expression to balance and optimize protein levels of regulators or metabolic enzymes. A key concept of synthetic biology is the development of modular parts that can be used in different contexts. Here, we have applied a computational multifactor design approach to generate de novo synthetic core promoters and 5' untranslated regions (UTRs) for yeast cells. In contrast to upstream cis-regulatory modules (CRMs), core promoters are typically not subject to specific regulation, making them ideal engineering targets for gene expression fine-tuning. 112 synthetic core promoter sequences were designed on the basis of the sequence/function relationship of natural core promoters, nucleosome occupancy and the presence of short motifs. The synthetic core promoters were fused to the Pichia pastoris AOX1 CRM, and the resulting activity spanned more than a 200-fold range (0.3% to 70.6% of the wild type AOX1 level). The top-ten synthetic core promoters with highest activity were fused to six additional CRMs (three in P. pastoris and three in Saccharomyces cerevisiae). Inducible CRM constructs showed significantly higher activity than constitutive CRMs, reaching up to 176% of natural core promoters. Comparing the activity of the same synthetic core promoters fused to different CRMs revealed high correlations only for CRMs within the same organism. These data suggest that modularity is maintained to some extent but only within the same organism. Due to the conserved role of eukaryotic core promoters, this rational design concept may be transferred to other organisms as a generic engineering tool.

  1. Polymorphic core promoter GA-repeats alter gene expression of the early embryonic developmental genes.

    PubMed

    Valipour, E; Kowsari, A; Bayat, H; Banan, M; Kazeminasab, S; Mohammadparast, S; Ohadi, M

    2013-12-01

    Protein complexes that bind to 'GAGA' DNA elements are necessary to replace nucleosomes to create a local chromatin environment that facilitates a variety of site-specific regulatory responses. Three to four elements are required for the disruption of a preassembled nucleosome. We have previously identified human protein-coding gene core promoters that are composed of exceptionally long GA-repeats. The functional implication of those GA-repeats is beginning to emerge in the core promoter of the human SOX5 gene, which is involved in multiple developmental processes. In the current study, we analyze the functional implication of GA-repeats in the core promoter of two additional genes, MECOM and GABRA3, whose expression is largely limited to embryogenesis. We report a significant difference in gene expression as a result of different alleles across those core promoters in the HEK-293 cell line. Across-species homology check for the GABRA3 GA-repeats revealed that those repeats are evolutionary conserved in mouse and primates (p<1 × 10(-8)). The MECOM core promoter GA-repeats are also conserved in numerous species, of which human has the longest repeat and complexity. We propose a novel role for GA-repeat core promoters to regulate gene expression in the genes involved in development and evolution.

  2. Identification of the core sequence elements in Penaeus stylirostris densovirus promoters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This manuscript describes the role of different core elements in the transcriptional activity of promoters in a marine parvovirus, Penaeus stylirostris densovirus (PstDNV) that infects shrimp. Although comprehensive information on the role of different elements in the promoters of several animal par...

  3. Allosteric transcriptional regulation via changes in the overall topology of the core promoter

    DOE PAGES

    Philips, Steven J.; Canalizo-Hernandez, Monica; Yildirim, Ilyas; ...

    2015-08-21

    Many transcriptional activators act at a distance from core promoter elements and work by recruiting RNA polymerase through protein-protein interactions. We show here how the prokaryotic regulatory protein CueR both represses and activates transcription by differentially modulating local DNA structure within the promoter. Structural studies reveal that the repressor state slightly bends the promoter DNA, precluding optimal RNA polymerase-promoter recognition. Upon binding a metal ion in the allosteric site, CueR switches into an activator conformation. It maintains all protein-DNA contacts but introduces torsional stresses that kink and undertwist the promoter, stabilizing an A-DNA-like conformation. Finally, these factors switch on andmore » off transcription by exerting dynamic control of DNA stereochemistry, reshaping the core promoter and making it a better or worse substrate for polymerase.« less

  4. TRANSCRIPTION. Allosteric transcriptional regulation via changes in the overall topology of the core promoter.

    PubMed

    Philips, Steven J; Canalizo-Hernandez, Monica; Yildirim, Ilyas; Schatz, George C; Mondragón, Alfonso; O'Halloran, Thomas V

    2015-08-21

    Many transcriptional activators act at a distance from core promoter elements and work by recruiting RNA polymerase through protein-protein interactions. We show here how the prokaryotic regulatory protein CueR both represses and activates transcription by differentially modulating local DNA structure within the promoter. Structural studies reveal that the repressor state slightly bends the promoter DNA, precluding optimal RNA polymerase-promoter recognition. Upon binding a metal ion in the allosteric site, CueR switches into an activator conformation. It maintains all protein-DNA contacts but introduces torsional stresses that kink and undertwist the promoter, stabilizing an A-form DNA-like conformation. These factors switch on and off transcription by exerting dynamic control of DNA stereochemistry, reshaping the core promoter and making it a better or worse substrate for polymerase.

  5. Allosteric transcriptional regulation via changes in the overall topology of the core promoter

    SciTech Connect

    Philips, Steven J.; Canalizo-Hernandez, Monica; Yildirim, Ilyas; Schatz, George C.; Mondragon, Alfonso; O'Halloran, Thomas V.

    2015-08-21

    Many transcriptional activators act at a distance from core promoter elements and work by recruiting RNA polymerase through protein-protein interactions. We show here how the prokaryotic regulatory protein CueR both represses and activates transcription by differentially modulating local DNA structure within the promoter. Structural studies reveal that the repressor state slightly bends the promoter DNA, precluding optimal RNA polymerase-promoter recognition. Upon binding a metal ion in the allosteric site, CueR switches into an activator conformation. It maintains all protein-DNA contacts but introduces torsional stresses that kink and undertwist the promoter, stabilizing an A-DNA-like conformation. Finally, these factors switch on and off transcription by exerting dynamic control of DNA stereochemistry, reshaping the core promoter and making it a better or worse substrate for polymerase.

  6. Detection of deep stratospheric intrusions by cosmogenic 35S

    PubMed Central

    Su, Lin; Shaheen, Robina; Fung, Jimmy C. H.; Thiemens, Mark H.

    2016-01-01

    The extent to which stratospheric intrusions on synoptic scales influence the tropospheric ozone (O3) levels remains poorly understood, because quantitative detection of stratospheric air has been challenging. Cosmogenic 35S mainly produced in the stratosphere has the potential to identify stratospheric air masses at ground level, but this approach has not yet been unambiguously shown. Here, we report unusually high 35S concentrations (7,390 atoms m−3; ∼16 times greater than annual average) in fine sulfate aerosols (aerodynamic diameter less than 0.95 µm) collected at a coastal site in southern California on May 3, 2014, when ground-level O3 mixing ratios at air quality monitoring stations across southern California (43 of 85) exceeded the recently revised US National Ambient Air Quality Standard (daily maximum 8-h average: 70 parts per billion by volume). The stratospheric origin of the significantly enhanced 35S level is supported by in situ measurements of air pollutants and meteorological variables, satellite observations, meteorological analysis, and box model calculations. The deep stratospheric intrusion event was driven by the coupling between midlatitude cyclones and Santa Ana winds, and it was responsible for the regional O3 pollution episode. These results provide direct field-based evidence that 35S is an additional sensitive and unambiguous tracer in detecting stratospheric air in the boundary layer and offer the potential for resolving the stratospheric influences on the tropospheric O3 level. PMID:27655890

  7. Detection of deep stratospheric intrusions by cosmogenic 35S.

    PubMed

    Lin, Mang; Su, Lin; Shaheen, Robina; Fung, Jimmy C H; Thiemens, Mark H

    2016-10-04

    The extent to which stratospheric intrusions on synoptic scales influence the tropospheric ozone (O3) levels remains poorly understood, because quantitative detection of stratospheric air has been challenging. Cosmogenic (35)S mainly produced in the stratosphere has the potential to identify stratospheric air masses at ground level, but this approach has not yet been unambiguously shown. Here, we report unusually high (35)S concentrations (7,390 atoms m(-3); ∼16 times greater than annual average) in fine sulfate aerosols (aerodynamic diameter less than 0.95 µm) collected at a coastal site in southern California on May 3, 2014, when ground-level O3 mixing ratios at air quality monitoring stations across southern California (43 of 85) exceeded the recently revised US National Ambient Air Quality Standard (daily maximum 8-h average: 70 parts per billion by volume). The stratospheric origin of the significantly enhanced (35)S level is supported by in situ measurements of air pollutants and meteorological variables, satellite observations, meteorological analysis, and box model calculations. The deep stratospheric intrusion event was driven by the coupling between midlatitude cyclones and Santa Ana winds, and it was responsible for the regional O3 pollution episode. These results provide direct field-based evidence that (35)S is an additional sensitive and unambiguous tracer in detecting stratospheric air in the boundary layer and offer the potential for resolving the stratospheric influences on the tropospheric O3 level.

  8. Detection of deep stratospheric intrusions by cosmogenic 35S

    NASA Astrophysics Data System (ADS)

    Lin, Mang; Su, Lin; Shaheen, Robina; Fung, Jimmy C. H.; Thiemens, Mark H.

    2016-10-01

    The extent to which stratospheric intrusions on synoptic scales influence the tropospheric ozone (O3) levels remains poorly understood, because quantitative detection of stratospheric air has been challenging. Cosmogenic 35S mainly produced in the stratosphere has the potential to identify stratospheric air masses at ground level, but this approach has not yet been unambiguously shown. Here, we report unusually high 35S concentrations (7,390 atoms m-3; ˜16 times greater than annual average) in fine sulfate aerosols (aerodynamic diameter less than 0.95 µm) collected at a coastal site in southern California on May 3, 2014, when ground-level O3 mixing ratios at air quality monitoring stations across southern California (43 of 85) exceeded the recently revised US National Ambient Air Quality Standard (daily maximum 8-h average: 70 parts per billion by volume). The stratospheric origin of the significantly enhanced 35S level is supported by in situ measurements of air pollutants and meteorological variables, satellite observations, meteorological analysis, and box model calculations. The deep stratospheric intrusion event was driven by the coupling between midlatitude cyclones and Santa Ana winds, and it was responsible for the regional O3 pollution episode. These results provide direct field-based evidence that 35S is an additional sensitive and unambiguous tracer in detecting stratospheric air in the boundary layer and offer the potential for resolving the stratospheric influences on the tropospheric O3 level.

  9. Evolutionary trend of exceptionally long human core promoter short tandem repeats.

    PubMed

    Ohadi, M; Mohammadparast, S; Darvish, H

    2012-10-01

    Short tandem repeats (STRs) are variable elements that play a significant role in genome evolution by creating and maintaining quantitative genetic variation. Because of their proximity to the +1 transcription start site (TSS) and polymorphic nature, core promoter STRs may be considered a novel source of variation across species. In a genome-scale analysis of the entire human protein-coding genes annotated in the GeneCards database (19,927), we analyze the prevalence and repeat numbers of different classes of core promoter STRs in the interval between -120 and +1 to the TSS. We also analyze the evolutionary trend of exceptionally long core promoter STRs of ≥6-repeats. 133 genes (~2%) had core promoter STRs of ≥6-repeats. In the majority of those genes, the STR motifs were found to be conserved across evolution. Di-nucleotide repeats had the highest representation in the human core promoter long STRs (72 genes). Tri- (52 genes), tetra-, penta-, and hexa-nucleotide STRs (9 genes) were also present in the descending prevalence. The majority of those genes (84 genes) revealed directional expansion of core promoter STRs from mouse to human. However, in a number of genes, the difference in average allele size across species was sufficiently small that there might be a constraint on the evolution of average allele size. Random drift of STRs from mouse to human was also observed in a minority of genes. Future work on the genes listed in the current study may further our knowledge into the potential importance of core promoter STRs in human evolution.

  10. Enhancer-core-promoter specificity separates developmental and housekeeping gene regulation.

    PubMed

    Zabidi, Muhammad A; Arnold, Cosmas D; Schernhuber, Katharina; Pagani, Michaela; Rath, Martina; Frank, Olga; Stark, Alexander

    2015-02-26

    Gene transcription in animals involves the assembly of RNA polymerase II at core promoters and its cell-type-specific activation by enhancers that can be located more distally. However, how ubiquitous expression of housekeeping genes is achieved has been less clear. In particular, it is unknown whether ubiquitously active enhancers exist and how developmental and housekeeping gene regulation is separated. An attractive hypothesis is that different core promoters might exhibit an intrinsic specificity to certain enhancers. This is conceivable, as various core promoter sequence elements are differentially distributed between genes of different functions, including elements that are predominantly found at either developmentally regulated or at housekeeping genes. Here we show that thousands of enhancers in Drosophila melanogaster S2 and ovarian somatic cells (OSCs) exhibit a marked specificity to one of two core promoters--one derived from a ubiquitously expressed ribosomal protein gene and another from a developmentally regulated transcription factor--and confirm the existence of these two classes for five additional core promoters from genes with diverse functions. Housekeeping enhancers are active across the two cell types, while developmental enhancers exhibit strong cell-type specificity. Both enhancer classes differ in their genomic distribution, the functions of neighbouring genes, and the core promoter elements of these neighbouring genes. In addition, we identify two transcription factors--Dref and Trl--that bind and activate housekeeping versus developmental enhancers, respectively. Our results provide evidence for a sequence-encoded enhancer-core-promoter specificity that separates developmental and housekeeping gene regulatory programs for thousands of enhancers and their target genes across the entire genome.

  11. Core promoter analysis of porcine Six1 gene and its regulation of the promoter activity by CpG methylation.

    PubMed

    Wu, Wangjun; Ren, Zhuqing; Liu, Honglin; Wang, Linjie; Huang, Ruihua; Chen, Jie; Zhang, Lin; Li, Pinghua; Xiong, Yuanzhu

    2013-10-25

    Six1, an evolutionary conserved transcription factor, has been shown to play an important role in organogenesis and diseases. However, no reports were shown to investigate its transcriptional regulatory mechanisms. In the present study, we first identified porcine Six1 gene core promoter region (+170/-360) using luciferase reporter assay system and found that promoter activities were significantly higher in the mouse myoblast C2C12 cells than that in the mouse fibroblast C3H10T1/2 cells, implying that Six1 promoter could possess muscle-specific characteristics. Moreover, our results showed that promoter activities of Six1 were decreased as induction of differentiation of C2C12 cells, which was accompanied by the down-regulation of mRNA expression of Six1 gene. In addition, we found that the DNA methylation of Six1 promoters in vitro obviously influences the promoter activities and the DNA methylation level of Six1 promoter core region was negatively correlated to Six1 gene expression in vivo. Taken together, we preliminarily clarified transcriptional regulatory mechanisms of Six1 gene, which should be useful for investigating its subtle transcriptional regulatory mechanisms in the future. On the other hand, based on Six1 involved in tumorigenesis, our data also provide a genetic foundation to control the generation of diseases via pursuing Six1 as therapeutic target gene.

  12. Understanding Antarctic sulfur cycle chemistry using cosmogenic 35S

    NASA Astrophysics Data System (ADS)

    Hill-Falkenthal, J.; Priyadarshi, A.; Savarino, J. P.; Thiemens, M. H.

    2011-12-01

    Sulfate aerosols have been recognized to possess strong light scattering abilities and act as cloud condensation nuclei, thus having an impact on Earth's climate and radiation budget. Improved understanding of the sulfate aerosol transport is needed for assessing its influences on climate. Cosmogenically produced 35S (half-life~87 days)(2) exists both in the gas and solid phases, thus making it ideal to trace the atmospheric processes of sulfate oxidation. Here, we present a yearlong sampling of sulfate aerosol in Antarctica with 35S measurements illustrating its boundary layer chemistry and stratospheric- tropospheric mixing. Samples were collected from Dome C station once a week from Jan 2010-Jan 2011. 35S activity in sulfate aerosols shows maximums in summer months between December and February and minimums in winter from June to August. Higher oxidative capacity of the atmosphere coupled with long range transport of mid-latitude air increases 35SO4 activity in the summer, whereas a lack of air mass mixing coupled with low oxidant concentration significantly decreases 35SO4 activity(1). Stratospheric/tropospheric exchange processes like tropopause folding could help explain a random spike in activity that deviates from the normal background activity. In the future, a box model calculation will be done to determine the contribution of stratospheric air mass transported downward during the exchange. The oxygen isotopes will also be measured to see the effect of stratospheric intrusion. References (1)Priyadarshi, A., G. Dominguez, J. Savarino, and M. Thiemens (2011), Cosmogenic 35S: A unique tracer to Antarctic atmospheric chemistry and the polar vortex, Geophys. Res. Lett., 38, L13808, doi:10.1029/2011/GL047469. (2)Lal, D., and B. Peters (1967), Cosmic ray produced radioactivity in the earth, Handb. Phys., 46, 551-612.

  13. Analytical method for measuring cosmogenic 35S in natural waters

    DOE PAGES

    Uriostegui, Stephanie H.; Bibby, Richard K.; Esser, Bradley K.; ...

    2015-05-18

    Here, cosmogenic sulfur-35 in water as dissolved sulfate (35SO4) has successfully been used as an intrinsic hydrologic tracer in low-SO4, high-elevation basins. Its application in environmental waters containing high SO4 concentrations has been limited because only small amounts of SO4 can be analyzed using current liquid scintillation counting (LSC) techniques. We present a new analytical method for analyzing large amounts of BaSO4 for 35S. We quantify efficiency gains when suspending BaSO4 precipitate in Inta-Gel Plus cocktail, purify BaSO4 precipitate to remove dissolved organic matter, mitigate interference of radium-226 and its daughter products by selection of high purity barium chloride, andmore » optimize LSC counting parameters for 35S determination in larger masses of BaSO4. Using this improved procedure, we achieved counting efficiencies that are comparable to published LSC techniques despite a 10-fold increase in the SO4 sample load. 35SO4 was successfully measured in high SO4 surface waters and groundwaters containing low ratios of 35S activity to SO4 mass demonstrating that this new analytical method expands the analytical range of 35SO4 and broadens the utility of 35SO4 as an intrinsic tracer in hydrologic settings.« less

  14. PARTNERSHIP BETWEEN CTSI AND BUSINESS SCHOOLS CAN PROMOTE BEST PRACTICES FOR CORE FACILITIES AND RESOURCES

    PubMed Central

    Reeves, Lilith; Dunn-Jensen, Linda M.; Baldwin, Timothy T.; Tatikonda, Mohan V.; Cornetta, Kenneth

    2013-01-01

    Biomedical research enterprises require a large number of core facilities and resources to supply the infrastructure necessary for translational research. Maintaining the financial viability and promoting efficiency in an academic environment can be particularly challenging for medical schools and universities. The Indiana Clinical and Translational Sciences Institute sought to improve core and service programs through a partnership with the Indiana University Kelley School of Business. The program paired teams of Masters of Business Administration students with cores and programs that self-identified the need for assistance in project management, financial management, marketing, or resource efficiency. The projects were developed by CTSI project managers and business school faculty using service-learning principles to ensure learning for students who also received course credit for their participation. With three years of experience, the program demonstrates a successful partnership that improves clinical research infrastructure by promoting business best practices and providing a valued learning experience for business students. PMID:23919365

  15. Partnership between CTSI and business schools can promote best practices for core facilities and resources.

    PubMed

    Reeves, Lilith; Dunn-Jensen, Linda M; Baldwin, Timothy T; Tatikonda, Mohan V; Cornetta, Kenneth

    2013-08-01

    Biomedical research enterprises require a large number of core facilities and resources to supply the infrastructure necessary for translational research. Maintaining the financial viability and promoting efficiency in an academic environment can be particularly challenging for medical schools and universities. The Indiana Clinical and Translational Sciences Institute sought to improve core and service programs through a partnership with the Indiana University Kelley School of Business. The program paired teams of Masters of Business Administration students with cores and programs that self-identified the need for assistance in project management, financial management, marketing, or resource efficiency. The projects were developed by CTSI project managers and business school faculty using service-learning principles to ensure learning for students who also received course credit for their participation. With three years of experience, the program demonstrates a successful partnership that improves clinical research infrastructure by promoting business best practices and providing a valued learning experience for business students.

  16. Optimised photocatalytic hydrogen production using core-shell AuPd promoters with controlled shell thickness.

    PubMed

    Jones, Wilm; Su, Ren; Wells, Peter P; Shen, Yanbin; Dimitratos, Nikolaos; Bowker, Michael; Morgan, David; Iversen, Bo B; Chutia, Arunabhiram; Besenbacher, Flemming; Hutchings, Graham

    2014-12-28

    The development of efficient photocatalytic routines for producing hydrogen is of great importance as society moves away from energy sources derived from fossil fuels. Recent studies have identified that the addition of metal nanoparticles to TiO2 greatly enhances the photocatalytic performance of these materials towards the reforming of alcohols for hydrogen production. The core-shell structured Au-Pd bimetallic nanoparticle supported on TiO2 has being of interest as it exhibited extremely high quantum efficiencies for hydrogen production. However, the effect of shell composition and thickness on photocatalytic performance remains unclear. Here we report the synthesis of core-shell structured AuPd NPs with the controlled deposition of one and two monolayers (ML) equivalent of Pd onto Au NPs by colloidal and photodeposition methods. We have determined the shell composition and thickness of the nanoparticles by a combination of X-ray absorption fine structure and X-ray photoelectron spectroscopy. Photocatalytic ethanol reforming showed that the core-shell structured Au-Pd promoters supported on TiO2 exhibit enhanced activity compared to that of monometallic Au and Pd as promoters, whilst the core-shell Au-Pd promoters containing one ML equivalent Pd provide the optimum reactivity.

  17. Variations in the core promoter/pre-core region in HBV genotype C in Japanese and Northern Vietnamese patients.

    PubMed

    Truong, Bui Xuan; Yano, Yoshihiko; Seo, Yasushi; Phuong, Tran Minh; Tanaka, Yasuhito; Kato, Hirotaka; Miki, Akira; Utsumi, Takako; Azuma, Takeshi; Trach, Nguyen Khanh; Mizokami, Masashi; Hayashi, Yoshitake; Kasuga, Masato

    2007-09-01

    Hepatitis B virus (HBV) subgenotypes Cs (C1) and Ce (C2) are common in East Asia. To investigate the genomic difference of HBV genotype C between two separated regions, 50 subgenotype Cs-infected Vietnamese and 70 subgenotype Ce-infected Japanese patients were enrolled for analysis. The patients were categorized to either a hepatocellular carcinoma group (HCC) or a non-HCC group including liver cirrhosis, chronic hepatitis, and asymptomatic carriers. HBV serology, HBV-DNA level, and variations in core promoter/pre-core region were examined. Phylogenetic analysis based on the full genome sequences and nucleotide sequences partly in the S gene and in the P gene revealed that all Japanese strains (70/70) were subgenotype Ce, and nearly all of the Vietnamese strains (50/51) were subgenotype Cs, excluding one subgenotype C5. C1858 and G1775 were common in the Vietnamese (64% and 40%) but not in the Japanese (0%). The prevalence of C/A1753 in Vietnamese was higher than that in the Japanese (32% vs. 17.1%), however the frequency of A1896 in the Japanese was significantly higher (32.9% vs. 12%, P < 0.05). Most of the Vietnamese patients with HCC had a high level of HBV-DNA, the Japanese HCC had a relatively low level. In the Vietnamese, C/A1753 and C1858 were associated closely with T1762A1764, higher HBV-DNA levels and higher HCC incidence. The multivariate analysis revealed that male, T1653 and C/A1753 were independent risk factors for HCC. The subgenotypes and unique mutations of HBV genotype C in the Vietnamese and Japanese differed, and C/A1753 and C1858 variants might play a role in the pathogenesis of liver disease in Vietnamese patients.

  18. Mutations in pre-core and basic core promoter regions of hepatitis B virus in chronic hepatitis B patients

    PubMed Central

    Wang, Xiao-Ling; Ren, Jian-Ping; Wang, Xue-Qing; Wang, Xiao-Hong; Yang, Shao-Fang; Xiong, Yi

    2016-01-01

    AIM: To investigate the frequency of mutations in pre-core (pre-C) and basic core promoter (BCP) regions of hepatitis B virus (HBV) from Shanxi Province, and the association between mutations and disease related indexes. METHODS: One hundred chronic hepatitis B patients treated at Shanxi Province Hospital of Traditional Chinese Medicine were included in this study. PCR-reverse dot blot hybridization and mismatch amplification mutation assay (MAMA)-PCR were used to detect the mutations in the HBV pre-C and BCP regions. HBV DNA content and liver function were compared between patients with mutant HBV pre-C and BCP loci and those with wild-type loci. The consistency between PCR-reverse dot blot hybridization and MAMA-PCR for detecting mutations in the HBV pre-C and BCP regions was assessed. RESULTS: Of the 100 serum samples detected, 9.38% had single mutations in the pre-C region, 29.17% had single mutations in the BCP region, 41.67% had mutations in both BCP and pre-C regions, and 19.79% had wild-type loci. The rates of BCP and pre-C mutations were 65.7% and 34.3%, respectively, in hepatitis B e antigen (HBeAg) positive patients, and 84.6% and 96.2%, respectively, in HBeAg negative patients. The rate of pre-C mutations was significantly higher in HBeAg negative patients than in HBeAg positive patients (χ2 = 26.62, P = 0.00), but there was no significant difference in the distribution of mutations in the BCP region between HBeAg positive and negative patients (χ2 = 2.43, P = 0.12). The presence of mutations in the pre-C (Wilcoxon W = 1802.5, P = 0.00) and BCP regions (Wilcoxon W = 2906.5, P = 0.00) was more common in patients with low HBV DNA content. Both AST and GGT were significantly higher in patients with mutant pre-C and BCP loci than in those with wild-type loci (P < 0.05). PCR-reverse dot blot hybridization and MAMA-PCR for detection of mutations in the BCP and pre-C regions had good consistency, and the Kappa values obtained were 0.91 and 0.58, respectively

  19. Development of vascular tissue and stress inducible hybrid-synthetic promoters through dof-1 motifs rearrangement.

    PubMed

    Ranjan, Rajiv; Dey, Nrisingha

    2012-07-01

    A Caulimovirus-based hybrid-promoter, EFCFS, was derived by fusing the distal region (-227 to -54, FUAS) of Figwort mosaic virus full-length transcript promoter (F20) with the core promoter (-151 to +12, FS3CP) domain of Figwort mosaic virus sub-genomic transcript promoter (FS3). The hybrid-promoter (EFCFS) showed enhanced activity compared to the CaMV35S, F20 and FS3 promoters; while it showed equivalent activity with that of the CAMV35S(2) promoter in both transient protoplast (Nicotiana tabacum cv. Xanthi Brad) and transgenic plants (Nicotiana tabacum; Samsun NN). Further, we have engineered the EFCFS promoter sequence by inserting additional copies of the stress-inducible 'AAAG' cis-motif (Dof-1) to generate a set of three hybrid-synthetic promoters namely; EFCFS-HS-1, EFCFS-HS-2 and EFCFS-HS-3-containing 10, 11 and 13 'AAAG' motif, respectively. Transgenic plants expressing these hybrid synthetic promoters coupled to the GUS reporter were developed and their transcriptional activities were compared with F20, FS3, 35S and 35S(2) promoters, respectively. The relative levels of uidA-mRNA accumulation in transgenic plants driven by above promoters individually were compared by qRT-PCR. Localization of GUS reporter activity in plant tissue was assayed by histochemical approach. CLSM-based study revealed that hybrid-synthetic promoters namely; EFCFS-HS-1, EFCFS-HS-2 and EFCFS-HS-3 showed enhanced activity in vascular tissue compared to the CaMV35S promoter. In the presence of abiotic stress elicitors, salicylic acid and jasmonic acid, the EFCFS-HS-1 promoters showed enhanced activity compared to the 35S promoter. Newly derived hybrid-synthetic promoter/s with enhanced activity and stress inducibility could become efficient tools for advancement of plant biotechnology.

  20. Genome-wide identification of human- and primate-specific core promoter short tandem repeats.

    PubMed

    Bushehri, A; Barez, M R Mashhoudi; Mansouri, S K; Biglarian, A; Ohadi, M

    2016-08-01

    Recent reports of a link between human- and primate-specific genetic factors and human/primate-specific characteristics and diseases necessitate genome-wide identification of those factors. We have previously reported core promoter short tandem repeats (STRs) of extreme length (≥6-repeats) that have expanded exceptionally in primates vs. non-primates, and may have a function in adaptive evolution. In the study reported here, we extended our study to the human STRs of ≥3-repeats in the category of penta and hexaucleotide STRs, across the entire human protein coding gene core promoters, and analyzed their status in several superorders and orders of vertebrates, using the Ensembl database. The ConSite software was used to identify the transcription factor (TF) sets binding to those STRs. STR specificity was observed at different levels of human and non-human primate (NHP) evolution. 73% of the pentanucleotide STRs and 68% of the hexanucleotide STRs were found to be specific to human and NHPs. AP-2alpha, Sp1, and MZF were the predominantly selected TFs (90%) binding to the human-specific STRs. Furthermore, the number of TF sets binding to a given STR was found to be a selection factor for that STR. Our findings indicate that selected STRs, the cognate binding TFs, and the number of TF set binding to those STRs function as switch codes at different levels of human and NHP evolution and speciation.

  1. Combined exposure to big endothelin-1 and mechanical loading in bovine sternal cores promotes osteogenesis.

    PubMed

    Meyer, Luisa A; Johnson, Michael G; Cullen, Diane M; Vivanco, Juan F; Blank, Robert D; Ploeg, Heidi-Lynn; Smith, Everett L

    2016-04-01

    Increased bone formation resulting from mechanical loading is well documented; however, the interactions of the mechanotransduction pathways are less well understood. Endothelin-1, a ubiquitous autocrine/paracrine signaling molecule promotes osteogenesis in metastatic disease. In the present study, it was hypothesized that exposure to big endothelin-1 (big ET1) and/or mechanical loading would promote osteogenesis in ex vivo trabecular bone cores. In a 2×2 factorial trial of daily mechanical loading (-2000με, 120cycles daily, "jump" waveform) and big ET1 (25ng/mL), 48 bovine sternal trabecular bone cores were maintained in bioreactor chambers for 23days. The bone cores' response to the treatment stimuli was assessed with percent change in core apparent elastic modulus (ΔEapp), static and dynamic histomorphometry, and prostaglandin E2 (PGE2) secretion. Two-way ANOVA with a post hoc Fisher's LSD test found no significant treatment effects on ΔEapp (p=0.25 and 0.51 for load and big ET1, respectively). The ΔEapp in the "no load + big ET1" (CE, 13±12.2%, p=0.56), "load + no big ET1" (LC, 17±3.9%, p=0.14) and "load + big ET1" (LE, 19±4.2%, p=0.13) treatment groups were not statistically different than the control group (CC, 3.3%±8.6%). Mineralizing surface (MS/BS), mineral apposition (MAR) and bone formation rates (BFR/BS) were significantly greater in LE than CC (p=0.037, 0.0040 and 0.019, respectively). While the histological bone formation markers in LC trended to be greater than CC (p=0.055, 0.11 and 0.074, respectively) there was no difference between CE and CC (p=0.61, 0.50 and 0.72, respectively). Cores in LE and LC had more than 50% greater MS/BS (p=0.037, p=0.055 respectively) and MAR (p=0.0040, p=0.11 respectively) than CC. The BFR/BS was more than two times greater in LE (p=0.019) and LC (p=0.074) than CC. The PGE2 levels were elevated at 8days post-osteotomy in all groups and the treatment groups remained elevated compared to the CC group on days 15

  2. Novel cofactors and TFIIA mediate functional core promoter selectivity by the human TAFII150-containing TFIID complex.

    PubMed

    Martinez, E; Ge, H; Tao, Y; Yuan, C X; Palhan, V; Roeder, R G

    1998-11-01

    TATA-binding protein-associated factors (TAFIIs) within TFIID control differential gene transcription through interactions with both activators and core promoter elements. In particular, TAFII150 contributes to initiator-dependent transcription through an unknown mechanism. Here, we address whether TAFIIs within TFIID are sufficient, in conjunction with highly purified general transcription factors (GTFs), for differential core promoter-dependent transcription by RNA polymerase II and whether additional cofactors are required. We identify the human homologue of Drosophila TAFII150 through cognate cDNA cloning and show that it is a tightly associated component of human TFIID. More importantly, we demonstrate that the human TAFII150-containing TFIID complex is not sufficient, in the context of all purified GTFs and RNA polymerase II, to mediate transcription synergism between TATA and initiator elements and initiator-directed transcription from a TAFII-dependent TATA-less promoter. Therefore, TAFII-promoter interactions are not sufficient for the productive core promoter-selective functions of TFIID. Consistent with this finding, we have partially purified novel cofactor activities (TICs) that potentiate the TAFII-mediated synergism between TATA and initiator elements (TIC-1) and TAFII-dependent transcription from TATA-less promoters (TIC-2 and -3). Furthermore, we demonstrate an essential function for TFIIA in TIC- and TAFII-dependent basal transcription from a TATA-less promoter. Our results reveal a parallel between the basal transcription activity of TAFIIs through core promoter elements and TAFII-dependent activator function.

  3. Development and Validation of a P-35S, T-nos, T-35S and P-FMV Tetraplex Real-time PCR Screening Method to Detect Regulatory Genes of Genetically Modified Organisms in Food.

    PubMed

    Eugster, Albert; Murmann, Petra; Kaenzig, Andre; Breitenmoser, Alda

    2014-10-01

    In routine analysis screening methods based on real-time PCR (polymerase chain reaction) are most commonly used for the detection of genetically modified (GM) plant material in food and feed. Screening tests are based on sequences frequently used for GM development, allowing the detection of a large number of GMOs (genetically modified organisms). Here, we describe the development and validation of a tetraplex real-time PCR screening assay comprising detection systems for the regulatory genes Cauliflower Mosaic Virus 35S promoter, Agrobacterium tumefaciens nos terminator, Cauliflower Mosaic Virus 35S terminator and Figwort Mosaic Virus 34S promoter. Three of the four primer and probe combinations have already been published elsewhere, whereas primers and probe for the 35S terminator have been developed in-house. Adjustment of primer and probe concentrations revealed a high PCR sensitivity with insignificant physical cross-talk between the four detection channels. The sensitivity of each PCR-system is sufficient to detect a GMO concentration as low as 0.05% of the containing respective element. The specificity of the described tetraplex is high when tested on DNA from GM maize, soy, rapeseed and tomato. We also demonstrate the robustness of the system by inter-laboratory tests. In conclusion, this method provides a sensitive and reliable screening procedure for the detection of the most frequently used regulatory elements present in GM crops either authorised or unauthorised for food.

  4. Ets transcription factors bind and transactivate the core promoter of the von Willebrand factor gene.

    PubMed

    Schwachtgen, J L; Janel, N; Barek, L; Duterque-Coquillaud, M; Ghysdael, J; Meyer, D; Kerbiriou-Nabias, D

    1997-12-18

    von Willebrand factor (vWF) gene expression is restricted to endothelial cells and megakaryocytes. Previous results demonstrated that basal transcription of the human vWF gene is mediated through a promoter located between base pairs -89 and +19 (cap site: +1) which is functional in endothelial and non endothelial cells. Two DNA repeats TTTCCTTT correlating with inverted consensus binding sites for the Ets family of transcription factors are present in the -56/-36 sequence. In order to analyse whether these DNA elements are involved in transcription, human umbilical vein endothelial cells (HUVEC), bovine calf pulmonary endothelial cell line (CPAE), HeLa and COS cells were transfected with constructs containing deletions of the -89/+19 fragment, linked to the chloramphenicol acetyl transferase (CAT) reporter gene. The -60/+19 region exhibits significant promoter activity in HUVEC and CPAE cells only. The -42/+19 fragment is not active. Mutations of the -60/+19 promoter fragment in the 5' (-56/-49) Ets binding site abolish transcription in endothelial cells whereas mutations in the 3' (-43/-36) site does not. The -60/-33 fragment forms three complexes with proteins from HUVEC nuclear extracts in electrophoretic mobility shift assay which are dependent on the presence of the 5' Ets binding site. Binding of recombinant Ets-1 protein to the -60/-33 fragment gives a complex which also depends on the 5' site. The -60/+19 vWF gene core promoter is transactivated in HeLa cells by cotransfecting with Ets-1 or Erg (Ets-related gene) expression plasmids. In contrast to the wild type construct, transcription of the 5' site mutants is not increased by these expressed proteins. The results indicate that the promoter activity of the -60/+19 region of the vWF gene depends on transcription factors of the Ets family of which several members like Ets-1, Ets-2 and Erg are expressed in endothelium. Cotransfection of Ets-1 and Erg expression plasmids is sufficient to induce the -60/+19 v

  5. Palindromic GOLGA8 core duplicons promote chromosome 15q13.3 microdeletion and evolutionary instability

    PubMed Central

    Antonacci, Francesca; Dennis, Megan Y.; Huddleston, John; Sudmant, Peter H.; Steinberg, Karyn Meltz; Rosenfeld, Jill A.; Miroballo, Mattia; Graves, Tina A.; Vives, Laura; Malig, Maika; Denman, Laura; Raja, Archana; Stuart, Andrew; Tang, Joyce; Munson, Brenton; Shaffer, Lisa G.; Amemiya, Chris T.; Wilson, Richard K.; Eichler, Evan E.

    2014-01-01

    Recurrent deletions of chromosome 15q13.3 associate with intellectual disability, schizophrenia, autism and epilepsy. To gain insight into its instability, we sequenced the region in patients, normal individuals and nonhuman primates. We discovered five structural configurations of the human chromosome 15q13.3 region ranging in size from 2 to 3 Mbp. These configurations arose recently (~0.5–0.9 million years ago) as a result of human-specific expansions of segmental duplications and two independent inversion events. All inversion breakpoints map near GOLGA8 core duplicons—a ~14 kbp primate-specific chromosome 15 repeat that became organized into larger palindromic structures. GOLGA8-flanked palindromes also demarcate the breakpoints of recurrent 15q13.3 microdeletions, the expansion of chromosome 15 segmental duplications in the human lineage, and independent structural changes in apes. The significant clustering (p=0.002) of breakpoints provides mechanistic evidence for the role of this core duplicon and its palindromic architecture in promoting evolutionary and disease-related instability of chromosome 15. PMID:25326701

  6. Hepatitis C Virus Core Protein Promotes miR-122 Destabilization by Inhibiting GLD-2

    PubMed Central

    Kim, Geon-Woo; Lee, Seung-Hoon; Cho, Hee; Kim, Minwoo; Shin, Eui-Cheol; Oh, Jong-Won

    2016-01-01

    The liver-specific microRNA miR-122, which has essential roles in liver development and metabolism, is a key proviral factor for hepatitis C virus (HCV). Despite its crucial role in the liver and HCV life cycle, little is known about the molecular mechanism of miR-122 expression regulation by HCV infection. Here, we show that the HCV core protein downregulates the abundance of miR-122 by promoting its destabilization via the inhibition of GLD-2, a non-canonical cytoplasmic poly(A) polymerase. The decrease in miR-122 expression resulted in the dysregulation of the known functions of miR-122, including its proviral activity for HCV. By high-throughput sequencing of small RNAs from human liver biopsies, we found that the 22-nucleotide (nt) prototype miR-122 is modified at its 3′ end by 3′-terminal non-templated and templated nucleotide additions. Remarkably, the proportion of miR-122 isomers bearing a single nucleotide tail of any ribonucleotide decreased in liver specimens from patients with HCV. We found that these single-nucleotide-tailed miR-122 isomers display increased miRNA activity and stability over the 22-nt prototype miR-122 and that the 3′-terminal extension is catalyzed by the unique terminal nucleotidyl transferase activity of GLD-2, which is capable of adding any single ribonucleotide without preference of adenylate to the miR-122 3′ end. The HCV core protein specifically inhibited GLD-2, and its interaction with GLD-2 in the cytoplasm was found to be responsible for miR-122 downregulation. Collectively, our results provide new insights into the regulatory role of the HCV core protein in controlling viral RNA abundance and miR-122 functions through miR-122 stability modulation. PMID:27366906

  7. LUC7L3/CROP inhibits replication of hepatitis B virus via suppressing enhancer II/basal core promoter activity

    PubMed Central

    Li, Yuan; Ito, Masahiko; Sun, Suofeng; Chida, Takeshi; Nakashima, Kenji; Suzuki, Tetsuro

    2016-01-01

    The core promoter of hepatitis B virus (HBV) genome is a critical region for transcriptional initiation of 3.5 kb, pregenome and precore RNAs and for the viral replication. Although a number of host-cell factors that potentially regulate the viral promoter activities have been identified, the molecular mechanisms of the viral gene expression, in particular, regulatory mechanisms of the transcriptional repression remain elusive. In this study, we identified LUC7 like 3 pre-mRNA splicing factor (LUC7L3, also known as hLuc7A or CROP) as a novel interacting partner of HBV enhancer II and basal core promoter (ENII/BCP), key elements within the core promoter, through the proteomic screening and found that LUC7L3 functions as a negative regulator of ENII/BCP. Gene silencing of LUC7L3 significantly increased expression of the viral genes and antigens as well as the activities of ENII/BCP and core promoter. In contrast, overexpression of LUC7L3 inhibited their activities and HBV replication. In addition, LUC7L3 possibly contributes to promotion of the splicing of 3.5 kb RNA, which may also be involved in negative regulation of the pregenome RNA level. This is the first to demonstrate the involvement of LUC7L3 in regulation of gene transcription and in viral replication. PMID:27857158

  8. Core-sigma interaction: probing the interaction of the bacteriophage T4 gene 55 promoter recognition protein with E.coli RNA polymerase core.

    PubMed Central

    Léonetti, J P; Wong, K; Geiduschek, E P

    1998-01-01

    The bacterial RNA polymerase sigma subunits are key participants in the early steps of RNA synthesis, conferring specificity of promoter recognition, facilitating promoter opening and promoter clearance, and responding to diverse transcriptional regulators. The T4 gene 55 protein (gp55), the sigma protein of the bacteriophage T4 late genes, is one of the smallest and most divergent members of this family. Protein footprinting was used to identify segments of gp55 that become buried upon binding to RNA polymerase core, and are therefore likely to constitute its interface with the core enzyme. Site-directed mutagenesis in two parts of this contact surface generated gene 55 proteins that are defective in polymerase-binding to different degrees. Alignment with the sequences of the sigma proteins and with a recently determined structure of a large segment of sigma70 suggests that the gp55 counterpart of sigma70 regions 2.1 and 2.2 is involved in RNA polymerase core binding, and that sigma70 and gp55 may be structurally similar in this region. The diverse phenotypes of the mutants implicate this region of gp55 in multiple aspects of sigma function. PMID:9482743

  9. Detection of nonauthorized genetically modified organisms using differential quantitative polymerase chain reaction: application to 35S in maize.

    PubMed

    Cankar, Katarina; Chauvensy-Ancel, Valérie; Fortabat, Marie-Noelle; Gruden, Kristina; Kobilinsky, André; Zel, Jana; Bertheau, Yves

    2008-05-15

    Detection of nonauthorized genetically modified organisms (GMOs) has always presented an analytical challenge because the complete sequence data needed to detect them are generally unavailable although sequence similarity to known GMOs can be expected. A new approach, differential quantitative polymerase chain reaction (PCR), for detection of nonauthorized GMOs is presented here. This method is based on the presence of several common elements (e.g., promoter, genes of interest) in different GMOs. A statistical model was developed to study the difference between the number of molecules of such a common sequence and the number of molecules identifying the approved GMO (as determined by border-fragment-based PCR) and the donor organism of the common sequence. When this difference differs statistically from zero, the presence of a nonauthorized GMO can be inferred. The interest and scope of such an approach were tested on a case study of different proportions of genetically modified maize events, with the P35S promoter as the Cauliflower Mosaic Virus common sequence. The presence of a nonauthorized GMO was successfully detected in the mixtures analyzed and in the presence of (donor organism of P35S promoter). This method could be easily transposed to other common GMO sequences and other species and is applicable to other detection areas such as microbiology.

  10. Exceptional expansion and conservation of a CT-repeat complex in the core promoter of PAXBP1 in primates.

    PubMed

    Mohammadparast, Saeid; Bayat, Hadi; Biglarian, Akbar; Ohadi, Mina

    2014-08-01

    Adaptive evolution may be linked with the genomic distribution and function of short tandem repeats (STRs). Proximity of the core promoter STRs to the +1 transcription start site (TSS), and their mutable nature are characteristics that highlight those STRs as a novel source of interspecies variation. The PAXBP1 gene (alternatively known as GCFC1) core promoter contains the longest STR identified in a Homo sapiens gene core promoter. Indeed, this core promoter is a stretch of four consecutive CT-STRs. In the current study, we used the Ensembl, NCBI, and UCSC databases to analyze the evolutionary trend and functional implication of this CT-STR complex in six major lineages across vertebrates, including primates, non-primate mammals, birds, reptiles, amphibians, and fish. We observed exceptional expansion (≥4-repeats) and conservation of this CT-STR complex across primates, except prosimians, Microcebus murinus and Otolemur garnettii (Fisher exact P<4.1×10(-7)). H. sapiens has the most complex STR formula, and longest repeats. Macaca mulatta and Callithrix jacchus monkeys have the simplest STR formulas, and shortest repeat numbers. CT≥4-repeats were not detected in non-primate lineages. Different length alleles across the PAXBP1 core promoter CT-STRs significantly altered gene expression in vitro (P<0.001, t-test). PAXBP1 has a crucial role in craniofacial development, myogenesis, and spine morphogenesis, properties that have been diverged between primates and non-primates. To our knowledge, this is the first instance of expansion and conservation of a STR complex co-occurring specifically with the primate lineage.

  11. Chemical synthesis of high specific-activity (/sup 35/S)adenosylhomocysteine

    SciTech Connect

    Stern, P.H.; Hoffman, R.M.

    1986-11-01

    The study of the family of transmethylases, critical to normal cellular function and often altered in cancer, can be facilitated by the availability of a high specific-activity S-adenosylhomocysteine. The authors report the two-step preparation of (/sup 35/S)adenosylhomocysteine from (/sup 35/S)methionine at a specific activity of 1420 Ci/mmol in an overall yield of 24% by a procedure involving demethylation of the (/sup 35/S)methionine to (/sup 35/S)homocysteine followed by condensation with 5'-chloro-5'-deoxyadenosine. The ease of the reactions, ready availability and low cost of the reagents and high specific-activity and stability of the product make the procedure an attractive one with many uses, and superior to current methodology.

  12. Strategies for Development of Functionally Equivalent Promoters with Minimum Sequence Homology for Transgene Expression in Plants: cis-Elements in a Novel DNA Context versus Domain Swapping1

    PubMed Central

    Bhullar, Simran; Chakravarthy, Suma; Advani, Sonia; Datta, Sudipta; Pental, Deepak; Burma, Pradeep Kumar

    2003-01-01

    The cauliflower mosaic virus 35S (35S) promoter has been extensively used for the constitutive expression of transgenes in dicotyledonous plants. The repetitive use of the same promoter is known to induce transgene inactivation due to promoter homology. As a way to circumvent this problem, we tested two different strategies for the development of synthetic promoters that are functionally equivalent but have a minimum sequence homology. Such promoters can be generated by (a) introducing known cis-elements in a novel or synthetic stretch of DNA or (b) “domain swapping,” wherein domains of one promoter can be replaced with functionally equivalent domains from other heterologous promoters. We evaluated the two strategies for promoter modifications using domain A (consisting of minimal promoter and subdomain A1) of the 35S promoter as a model. A set of modified 35S promoters were developed whose strength was compared with the 35S promoter per se using β-glucuronidase as the reporter gene. Analysis of the expression of the reporter gene in transient assay system showed that domain swapping led to a significant fall in promoter activity. In contrast, promoters developed by placing cis-elements in a novel DNA context showed levels of expression comparable with that of the 35S. Two promoter constructs Mod2A1T and Mod3A1T were then designed by placing the core sequences of minimal promoter and subdomain A1 in divergent DNA sequences. Transgenics developed in tobacco (Nicotiana tabacum) with the two constructs and with 35S as control were used to assess the promoter activity in different tissues of primary transformants. Mod2A1T and Mod3A1T were found to be active in all of the tissues tested, at levels comparable with that of 35S. Further, the expression of the Mod2A1T promoter in the seedlings of the T1 generation was also similar to that of the 35S promoter. The present strategy opens up the possibility of creating a set of synthetic promoters with minimum sequence

  13. Widespread Use of TATA Elements in the Core Promoters for RNA Polymerases III, II, and I in Fission Yeast

    PubMed Central

    Hamada, Mitsuhiro; Huang, Ying; Lowe, Todd M.; Maraia, Richard J.

    2001-01-01

    In addition to directing transcription initiation, core promoters integrate input from distal regulatory elements. Except for rare exceptions, it has been generally found that eukaryotic tRNA and rRNA genes do not contain TATA promoter elements and instead use protein-protein interactions to bring the TATA-binding protein (TBP), to the core promoter. Genomewide analysis revealed TATA elements in the core promoters of tRNA and 5S rRNA (Pol III), U1 to U5 snRNA (Pol II), and 37S rRNA (Pol I) genes in Schizosaccharomyces pombe. Using tRNA-dependent suppression and other in vivo assays, as well as in vitro transcription, we demonstrated an obligatory requirement for upstream TATA elements for tRNA and 5S rRNA expression in S. pombe. The Pol III initiation factor Brf is found in complexes with TFIIIC and Pol III in S. pombe, while TBP is not, consistent with independent recruitment of TBP by TATA. Template commitment assays are consistent with this and confirm that the mechanisms of transcription complex assembly and initiation by Pol III in S. pombe differ substantially from those in other model organisms. The results were extended to large-rRNA synthesis, as mutation of the TATA element in the Pol I promoter also abolishes rRNA expression in fission yeast. A survey of other organisms' genomes reveals that a substantial number of eukaryotes may use widespread TATAs for transcription. These results indicate the presence of TATA-unified transcription systems in contemporary eukaryotes and provide insight into the residual need for TBP by all three Pols in other eukaryotes despite a lack of TATA elements in their promoters. PMID:11564871

  14. Structure of newly synthesized (/sup 35/S)-proteoglycans and (/sup 35/S)-proteoglycan turnover products of cartilage explant cultures from dogs with experimental osteoarthritis

    SciTech Connect

    Carney, S.L.; Billingham, M.E.; Muir, H.; Sandy, J.D.

    1985-01-01

    The structure of newly synthesized proteoglycans from explant cultures of cartilage from joints subjected to transection of the anterior cruciate ligament (osteoarthritic) and from normal (non- or sham-operated) joints was examined. The structure of the products of proteoglycan turnover was also examined using explants of normal and osteoarthritic cartilage maintained in culture for a 48 h chase period. The findings were as follows: Newly synthesized (/sup 35/S)-proteoglycans extracted from cartilage explants from osteoarthritic joints whether examined 3 weeks, 3 months, or 6 months after surgery were larger than those from corresponding normal cartilage. This can be explained by the synthesis in osteoarthritic cartilage of abnormally long chondroitin sulfate chains on newly synthesised proteoglycans. The extracts also contained a newly formed small proteoglycan species that was unable to interact with hyaluronic acid. The proportion of this species was higher in osteoarthritic cartilage compared with normal, examined 3 weeks after surgery, but was generally absent from cartilage obtained 3 and 6 months after surgery. Compared with controls, a smaller proportion of the (/sup 35/S)-proteoglycans released into the maintenance medium of explant cultures of osteoarthritic cartilage during a 48 h chase period was able to interact with hyaluronic acid. However, although furnished with longer (/sup 35/S)-glycosaminoglycan chains, these proteoglycans were smaller than those from control explants.

  15. Transcription initiation in vivo without classical transactivators: DNA kinks flanking the core promoter of the housekeeping yeast adenylate kinase gene, AKY2, position nucleosomes and constitutively activate transcription.

    PubMed

    Angermayr, Michaela; Oechsner, Ulrich; Gregor, Kerstin; Schroth, Gary P; Bandlow, Wolfhard

    2002-10-01

    The housekeeping gene of the major adenylate kinase in Saccharomyces cerevisiae (AKY2, ADK1) is constitutively transcribed at a moderate level. The promoter has been dissected in order to define elements that effect constitutive transcription. Initiation of mRNA synthesis at the AKY2 promoter is shown to be mediated by a non-canonic core promoter, (TA)(6). Nucleotide sequences 5' of this element only marginally affect transcription suggesting that promoter activation can dispense with transactivators and essentially involves basal transcription. We show that the core promoter of AKY2 is constitutively kept free of nucleosomes. Analyses of permutated AKY2 promoter DNA revealed the presence of bent DNA. DNA structure analysis by computer and by mutation identified two kinks flanking an interstitial stretch of 65 bp of moderately bent core promoter DNA. Kinked DNA is likely incompatible with packaging into nucleosomes and responsible for positioning nucleosomes at the flanks allowing unimpeded access of the basal transcription machinery to the core promoter. The data show that in yeast, constitutive gene expression can dispense with classical transcriptional activator proteins, if two prerequisites are met: (i) the core promoter is kept free of nucleosomes; this can be due to structural properties of the DNA as an alternative to chromatin remodeling factors; and (ii) the core promoter is pre-bent to allow a high rate of basal transcription initiation.

  16. Promoting integration of genetics core competencies into entry-level nursing curricula.

    PubMed

    Read, Catherine Y; Dylis, Ann M; Mott, Sandra R; Fairchild, Nancy J

    2004-08-01

    Nurse educators must respond to the growing need to teach genetics content in undergraduate nursing curricula. Recently developed genetics core competencies can be used to guide curriculum assessment and planning. This article describes a 5-year effort to integrate genetics education into a baccalaureate nursing curriculum and provides the results of a curriculum survey based on published genetics core competencies.

  17. Analysis of core promoter sequences located downstream from the TATA element in the hsp70 promoter from Drosophila melanogaster.

    PubMed

    Wu, C H; Madabusi, L; Nishioka, H; Emanuel, P; Sypes, M; Arkhipova, I; Gilmour, D S

    2001-03-01

    TFIID recognizes multiple sequence elements in the hsp70 promoter of Drosophila. Here, we investigate the function of sequences downstream from the TATA element. A mutation in the initiator was identified that caused an eightfold reduction in binding of TFIID and a fourfold reduction in transcription in vitro. Another mutation in the +24 to +29 region was somewhat less inhibitory, but a mutation in the +14 to +19 region had essentially no effect. The normal promoter and the mutants in the initiator and the +24 to +29 region were transformed into flies by P element-mediated transformation. The initiator mutation reduced expression an average of twofold in adult flies, whereas the mutation in the +24 to +29 region had essentially no effect. In contrast, a promoter combining the two mutations was expressed an average of sixfold less than the wild type. The results suggest that the initiator and the +24 to +29 region could serve overlapping functions in vivo. Protein-DNA cross-linking was used to identify which subunits of TFIID contact the +24 to +29 region and the initiator. No specific subunits were found to cross-link to the +24 to +29 region. In contrast, the initiator cross-linked exclusively to dTAF230. Remarkably, dTAF230 cross-links approximately 10 times more efficiently to the nontranscribed strand than to the transcribed strand at the initiator.

  18. The CEBPA gene is down-regulated in acute promyelocytic leukemia and its upstream promoter, but not the core promoter, is highly methylated

    PubMed Central

    Santana-Lemos, Bárbara Amélia; de Lima Lange, Ana Paula Alencar; de Lira Benício, Mariana Tereza; da Silva José, Thiago Donizete; Lucena-Araújo, Antônio Roberto; Krause, Alexandre; Thomé, Carolina Hassibe; Rego, Eduardo Magalhães

    2011-01-01

    Impairment of CCAAT Enhancer Binding Protein alpha (CEBPA) function is a common finding in acute myeloid leukemia; nevertheless, its relevance for acute promyelocytic leukemia pathogenesis is unclear. We analyzed the expression and assessed the methylation status of the core and upstream promoters of CEBPA in acute promyelocytic leukemia at diagnosis. Patients with acute promyelocytic leukemia (n=18) presented lower levels of CEBPA expression compared to healthy controls (n=5), but higher levels than those in acute myeloid leukemia with t(8;21) (n=9) and with inv(16) (n=5). Regarding the core promoter, we detected no methylation in 39 acute promyelocytic leukemia samples or in 8 samples from controls. In contrast, analysis of the upstream promoter showed methylation in 37 of 39 samples, with 17 patients showing methylation levels over 30%. Our results corroborate data obtained in animal models showing that CEBPA is down-regulated in acute promyelocytic leukemia stem cells and suggest that epigenetic mechanisms may be involved. PMID:21134977

  19. 5' termini of poliovirus RNA: difference between virion and nonencapsidated 35S RNA.

    PubMed Central

    Fernandez-Muñoz, R; Lavi, U

    1977-01-01

    Poliovirus cytoplasmic, nonencapsidated 35S RNA yields approximately one pUp per molecule upon T2 RNase digestion, indicating that this RNA has the same 5' end as the polyribosome-associated viral RNA fraction. Double-stranded, replicative form RNA after the same treatment yielded approximately four pNp structures per molecule, 65% of which was pUp. In contrast, the 35S RNA from mature virions contained no detectable pNp, indicating that the 5' end of the virion RNA is different from that of the nonencapsidated RNA. None of the above molecules contained pppNp, ppNp, or GpppNp structures present in host mRNA. The virion RNA molecules, as we have shown previously for thenonencapsidated 35S viral RNA (Fernandez-Muñoz and Darnell, 1976), is not labeled with [methyl-3H]methionine. PMID:189096

  20. FANCD2, FANCJ and BRCA2 cooperate to promote replication fork recovery independently of the Fanconi Anemia core complex.

    PubMed

    Raghunandan, Maya; Chaudhury, Indrajit; Kelich, Stephanie L; Hanenberg, Helmut; Sobeck, Alexandra

    2015-01-01

    Fanconi Anemia (FA) is an inherited multi-gene cancer predisposition syndrome that is characterized on the cellular level by a hypersensitivity to DNA interstrand crosslinks (ICLs). To repair these lesions, the FA pathway proteins are thought to act in a linear hierarchy: Following ICL detection, an upstream FA core complex monoubiquitinates the central FA pathway members FANCD2 and FANCI, followed by their recruitment to chromatin. Chromatin-bound monoubiquitinated FANCD2 and FANCI subsequently coordinate DNA repair factors including the downstream FA pathway members FANCJ and FANCD1/BRCA2 to repair the DNA ICL. Importantly, we recently showed that FANCD2 has additional independent roles: it binds chromatin and acts in concert with the BLM helicase complex to promote the restart of aphidicolin (APH)-stalled replication forks, while suppressing the firing of new replication origins. Here, we show that FANCD2 fulfills these roles independently of the FA core complex-mediated monoubiquitination step. Following APH treatment, nonubiquitinated FANCD2 accumulates on chromatin, recruits the BLM complex, and promotes robust replication fork recovery regardless of the absence or presence of a functional FA core complex. In contrast, the downstream FA pathway members FANCJ and BRCA2 share FANCD2's role in replication fork restart and the suppression of new origin firing. Our results support a non-linear FA pathway model at stalled replication forks, where the nonubiquitinated FANCD2 isoform - in concert with FANCJ and BRCA2 - fulfills a specific function in promoting efficient replication fork recovery independently of the FA core complex.

  1. Development of an efficient bi-directional promoter with tripartite enhancer employing three viral promoters.

    PubMed

    Patro, Sunita; Maiti, Indu B; Dey, Nrisingha

    2013-02-10

    We have developed a novel bi-directional promoter (FsFfCBD) by placing two heterogeneous core-promoters from the Figwort mosaic virus sub-genomic transcript promoter (FsCP, -69 to +31) and Cauliflower mosaic virus 35S promoter (CCP, -89 to +1) respectively on upstream (5') and downstream (3') ends of a tri-hybrid enhancer (FsEFfECE), in reverse orientation. The FsEFfECE domain encompasses three heterologous enhancer fragments from Figwort mosaic virus sub-genomic transcript promoter (FsE, 101 bp, -70 to -170), Figwort mosaic virus full-length transcript promoter (FfE, 196 bp, -249 to -54) and Cauliflower mosaic virus 35S promoter (CE, 254 bp, -343 to -90). The bi-directional nature of the FsFfCBD promoter (coupled to GFP and GUS) was established both in transient systems (onion epidermal cells and tobacco protoplasts) and transgenic plant (Nicotiana tabacum samsun NN) by monitoring the simultaneous expression of GFP and GUS employing fluorescence (for GFP) and biochemical (for GUS) based assays. In transgenic plants, the FsFfCBD promoter was found to be 6.8 and 2.5 times stronger than two parent promoters; Fs and FfC respectively. The bi-directional compound promoter FsFfCBD, composed of three heterologous enhancers with enhanced activity could become a valuable additional tool for efficient plant metabolic engineering and molecular pharming.

  2. ((35)S)sulfate incorporation into glomerular basement membrane glycosaminoglycans is decreased in experimental diabetes

    SciTech Connect

    Cohen, M.P.; Surma, M.L.

    1981-11-01

    Isolated rat renal glomeruli incorporate radioactive sulfate into glycosaminoglycans, which are integral components of the glomerular basement membrane. Cellulose acetate electrophoresis and specific enzymatic sensitivities of glycosaminoglycans prepared after pronase digestion of purified glomerular basement membrane indicate the presence of heparan sulfate. We examined the effect of experimental diabetes on the incorporation of ((35)S)-sulfate into glycosaminoglycans deposited into newly synthesized glomerular basement membrane in vitro. Basement membranes were purified from glomeruli isolated from normal and streptozotocin-diabetic rats after incubation for 2 hr with radiolabeled sulfate and then were subjected to pronase digestion for isolation of the glycosaminoglycans. ((35)S) incorporation into basement membrane glycosaminoglycans was significantly decreased in glomeruli from diabetic animals. The addition of insulin (100 micron U/ml) in vitro did not affect ((35)S) incorporation into glycosaminoglycans of the glomerular basement membranes in normal or diabetic glomeruli. High glucose concentration (5 vs. 20 mM) was without effect in short-term incubations of glomeruli from normal animals. The results indicate that experimental diabetes influences ((35)S) sulfate incorporation into glomerular basement membrane glycosaminoglycans and suggest that decreased heparan sulfate production and/or sulfation may contribute to the increased permeability of the glomerular basement membrane in diabetes.

  3. Promoting utilization of Saccharum spp. genetic resources through genetic diversity analysis and core collection construction.

    PubMed

    Nayak, Spurthi N; Song, Jian; Villa, Andrea; Pathak, Bhuvan; Ayala-Silva, Tomas; Yang, Xiping; Todd, James; Glynn, Neil C; Kuhn, David N; Glaz, Barry; Gilbert, Robert A; Comstock, Jack C; Wang, Jianping

    2014-01-01

    Sugarcane (Saccharum spp.) and other members of Saccharum spp. are attractive biofuel feedstocks. One of the two World Collections of Sugarcane and Related Grasses (WCSRG) is in Miami, FL. This WCSRG has 1002 accessions, presumably with valuable alleles for biomass, other important agronomic traits, and stress resistance. However, the WCSRG has not been fully exploited by breeders due to its lack of characterization and unmanageable population. In order to optimize the use of this genetic resource, we aim to 1) genotypically evaluate all the 1002 accessions to understand its genetic diversity and population structure and 2) form a core collection, which captures most of the genetic diversity in the WCSRG. We screened 36 microsatellite markers on 1002 genotypes and recorded 209 alleles. Genetic diversity of the WCSRG ranged from 0 to 0.5 with an average of 0.304. The population structure analysis and principal coordinate analysis revealed three clusters with all S. spontaneum in one cluster, S. officinarum and S. hybrids in the second cluster and mostly non-Saccharum spp. in the third cluster. A core collection of 300 accessions was identified which captured the maximum genetic diversity of the entire WCSRG which can be further exploited for sugarcane and energy cane breeding. Sugarcane and energy cane breeders can effectively utilize this core collection for cultivar improvement. Further, the core collection can provide resources for forming an association panel to evaluate the traits of agronomic and commercial importance.

  4. Promoting Utilization of Saccharum spp. Genetic Resources through Genetic Diversity Analysis and Core Collection Construction

    PubMed Central

    Pathak, Bhuvan; Ayala-Silva, Tomas; Yang, Xiping; Todd, James; Glynn, Neil C.; Kuhn, David N.; Glaz, Barry; Gilbert, Robert A.; Comstock, Jack C.; Wang, Jianping

    2014-01-01

    Sugarcane (Saccharum spp.) and other members of Saccharum spp. are attractive biofuel feedstocks. One of the two World Collections of Sugarcane and Related Grasses (WCSRG) is in Miami, FL. This WCSRG has 1002 accessions, presumably with valuable alleles for biomass, other important agronomic traits, and stress resistance. However, the WCSRG has not been fully exploited by breeders due to its lack of characterization and unmanageable population. In order to optimize the use of this genetic resource, we aim to 1) genotypically evaluate all the 1002 accessions to understand its genetic diversity and population structure and 2) form a core collection, which captures most of the genetic diversity in the WCSRG. We screened 36 microsatellite markers on 1002 genotypes and recorded 209 alleles. Genetic diversity of the WCSRG ranged from 0 to 0.5 with an average of 0.304. The population structure analysis and principal coordinate analysis revealed three clusters with all S. spontaneum in one cluster, S. officinarum and S. hybrids in the second cluster and mostly non-Saccharum spp. in the third cluster. A core collection of 300 accessions was identified which captured the maximum genetic diversity of the entire WCSRG which can be further exploited for sugarcane and energy cane breeding. Sugarcane and energy cane breeders can effectively utilize this core collection for cultivar improvement. Further, the core collection can provide resources for forming an association panel to evaluate the traits of agronomic and commercial importance. PMID:25333358

  5. Astonishing 35S rDNA diversity in the gymnosperm species Cycas revoluta Thunb.

    PubMed

    Wang, Wencai; Ma, Lu; Becher, Hannes; Garcia, Sònia; Kovarikova, Alena; Leitch, Ilia J; Leitch, Andrew R; Kovarik, Ales

    2016-09-01

    In all eukaryotes, the highly repeated 35S ribosomal DNA (rDNA) sequences encoding 18S-5.8S-26S ribosomal RNA (rRNA) typically show high levels of intragenomic uniformity due to homogenisation processes, leading to concerted evolution of 35S rDNA repeats. Here, we compared 35S rDNA divergence in several seed plants using next generation sequencing and a range of molecular and cytogenetic approaches. Most species showed similar 35S rDNA homogeneity indicating concerted evolution. However, Cycas revoluta exhibits an extraordinary diversity of rDNA repeats (nucleotide sequence divergence of different copies averaging 12 %), influencing both the coding and non-coding rDNA regions nearly equally. In contrast, its rRNA transcriptome was highly homogeneous suggesting that only a minority of genes (<20 %) encode functional rRNA. The most common SNPs were C > T substitutions located in symmetrical CG and CHG contexts which were also highly methylated. Both functional genes and pseudogenes appear to cluster on chromosomes. The extraordinary high levels of 35S rDNA diversity in C. revoluta, and probably other species of cycads, indicate that the frequency of repeat homogenisation has been much lower in this lineage, compared with all other land plant lineages studied. This has led to the accumulation of methylation-driven mutations and pseudogenisation. Potentially, the reduced homology between paralogs prevented their elimination by homologous recombination, resulting in long-term retention of rDNA pseudogenes in the genome.

  6. LDL particle core enrichment in cholesteryl oleate increases proteoglycan binding and promotes atherosclerosis[S

    PubMed Central

    Melchior, John T.; Sawyer, Janet K.; Kelley, Kathryn L.; Shah, Ramesh; Wilson, Martha D.; Hantgan, Roy R.; Rudel, Lawrence L.

    2013-01-01

    Several studies in humans and animals suggest that LDL particle core enrichment in cholesteryl oleate (CO) is associated with increased atherosclerosis. Diet enrichment with MUFAs enhances LDL CO content. Steroyl O-acyltransferase 2 (SOAT2) is the enzyme that catalyzes the synthesis of much of the CO found in LDL, and gene deletion of SOAT2 minimizes CO in LDL and protects against atherosclerosis. The purpose of this study was to test the hypothesis that the increased atherosclerosis associated with LDL core enrichment in CO results from an increased affinity of the LDL particle for arterial proteoglycans. ApoB-100-only Ldlr−/− mice with and without Soat2 gene deletions were fed diets enriched in either cis-MUFA or n-3 PUFA, and LDL particles were isolated. LDL:proteogylcan binding was measured using surface plasmon resonance. Particles with higher CO content consistently bound with higher affinity to human biglycan and the amount of binding was shown to be proportional to the extent of atherosclerosis of the LDL donor mice. The data strongly support the thesis that atherosclerosis was induced through enhanced proteoglycan binding of LDL resulting from LDL core CO enrichment. PMID:23804810

  7. Sequence homology requirements for transcriptional silencing of 35S transgenes and post-transcriptional silencing of nitrite reductase (trans)genes by the tobacco 271 locus.

    PubMed

    Thierry, D; Vaucheret, H

    1996-12-01

    The transgene locus of the tobacco plant 271 (271 locus) is located on a telomere and consists of multiple copies of a plasmid carrying an NptII marker gene driven by the cauliflower mosaic virus (CaMV) 19S promoter and the leaf-specific nitrite reductase Nii1 cDNA cloned in the antisense orientation under the control of the CaMV 35S promoter. Previous analysis of gene expression in leaves has shown that this locus triggers both post-transcriptional silencing of the host leaf-specific Nii genes and transcriptional silencing of transgenes driven by the 19S or 35S promoter irrespective of their coding sequence and of their location in the genome. In this paper we show that silencing of transgenes carrying Nii1 sequences occurs irrespective of the promoter driving their expression and of their location within the genome. This phenomenon occurs in roots as well as in leaves although root Nii genes share only 84% identity with leaf-specific Nii1 sequences carried by the 271 locus. Conversely, transgenes carrying the bean Nii gene (which shares 76% identity with the tobacco Nii1 gene) escape silencing by the 271 locus. We also show that transgenes driven by the figwort mosaic virus 34S promoter (which shares 63% identity with the 35S promoter) also escape silencing by the 271 locus. Taken together, these results indicate that a high degree of sequence similarity is required between the sequences of the silencing locus and of the target (trans)genes for both transcriptional and post-transcriptional silencing.

  8. Promoting the Use of Common Oat Genetic Resources through Diversity Analysis and Core Collection Construction.

    PubMed

    Boczkowska, Maja; Łapiński, Bogusław; Kordulasińska, Izabela; Dostatny, Denise F; Czembor, Jerzy H

    2016-01-01

    The assessment of diversity and population structure and construction of a core collection is beneficial for the efficient use and management of germplasm. A unique collection of common oat landraces, cultivated in the temperate climate of central Europe until the end of the twentieth century, is preserved in the Polish gene bank. It consists of 91 accessions that have never been used in breeding programs. In order to optimise the use of this genetic resource, we aimed to: (1) determine genetic and agro-morphological diversity, (2) identify internal genetic variation of the tested accessions, (3) form a core collection and (4) recognise the accessions useful for breeding programs or re-release for cultivation. The collection was screened using ISSR markers (1520 loci) and eight agro-morphological traits. Uniquely, we performed molecular studies based on 24 individuals of every accession instead of bulk samples. Therefore, assessment of the degree of diversity within each population and the identification of overlapping gene pools were possible. The observed internal diversity (Nei unbiased coefficient) was in the range of 0.17-0.31. Based on combined genetic and agro-morphological data, we established the core collection composed of 21 landraces. Due to valuable compositions of important traits, some accessions were also identified as useful for breeding programs. The population structure and principal coordinate analysis revealed two major clusters. Based on the previous results, the accessions classified within the smaller one were identified as obsolete varieties instead of landraces. Our results show that the oat landraces are, in general, resistant to local races of diseases, well adapted to local conditions and, in some cases, yielding at the level of modern varieties. Therefore, in situ conservation of the landraces in the near future may be satisfactory for both farmers and researchers in terms of the genetic resources preservation.

  9. Promoting the Use of Common Oat Genetic Resources through Diversity Analysis and Core Collection Construction

    PubMed Central

    Łapiński, Bogusław; Kordulasińska, Izabela; Dostatny, Denise F.; Czembor, Jerzy H.

    2016-01-01

    The assessment of diversity and population structure and construction of a core collection is beneficial for the efficient use and management of germplasm. A unique collection of common oat landraces, cultivated in the temperate climate of central Europe until the end of the twentieth century, is preserved in the Polish gene bank. It consists of 91 accessions that have never been used in breeding programs. In order to optimise the use of this genetic resource, we aimed to: (1) determine genetic and agro-morphological diversity, (2) identify internal genetic variation of the tested accessions, (3) form a core collection and (4) recognise the accessions useful for breeding programs or re-release for cultivation. The collection was screened using ISSR markers (1520 loci) and eight agro-morphological traits. Uniquely, we performed molecular studies based on 24 individuals of every accession instead of bulk samples. Therefore, assessment of the degree of diversity within each population and the identification of overlapping gene pools were possible. The observed internal diversity (Nei unbiased coefficient) was in the range of 0.17–0.31. Based on combined genetic and agro-morphological data, we established the core collection composed of 21 landraces. Due to valuable compositions of important traits, some accessions were also identified as useful for breeding programs. The population structure and principal coordinate analysis revealed two major clusters. Based on the previous results, the accessions classified within the smaller one were identified as obsolete varieties instead of landraces. Our results show that the oat landraces are, in general, resistant to local races of diseases, well adapted to local conditions and, in some cases, yielding at the level of modern varieties. Therefore, in situ conservation of the landraces in the near future may be satisfactory for both farmers and researchers in terms of the genetic resources preservation. PMID:27959891

  10. Analysis of HBV genotype, drug resistant mutations, and pre-core/basal core promoter mutations in Korean patients with acute hepatitis B.

    PubMed

    Lee, Jong Ho; Hong, Sun Pyo; Jang, Eun Sun; Park, Sang Jong; Hwang, Seong Gyu; Kang, Sook-Kyoung; Jeong, Sook-Hyang

    2015-06-01

    Acute hepatitis B, caused by hepatitis B virus (HBV) strains with drug resistant mutations or pre-core/basal core promoter (PC/BCP) mutations, is a public health concern, because this infection is often associated with poor disease outcome or difficulty in therapeutic choice. The HBV genotype, the prevalence of drug resistant mutations, and PC/BCP mutations in Korean patients with acute hepatitis B were studied. From 2006 to 2008, 36 patients with acute hepatitis B were enrolled prospectively in four general hospitals. Among them, 20 showed detectable HBV DNA (median value was 4.8 log copies/mL). HBV genotyping and analysis of HBV mutations that conferred resistance against lamivudine, adefovir, or entecavir and of PC/BCP mutations were performed using highly sensitive restriction fragment mass polymorphism (RFMP) analysis. All 20 patients were infected with HBV genotype C, which causes almost all cases of chronic hepatitis B in Korea. No patient showed mutations that conferred resistance against lamivudine (L180M, M204V/I), adefovir (A181T, N236S), or entecavir (I169M, A184T/V, S202I/G, M250V/I/L). However, four patients had BCP mutations, and two had PC mutations. Platelet counts were significantly lower in the four patients with PC/BCP mutations compared to those with wild type. In this study, all acute hepatitis B patients had genotype C HBV strains with no drug resistant mutations. However, 20% showed PC/BCP mutations. This highlights the need for further study on the significance of PC/BCP mutations.

  11. The Core Promoter and Redox-sensitive Cis-elements as Key Targets for Inactivation of the Lysyl Oxidase Gene by Cadmium

    PubMed Central

    Li, Jianmin; Cheng, Guang; Zheng, Maoguen; Zhao, Yinzhi; Zhou, Jing; Li, Wande

    2015-01-01

    Exposure of humans to cadmium (Cd) either from environmental contamination or from cigarette smoke, often induces lung emphysema and cancers. Lysyl oxidase (LOX), a copper-dependent enzyme essential for crosslinking of the extracellular matrix, displays antagonistic effects on emphysema and cancer pathogenesis. Our previous studies showed down-regulation of LOX in Cd-resistant (CdR) rat fetal lung fibroblasts (RFL6) derived from parental cells via long-term Cd exposure. The cloned rat LOX gene promoter −804/−1 (relative to ATG) with the maximal promoter activity contains the Inr-DPE core promoter, putative NFI binding sites, metal response elements (MRE) and antioxidant response elements (ARE). ChIP assays reported here further characterize the rat LOX gene promoter in response to Cd. CdR cells exhibited enhanced methylation of CpG at the LOX core promoter region and reduced activities of the NFI binding sites and MRE, but increased activity of the ARE in a dose-dependent manner. The collective effect of Cd on the LOX promoter is trans-inhibition of the LOX gene as shown by suppression of histone H3 acetylation in the LOX core promoter region. Thus, the LOX core promoter and redox-sensitive cis-elements are key Cd targets for down-regulation of LOX relevant to mechanisms for Cd-induced emphysema and lung cancers. PMID:25741534

  12. Replacement of the human cytomegalovirus promoter with fish enhancer and core elements to control the expression of the G gene of viral haemorrhagic septicemia virus (VHSV).

    PubMed

    Martinez-Lopez, A; Chinchilla, B; Encinas, P; Gomez-Casado, E; Estepa, A; Coll, J M

    2012-12-15

    This work explores some of the possibilities to replace human cytomegalovirus (CMV) core and/or enhancer promoter control elements to create new expression vectors for use with fish. The work is relevant to fish vaccination, since DNA vaccines use eukaryotic expression plasmids controlled by the human cytomegalovirus (CMV) promoter to be effective against novirhabdoviruses, such as viral haemorrhagic septicemia virus (VHSV), one of the most devastating fish viral European diseases. To reduce possible homologous recombination with fish genome, core and enhancer sequences from fish origin, such as trout interferon-inducible myxovirus protein (Mx), zebrafish retrovirus long terminal repeat (LTR) and carp β-actin (AE6), were combined with those of CMV to design alternative hybrid promoters. The substitution of CMV core and/or enhancer with the corresponding elements of Mx or the LTR core maintained a similar in vitro protein G expression level than that obtained by using the CMV promoter. Vectors using the dsRNA-inducible Mx enhancer followed either by the LTR or the AE6 cores showed the highest in vitro protein G expression levels. Furthermore, synthetic constructs using the Mx enhancer maintained their polyI:C induction capabilities despite the core used. Some of these hybrid promoters might contribute to the development of all-fish-vectors for DNA vaccines while others might be useful for more basic studies.

  13. The missing flux in a 35S budget for the soils of a small polluted catchment

    USGS Publications Warehouse

    Novak, M.; Michel, R.L.; Prechova, E.; Stepanova, M.

    2004-01-01

    A combination of cosmogenic and artificial 35S was used to assess the movement of sulfur in a steep Central European catchment affected by spruce die-back. The Jezer??i?? catchment, Krus??ne?? Hory Mts. (Czech Republic) is characterized by a large disproportion between atmospheric S input and S output via stream discharge, with S output currently exceeding S input three times. A relatively high natural concentration of cosmogenic 35S (42 mBq L-1) was found in atmospheric deposition into the catchment in winter and spring of 2000. In contrast, stream discharge contained only 2 mBq L-1. Consequently, more than 95% of the deposited S is cycled or retained within the catchment for more than several months, while older S is exported via surface water. In spring, when the soil temperature is above 0 ??C, practically no S from instantaneous rainfall is exported, despite the steepness of the slopes and the relatively short mean residence time of water in the catchment (6.5 months). Sulfur cycling in the soil includes not just adsorption of inorganic sulfate and biological uptake, but also volatilization of S compounds back into the atmosphere. Laboratory incubations of an Orthic Podzol from Jezer??i?? spiked with h 720 kBq of artificial 35S showed a 20% loss of the spike within 18 weeks under summer conditions. Under winter conditions, the 35S loss was insignificant (< 5%). This missing S flux was interpreted as volatilized hydrogen sulfide resulting from intermittent dissimilatory bacterial sulfate reduction. The missing S flux is comparable to the estimated uncertainty in many catchment S mass balances (??10%), or even larger, and should be considered in constructing these mass balances. In severely polluted forest catchments, such as Jezer??i??, sulfur loss to volatilization may exceed 13 kg ha-1 a-1, which is more than the current total atmospheric S input in large parts of North America and Europe. ?? 2004 Kluwer Academic Publishers.

  14. The Core Apoptotic Executioner Proteins CED-3 and CED-4 Promote Initiation of Neuronal Regeneration in Caenorhabditis elegans

    PubMed Central

    Reina, Christopher P.; Hulme, S. Elizabeth; Shevkoplyas, Sergey S.; Slone, R. Daniel; Xue, Jian; Qiao, Yujie; Weisberg, Sarah; Roodhouse, Kevin; Sun, Lin; Whitesides, George M.; Samuel, Aravinthan; Driscoll, Monica

    2012-01-01

    A critical accomplishment in the rapidly developing field of regenerative medicine will be the ability to foster repair of neurons severed by injury, disease, or microsurgery. In C. elegans, individual visualized axons can be laser-cut in vivo and neuronal responses to damage can be monitored to decipher genetic requirements for regeneration. With an initial interest in how local environments manage cellular debris, we performed femtosecond laser axotomies in genetic backgrounds lacking cell death gene activities. Unexpectedly, we found that the CED-3 caspase, well known as the core apoptotic cell death executioner, acts in early responses to neuronal injury to promote rapid regeneration of dissociated axons. In ced-3 mutants, initial regenerative outgrowth dynamics are impaired and axon repair through reconnection of the two dissociated ends is delayed. The CED-3 activator, CED-4/Apaf-1, similarly promotes regeneration, but the upstream regulators of apoptosis CED-9/Bcl2 and BH3-domain proteins EGL-1 and CED-13 are not essential. Thus, a novel regulatory mechanism must be utilized to activate core apoptotic proteins for neuronal repair. Since calcium plays a conserved modulatory role in regeneration, we hypothesized calcium might play a critical regulatory role in the CED-3/CED-4 repair pathway. We used the calcium reporter cameleon to track in vivo calcium fluxes in the axotomized neuron. We show that when the endoplasmic reticulum calcium-storing chaperone calreticulin, CRT-1, is deleted, both calcium dynamics and initial regenerative outgrowth are impaired. Genetic data suggest that CED-3, CED-4, and CRT-1 act in the same pathway to promote early events in regeneration and that CED-3 might act downstream of CRT-1, but upstream of the conserved DLK-1 kinase implicated in regeneration across species. This study documents reconstructive roles for proteins known to orchestrate apoptotic death and links previously unconnected observations in the vertebrate

  15. Quantifying groundwater travel time near managed recharge operations using 35S as an intrinsic tracer

    NASA Astrophysics Data System (ADS)

    Urióstegui, Stephanie H.; Bibby, Richard K.; Esser, Bradley K.; Clark, Jordan F.

    2016-12-01

    Identifying groundwater retention times near managed aquifer recharge (MAR) facilities is a high priority for managing water quality, especially for operations that incorporate recycled wastewater. To protect public health, California guidelines for Groundwater Replenishment Reuse Projects require a minimum 2-6 month subsurface retention time for recycled water depending on the level of disinfection, which highlights the importance of quantifying groundwater travel times on short time scales. This study developed and evaluated a new intrinsic tracer method using the naturally occurring radioisotope sulfur-35 (35S). The 87.5 day half-life of 35S is ideal for investigating groundwater travel times on the <1 year timescale of interest to MAR managers. Natural concentrations of 35S found in water as dissolved sulfate (35SO4) were measured in source waters and groundwater at the Rio Hondo Spreading Grounds in Los Angeles County, CA, and Orange County Groundwater Recharge Facilities in Orange County, CA. 35SO4 travel times are comparable to travel times determined by well-established deliberate tracer studies. The study also revealed that 35SO4 in MAR source water can vary seasonally and therefore careful characterization of 35SO4 is needed to accurately quantify groundwater travel time. More data is needed to fully assess whether or not this tracer could become a valuable tool for managers.

  16. Quantifying groundwater travel time near managed recharge operations using 35S as an intrinsic tracer

    DOE PAGES

    Urióstegui, Stephanie H.; Bibby, Richard K.; Esser, Bradley K.; ...

    2016-04-23

    By identifying groundwater retention times near managed aquifer recharge (MAR) facilities is a high priority for managing water quality, especially for operations that incorporate recycled wastewater. In order to protect public health, California guidelines for Groundwater Replenishment Reuse Projects require a minimum 2–6 month subsurface retention time for recycled water depending on the level of disinfection, which highlights the importance of quantifying groundwater travel times on short time scales. This study developed and evaluated a new intrinsic tracer method using the naturally occurring radioisotope sulfur-35 (35S). The 87.5 day half-life of 35S is ideal for investigating groundwater travel times onmore » the <1 year timescale of interest to MAR managers. Natural concentrations of 35S found in water as dissolved sulfate (35SO4) were measured in source waters and groundwater at the Rio Hondo Spreading Grounds in Los Angeles County, CA, and Orange County Groundwater Recharge Facilities in Orange County, CA. 35SO4 travel times are comparable to travel times determined by well-established deliberate tracer studies. The study also revealed that 35SO4 in MAR source water can vary seasonally and therefore careful characterization of 35SO4 is needed to accurately quantify groundwater travel time. But, more data is needed to fully assess whether or not this tracer could become a valuable tool for managers.« less

  17. Quantifying groundwater travel time near managed recharge operations using 35S as an intrinsic tracer

    SciTech Connect

    Urióstegui, Stephanie H.; Bibby, Richard K.; Esser, Bradley K.; Clark, Jordan F.

    2016-04-23

    By identifying groundwater retention times near managed aquifer recharge (MAR) facilities is a high priority for managing water quality, especially for operations that incorporate recycled wastewater. In order to protect public health, California guidelines for Groundwater Replenishment Reuse Projects require a minimum 2–6 month subsurface retention time for recycled water depending on the level of disinfection, which highlights the importance of quantifying groundwater travel times on short time scales. This study developed and evaluated a new intrinsic tracer method using the naturally occurring radioisotope sulfur-35 (35S). The 87.5 day half-life of 35S is ideal for investigating groundwater travel times on the <1 year timescale of interest to MAR managers. Natural concentrations of 35S found in water as dissolved sulfate (35SO4) were measured in source waters and groundwater at the Rio Hondo Spreading Grounds in Los Angeles County, CA, and Orange County Groundwater Recharge Facilities in Orange County, CA. 35SO4 travel times are comparable to travel times determined by well-established deliberate tracer studies. The study also revealed that 35SO4 in MAR source water can vary seasonally and therefore careful characterization of 35SO4 is needed to accurately quantify groundwater travel time. But, more data is needed to fully assess whether or not this tracer could become a valuable tool for managers.

  18. Predominance of precore mutations and clinical significance of basal core promoter mutations in chronic hepatitis B virus infection in Indonesia.

    PubMed

    Juniastuti; Utsumi, Takako; Aksono, Eduardus Bimo; Yano, Yoshihiko; Soetjipto; Hayashi, Yoshitake; Hotta, Hak; Rantam, Fedik Abdul; Kusumobroto, Hernomo Ontoseno; Lusida, Maria Inge

    2013-07-01

    Chronic hepatitis B virus (HBV) infection is a major health problem worldwide, with a particularly high prevalence in the Asian-Pacific region. During chronic hepatitis B virus (HBV) infection, mutations commonly occur in the basal core promoter (BCP) and precore (PC) regions of HBV, affecting HBeAg expression, particularly following HBeAg serocon-version. Mutations in the B- and T-cell epitopes of the HBV core have also been observed during disease progression. The clinical significance of HBV genome variability has been demonstrated, however the results are a subject of controversy. Considering the characteristics of the virus associated with geographical location, the profiles of BCP, PC and core mutations and their clinical implications in patients with chronic HBV infection in Surabaya, Indonesia, were investigated. The BCP, PC and core mutations and HBV genotypes were detected by direct sequencing. The HBeAg/anti-HBe status and HBV DNA levels were also assessed. This study enrolled 10 patients with chronic HBV infection (UC) from Dr Soetomo General Hospital and Indonesian Red Cross, Surabaya, East Java, Indonesia, 10 patients with chronic hepatitis B and liver cirrhosis (LC) and 4 patients with chronic hepatitis B and hepatocellular carcinoma (HCC) from Dr Soetomo General Hospital. The PC mutation A1896 was predominant in all the groups (60-100%), together with the PC variant T1858, which was associated with HBV genotype B. The number of detected core mutations (Thr/Ser130) was higher in HCC patients (50%). However, the BCP mutations T1762/A1764 were predominant in LC patients (50-60%). The LC and HCC patients carried HBV isolates with additional mutations, at least at BCP or PC, mainly following HBeAg seroconversion. In the majority of anti-HBe-positive samples, the BCP T1762/A1764 mutations were associated with a high viral load, regardless of the PC 1896 status. In conclusion, the PC mutations were found to be predominant in all the groups. However, the

  19. Predominance of precore mutations and clinical significance of basal core promoter mutations in chronic hepatitis B virus infection in Indonesia

    PubMed Central

    JUNIASTUTI; UTSUMI, TAKAKO; AKSONO, EDUARDUS BIMO; YANO, YOSHIHIKO; SOETJIPTO; HAYASHI, YOSHITAKE; HOTTA, HAK; RANTAM, FEDIK ABDUL; KUSUMOBROTO, HERNOMO ONTOSENO; LUSIDA, MARIA INGE

    2013-01-01

    Chronic hepatitis B virus (HBV) infection is a major health problem worldwide, with a particularly high prevalence in the Asian-Pacific region. During chronic hepatitis B virus (HBV) infection, mutations commonly occur in the basal core promoter (BCP) and precore (PC) regions of HBV, affecting HBeAg expression, particularly following HBeAg serocon-version. Mutations in the B- and T-cell epitopes of the HBV core have also been observed during disease progression. The clinical significance of HBV genome variability has been demonstrated, however the results are a subject of controversy. Considering the characteristics of the virus associated with geographical location, the profiles of BCP, PC and core mutations and their clinical implications in patients with chronic HBV infection in Surabaya, Indonesia, were investigated. The BCP, PC and core mutations and HBV genotypes were detected by direct sequencing. The HBeAg/anti-HBe status and HBV DNA levels were also assessed. This study enrolled 10 patients with chronic HBV infection (UC) from Dr Soetomo General Hospital and Indonesian Red Cross, Surabaya, East Java, Indonesia, 10 patients with chronic hepatitis B and liver cirrhosis (LC) and 4 patients with chronic hepatitis B and hepatocellular carcinoma (HCC) from Dr Soetomo General Hospital. The PC mutation A1896 was predominant in all the groups (60–100%), together with the PC variant T1858, which was associated with HBV genotype B. The number of detected core mutations (Thr/Ser130) was higher in HCC patients (50%). However, the BCP mutations T1762/A1764 were predominant in LC patients (50–60%). The LC and HCC patients carried HBV isolates with additional mutations, at least at BCP or PC, mainly following HBeAg seroconversion. In the majority of anti-HBe-positive samples, the BCP T1762/A1764 mutations were associated with a high viral load, regardless of the PC 1896 status. In conclusion, the PC mutations were found to be predominant in all the groups. However

  20. TAF4/4b·TAF12 Displays a Unique Mode of DNA Binding and Is Required for Core Promoter Function of a Subset of Genes*

    PubMed Central

    Gazit, Kfir; Moshonov, Sandra; Elfakess, Rofa; Sharon, Michal; Mengus, Gabrielle; Davidson, Irwin; Dikstein, Rivka

    2009-01-01

    The major core promoter-binding factor in polymerase II transcription machinery is TFIID, a complex consisting of TBP, the TATA box-binding protein, and 13 to 14 TBP-associated factors (TAFs). Previously we found that the histone H2A-like TAF paralogs TAF4 and TAF4b possess DNA-binding activity. Whether TAF4/TAF4b DNA binding directs TFIID to a specific core promoter element or facilitates TFIID binding to established core promoter elements is not known. Here we analyzed the mode of TAF4b·TAF12 DNA binding and show that this complex binds DNA with high affinity. The DNA length required for optimal binding is ∼70 bp. Although the complex displays a weak sequence preference, the nucleotide composition is less important than the length of the DNA for high affinity binding. Comparative expression profiling of wild-type and a DNA-binding mutant of TAF4 revealed common core promoter features in the down-regulated genes that include a TATA-box and an Initiator. Further examination of the PEL98 gene from this group showed diminished Initiator activity and TFIID occupancy in TAF4 DNA-binding mutant cells. These findings suggest that DNA binding by TAF4/4b-TAF12 facilitates the association of TFIID with the core promoter of a subset of genes. PMID:19635797

  1. Core promoter-specific gene regulation: TATA box selectivity and Initiator-dependent bi-directionality of serum response factor-activated transcription.

    PubMed

    Xu, Muyu; Gonzalez-Hurtado, Elsie; Martinez, Ernest

    2016-04-01

    Gene-specific activation by enhancers involves their communication with the basal RNA polymerase II transcription machinery at the core promoter. Core promoters are diverse and may contain a variety of sequence elements such as the TATA box, the Initiator (INR), and the downstream promoter element (DPE) recognized, respectively, by the TATA-binding protein (TBP) and TBP-associated factors of the TFIID complex. Core promoter elements contribute to the gene selectivity of enhancers, and INR/DPE-specific enhancers and activators have been identified. Here, we identify a TATA box-selective activating sequence upstream of the human β-actin (ACTB) gene that mediates serum response factor (SRF)-induced transcription from TATA-dependent but not INR-dependent promoters and requires the TATA-binding/bending activity of TBP, which is otherwise dispensable for transcription from a TATA-less promoter. The SRF-dependent ACTB sequence is stereospecific on TATA promoters but activates in an orientation-independent manner a composite TATA/INR-containing promoter. More generally, we show that SRF-regulated genes of the actin/cytoskeleton/contractile family tend to have a TATA box. These results suggest distinct TATA-dependent and INR-dependent mechanisms of TFIID-mediated transcription in mammalian cells that are compatible with only certain stereospecific combinations of activators, and that a TBP-TATA binding mechanism is important for SRF activation of the actin/cytoskeleton-related gene family.

  2. Prevalence and Characteristics of Basal Core Promoter Mutations in Iran and its Correlation with Acute and Chronic Hepatitis B Infection

    PubMed Central

    Nasrollaheian, Sadegh; Farshidfar, Gholamreza; Kheirabad, Ali Kargar; Gouklani, Hamed

    2016-01-01

    Introduction Manifestations of HBV infection differ in chronic and acute phases. Therefore, identifying the determinants such as mutations has a vital role in the treatment of the disease. A dual transversion in the basal core promoter (BCP) region is common among HBV patients. Thus, the present study was conducted with the objective of determining the prevalence of basal core promoter (BCP) mutations and its correlation with the outcome of HBV infection. Method In this cross-sectional study, samples were obtained from 182 Iranian HBsAg positive patients who were admitted to the Bandar Abbas Blood Transfusion Organization in 2012 and 2013. They were screened by ELISA test using commercial kits to detect serological marker anti-HBc IgM for distinct chronic hepatitis from acute infection. Thereafter, the extracted DNA was used for determination of the BCP mutations by PCR-RFLP technique. Data analyses were performed with SPSS 12 by Mann–Whitney U test, Fisher’s exact probability test, and t-test. Results BCP mutations were observed in 15 samples (8.24%) of the study population, and serological tests determined that, among the BCP mutants, one sample (6.67%) was HBeAg positive, 14 samples (93.33%) were HBeAg negative, and four samples (2.2%) were positive for anti-HBc IgM test. Data analysis indicated a statistically significant association between BCP mutations and acute hepatitis (p=0.002). However, no relationship was detected between the prevalences of the BCP mutations and gender of subjects (p>0.567). Conclusions The prevalence of BCP variants was low in the south of Iran, and this mutation can lead to acute phase of viral hepatitis. PMID:28163866

  3. New metabolic labelling medium for Trichomonas vaginalis and Tritrichomonas foetus using 35S methionine

    SciTech Connect

    Torian, B.E.; Kenny, G.E.

    1986-04-01

    A metabolic labelling medium was devised for Trichomonas vaginalis and Tritrichomonas foetus utilizing 35S methionine. T. vaginalis cultured for 24h in the medium took up approximately 27% of the available label and increased greater than two fold in number. Counts per microgram of protein were 32,555 +/- 10% between different strains or identical strains in different labelling runs. T. foetus took up approximately 5% of the available label and increased greater than two fold in 24h. This resulted in specific labelling of 12,704 cpm/ug protein +/- 10% between different runs with the same strain.

  4. Promotion

    PubMed Central

    Alam, Hasan B.

    2013-01-01

    This article gives an overview of the promotion process in an academic medical center. A description of different promotional tracks, tenure and endowed chairs, and the process of submitting an application is provided. Finally, some practical advice about developing skills and attributes that can help with academic growth and promotion is dispensed. PMID:24436683

  5. Hypocretin stimulates [(35)S]GTP gamma S binding in Hcrtr 2-transfected cell lines and in brain homogenate.

    PubMed

    Shiba, T; Ozu, M; Yoshida, Y; Mignot, E; Nishino, S

    2002-06-14

    In vitro functional analyses of hypocretin/orexin receptor systems were performed using [(125)I]hypocretin radioreceptor and hypocretin-stimulated [(35)S]GTP gamma S binding assay in cell lines expressing human or canine (wild-type and narcoleptic-mutation) hypocretin receptor 2 (Hcrtr 2). Hypocretin-2 stimulated [(35)S]GTP gamma S binding in human and canine Hcrtr 2 expressing cell lines, while cell lines expressing the mutated canine Hcrtr 2 did not exhibit specific binding for [(125)I]hypocretin or hypocretin-stimulated [(35)S]GTP gamma S. In rat brain homogenates, regional specific hypocretin-stimulated [(35)S]GTP gamma S binding was also observed. Hypocretin-stimulated [(35)S]GTP gamma S binding, may thus be a useful functional assay for hypocretin receptors in both cell lines and brain tissue homogenates.

  6. Green Infrastructure Research Promotes Students' Deeper Interest in Core Courses of a Water Resources Program

    NASA Astrophysics Data System (ADS)

    Yerk, W.; Montalto, F. A.; Foti, R.

    2015-12-01

    As one of most innovative among low impact development technologies, Green Infrastructure (GI) is a new technology that presents a range of potential research opportunities. Inherently linked to sustainability, urban quality of life, resilience, and other such topics, GI also represents a unique opportunity to highlight the social relevance of practical STEM research to undergraduate students. The nature of research on urban GI, in fact, as well as the accessibility of the GI sites, allows students to combine hands-on experience with theoretical work. Furthermore, the range of scales of the projects is such that they can be managed within a single term, but does not preclude longer engagement. The Sustainable Water Resource Engineering lab at Drexel University is engaged in two types of GI research outside the classroom. One type is a research co-op research internship. The second is a selective university-wide faculty-mentored summer scholarship STAR (Students Tackling Advanced Research) specifically designed for freshmen. The research projects we developed for those curricula can be accomplished by undergraduate students, but also address a larger research need in this emerging field. The research tasks have included identifying and calibrating affordable instruments, designing and building experimental setups, and monitoring and evaluating performance of GI sites. The work also promoted deeper understanding of the hydrological processes and initiated learning beyond the students' current curricula. The practice of the Lab's research being embedded into the educational process receives positive feedback from the students and achieves meaningful and long-lasting learning objectives. The experience helps students to students acquire hands-on experience, improves their metacognition and evidence-based inquiring into real-world problems, and further advances decision-making and communication skills.

  7. HBV core promoter mutations and AKT upregulate S-phase kinase-associated protein 2 to promote postoperative hepatocellular carcinoma progression

    PubMed Central

    Chen, Lubiao; Gu, Lin; Gu, Yurong; Wang, Hongbo; Deng, Meihai; Stamataki, Zania; Oo, Ye Htun; Huang, Yuehua

    2016-01-01

    Mutations in the hepatitis B virus (HBV) core promoter (CP) have been shown to be associated with hepatocellular carcinoma (HCC). The CP region overlaps HBV X gene, which activates AKT to regulate hepatocyte survival. However, the cooperation between these two cascades in HCC progression remains poorly understood. Here, we assayed virological factors and AKT expression in liver tissues from 56 HCC patients with better prognoses (BHCC, ≥5-year survival) and 58 with poor prognoses (PHCC, <5-year survival) after partial liver resection. Results showed double mutation A1762T/G1764A (TA) combined with other mutation(s) (TACO) in HBV genome and phosphorylated AKT (pAKT) were more common in PHCC than BHCC. TACO and pAKT levels correlated with proliferation and microvascularization but inversely correlated with apoptosis in HCC samples. These were more pronounced when TACO and pAKT co-expressed. Levels of p21 and p27 were decreased in TACO or pAKT overexpressing HCC due to SKP2 upregulation. Levels of E2F1 and both mRNA and protein of SKP2 were increased in TACO expressing HCC. Levels of 4EBP1/2 decreased and SKP2 mRNA level remained constant in pAKT-overexpressing HCC. Therefore, TACO and AKT are two independent predictors of postoperative survival in HCC. Their co-target, SKP2 may be a diagnostic or therapeutic marker. PMID:27779207

  8. Some dipeptides reverse the inhibitory effect of GABA on /sup 35/S-TBPS binding

    SciTech Connect

    Squires, R.F.; Saederup, E.

    1987-05-01

    All known GABA-A receptor blocker reverse the inhibitory effect of GABA on /sup 35/S-t-butylphosphorothionate (TBPS) binding to rat brain membranes in vitro. This system has already been used to identify several novel GABA antagonists. The authors now report that 12 out of 52 dipeptides tested (all containing L-amino acids), at 1 mM, significantly reverse the inhibitory effect of 1 ..mu..M GABA, which inhibits specific /sup 35/S-TBPS binding about 60%. Most of the active dipeptides contain an aromatic and a basic amino acid. Tryptophan usually conferred greater activity than phe or tyr, while arg usually conferred greater activity than lys or his. Several larger peptides containing the HFRW sequence found in ACTH were also GABA antagonists; ACTH(1-24), ACTH(1-18), ACTH(1-13), ACTH(4-10) and ..gamma..-MSH while ACTH(11-24) was inactive. The excitatory effects of these later peptides may be in part due to blockade of GABA-A receptors.

  9. Comparative labelling of rat epididymal spermatozoa by intratesticularly administered 65ZnCl2 and [35S]cysteine.

    PubMed

    Calvin, H I

    1981-01-01

    Spermatozoa of rats injected intratesticularly with 20 muCi65ZnCl2 and 10 muCi [35S]cysteine were collected from the caput and cauda of the epididymis at 2, 6, 10, 14, 18, 22 and 28 days after injection. The highest specific activities with respect to each isotope were observed in spermatozoa from the caput on Day 10. Maximal levels in spermatozoa from the cauda were obtained on Days 14 and 18 for 35S and Day 18 for 65Zn. Estimation of the relative behaviour of 65Zn and 35S by calculation of 65Zn/35S ratios suggests that: (1) 35S associated with spermatozoa arrived in the epididymis slightly in advance of 65Zn and was therefore probably incorporated more readily into proteins of very late spermatids; (2) approximately 60% of 65Zn was lost from spermatozoa and 75% from isolated sperm heads during transit from caput to cauda, assuming total retention of 35S; and (3) retention of 65Zn by the seminiferous epithelium was superior to that of [35S]cysteine, as indicated by increasing 65Zn/35S ratios following the days of peak specific activity in both caput and cauda epididymidal spermatozoa. Only small percentages of either isotope were recovered in isolated sperm heads, suggesting that the primary sites of labelling were in the sperm tail. Superior retention of 65Zn by testis was confirmed by increasing 65Zn/35S ratios in individual fractions of testicular homogenates between 2 and 10 days after injection. In addition, both isotopes appeared to be transferred from the testis cytosol to particulate material during this period.

  10. Environmental regulation of leaf colour in red 35S:PAP1 Arabidopsis thaliana.

    PubMed

    Rowan, Daryl D; Cao, Mingshu; Lin-Wang, Kui; Cooney, Janine M; Jensen, Dwayne J; Austin, Paul T; Hunt, Martin B; Norling, Cara; Hellens, Roger P; Schaffer, Robert J; Allan, Andrew C

    2009-01-01

    * High-temperature, low-light (HTLL) treatment of 35S:PAP1 Arabidopsis thaliana over-expressing the PAP1 (Production of Anthocyanin Pigment 1) gene results in reversible reduction of red colouration, suggesting the action of additional anthocyanin regulators. High-performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LCMS) and Affimetrix-based microarrays were used to measure changes in anthocyanin, flavonoids, and gene expression in response to HTLL. * HTLL treatment of control and 35S:PAP1 A. thaliana resulted in a reversible reduction in the concentrations of major anthocyanins despite ongoing over-expression of the PAP1 MYB transcription factor. Twenty-one anthocyanins including eight cis-coumaryl esters were identified by LCMS. The concentrations of nine anthocyanins were reduced and those of three were increased, consistent with a sequential process of anthocyanin degradation. Analysis of gene expression showed down-regulation of flavonol and anthocyanin biosynthesis and of transport-related genes within 24 h of HTLL treatment. No catabolic genes up-regulated by HTLL were found. * Reductions in the concentrations of anthocyanins and down-regulation of the genes of anthocyanin biosynthesis were achieved by environmental manipulation, despite ongoing over-expression of PAP1. Quantitative PCR showed reduced expression of three genes (TT8, TTG1 and EGL3) of the PAP1 transcriptional complex, and increased expression of the potential transcriptional repressors AtMYB3, AtMYB6 and AtMYBL2 coincided with HTLL-induced down-regulation of anthocyanin biosynthesis. * HTLL treatment offers a model system with which to explore anthocyanin catabolism and to discover novel genes involved in the environmental control of anthocyanins.

  11. An exceptionally long CA-repeat in the core promoter of SCGB2B2 links with the evolution of apes and Old World monkeys.

    PubMed

    Nikkhah, M; Rezazadeh, M; Khorram Khorshid, H R; Biglarian, A; Ohadi, M

    2016-01-15

    We have recently reported a genome-scale catalog of human protein-coding genes that contain "exceptionally long" STRs (≥6-repeats) in their core promoter, which may be of selective advantage in this species. At the top of that list, SCGB2B2 (also known as SCGBL), contains one of the longest CA-repeat STRs identified in a human gene core promoter, at 25-repeats. In the study reported here, we analyzed the conservation status of this CA-STR across evolution. The functional implication of this STR to alter gene expression activity was also analyzed in the HEK-293 cell line. We report that the SCGB2B2 core promoter CA-repeat reaches exceptional lengths, ranging from 9- to 25-repeats, across Apes (Hominoids) and the Old World monkeys (CA>2-repeats were not detected in any other species). The longest CA-repeats and highest identity in the SCGB2B2 protein sequence were observed between human and bonobo. A trend for increased gene expression activity was observed from the shorter to the longer CA-repeats (p<0.009), and the CA-repeat increased gene expression activity, per se (p<0.02). We propose that the SCGB2B2 gene core promoter CA-repeat functions as an expression code for the evolution of Apes and the Old World monkeys.

  12. Novel core promoter elements in the oomycete pathogen Phytophthora infestans and their influence on expression detected by genome-wide analysis

    PubMed Central

    2013-01-01

    Background The core promoter is the region flanking the transcription start site (TSS) that directs formation of the pre-initiation complex. Core promoters have been studied intensively in mammals and yeast, but not in more diverse eukaryotes. Here we investigate core promoters in oomycetes, a group within the Stramenopile kingdom that includes important plant and animal pathogens. Prior studies of a small collection of genes proposed that oomycete core promoters contain a 16 to 19 nt motif bearing an Initiator-like sequence (INR) flanked by a novel sequence named FPR, but this has not been extended to whole-genome analysis. Results We used expectation maximization to find over-represented motifs near TSSs of Phytophthora infestans, the potato blight pathogen. The motifs corresponded to INR, FPR, and a new element found about 25 nt downstream of the TSS called DPEP. TATA boxes were not detected. Assays of DPEP function by mutagenesis were consistent with its role as a core motif. Genome-wide searches found a well-conserved combined INR+FPR in only about 13% of genes after correcting for false discovery, which contradicted prior reports that INR and FPR are found together in most genes. INR or FPR were found alone near TSSs in 18% and 7% of genes, respectively. Promoters lacking the motifs had pyrimidine-rich regions near the TSS. The combined INR+FPR motif was linked to higher than average mRNA levels, developmentally-regulated transcription, and functions related to plant infection, while DPEP and FPR were over-represented in constitutively-expressed genes. The INR, FPR, and combined INR+FPR motifs were detected in other oomycetes including Hyaloperonospora arabidopsidis, Phytophthora sojae, Pythium ultimum, and Saprolegnia parasitica, while DPEP was found in all but S. parasitica. Only INR seemed present in a non-oomycete stramenopile. Conclusions The absence of a TATA box and presence of novel motifs show that the oomycete core promoter is diverged from that of

  13. CAPS-1 promotes fusion competence of stationary dense-core vesicles in presynaptic terminals of mammalian neurons

    PubMed Central

    Farina, Margherita; van de Bospoort, Rhea; He, Enqi; Persoon, Claudia M; van Weering, Jan RT; Broeke, Jurjen H; Verhage, Matthijs; Toonen, Ruud F

    2015-01-01

    Neuropeptides released from dense-core vesicles (DCVs) modulate neuronal activity, but the molecules driving DCV secretion in mammalian neurons are largely unknown. We studied the role of calcium-activator protein for secretion (CAPS) proteins in neuronal DCV secretion at single vesicle resolution. Endogenous CAPS-1 co-localized with synaptic markers but was not enriched at every synapse. Deletion of CAPS-1 and CAPS-2 did not affect DCV biogenesis, loading, transport or docking, but DCV secretion was reduced by 70% in CAPS-1/CAPS-2 double null mutant (DKO) neurons and remaining fusion events required prolonged stimulation. CAPS deletion specifically reduced secretion of stationary DCVs. CAPS-1-EYFP expression in DKO neurons restored DCV secretion, but CAPS-1-EYFP and DCVs rarely traveled together. Synaptic localization of CAPS-1-EYFP in DKO neurons was calcium dependent and DCV fusion probability correlated with synaptic CAPS-1-EYFP expression. These data indicate that CAPS-1 promotes fusion competence of immobile (tethered) DCVs in presynaptic terminals and that CAPS-1 localization to DCVs is probably not essential for this role. DOI: http://dx.doi.org/10.7554/eLife.05438.001 PMID:25719439

  14. CAPS-1 promotes fusion competence of stationary dense-core vesicles in presynaptic terminals of mammalian neurons.

    PubMed

    Farina, Margherita; van de Bospoort, Rhea; He, Enqi; Persoon, Claudia M; van Weering, Jan R T; Broeke, Jurjen H; Verhage, Matthijs; Toonen, Ruud F

    2015-02-26

    Neuropeptides released from dense-core vesicles (DCVs) modulate neuronal activity, but the molecules driving DCV secretion in mammalian neurons are largely unknown. We studied the role of calcium-activator protein for secretion (CAPS) proteins in neuronal DCV secretion at single vesicle resolution. Endogenous CAPS-1 co-localized with synaptic markers but was not enriched at every synapse. Deletion of CAPS-1 and CAPS-2 did not affect DCV biogenesis, loading, transport or docking, but DCV secretion was reduced by 70% in CAPS-1/CAPS-2 double null mutant (DKO) neurons and remaining fusion events required prolonged stimulation. CAPS deletion specifically reduced secretion of stationary DCVs. CAPS-1-EYFP expression in DKO neurons restored DCV secretion, but CAPS-1-EYFP and DCVs rarely traveled together. Synaptic localization of CAPS-1-EYFP in DKO neurons was calcium dependent and DCV fusion probability correlated with synaptic CAPS-1-EYFP expression. These data indicate that CAPS-1 promotes fusion competence of immobile (tethered) DCVs in presynaptic terminals and that CAPS-1 localization to DCVs is probably not essential for this role.

  15. Simple Method for High-Sensitivity Determination of Cosmogenic (35)S in Snow and Water Samples Collected from Remote Regions.

    PubMed

    Lin, Mang; Wang, Kun; Kang, Shichang; Thiemens, Mark H

    2017-03-15

    Cosmogenic (35)S is useful in understanding a wide variety of chemical and physical processes in the atmosphere, the hydrosphere, and the cryosphere. The 87.4-day half-life and the ubiquity of sulfur in natural environments renders it an ideal tracer of many phenomena. Measurements of (35)S in snow and water samples are scarce as existing analytical methods require a large volume of sample (>20 L) due to their high analytical activity background and low counting efficiency. Here, we present a new set of snow/water sample collecting and handling procedures for high-sensitivity determination of cosmogenic (35)S using a low-level liquid scintillation spectrometer. Laboratory experiments using diluted (35)S standards (with activities of <5 disintegrations per minute) showed a (35)S recovery percentage of ∼95%, demonstrating a relatively small deviation from the true value. Using this method, we successfully measured (35)S in ∼1 L of fresh snow sample collected from a glacier on the Tibetan Plateau to be 47 ± 7 mBq/L. On the basis of (35)S activities in 9 natural samples measured in this study, a first proof-of-concept approximation for age determinations and source attributions was presented. This new method will provide a powerful tool in studying (35)S in small volumes of snow and water samples, especially those from remote but climatically important regions such as the polar regions and the Tibetan Plateau and Himalayas. The measurements are particularly important as the radioactive sulfur provides an actual clock of glacial melting processes. With the growing rate of glacial loss, the need for measurements from remote locations becomes all the more important.

  16. Use of natural 35S to trace sulphate cycling in small lakes, Flattops Wilderness Area, Colorado, U.S.A.

    USGS Publications Warehouse

    Michel, Robert L.; Turk, John T.; Campbell, Donald H.; Mast, M. Alisa

    2002-01-01

    Measurements of the cosmogenically-produced 35S, a radioisotope of sulphur (t1/2 = 87 days), are reported for the Ned Wilson Lake watershed in Colorado. The watershed contains two small lakes and a flowing spring presumed to be representative of local ground water. The watershed is located in the Flattops Wilderness Area and the waters in the system have low alkalinity, making them sensitive to increases in acid and sulphate deposition. Time series of 35S measurements were made during the summers of 1995 and 1996 (July–September) at all three sites. The system is dominated by melting snow and an initial concentration of 16–20 mBq L-1was estimated for snowmelt based on a series of snow samples collected in the Rocky Mountains. The two lakes had large initial 35S concentrations in July, indicating that a large fraction of the lake water and sulphate was introduced by meltwater from that year's snowpack. In 1995 and 1996, 35S concentrations decreased more rapidly than could be accounted for by decay, indicating that other processes were affecting 35S concentrations. The most likely explanation is that exchange with sediments or the biota was removing 35S from the lake and replacing it with older sulphate devoid of 35S. In September of 1995 and 1996, 35S concentrations increased, suggesting that atmospheric deposition is important in the sulphate flux of these lakes in late summer. Sulphur-35 concentrations in the spring water were highly variable but never higher than 3.6 mBq L-1 and averaged 2 mBq L-1. Using a simple mixing model, it was estimated that 75% of the spring water was derived from precipitation of previous years.

  17. Variability in the Precore and Core Promoter Regions of HBV Strains in Morocco: Characterization and Impact on Liver Disease Progression

    PubMed Central

    Kitab, Bouchra; Essaid El Feydi, Abdellah; Afifi, Rajaa; Trepo, Christian; Benazzouz, Mustapha; Essamri, Wafaa; Zoulim, Fabien; Chemin, Isabelle; Alj, Hanane Salih; Ezzikouri, Sayeh; Benjelloun, Soumaya

    2012-01-01

    Background Hepatitis B virus (HBV) is one of the most common human pathogens that cause aggressive hepatitis and advanced liver disease (AdLD), including liver cirrhosis and Hepatocellular Carcinoma. The persistence of active HBV replication and liver damage after the loss of hepatitis B e antigen (HBeAg) has been frequently associated with mutations in the pre-core (pre-C) and core promoter (CP) regions of HBV genome that abolish or reduce HBeAg expression. The purpose of this study was to assess the prevalence of pre-C and CP mutations and their impact on the subsequent course of liver disease in Morocco. Methods/Principal Findings A cohort of 186 patients with HBeAg-negative chronic HBV infection was studied (81 inactive carriers, 69 with active chronic hepatitis, 36 with AdLD). Pre-C and CP mutations were analyzed by PCR-direct sequencing method. The pre-C stop codon G1896A mutation was the most frequent (83.9%) and was associated with a lower risk of AdLD development (OR, 0.4; 95% CI, 0.15–1.04; p = 0.04). HBV-DNA levels in patients with G1896A were not significantly different from the other patients carrying wild-type strains (p = 0.84). CP mutations C1653T, T1753V, A1762T/G1764A, and C1766T/T1768A were associated with higher HBV-DNA level and increased liver disease severity. Multiple logistic regression analysis showed that older age (≥40 years), male sex, high viral load (>4.3 log10 IU/mL) and CP mutations C1653T, T1753V, A1762T/G1764A, and C1766T/T1768A were independent risk factors for AdLD development. Combination of these mutations was significantly associated with AdLD (OR, 7.52; 95% CI, 4.8–8; p<0.0001). Conclusions This study shows for the first time the association of HBV viral load and CP mutations with the severity of liver disease in Moroccan HBV chronic carriers. The examination of CP mutations alone or in combination could be helpful for prediction of the clinical outcome. PMID:22905181

  18. Hepatitis B virus genotype C isolates with wild-type core promoter sequence replicate less efficiently than genotype B isolates but possess higher virion secretion capacity.

    PubMed

    Qin, Yanli; Tang, Xiaoli; Garcia, Tamako; Hussain, Munira; Zhang, Jiming; Lok, Anna; Wands, Jack; Li, Jisu; Tong, Shuping

    2011-10-01

    Infection by hepatitis B virus (HBV) genotype C is associated with a prolonged viremic phase, delayed hepatitis B e antigen (HBeAg) seroconversion, and an increased incidence of liver cirrhosis and hepatocellular carcinoma compared with genotype B infection. Genotype C is also associated with the more frequent emergence of core promoter mutations, which increase genome replication and are independently associated with poor clinical outcomes. We amplified full-length HBV genomes from serum samples from Chinese and U. S. patients with chronic HBV infection and transfected circularized genome pools or dimeric constructs of individual clones into Huh7 cells. The two genotypes could be differentiated by Western blot analysis due to the reactivities of M and L proteins toward a monoclonal pre-S2 antibody and slightly different S-protein mobilities. Great variability in replication capacity was observed for both genotypes. The A1762T/G1764A core promoter mutations were prevalent in genotype C isolates and correlated with increased replication capacity, while the A1752G/T mutation frequently found in genotype B isolates correlated with a low replication capacity. Importantly, most genotype C isolates with wild-type core promoter sequence replicated less efficiently than the corresponding genotype B isolates due to less efficient transcription of the 3.5-kb RNA. However, genotype C isolates often displayed more efficient virion secretion. We propose that the low intracellular levels of viral DNA and core protein of wild-type genotype C delay immune clearance and trigger the subsequent emergence of A1762T/G1764A core promoter mutations to upregulate replication; efficient virion secretion compensates for the low replication capacity to ensure the establishment of persistent infection by genotype C.

  19. Transfer of [3H]estrone-[35S]sulfate across guinea pig fetal membranes.

    PubMed

    Goldhawk, D E; Hobkirk, R

    1998-10-01

    The possible role of fetal membrane deconjugating activity in the movement of a charged steroid conjugate between fetal and maternal compartments was investigated. The ability of amnion and chorion laeve to transfer [3H]estrone-[35S]sulfate was assessed in both orientations of guinea pig tissue at 45 days and near parturition. While early amnion was impermeable, late tissue transferred approximately 50% (w/w) of the substrate in a bidirectional process that was non-saturable and independent of either deconjugation or ATP. Transfer across early chorion was similar to late amnion. Saturation curves from each tissue were superimposable, as were those of the time course. Transfer across both early and late chorion proceeded in the absence of deconjugation, with no effect of tissue orientation or ATP depletion. However, late chorion exhibited a decrease in estrone-sulfate transfer, as verified by concentration dependency and time course analyses, though transport across the tissue remained non-saturable. The results in amnion were congruous with the presence and absence of tight junctions in the epithelium of early and late tissue, respectively. However, sulfoconjugate transfer across early chorion proceeded in the presence of a paracellular barrier, suggesting specialized regulation of the transport process which extended late into gestation.

  20. Determination of {sup 35}S in radioisotope wastes by a wet oxidation

    SciTech Connect

    Lee, Heung N.; Sang-Hoon Kang; Hong Joo Ahn; Kwang Yong Jee; Wook Hyun Sohn

    2007-07-01

    The oxidation studies of a sulfur to a sulfate ion by various oxy-halide oxidants in organic (thiourea, methionine) and inorganic (sulfate, thiophosphate) compounds were carried out in an acidic solution. The optimized result of the oxidation reaction was obtained when a bromate compound (BrO{sub 3}{sup -}) as an oxidant and a 3 M HNO{sub 3} solvent. The chemical yield for the oxidation of the organic and inorganic sulfur compounds to a sulfate ion was monitored as 80% for thiophosphate, 87% for methionine, and 100% for thiourea and sulfate within 5% RSD. The oxidation of thiourea required at least 1.6 equivalents of the bromate in an acidic solution. In the case of the oxidation of methionine and thiophosphate, the oxidation yield was above 80% if the bromate was used at 20 times that of the substrates. The chemical yield in the paper sample (WypAll) exceeded 100% because of its background sulfur contents (910 ppm). The sulfate ion was quantitatively measured by using GPC and/or LSC counting of 3 S followed by precipitates of BaSO{sub 4}. The interfering nuclides ({sup 14}C, {sup 32}P) were removed in an acidic condition. The minimum detectable activity (MDA) of {sup 35}S was found to be 0.1 Bq/g. (authors)

  1. Transformation of Lesquerella fendleri with the new binary vector pGPro4-35S

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crop genetic engineering requires the use of various promoters to control the expression of introduced transgenes. Some of the binary vectors currently available for promoter characterization in dicotyledonous plants have pitfalls due to their construction, such as containing a selectable marker ca...

  2. Can a functionalized phosphine ligand promote room temperature luminescence of the [Ru(bpy)(tpy)]2+ core?

    PubMed

    Lebon, Emilie; Bastin, Stéphanie; Sutra, Pierre; Vendier, Laure; Piau, Rémi E; Dixon, Isabelle M; Boggio-Pasqua, Martial; Alary, Fabienne; Heully, Jean-Louis; Igau, Alain; Juris, Alberto

    2012-01-18

    Unexpected room temperature luminescence is observed and rationalized by highly challenging excited state calculations for a functionalized phosphine ligand coordinated on the [Ru(bpy)(tpy)](2+) core.

  3. Modulation of [(35)S]GTPgammaS binding to chinese hamster ovary cell membranes by D(2(short)) dopamine receptors.

    PubMed

    Terasmaa, A; Finnman, U B; Owman, C; Ferré, S; Fuxe, K; Rinken, A

    2000-02-18

    Rat dopamine D(2short) expressed in Chinese hamster ovary (CHO) cells were characterized by means of activation of [(35)S]-guanosine 5'-O-(gamma-thiotriphosphate) ([(35)S]GTPgammaS) binding and inhibition of [(3)H]raclopride binding. Among 18 dopaminergic ligands studied dopamine, NPA, apomorphine and quinpirole were full agonists in activation of [(35)S]GTPgammaS binding, while seven ligands were partial agonists with efficacies from 16 to 69% of the effect of dopamine and seven ligands were antagonists having no effect on the basal level of [(35)S]GTPgammaS binding, but inhibited dopamine-dependent activation in a dose-response manner. Despite the different efficacies, the potencies of all 18 ligands to modulate [(35)S]GTPgammaS binding revealed a good correlation with their potencies to inhibit [(3)H]raclopride binding in the CHO cell membranes. This indicates that the binding of the ligand to the receptor determines its potency, but has no direct correlation with its intrinsic activity.

  4. Human differentiation-related gene NDRG1 is a Myc downstream-regulated gene that is repressed by Myc on the core promoter region.

    PubMed

    Zhang, Jian; Chen, Suning; Zhang, Wei; Zhang, Jing; Liu, Xinping; Shi, Hai; Che, Honglei; Wang, Weizhong; Li, Fuyang; Yao, Libo

    2008-07-01

    N-Myc downstream-regulated gene 1 (ndrg1) is up-regulated in N-Myc knockout mouse embryos. The human NDRG family consists of 4 highly homologous members and human Ndrg1 exhibits approximately 94% homology with mouse ndrg1. However, the regulatory mechanism of NDRG1 via Myc repression is as yet unknown. We previously identified human NDRG2 and demonstrated that this gene is transcriptionally down-regulated by Myc via Miz-1-dependent interaction with the core promoter region of NDRG2. Here, we provide evidence that human NDRG1 is regulated by Myc in a manner similar to NDRG2. We found that Ndrg1 expression levels were enhanced as Myc expression declined in differentiated cells, but were down-regulated following Myc induction. The data revealed that both N-Myc and c-Myc can repress human NDRG1 at the transcriptional level. We further determined that the core promoter region of human NDRG1 is required for Myc repression, and verified the interaction of Myc with the core promoter region. However, the presence of the protein synthesis inhibitor cycloheximide could reverse the repression of Myc, indicating the indirect repression of human NDRG1 by Myc. Moreover, we found that c-Myc-mediated repression can be inhibited by TSA, an HDACs inhibitor, which suggests the involvement of HDACs in the repression process. Taken together, our results demonstrate that, in common with NDRG2, human NDRG1 can be indirectly transcriptionally down-regulated by Myc via interaction with the NDRG1 core promoter.

  5. Investigations into the origin of the spurious 17 keV neutrino signal observed in35S beta decay

    NASA Astrophysics Data System (ADS)

    Bowler, M. G.; Jelley, N. A.

    1995-09-01

    An exhaustive study has been made of the β spectrum of35S, recorded with a Si(Li) detector. The object was to identify the origin of a distortion in the35S β spectrum some 17 keV below the end point, reported over three years ago and interpreted then as evidence for a 17 keV neutrino. Measurements with different source-detector spacings and with varied collimation have shown that there is a long range curvature in the Kurie plot which is a sensitive function of configuration, but the principal origin of the distortion is energy loss in the35S sources. The35S sources, prepared by chemical adsorption of Ba35SO4 on a gold substrate, are clumped and locally thick. Electrons near the end point lose ˜0.3 keV in the source material and if this is taken into account the spectra are well fitted without any admixture of 17 keV neutrino. The source thickness has been investigated with a proton microprobe and determined from both source tilting and the yield of barium K X-rays; these studies are discussed in detail. The uncertainties in and justification for the form of the electron response function employed are also thoroughly discussed. If there is no systematic error common to the majority of 14 independent sets of35S data, the admixture of 17 keV neutrino is <10-3 (95% CL). A simple search for a kink at 150 keV in the combined data from all 14 runs yielded a limit of 1.8×10-3 (95% CL). The end point of the35S β spectrum is found to be 167.60±0.05 keV.

  6. Metabolism of 35S- and 14C-labeled 1-methyl-2-mercaptoimidazole in vitro and in vivo.

    PubMed

    Taurog, A; Dorris, M L; Guziec, F S

    1989-01-01

    We previously described an in vitro incubation system for studying the mechanism of inhibition of thyroid peroxidase (TPO)-catalyzed iodination by the antithyroid drug 1-methyl-2-mercaptoimidazole (MMI). Inhibition of iodination in this system may be reversible or irreversible, depending on the relative concentrations of iodide and MMI and on the TPO concentration. Metabolism of the drug occurs under both conditions, and in the present investigation we used 35S- and 14C-labeled MMI together with reverse phase HPLC to examine the metabolic products associated with reversible and irreversible inhibition of iodination by MMI. Under conditions of reversible inhibition, MMI was rapidly metabolized and disappeared completely from the incubation mixture. With [35S]MMI, the earliest detectable 35S-labeled product was MMI disulfide, which reached a peak after a few minutes and then declined to undetectable levels. Coincident with the decrease in disulfide was the appearance of two 35S peaks, the major one corresponding to sulfate/sulfite, and the other to a component eluting at 7.5 min. Similar results were obtained for the disulfide and for the 7.5 min metabolite with [14C]MMI. The major 14C-labeled metabolite containing no S appeared to be 1-methylimidazole. Under conditions of irreversible inhibition, MMI disulfide was also the earliest detectable 35S-labeled metabolite. However, MMI decreased more slowly, and after reaching a nadir at about 6 min returned gradually to a level about halfway between the initial and the minimum value. The reformation of MMI appeared to involve the nonenzymatic disproportionation of MMI disulfide. Formation of the 7.5 min peak was also observed, but there was no formation of sulfate/sulfite. The difference in metabolic pattern between the reversible and irreversible conditions is primarily related to the rapid inactivation of TPO that occurs under irreversible conditions. The metabolism of [35S]MMI in thyroids of rats injected with the

  7. Forming the phosphate layer in reconstituted horse spleen ferritin and the role of phosphate in promoting core surface redox reactions.

    PubMed

    Johnson, J L; Cannon, M; Watt, R K; Frankel, R B; Watt, G D

    1999-05-18

    Apo horse spleen ferritin (apo HoSF) was reconstituted to various core sizes (100-3500 Fe3+/HoSF) by depositing Fe(OH)3 within the hollow HoSF interior by air oxidation of Fe2+. Fe2+ and phosphate (Pi) were then added anaerobically at a 1:4 ratio, and both Fe2+ and Pi were incorporated into the HoSF cores. The resulting Pi layer consisted of Fe2+ and Pi at about a 1:3 ratio which is strongly attached to the reconstituted ferritin mineral core surface and is stable even after air oxidation of the bound Fe2+. The total amount of Fe2+ and Pi bound to the iron core surface increases as the core volume increases up to a maximum near 2500 iron atoms, above which the size of the Pi layer decreases with increasing core size. Mössbauer spectroscopic measurements of the Pi-reconstituted HoSF cores using 57Fe2+ show that 57Fe3+ is the major species present under anaerobic conditions. This result suggests that the incoming 57Fe2+ undergoes an internal redox reaction to form 57Fe3+ during the formation of the Pi layer. Addition of bipyridine removes the 57Fe3+ bound in the Pi layer as [57Fe(bipy)3]2+, showing that the bound 57Fe2+ has not undergone irreversible oxidation. This result is related to previous studies showing that 57Fe2+ bound to native core is reversibly oxidized under anaerobic conditions in native holo bacterial and HoSF ferritins. Attempts to bury the Pi layer of native or reconstituted HoSF by adding 1000 additional iron atoms were not successful, suggesting that after its formation, the Pi layer "floats" on the developing iron mineral core.

  8. The global regulator Ncb2 escapes from the core promoter and impacts transcription in response to drug stress in Candida albicans.

    PubMed

    Shariq, Mohd; Dhamgaye, Sanjiveeni; Nair, Remya; Goyal, Neha; Jain, Vaibhav; Mukhopadhyay, Arnab; Mondal, Alok K; Mukhopadhyay, Gauranga; Prasad, Rajendra

    2017-04-06

    Ncb2, the β subunit of NC2 complex, a heterodimeric regulator of transcription was earlier shown to be involved in the activated transcription of CDR1 gene in azole resistant isolate (AR) of Candida albicans. This study examines its genome-wide role by profiling Ncb2 occupancy between genetically matched pair of azole sensitive (AS) and AR clinical isolates. A comparison of Ncb2 recruitment between the two isolates displayed that 29 genes had higher promoter occupancy of Ncb2 in the AR isolate. Additionally, a host of genes exhibited exclusive occupancy of Ncb2 at promoters of either AR or AS isolate. The analysis also divulged new actors of multi-drug resistance, whose transcription was activated owing to the differential occupancy of Ncb2. The conditional, sequence-specific positional escape of Ncb2 from the core promoter in AS isolate and its preferential recruitment to the core promoter of certain genes in AR isolates was most noteworthy means of transcription regulation. Together, we show that positional rearrangement of Ncb2 resulting in either activation or repression of gene expression in response to drug-induced stress, represents a novel regulatory mechanism that opens new opportunities for therapeutic intervention to prevent development of drug tolerance in C. albicans cells.

  9. The global regulator Ncb2 escapes from the core promoter and impacts transcription in response to drug stress in Candida albicans

    PubMed Central

    Shariq, Mohd; Dhamgaye, Sanjiveeni; Nair, Remya; Goyal, Neha; Jain, Vaibhav; Mukhopadhyay, Arnab; Mondal, Alok K.; Mukhopadhyay, Gauranga; Prasad, Rajendra

    2017-01-01

    Ncb2, the β subunit of NC2 complex, a heterodimeric regulator of transcription was earlier shown to be involved in the activated transcription of CDR1 gene in azole resistant isolate (AR) of Candida albicans. This study examines its genome-wide role by profiling Ncb2 occupancy between genetically matched pair of azole sensitive (AS) and AR clinical isolates. A comparison of Ncb2 recruitment between the two isolates displayed that 29 genes had higher promoter occupancy of Ncb2 in the AR isolate. Additionally, a host of genes exhibited exclusive occupancy of Ncb2 at promoters of either AR or AS isolate. The analysis also divulged new actors of multi-drug resistance, whose transcription was activated owing to the differential occupancy of Ncb2. The conditional, sequence-specific positional escape of Ncb2 from the core promoter in AS isolate and its preferential recruitment to the core promoter of certain genes in AR isolates was most noteworthy means of transcription regulation. Together, we show that positional rearrangement of Ncb2 resulting in either activation or repression of gene expression in response to drug-induced stress, represents a novel regulatory mechanism that opens new opportunities for therapeutic intervention to prevent development of drug tolerance in C. albicans cells. PMID:28383050

  10. Endosperm protein synthesis and L-(/sup 35/S)methionine incorporation in maize kernels cultured in vitro

    SciTech Connect

    Cully, D.E.; Gengenbach, B.G.; Smith, J.A.; Rubenstein, I.; Connely, J.A.; Park, W.D.

    1984-02-01

    This study was conducted to examine protein synthesis and L-(/sup 35/S)methionine incorporation into the endosperm of Zea mays L. kernels developing in vitro. Two-day-old kernels of the inbred line W64A were placed in culture on a defined medium containing 10 microCuries L-(/sup 35/S)methionine per milliliter (13 milliCuries per millimole) and harvested at 10, 15, 20, 25, 30, 35, and 40 days after pollination. Cultured kernels attained a final endosperm mass of 120 milligrams compared to 175 milligrams for field-grown controls. Field and cultured kernels had similar concentrations (microgram per milligram endosperm for total protein, albumin plus globulin, zein, and glutelin fractions at most kernel ages. Sodium, dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing patterns for endosperm proteins were similar for field and cultured kernels throughout development. By 15 days, over 70% of the L-(/sup 35/S)methionine taken up was present in endosperm proteins. Label incorporation visualized by fluorography generally followed the protein intensity of the stained gels. The high methionine content, low molecular weight zeins (i.e. 15 and 9 kilodaltons) were highly labeled. All of the radioactivity in hydrolyzed zein samples was recovered in the methionine peak indicating minimal conversion to L-(/sup 35/S)cysteine. The procedure described here is suitable for long term culture and labeling experiments in which continued kernel development is required.

  11. [Age peculiarities of the intake dynamics of (35S)thiamine and its phosphoric esters administered parenterally into rat organs].

    PubMed

    Rozanov, A Ia; Karpov, L M

    1981-01-01

    The maximal intake of [35S]thiamine for the first hours followed administration of its physiological dose (150 mumol/kg) into the blood small intestine, kidneys, liver, myocardium and brain grows in ontogenesis by 55-60, 25-30, 80-110, 25-40, 15-30, 5-12%. This evidences for a more pronounced thiamine lack in old animals as compared to the young ones. After injection of labelled thiamine diphosphate the increment of the vitamin B1 total amount is the highest in the kidneys and small intestine of old animals. A higher increment of the vitamin B1 total amount in tissues of old rats after the labelled thiamine injection may be explained by a delayed intensity of its renewal deficiency. [35S]thiamine phosphate and [35S]thiamine diphosphate especially enter all organs, except for the liver, more intensively than [35S]thiamine (their amount is by 25-40% higher in all age groups).

  12. Electrophoresis of /sup 35/S-labeled proteoglycans of polyacrylamide-agarose composite gels and their visualization by fluorography

    SciTech Connect

    Carney, S.L.; Bayliss, M.T.; Collier, J.M.; Muir, H.

    1986-01-01

    Techniques for the electrophoresis of /sup 35/S-labeled proteoglycans on polyacrylamide-agarose gel slabs and subsequent fixation, impregnation, and fluorography of such electrophoretograms have been developed. The procedure permits the examination of newly synthesized proteoglycan subspecies using a rapid technique, previously unavailable for these labeled molecules.

  13. Development of Useful Recombinant Promoter and Its Expression Analysis in Different Plant Cells Using Confocal Laser Scanning Microscopy

    PubMed Central

    Kumar, Deepak; Sahoo, Dipak K.; Maiti, Indu B.; Dey, Nrisingha

    2011-01-01

    Background Designing functionally efficient recombinant promoters having reduced sequence homology and enhanced promoter activity will be an important step toward successful stacking or pyramiding of genes in a plant cell for developing transgenic plants expressing desired traits(s). Also basic knowledge regarding plant cell specific expression of a transgene under control of a promoter is crucial to assess the promoter's efficacy. Methodology/Principal Findings We have constructed a set of 10 recombinant promoters incorporating different up-stream activation sequences (UAS) of Mirabilis mosaic virus sub-genomic transcript (MS8, -306 to +27) and TATA containing core domains of Figwort mosaic virus sub-genomic transcript promoter (FS3, −271 to +31). Efficacies of recombinant promoters coupled to GUS and GFP reporter genes were tested in tobacco protoplasts. Among these, a 369-bp long hybrid sub-genomic transcript promoter (MSgt-FSgt) showed the highest activity in both transient and transgenic systems. In a transient system, MSgt-FSgt was 10.31, 2.86 and 2.18 times more active compared to the CaMV35S, MS8 and FS3 promoters, respectively. In transgenic tobacco (Nicotiana tabaccum, var. Samsun NN) and Arabidopsis plants, the MSgt-FSgt hybrid promoter showed 14.22 and 7.16 times stronger activity compared to CaMV35S promoter respectively. The correlation between GUS activity and uidA-mRNA levels in transgenic tobacco plants were identified by qRT-PCR. Both CaMV35S and MSgt-FSgt promoters caused gene silencing but the degree of silencing are less in the case of the MSgt-FSgt promoter compared to CaMV35S. Quantification of GUS activity in individual plant cells driven by the MSgt-FSgt and the CaMV35S promoter were estimated using confocal laser scanning microscopy and compared. Conclusion and Significance We propose strong recombinant promoter MSgt-FSgt, developed in this study, could be very useful for high-level constitutive expression of transgenes in a wide variety

  14. Simvastatin inhibits the core promoter of the TXNRD1 gene and lowers cellular TrxR activity in HepG2 cells.

    PubMed

    Ekström, Lena; Johansson, Maria; Monostory, Katalin; Rundlöf, Anna-Klara; Arnér, Elias S J; Björkhem-Bergman, Linda

    2013-01-04

    Thioredoxin reductase 1 (TrxR1) is a selenocysteine-containing redox-active enzyme that is thought to be important during carcinogenesis. We have recently shown that treatment with statins, HMGCoA reductase inhibitors, reduces the levels of TrxR1 in liver of both rat and human. The reduced TrxR1 levels were correlated with inhibited hepatocarcinogenesis in a rat model. The aim of the present study was to investigate if statins affect the activity of the human TXNRD1 core promoter, which guides expression of TrxR1, and if the effects by statins on TrxR1 expression in liver could be reproduced in a cellular model system. We found that simvastatin and fluvastatin decreased cellular TrxR activity in cultured human liver-derived HepG2 cells with approximately 40% (p<0.05). Simvastatin, but not fluvastatin or atorvastatin, also reduced the TXNRD1 promoter activity in HepG2 cells by 20% (p<0.01). In line with this result, TrxR1 mRNA levels decreased with about 25% in non-transfected HepG2 cells upon treatment with simvastatin (p<0.01). Concomitant treatment with mevalonate could not reverse these effects of simvastatin, indicating that other mechanisms than HMGCoA reductase inhibition was involved. Also, simvastatin did not inhibit sulforaphane-derived stimulation of the TXNRD1 core promoter activity, suggesting that the inhibition by simvastatin was specific for basal and not Nrf2-activated TrxR1 expression. In contrast to simvastatin, the two other statins tested, atorvastatin or fluvastatin, did not influence the TrxR1 mRNA levels. Thus, our results reveal a simvastatin-specific reduction of cellular TrxR1 levels that at least in part involves direct inhibitory effects on the basal activity of the core promoter guiding TrxR1 expression.

  15. Insertion of core CpG island element into human CMV promoter for enhancing recombinant protein expression stability in CHO cells.

    PubMed

    Mariati; Yeo, Jessna H M; Koh, Esther Y C; Ho, Steven C L; Yang, Yuansheng

    2014-01-01

    The human cytomegalovirus promoter (hCMV) is susceptible to gene silencing in CHO cells, most likely due to epigenetic events, such as DNA methylation and histone modifications. The core CpG island element (IE) from the hamster adenine phosphoribosyltransferase gene has been shown to prevent DNA methylation. A set of modified hCMV promoters was developed by inserting one or two copies of IE in either forward or reverse orientations either upstream of the hCMV enhancer, between the enhancer and core promoter (CP), or downstream of the CP. The modified hCMV with one copy of IE inserted between the enhancer and core promoter in reverse orientation (MR1) was most effective at enhancing expression stability without compromising expression level when compared with the wild-type (WT) hCMV. A third of 18 EGFP expressing clones generated using MR1 retained 70% of their starting expression level after 8 weeks of culture in the absence of selection pressure, while none of 18 WT hCMV generated clones had expression above 50%. MR1 also improved antibody expression stability of methotrexate (MTX) amplified CHO cell lines. Stably transfected pools generated using MR1 maintained 62% of their original monoclonal antibody titer after 8 weeks of culture in the absence of MTX, compared to only 37% for WT hCMV pools. Low levels of CpG methylation within both WT hCMV and MR1 were observed in all the analyzed cell lines and the methylation levels did not correlate to the expression stability, suggesting IE enhances expression stability by other mechanisms other than preventing methylation.

  16. Domains of Core Competency, Standards, and Quality Assurance for Building Global Capacity in Health Promotion: The Galway Consensus Conference Statement

    ERIC Educational Resources Information Center

    Allegrante, John P.; Barry, Margaret M.; Airhihenbuwa, Collins O.; Auld, M. Elaine; Collins, Janet L.; Lamarre, Marie-Claude; Magnusson, Gudjon; McQueen, David V.; Mittelmark, Maurice B.

    2009-01-01

    This paper reports the outcome of the Galway Consensus Conference, an effort undertaken as a first step toward international collaboration on credentialing in health promotion and health education. Twenty-nine leading authorities in health promotion, health education, and public health convened a 2-day meeting in Galway, Ireland, during which the…

  17. Metabolism of /sup 35/S- and /sup 14/C-labeled 1-methyl-2-mercaptoimidazole in vitro and in vivo

    SciTech Connect

    Taurog, A.; Dorris, M.L.; Guziec, F.S. Jr.

    1989-01-01

    We previously described an in vitro incubation system for studying the mechanism of inhibition of thyroid peroxidase (TPO)-catalyzed iodination by the antithyroid drug 1-methyl-2-mercaptoimidazole (MMI). Inhibition of iodination in this system may be reversible or irreversible, depending on the relative concentrations of iodide and MMI and on the TPO concentration. Metabolism of the drug occurs under both conditions, and in the present investigation we used 35S- and 14C-labeled MMI together with reverse phase HPLC to examine the metabolic products associated with reversible and irreversible inhibition of iodination by MMI. Under conditions of reversible inhibition, MMI was rapidly metabolized and disappeared completely from the incubation mixture. With (35S)MMI, the earliest detectable 35S-labeled product was MMI disulfide, which reached a peak after a few minutes and then declined to undetectable levels. Coincident with the decrease in disulfide was the appearance of two 35S peaks, the major one corresponding to sulfate/sulfite, and the other to a component eluting at 7.5 min. Similar results were obtained for the disulfide and for the 7.5 min metabolite with (14C)MMI. The major 14C-labeled metabolite containing no S appeared to be 1-methylimidazole. Under conditions of irreversible inhibition, MMI disulfide was also the earliest detectable 35S-labeled metabolite. However, MMI decreased more slowly, and after reaching a nadir at about 6 min returned gradually to a level about halfway between the initial and the minimum value. The reformation of MMI appeared to involve the nonenzymatic disproportionation of MMI disulfide. Formation of the 7.5 min peak was also observed, but there was no formation of sulfate/sulfite.

  18. Heparan Sulfate Proteoglycans Containing a Glypican 5 Core and 2-O-Sulfo-iduronic Acid Function as Sonic Hedgehog Co-receptors to Promote Proliferation*♦

    PubMed Central

    Witt, Rochelle M.; Hecht, Marie-Lyn; Pazyra-Murphy, Maria F.; Cohen, Samuel M.; Noti, Christian; van Kuppevelt, Toin H.; Fuller, Maria; Chan, Jennifer A.; Hopwood, John J.; Seeberger, Peter H.; Segal, Rosalind A.

    2013-01-01

    Sonic Hedgehog (Shh) signaling is crucial for growth, cell fate determination, and axonal guidance in the developing nervous system. Although the receptors Patched (Ptch1) and Smoothened (Smo) are required for Shh signaling, a number of distinct co-receptors contribute to these critical responses to Shh. Several membrane-embedded proteins such as Boc, Cdo, and Gas1 bind Shh and promote signaling. In addition, heparan sulfate proteoglycans (HSPGs) have also been implicated in the initiation of Shh responses. However, the attributes of HSPGs that function as co-receptors for Shh have not yet been defined. Here, we identify HSPGs containing a glypican 5 core protein and 2-O-sulfo-iduronic acid residues at the nonreducing ends of the glycans as co-receptors for Shh. These HSPG co-receptors are expressed by cerebellar granule cell precursors and promote Shh binding and signaling. At the subcellular level, these HSPG co-receptors are located adjacent to the primary cilia that act as Shh signaling organelles. Thus, Shh binds to HSPG co-receptors containing a glypican 5 core and 2-O-sulfo-iduronic acid to promote neural precursor proliferation. PMID:23867465

  19. Heparan sulfate proteoglycans containing a glypican 5 core and 2-O-sulfo-iduronic acid function as Sonic Hedgehog co-receptors to promote proliferation.

    PubMed

    Witt, Rochelle M; Hecht, Marie-Lyn; Pazyra-Murphy, Maria F; Cohen, Samuel M; Noti, Christian; van Kuppevelt, Toin H; Fuller, Maria; Chan, Jennifer A; Hopwood, John J; Seeberger, Peter H; Segal, Rosalind A

    2013-09-06

    Sonic Hedgehog (Shh) signaling is crucial for growth, cell fate determination, and axonal guidance in the developing nervous system. Although the receptors Patched (Ptch1) and Smoothened (Smo) are required for Shh signaling, a number of distinct co-receptors contribute to these critical responses to Shh. Several membrane-embedded proteins such as Boc, Cdo, and Gas1 bind Shh and promote signaling. In addition, heparan sulfate proteoglycans (HSPGs) have also been implicated in the initiation of Shh responses. However, the attributes of HSPGs that function as co-receptors for Shh have not yet been defined. Here, we identify HSPGs containing a glypican 5 core protein and 2-O-sulfo-iduronic acid residues at the nonreducing ends of the glycans as co-receptors for Shh. These HSPG co-receptors are expressed by cerebellar granule cell precursors and promote Shh binding and signaling. At the subcellular level, these HSPG co-receptors are located adjacent to the primary cilia that act as Shh signaling organelles. Thus, Shh binds to HSPG co-receptors containing a glypican 5 core and 2-O-sulfo-iduronic acid to promote neural precursor proliferation.

  20. Detailed analysis of Helicobacter pylori Fur-regulated promoters reveals a Fur box core sequence and novel Fur-regulated genes.

    PubMed

    Pich, Oscar Q; Carpenter, Beth M; Gilbreath, Jeremy J; Merrell, D Scott

    2012-06-01

    In Helicobacter pylori, iron balance is controlled by the Ferric uptake regulator (Fur), an iron-sensing repressor protein that typically regulates expression of genes implicated in iron transport and storage. Herein, we carried out extensive analysis of Fur-regulated promoters and identified a 7-1-7 motif with dyad symmetry (5'-TAATAATnATTATTA-3'), which functions as the Fur box core sequence of H. pylori. Addition of this sequence to the promoter region of a typically non-Fur regulated gene was sufficient to impose Fur-dependent regulation in vivo. Moreover, mutation of this sequence within Fur-controlled promoters negated regulation. Analysis of the H. pylori chromosome for the occurrence of the Fur box established the existence of well-conserved Fur boxes in the promoters of numerous known Fur-regulated genes, and revealed novel putative Fur targets. Transcriptional analysis of the new candidate genes demonstrated Fur-dependent repression of HPG27_51, HPG27_52, HPG27_199, HPG27_445, HPG27_825 and HPG27_1063, as well as Fur-mediated activation of the cytotoxin associated gene A, cagA (HPG27_507). Furthermore, electrophoretic mobility shift assays confirmed specific binding of Fur to the promoters of each of these genes. Future experiments will determine whether loss of Fur regulation of any of these particular genes contributes to the defects in colonization exhibited by the H. pylori fur mutant.

  1. Interaction of the Transcription Start Site Core Region and Transcription Factor YY1 Determine Ascorbate Transporter SVCT2 Exon 1a Promoter Activity

    PubMed Central

    Qiao, Huan; May, James M.

    2012-01-01

    Transcription of the ascorbate transporter, SVCT2, is driven by two distinct promoters in exon 1 of the transporter sequence. The exon 1a promoter lacks a classical transcription start site and little is known about regulation of promoter activity in the transcription start site core (TSSC) region. Here we present evidence that the TSSC binds the multifunctional initiator-binding protein YY1. Electrophoresis shift assays using YY1 antibody showed that YY1 is present as one of two major complexes that specifically bind to the TSSC. The other complex contains the transcription factor NF-Y. Mutations in the TSSC that decreased YY1 binding also impaired the exon 1a promoter activity despite the presence of an upstream activating NF-Y/USF complex, suggesting that YY1 is involved in the regulation of the exon 1a transcription. Furthermore, YY1 interaction with NF-Y and/or USF synergistically enhanced the exon 1a promoter activity in transient transfections and co-activator p300 enhanced their synergistic activation. We propose that the TSSC plays a vital role in the exon 1a transcription and that this function is partially carried out by the transcription factor YY1. Moreover, co-activator p300 might be able to synergistically enhance the TSSC function via a “bridge” mechanism with upstream sequences. PMID:22532872

  2. Professional Development for Promoting 21st Century Skills and Common Core State Standards in Foreign Language and Social Studies Classrooms

    ERIC Educational Resources Information Center

    Beriswill, Joanne Elizabeth; Bracey, Pamela Scott; Sherman-Morris, Kathleen; Huang, Kun; Lee, Sang Joon

    2016-01-01

    To help satisfy the pressing need for technology-related professional development for in-service teachers, the Global Academic Essentials Teacher Institute (GAETI) was implemented to provide in-service foreign language and social studies teachers with content, pedagogy, and technology explorations centered on the teaching of the Common Core State…

  3. Toward core inter-professional health promotion competencies to address the non-communicable diseases and their risk factors through knowledge translation: Curriculum content assessment

    PubMed Central

    2014-01-01

    Background To increase the global impact of health promotion related to non-communicable diseases, health professionals need evidence-based core competencies in health assessment and lifestyle behavior change. Assessment of health promotion curricula by health professional programs is a first step. Such program assessment is a means of 1. demonstrating collective commitment across health professionals to prevent non-communicable diseases; 2. addressing the knowledge translation gap between what is known about non-communicable diseases and their risk factors consistent with ‘best’ practice; and, 3. establishing core health-based competencies in the entry-level curricula of established health professions. Discussion Consistent with the World Health Organization’s definition of health (i.e., physical, emotional and social wellbeing) and the Ottawa Charter, health promotion competencies are those that support health rather than reduce signs and symptoms primarily. A process algorithm to guide the implementation of health promotion competencies by health professionals is described. The algorithm outlines steps from the initial assessment of a patient’s/client’s health and the indications for health behavior change, to the determination of whether that health professional assumes primary responsibility for implementing health behavior change interventions or refers the patient/client to others. An evidence-based template for assessment of the health promotion curriculum content of health professional education programs is outlined. It includes clinically-relevant behavior change theory; health assessment/examination tools; and health behavior change strategies/interventions that can be readily integrated into health professionals’ practices. Summary Assessment of the curricula in health professional education programs with respect to health promotion competencies is a compelling and potentially cost-effective initial means of preventing and reversing non

  4. Specific mutations of basal core promoter are associated with chronic liver disease in hepatitis B virus subgenotype D1 prevalent in Turkey.

    PubMed

    Sunbul, Mustafa; Sugiyama, Masaya; Kurbanov, Fuat; Leblebicioglu, Hakan; Khan, Anis; Elkady, Abeer; Tanaka, Yasuhito; Mizokami, Masashi

    2013-02-01

    The role of hepatitis B virus (HBV) genetics in the clinical manifestations of infection is being increasingly recognized. Genotype D is one of eight currently recognized major HBV genotypes. The virus is ubiquitous worldwide, but shows different features in different regions. One hundred and ninety-eight patients with chronic HBV infection were enrolled in this study, 38 of whom had been diagnosed with cirrhosis of the liver and/or hepatocellular carcinoma. HBV DNA was isolated from the patients' blood samples and the entire genome and/or the basal core promoter/core promoter region sequenced. Phylogenetic analysis of the complete genomes revealed that subgenotype D1 is the most prevalent subgenotype in Turkey, but there was no definite phylogenetic grouping according to geography for isolates from different regions within Turkey, or for isolates in Turkey relative to other parts of the world. Turkish isolates tended to be genetically similar to European and central Asian isolates. Overall, HBV-infection in Turkey appears to be characterized by early HBeAg seroconversion, a high incidence of the A1896 core promoter mutation and a small viral load. Genotype D characteristic mutations A1757 and T1764/G1766 were found in the BCP region. T1773 was associated with T1764/G1766 and a larger viral load. In conclusion, infection with HBV genotype D in Turkey has a similar clinical outcome to that of Europe and central Asia. Genotypic mutations in genotype D may be linked with disease prognosis in Turkey, but further studies with higher sample numbers and balanced clinical groups are needed to confirm this.

  5. The core planar cell polarity gene prickle interacts with flamingo to promote sensory axon advance in the Drosophila embryo.

    PubMed

    Mrkusich, Eli M; Flanagan, Dustin J; Whitington, Paul M

    2011-10-01

    The atypical cadherin Drosophila protein Flamingo and its vertebrate homologues play widespread roles in the regulation of both dendrite and axon growth. However, little is understood about the molecular mechanisms that underpin these functions. Whereas flamingo interacts with a well-defined group of genes in regulating planar cell polarity, previous studies have uncovered little evidence that the other core planar cell polarity genes are involved in regulation of neurite growth. We present data in this study showing that the planar cell polarity gene prickle interacts with flamingo in regulating sensory axon advance at a key choice point - the transition between the peripheral nervous system and the central nervous system. The cytoplasmic tail of the Flamingo protein is not required for this interaction. Overexpression of another core planar cell polarity gene dishevelled produces a similar phenotype to prickle mutants, suggesting that this gene may also play a role in regulation of sensory axon advance.

  6. Incorporation of (/sup 35/S)sulfate in normal and neoplastic rat pancreatic acinar cells in relationship to cytodifferentiation

    SciTech Connect

    Kanwar, Y.S.; Rao, M.S.; Longnecker, D.S.; Reddy, J.K.

    1984-11-01

    The rates of (/sup 35/S)sulfate incorporation in highly differentiated acinar cells from normal pancreas, moderately differentiated cells of nafenopin-induced transplantable pancreatic carcinoma, and poorly differentiated cells from azaserine-induced transplantable pancreatic carcinoma were examined in an attempt to determine if sulfation is a property of acinar cells with well-developed secretory granules. The cells were dissociated, pulsed with (/sup 35/S)sulfate (specific activity, approximately 1000 Ci/mmol) for 10 and 60 min, and chased with medium containing 100 X excess of cold inorganic sulfate for 0, 15, 60, and 120 min. The cells were then processed for determining their pool size and light and electron microscopic autoradiography. No significant differences among their pool sizes were observed. However, the light microscopic autoradiograms revealed the (/sup 35/S)sulfate incorporation as follows: azaserine-induced transplantable pancreatic carcinoma greater than nafenopin-induced transplantable pancreatic carcinoma greater than normal pancreas. Electron microscopic autoradiograms revealed similar trends. The grain densities (concentration of radiation) were highest in the Golgi regions immediately postpulse (0 min) and gradually shifted toward the secretory granules over a 120-min period. In addition, the grain density values of the secretory granule-rich cells of nafenopin-induced transplantable pancreatic carcinoma were relatively similar to the cells of normal pancreas, whereas the grain density values of secretory granule-deficient cells from this tumor were similar to those of poorly differentiated neoplastic cells of azaserine-induced transplantable pancreatic carcinoma. These results show that poorly differentiated neoplastic cells incorporate more (/sup 35/S)sulfate than do the well-differentiated cells, but the reasons for this unexpected differential incorporation are at present unknown.

  7. Real-time polymerase chain reaction detection of cauliflower mosaic virus to complement the 35S screening assay for genetically modified organisms.

    PubMed

    Cankar, Katarina; Ravnikar, Maja; Zel, Jana; Gruden, Kristina; Toplak, Natasa

    2005-01-01

    Labeling of genetically modified organisms (GMOs) is now in place in many countries, including the European Union, in order to guarantee the consumer's choice between GM and non-GM products. Screening of samples is performed by polymerase chain reaction (PCR) amplification of regulatory sequences frequently introduced into genetically modified plants. Primers for the 35S promoter from Cauliflower mosaic virus (CaMV) are those most frequently used. In virus-infected plants or in samples contaminated with plant material carrying the virus, false-positive results can consequently occur. A system for real-time PCR using a TaqMan minor groove binder probe was designed that allows recognition of virus coat protein in the sample, thus allowing differentiation between transgenic and virus-infected samples. We measured the efficiency of PCR amplification, limits of detection and quantification, range of linearity, and repeatability of the assay in order to assess the applicability of the assay for routine analysis. The specificity of the detection system was tested on various virus isolates and plant species. All 8 CaMV isolates were successfully amplified using the designed system. No cross-reactivity was detected with DNA from 3 isolates of the closely related Carnation etched ring virus. Primers do not amplify plant DNA from available genetically modified maize and soybean lines or from different species of Brassicaceae or Solanaceae that are natural hosts for CaMV. We evaluated the assay for different food matrixes by spiking CaMV DNA into DNA from food samples and have successfully amplified CaMV from all samples. The assay was tested on rapeseed samples from routine GMO testing that were positive in the 35S screening assay, and the presence of the virus was confirmed.

  8. The 35S U5 snRNP Is Generated from the Activated Spliceosome during In vitro Splicing

    PubMed Central

    2015-01-01

    Primary gene transcripts of eukaryotes contain introns, which are removed during processing by splicing machinery. Biochemical studies In vitro have identified a specific pathway in which introns are recognised and spliced out. This occurs by progressive formation of spliceosomal complexes designated as E, A, B, and C. The composition and structure of these spliceosomal conformations have been characterised in many detail. In contrast, transitions between the complexes and the intermediates of these reactions are currently less clear. We have previously isolated a novel 35S U5 snRNP from HeLa nuclear extracts. The protein composition of this particle differed from the canonical 20S U5 snRNPs but was remarkably similar to the activated B* spliceosomes. Based on this observation we have proposed a hypothesis that 35S U5 snRNPs represent a dissociation product of the spliceosome after both transesterification reactions are completed. Here we provide experimental evidence that 35S U5 snRNPs are generated from the activated B* spliceosomes during In vitro splicing. PMID:26020933

  9. Handling of L-(/sup 35/S)cystine by cysteamine-pretreated cystinotic and normal fibroblasts

    SciTech Connect

    States, B.; Lee, J.; Segal, S.

    1983-02-01

    In short incubations with 0.1 mM L-(/sup 35/S)cystine in phosphate-buffered saline medium, and long incubations with label in complete minimum Eagle's medium with Earle salts, cystine-depleted cystinotic cells reaccumulate labeled cystine more rapidly than pretreated normal cells. Cysteamine pretreatment of both normal and cystinotic cells resulted in an initial increased conversion of exogenous cystine to intracellular cysteine. In 24-h incubations in complete medium, cysteamine-pretreated cells showed enhanced conversion of 0.1 mM L-(/sup 35/S)cystine to cysteine and reduced glutathione. Addition of cycloheximide to the incubation media decreased the incorporation of /sup 35/S into cellular protein by more than 90% but did not affect the accumulation of intracellular labeled cystine in cystinotic cells. Therefore, the incorporation and release of cystine from protein is not an obligatory source of accumulated cystine and researchers speculate that there may be early extralysosomal entrapment of cystine in cystinotic cells.

  10. Cloning of the 5' regulatory regions and functional characterization of the core promoters of ovine PLAU (u-PA) and SERPIN1 (PAI-1).

    PubMed

    Lampidonis, A D; Theodorou, G; Pecorini, C; Rebucci, R; Baldi, A; Politis, I

    2011-12-01

    The activation of plasminogen plays a crucial role in various extracellular proteolytic events (fibrinolysis, cell migration, ovulation and involution of the mammary gland). In the present study we describe the isolation of the 5' proximal and distal promoter regions of ovine PLAU (urokinase plasminogen activator, u-PA) and SERPIN1 (plasminogen activator inhibitor 1, PAI-1) genes for the first time in ruminants. Analysis of the 5.645kb 5'-flanking region of u-PA revealed a putative TATA-less promoter. In contrast the isolated 2.787kb 5'-flanking region of PAI-1 included a TATA-box. It should be noted that both genes lack the initiator motif around the transcription start site. The two genes share a number of transcription factor binding sites, namely Nuclear Factor-kappa B, Stimulating Protein 1 and Activating protein 1, suggesting co-expression of the two genes. Moreover, additional, not shared, transcription factor binding sites were identified in u-PA and PAI-1. More important of these are the cis-regulatory elements for plasminogen activator inhibitor 2 located in the distal promoter region of u-PA, suggesting an involvement of the other specific inhibitor in the regulation of ovine u-PA gene expression, and the three stress response elements sites present in the proximal and distal promoter of PAI-1. Different genomic fragments of the two 5' flanking regions were directionally cloned into a suitable reporter vector upstream of a promoter-less luciferase gene. Transient transfection into bovine mammary epithelial (BME-UV) cells demonstrated that the regions of -384/+27 and -382/+22 for the u-PA and PAI-1genes, respectively, potentially function as core promoter regions.

  11. Neuroanatomical mapping of juvenile rat brain regions with prominent basal signal in [(35)S]GTPgammaS autoradiography.

    PubMed

    Aaltonen, Niina; Palomäki, Ville A B; Lecklin, Anne; Laitinen, Jarmo T

    2008-03-01

    [(35)S]GTPgammaS autoradiography represents a powerful functional approach to detect receptor-dependent G(i/o) protein activity in anatomically defined brain structures. Inherent to this technique, however, is the notable basal signal evident in several brain regions in the absence of receptor stimulation by exogenously added agonist. In the rat brain, much of this basal labelling derives from tonic activation of adenosine A(1) and lysophosphatidic acid LPA(1) receptors in the gray and white matter regions, respectively. Despite the elimination of the two receptor activities, prominent basal [(35)S]GTPgammaS labelling is still evident in discrete brain structures, possibly reflecting regional enrichment of G(i/o) and/or constitutive receptor activity or the presence of still unknown endogenous ligands activating their orphan receptors. Here, the anatomical distribution of the enhanced basal signal was systematically mapped in brain sections of 4-week-old male Wistar rats. Regions with prominent basal [(35)S]GTPgammaS labelling represented neuroanatomically distinct structures, in particular various thalamic and hypothalamic nuclei. For instance, the paraventricular thalamic nucleus, the bed nucleus of the stria terminalis and the subfornical organ were highly labelled, as were the periaqueductal gray and the nucleus of the solitary tract. Pre-treatment with N-ethylmaleimide (NEM), an alkylating agent preventing all known receptor-driven G protein activity in cryostat sections markedly decreased the basal binding in all examined regions. In preliminary screening, selective antagonists for various brain-enriched G(i/o)-coupled receptors failed to suppress the basal signal in any of the studied regions.

  12. (/sup 35/S)autoradiographic study of sulfated GAG accumulation and turnover in embryonic mouse tooth germs

    SciTech Connect

    Lau, E.C.; Boukari, A.; Arechaga, J.; Osman, M.; Ruch, J.V.

    1983-01-01

    The accumulation of sulfated glycosaminoglycans(GAG) in embryonic mouse molars before, during, and after terminal differentiation of odontoblasts was localized by (/sup 35/S)autoradiography combined with the use of chondroitin ABC lyase. Much more sulfated GAG were accumulated in the dental papilla than in the dental epithelium. High incorporation of (/sup 35/S)sulfate occurred at the epithelio-mesenchymal junction, which is the site of dental basement membrane and predentin. Before terminal differentiation of odontoblasts, the distribution of sulfated GAG was uniform at the basement membrane. After the onset of terminal differentiation of odontoblasts, much more sulfated GAG accumulated at the tip of principal cusps than at the apical (inferior) parts of cusps, and sulfated GAG were then found to be degraded more rapidly at the epithelio-mesenchymal junction than at other parts of the tooth germ. Thus regional variation in the rate of degradation of GAG exists in the tooth germs. Trypsin-isolated dental epithelia cultured in vitro synthesized a new basement membrane that could be labeled with (/sup 3/H)glucosamine but not with /sup 35/SO4(-2). The epithelial-derived basal lamina contains little or no sulfatated GAG.

  13. Effect of metallothionein core promoter region polymorphism on cadmium, zinc and copper levels in autopsy kidney tissues from a Turkish population

    SciTech Connect

    Kayaalti, Zeliha; Mergen, Goerkem; Soeylemezoglu, Tuelin

    2010-06-01

    Metallothioneins (MTs) are metal-binding, low molecular weight proteins and are involved in pathophysiological processes like metabolism of essential metals, metal ion homeostasis and detoxification of heavy metals. Metallothionein expression is induced by various heavy metals especially cadmium, mercury and zinc; MTs suppress toxicity of heavy metals by binding themselves to these metals. The aim of this study was to investigate the association between the - 5 A/G metallothionein 2A (MT2A) single nucleotide polymorphism (SNP) and Cd, Zn and Cu levels in the renal cortex from autopsy cases. MT2A core promoter region - 5 A/G SNP was analyzed by PCR-RFLP method using 114 autopsy kidney tissues and the genotype frequencies of this polymorphism were found as 87.7% homozygote typical (AA), 11.4% heterozygote (AG) and 0.9% homozygote atypical (GG). In order to assess the Cd, Zn and Cu levels in the same autopsy kidney tissues, a dual atomic absorption spectrophotometer system was used and the average levels of Cd, Zn and Cu were measured as 95.54 {+-} 65.58 {mu}g/g, 181.20 {+-} 87.72 {mu}g/g and 17.14 {+-} 16.28 {mu}g/g, respectively. As a result, no statistical association was found between the - 5 A/G SNP in the MT2A gene and the Zn and Cu levels in the renal cortex (p > 0.05), but considerably high accumulation of Cd was monitored for individuals having AG (151.24 {+-} 60.21 {mu}g/g) and GG genotypes (153.09 {mu}g/g) compared with individuals having AA genotype (87.72 {+-} 62.98 {mu}g/g) (p < 0.05). These results show that the core promoter region polymorphism of metallothionein 2A increases the accumulation of Cd in human renal cortex.

  14. Effects of hepatitis B virus precore and basal core promoter mutations on the expression of viral antigens: genotype B vs C.

    PubMed

    Liu, C-J; Cheng, H-R; Chen, C-L; Chen, T-C; Tseng, T-C; Wang, Z-L; Chen, P-J; Liu, C-H; Chen, D-S; Kao, J-H

    2011-10-01

    Hepatitis B virus (HBV) genotypes/mutants are known to affect natural outcomes. The virologic differences among HBV genotype, precore and basal core promoter (BCP) mutations were investigated. HBV strains were isolated from 18 hepatitis B e antigen (HBeAg)-positive patients (nine genotype B and nine genotype C). All had precore and BCP wild-type sequences. After cloning of full-length HBV genome, the effects of viral genotype, precore and BCP mutations singly or additively on the expression of viral DNA and antigens were investigated by mutagenesis and transfection assays in Huh7 cells. Significant findings included the following: (i) expression of intracellular core protein increased when precore or BCP mutation was introduced in genotype C strains; (ii) expression of intracellular surface protein was lower in genotype C precore wild-type strain compared with genotype B; (iii) precore mutation was associated with a lower extracellular expression level of HBV DNA; (iv) secretion of hepatitis B surface antigen in genotype C was lower than that in genotype B; and (v) secretion of HBeAg in genotype B was lower than that in genotype C. No additive effect was observed by combining precore and BCP mutations. Hence, HBV genotype and precore/BCP mutations correlate with intrahepatic expression of viral antigens in vitro.

  15. Mutation analysis of TRPS1 gene including core promoter, 5'UTR, and 3'UTR regulatory sequences with insight into their organization.

    PubMed

    Solc, Roman; Klugerova, Michaela; Vcelak, Josef; Baxova, Alice; Kuklik, Miloslav; Vseticka, Jan; Beharka, Rastislav; Hirschfeldova, Katerina

    2017-01-01

    The TRPS1 protein is a potent regulator of proliferation, differentiation, and apoptosis. The TRPS1 gene aberrations are strongly associated with rare trichorhinophalangeal syndrome (TRPS) development. We have conducted MLPA analysis to capture deletion within the crucial 8q24.1 chromosomal region in combination with mutation analysis of TRPS1 gene including core promoter, 5'UTR, and 3'UTR sequences in nine TRPS patients. Low complexity or extent of untranslated regulatory sequences avoided them from analysis in previous studies. Amplicon based next generation sequencing used in our study bridge over these technical limitations. Finally, we have made extended in silico analysis of TRPS1 gene regulatory sequences organization. Single contiguous deletion and an intragenic deletion intervening several exons were detected. Mutation analysis revealed five TRPS1 gene aberrations (two structural rearrangements, two nonsense mutations, and one missense substitution) reaching the overall detection rate of 78%. Several polymorphic variants were detected within the analysed regulatory sequences but without proposed pathogenic effect. In silico analysis suggested alternative promoter usage and diverse expression effectivity for different TRPS1 transcripts. Haploinsufficiency of TRPS1 gene was responsible for most of the TRPS phenotype. Structure of TRPS1 gene regulatory sequences is indicative of generally low single allele expression and its tight control.

  16. Synthesis, Binding and Antiviral Properties of Potent Core-Extended Naphthalene Diimides Targeting the HIV-1 Long Terminal Repeat Promoter G-Quadruplexes

    PubMed Central

    2015-01-01

    We have previously reported that stabilization of the G-quadruplex structures in the HIV-1 long terminal repeat (LTR) promoter suppresses viral transcription. Here we sought to develop new G-quadruplex ligands to be exploited as antiviral compounds by enhancing binding toward the viral G-quadruplex structures. We synthesized naphthalene diimide derivatives with a lateral expansion of the aromatic core. The new compounds were able to bind/stabilize the G-quadruplex to a high extent, and some of them displayed clear-cut selectivity toward the viral G-quadruplexes with respect to the human telomeric G-quadruplexes. This feature translated into low nanomolar anti-HIV-1 activity toward two viral strains and encouraging selectivity indexes. The selectivity depended on specific recognition of LTR loop residues; the mechanism of action was ascribed to inhibition of LTR promoter activity in cells. This is the first example of G-quadruplex ligands that show increased selectivity toward the viral G-quadruplexes and display remarkable antiviral activity. PMID:26599611

  17. CNS depressants accelerate the dissociation of /sup 35/S-TBPS binding and GABA enhances their displacing potencies

    SciTech Connect

    Maksay, G.; Ticku, M.K.

    1988-01-01

    The specific binding of /sup 35/S-t-butylbicyclophosphorothionate (TBPS) was studied in synaptosomal membranes of rat cerebral cortex. The displacing potencies of eleven CNS depressants and three convulsants were determined in the presence of 1 /sup +/M GABA and 10 nM R 5135. GABA enhanced the displacing potencies of depressants of most diverse chemical structures: diaryltriazine (LY 81067), pyrazolopyridine (etazolate), cinnamide, glutarimide, 2,3-benzodiazepine (tofizopam) and alcohol derivatives, barbiturates, (+)etomidate, methaqualone and meprobamate. In contrast, the IC/sub 50/ values of convulsants (picrotoxinin, pentetrazol and the barbiturate enantiomer S(+)MPPB) were not significantly affected. The depressants accelerated either basal or GABA-augmented dissociation of /sup 35/-TBPS mainly by increasing the contribution of its rapid first phase.

  18. Use Of Cosmogenic 35S To Trace The Uptake Process Of SO2 In Aerosols In The Atmosphere

    NASA Astrophysics Data System (ADS)

    Abramian, A.; Corbin, A.

    2008-12-01

    Environmental issues, such as acid rain and global warming, are linked to increased sulfur emissions and sulfate production in the atmosphere. Sulfate aerosol particles act as cloud condensation nuclei and can reduce the greenhouse effect by the indirect effect. Our understanding of the chemical and photochemical processes that govern the chemical transformations and transport of sulfur compounds in the atmosphere is still incomplete due to the complex, multivalent nature of sulfur and uncertainties in aerosol chemistry and transport (particularly trans-oceanic). We explore the use of cosmogenically produced 35S (half-life~87 days) to trace the uptake of SO2 gas into aerosols, as a function of aerosol size, in two different environments by simultaneously collecting and measuring [35SO42- ]and [35SO2]. These measurements can in turn be used to understand the time scales of SO2 oxidation to SO42-, aerosol 'age' and boundary layer dynamics. Aerosol samples are collected on glass fiber filters twice a week at Scripps Institute of Oceanography Pier in La Jolla, CA and the San Fernando Valley, CA for a 21-day period. SO2 (g) was collected on KOH impregnated filters placed after a 4-stage aerosol filter stack. We present preliminary results for both fine and coarse aerosol sulfate [35SO4] as well as [35SO2]. These measurements were done using low-noise liquid scintillation spectroscopy. By measuring the activity of each sample repeatedly over a period of 100 days, the exponential decay of 35S was observed, confirming the identity of the radioactive signal. The coastal and inland measurements are compared and implications for the atmospheric chemistry of SO2 and SO4 are discussed. Finally, we assess the potential of using [35SO4]/[nss-SO4] as a tracer of primary sulfate and trans-oceanic transport by coupling the measurements of the cation (Na+, Ca2+, K+, Mg2+, NH4+) and anion (Cl, NO3, SO4) concentrations in the aerosols.

  19. Both the constitutive Cauliflower Mosaic Virus 35S and tissue-specific AGAMOUS enhancers activate transcription autonomously in Arabidopsis thaliana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The presence of multiple enhancers and promoters within a single vector often provokes complicated mutual interaction and crosstalk, thereby, altering promoter specificity, which causes serious problems for precisely engineering gene function and agronomic traits in transgenic plants. Enhancer elem...

  20. Application of a Novel, Rapid, and Sensitive Oligonucleotide Ligation Assay for Detection of Cancer-Predicting Mutations in the Precore and Basal Core Promoter of Hepatitis B Virus▿

    PubMed Central

    Mendy, M. E.; Kaye, S.; Le Roux, E.; Kirk, G. D.; Jeng-Barry, A.; McConkey, S.; Cotten, M.; Kuniholm, M. H.; Leligdowicz, A.; Hainaut, P.; Rowland-Jones, S.; Whittle, H.

    2008-01-01

    Hepatocellular carcinoma (HCC) and cirrhosis are important causes of mortality worldwide. Persistent hepatitis B virus (HBV) infection is a major cause of these diseases. Double mutations in the basal core promoter (BCP) (A1762T and G1764A) and precore (pre-C) (G1896A) regions of the virus are associated with progression to HCC. The current study is aimed at developing a simple method for screening and detecting BCP and pre-C mutations in HBV carriers. We have developed and validated an oligonucleotide ligation assay (OLA) to detect point mutations in the HBV core gene. We have applied OLA methods to samples from HBV-infected carriers recruited from the Gambia Liver Cancer Study (GLCS) comprising asymptomatic HBsAg carriers, patients with cirrhosis, and patients with HCC. We observed an 89.3% and 95.8% concordance between the OLA and DNA sequencing for BCP and pre-C mutations, respectively. OLA detected the mutations in single-strain infections and in infections with mixtures of wild-type and mutant viruses under conditions where sequencing detected only the single dominant strains. BCP mutations were detected in 75.7% of patients with advanced liver disease (cirrhosis/HCC) compared to 47.6% of asymptomatic carriers, while pre-C mutations were detected in 34.5% of advanced liver disease patients and in 47.6% of asymptomatic HBsAg carriers. There was a significant association between the presence of BCP mutations and advanced liver disease. In conclusion, OLA is a simple, economical, and reliable assay for detection of pre-C and BCP mutations. Its application can lead to improvement in diagnosis and clinical care in regions where HBV is endemic. PMID:18508941

  1. Clinical instructors' perception of a faculty development programme promoting postgraduate year-1 (PGY1) residents' ACGME six core competencies: a 2-year study.

    PubMed

    Lee, Fa-Yauh; Yang, Ying-Ying; Hsu, Hui-Chi; Chuang, Chiao-Lin; Lee, Wei-Shin; Chang, Ching-Chih; Huang, Chia-Chang; Chen, Jaw-Wen; Cheng, Hao-Min; Jap, Tjin-Shing

    2011-01-01

    Objective The six core competencies designated by Accreditation Council for Graduate Medical Education (ACGME) are essential for establishing a patient centre holistic medical system. The authors developed a faculty programme to promote the postgraduate year 1 (PGY(1)) resident, ACGME six core competencies. The study aims to assess the clinical instructors' perception, attitudes and subjective impression towards the various sessions of the 'faculty development programme for teaching ACGME competencies.' Methods During 2009 and 2010, 134 clinical instructors participated in the programme to establish their ability to teach and assess PGY(1) residents about ACGME competencies. Results The participants in the faculty development programme reported that the skills most often used while teaching were learnt during circuit and itinerant bedside, physical examination teaching, mini-clinical evaluation exercise (mini-CEX) evaluation demonstration, training workshop and videotapes of 'how to teach ACGME competencies.' Participants reported that circuit bedside teaching and mini-CEX evaluation demonstrations helped them in the interpersonal and communication skills domain, and that the itinerant teaching demonstrations helped them in the professionalism domain, while physical examination teaching and mini-CEX evaluation demonstrations helped them in the patients' care domain. Both the training workshop and videotape session increase familiarity with teaching and assessing skills. Participants who applied the skills learnt from the faculty development programme the most in their teaching and assessment came from internal medicine departments, were young attending physician and had experience as PGY(1) clinical instructors. Conclusions According to the clinical instructors' response, our faculty development programme effectively increased their familiarity with various teaching and assessment skills needed to teach PGY(1) residents and ACGME competencies, and these clinical

  2. Saturable binding of /sup 35/S-t-butylbicyclophosphorothionate to the sites linked to the GABA receptor and the interaction with gabaergic agents

    SciTech Connect

    Wong, D.T.; Threlkeld, P.G.; Bymaster, F.P.; Squires, R.F.

    1984-02-27

    /sup 35/S-t-Butylbicyclophosphorothionate (/sup 35/S-TBPS) binds in a concentration-saturable manner to specific sites on membranes from rat cerebral cortex. Using a filtration assay at 25/sup 0/C, in 250 mM NaCl, specific binding of /sup 35/S-TBPS constitutes about 84 to 94 percent of total binding, depending on radioligand concentrations. /sup 35/S-TBPS binding is optimal in the presence of NaCl or NaBr and substantially less in the presence of NaI or NaF. It is sensitive to the treatment with 0.05 percent Triton X-100 but not to repeated freezing and thawing, procedures which increase /sup 3/H-GABA binding. Pharmacological studies show that /sup 35/S-TBPS binding is strongly inhibited by GABA-A receptor agonists (e.g., GABA and muscimol) and by the noncompetitive antagonist, picrotoxin, but not the competitive antagonist, bicuculline. Compounds which enhance binding of radioactive GABA and benzodiazepines, such as the pyrazolopyridines, cartazolate and tracazolate, and a diaryltriazine, LY81067, are also potent inhibitors of /sup 35/S-TBPS binding, with LY81067 being the most effective. The effects of GABA, picrotoxin and LY81067 on the saturable binding of /sup 35/S-TBPS in cortical membranes are compared. The present findings are consistent with the interpretation that /sup 35/S-TBPS bind at or near the picrotoxin-sensitive anion recognition sites of the GABA/benzodiazepine/picrotoxin receptor complex.

  3. Specific mutations in the enhancer II/core promoter/precore regions of hepatitis B virus subgenotype C2 in Korean patients with hepatocellular carcinoma.

    PubMed

    Kim, Ja Kyung; Chang, Hye Young; Lee, Jung Min; Baatarkhuu, Oidov; Yoon, Young Joon; Park, Jun Yong; Kim, Do Young; Han, Kwang-Hyub; Chon, Chae Yoon; Ahn, Sang Hoon

    2009-06-01

    Recently, hepatitis B virus (HBV) genotypes and mutations have been reported to be related to hepatocellular carcinoma (HCC). This cross-sectional case-control study examined the relationship between HCC and mutations in the enhancer II/core promoter and precore regions of HBV by comparing 135 Korean HCC patients infected with HBV genotype C2 (HBV/C2; HCC group) with 135 age-, sex-, and hepatitis B e antigen (HBeAg) status-matched patients without HCC (non- HCC group). Age and sex were also matched between HBeAg-positive and -negative patients. The prevalence of T1653, A1689, V1753, T1762/A1764, T1846, A1850, C1858, and A1896 mutations was evaluated in this population. The prevalence of the T1653 mutation in the box alpha region, the T1689 [corrected] mutation in between the box alpha and beta regions, and the T1762/A1764 mutations in the basal core promoter region was significantly higher in the HCC group compared to the non-HCC group (8.9% vs. 2.2%, P = 0.017; 19.3% vs. 4.4%, P < 0.001; and 60.7% vs. 22.2%; P < 0.001). Among HBeAg-negative patients, the frequency of the T1653 mutation was higher in the HCC group. Regardless of HBeAg status, the prevalence of the T1689, [corrected] and T1762/A1764 mutations was higher in the HCC group than in the non-HCC group. However, no association was observed between mutations in the precore region and HCC. Upon multivariate analysis, the presence of the T1653, T1689, [corrected] and T1762/A1764 mutations was an independent predictive factor for HCC. The addition of the T1653 or T1689 [corrected] mutation to T1762/A1764 increased the risk of HCC. In conclusion, the T1653, T1689, [corrected] and/or T1762/A1764 mutations were associated with the development of HCC in Korean patients infected with HBV/C2.

  4. Saturable binding of /sup 35/S-t-butylbicyclophosphorothionate to the sites linked to the GABA receptor and the interaction with gabaergic agents

    SciTech Connect

    Wong, D.T.; Threlkeld, P.G.; Bymaster, F.P.; Squires, R.F.

    1984-02-27

    /sup 35/-S-t-Butylbicyclophosphorothionate (/sup 35/S-TBPS) binds in a concentration-saturable manner to specific sites on membranes from rat cerebral cortex. Using a filtration assay at 25/sup 0/C, in 250 mM NaCl, specific binding of /sup 35/S-TBPS constitutes about 84 to 94 percent of total binding, depending on radioligand concentrations. /sup 35/S-TBPS binding is optimal in the presence of NaCl or NaBr and substantially less in the presence of NaI or NaF. It is sensitive to the treatment with 0.05 percent Triton X-100 but not to repeated freezing and thawing, procedures which increase /sup 3/H-GABA binding. Pharmacological studies show that /sup 35/S-TBPS binding is strongly inhibited by GABA-A receptor agonists (e.g., GABA and muscimol) and by the noncompetitive antagonist, picrotoxin, but not the competitive antagonist, bicuculline. Compounds which enhance binding of radioactive GABA and benzodiazepines, such as the pyrazolopyridines, cartazolate and trazolate, and a diaryl-triazine, LY81067, are also potent inhibitors of /sup 35/S-TBPS binding, with LY81067 being the most effective. The effects of GABA, picrotoxin

  5. A 1-kb bacteriophage lambda fragment functions as an insulator to effectively block enhancer-promoter interactions in Arabidopsis thaliana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 35S cauliflower mosaic virus (CaMV) promoter contains an enhancer element that is able to override the tissue-, organ- and developmental-stage specificity of nearby promoters. Consequently, the precise control of transgene expression in transgenic plants, which often contain the 35S CaMV promot...

  6. Stronger enhancer II/core promoter activities of hepatitis B virus isolates of B2 subgenotype than those of C2 subgenotype

    PubMed Central

    Qin, Yanli; Zhou, Xueshi; Jia, Haodi; Chen, Chaoyang; Zhao, Weifeng; Zhang, Jiming; Tong, Shuping

    2016-01-01

    Hepatitis B virus (HBV) genotype C causes prolonged chronic infection and increased risk for liver cancer than genotype B. Our previous work revealed lower replication capacity of wild-type genotype C2 than B2 isolates. HBV DNA replication is driven by pregenomic RNA, which is controlled by core promoter (CP) and further augmented by enhancer I (ENI) and enhancer II (ENII). DNA fragments covering these regulatory elements were amplified from B2 and C2 isolates to generate luciferase reporter constructs. As ENII is fully embedded in CP, we inserted HBV DNA fragments in the sense orientation to determine their combined activities, and in the antisense orientation to measure enhancer activities alone. Genotype B2 isolates displayed higher ENI+ENII+CP, ENII+CP, and ENII activities, but not ENI or ENI+ENII activity, than C2 isolates. The higher ENII+CP activity was partly attributable to 4 positions displaying genotype-specific variability. Exchanging CP region was sufficient to revert the replication phenotypes of several B2 and C2 clones tested. These results suggest that a weaker ENII and/or CP at least partly accounts for the lower replication capacities of wild-type C2 isolates, which could drive the subsequent acquisition of CP mutations. Such mutations increase genome replication and are implicated in liver cancer development. PMID:27461034

  7. Stronger enhancer II/core promoter activities of hepatitis B virus isolates of B2 subgenotype than those of C2 subgenotype.

    PubMed

    Qin, Yanli; Zhou, Xueshi; Jia, Haodi; Chen, Chaoyang; Zhao, Weifeng; Zhang, Jiming; Tong, Shuping

    2016-07-27

    Hepatitis B virus (HBV) genotype C causes prolonged chronic infection and increased risk for liver cancer than genotype B. Our previous work revealed lower replication capacity of wild-type genotype C2 than B2 isolates. HBV DNA replication is driven by pregenomic RNA, which is controlled by core promoter (CP) and further augmented by enhancer I (ENI) and enhancer II (ENII). DNA fragments covering these regulatory elements were amplified from B2 and C2 isolates to generate luciferase reporter constructs. As ENII is fully embedded in CP, we inserted HBV DNA fragments in the sense orientation to determine their combined activities, and in the antisense orientation to measure enhancer activities alone. Genotype B2 isolates displayed higher ENI+ENII+CP, ENII+CP, and ENII activities, but not ENI or ENI+ENII activity, than C2 isolates. The higher ENII+CP activity was partly attributable to 4 positions displaying genotype-specific variability. Exchanging CP region was sufficient to revert the replication phenotypes of several B2 and C2 clones tested. These results suggest that a weaker ENII and/or CP at least partly accounts for the lower replication capacities of wild-type C2 isolates, which could drive the subsequent acquisition of CP mutations. Such mutations increase genome replication and are implicated in liver cancer development.

  8. Comparison of 35S and biotin as labels for in situ hybridization: Use of an HPV model system

    SciTech Connect

    Unger, E.R.; Hammer, M.L.; Chenggis, M.L. )

    1990-01-01

    Colorimetric in situ hybridization is a method of potential importance in diagnosis and research. The largest criticism of the method has been a perceived loss of sensitivity compared with autoradiographic techniques. Our more positive experience with automation of colorimetric in situ hybridization led us to undertake a direct comparison of the sensitivity of 35S- and biotin-labeled probes. Serial sections of formalin-fixed, paraffin-embedded cell pellets from four human cervical carcinoma cell lines with known copies of HPV (CaSki, 400-600 copies HPV 16; HeLa, 10-50 copies HPV 18; SiHa, 1-2 copies HPV 16; HTB31, no known copies HPV) were hybridized with protocols optimized for autoradiographic or colorimetric detection. Both methods gave comparable results, with differences in each technique seen at the limits of sensitivity. The 1-2 copies of HPV 16 per SiHa cell can be detected with both methods; however, grain counting is required for interpretation of the autoradiographic result. This degree of sensitivity for colorimetric in situ hybridization in formalin-fixed, paraffin-embedded material is achieved through careful optimization of probe size and labeling, adequate tissue digestion, and removal of background. Autoradiography may be preferred in situations where quantitation is required, but colorimetric detection retains the advantages of speed, potential for automation, and improved localization of signal with comparable sensitivity.

  9. Superconductivity versus structural phase transition in the closely related Bi2Rh3.5S2 and Bi2Rh3S2

    DOE PAGES

    Kaluarachchi, Udhara S.; Xie, Weiwei; Lin, Qisheng; ...

    2015-05-19

    Single crystals of Bi2Rh3S2 and Bi2Rh3.5S2 were synthesized by solution growth, and the crystal structures and thermodynamic and transport properties of both compounds were studied. In the case of Bi2Rh3S2, a structural first-order transition at around 165 K is identified by single-crystal diffraction experiments, with clear signatures visible in resistivity, magnetization, and specific heat data. No superconducting transition for Bi2Rh3S2 was observed down to 0.5 K. In contrast, no structural phase transition at high temperature was observed for Bi2Rh3.5S2; however, bulk superconductivity with a critical temperature, Tc ≈ 1.7 K, was observed. The Sommerfeld coefficient γ and the Debye temperaturemore » (ΘD) were found to be 9.41 mJ mol–1K–2 and 209 K, respectively, for Bi2Rh3S2, and 22 mJ mol–1K–2 and 196 K, respectively, for Bi2Rh3.5S2. As a result, the study of the specific heat in the superconducting state of Bi2Rh3.5S2 suggests that Bi2Rh3.5S2 is a weakly coupled, BCS superconductor.« less

  10. The effect of metallothionein 2A core promoter region single-nucleotide polymorphism on accumulation of toxic metals in sinonasal inverted papilloma tissues

    SciTech Connect

    Starska, Katarzyna; Bryś, Magdalena; Forma, Ewa; Lewy-Trenda, Iwona; Danilewicz, Marian; Krześlak, Anna

    2015-06-15

    Metallothioneins (MTs) are intracellular thiol-rich heavy metal-binding proteins which join trace metal ions protecting cells against heavy metal toxicity and regulate metal distribution and donation to various enzymes and transcription factors. The goal of this study was to identify the − 5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene, and to investigate its effect on allele-specific gene expression and Cd, Zn, Cu and Ni content in sinonasal inverted papilloma tissue (IP), with non-cancerous sinonasal mucosa (NCM) as a control. The MT2A promoter region − 5 A/G SNP was identified by restriction fragment length polymorphism using 117 IP and 132 NCM. MT2A gene analysis was performed by quantitative real-time PCR. Metal levels were analyzed by flame atomic absorption spectrometry. The frequency of A allele carriage was 99.2% and 100% in IP and NCM, respectively. The G allele carriage was detected in 23.9% of IP and in 12.1% of the NCM samples. As a result, a significant association of − 5 A/G SNP in MT2A gene with mRNA expression in both groups was determined. A significant association was identified between the − 5 A/G SNP in the MT2A gene with mRNA expression in both groups. A highly significant association was detected between the rs28366003 genotype and Cd and Zn content in IP. Furthermore, significant differences were identified between A/A and A/G genotype with regard to the type of metal contaminant. The Spearman rank correlation results showed the MT2A gene expression and both Cd and Cu levels were negatively correlated. The results obtained in this study suggest that the − 5 A/G SNP in the MT2A gene may have an effect on allele-specific gene expression and toxic metal accumulation in sinonasal inverted papilloma. - Highlights: • MT2A gene expression and metal content in sinonasal inverted papilloma tissues • Association between SNP (rs28366003) and expression of MT2A • Significant

  11. RNase MRP is required for entry of 35S precursor rRNA into the canonical processing pathway.

    PubMed

    Lindahl, Lasse; Bommankanti, Ananth; Li, Xing; Hayden, Lauren; Jones, Adrienne; Khan, Miriam; Oni, Tolulope; Zengel, Janice M

    2009-07-01

    RNase MRP is a nucleolar RNA-protein enzyme that participates in the processing of rRNA during ribosome biogenesis. Previous experiments suggested that RNase MRP makes a nonessential cleavage in the first internal transcribed spacer. Here we report experiments with new temperature-sensitive RNase MRP mutants in Saccharomyces cerevisiae that show that the abundance of all early intermediates in the processing pathway is severely reduced upon inactivation of RNase MRP. Transcription of rRNA continues unabated as determined by RNA polymerase run-on transcription, but the precursor rRNA transcript does not accumulate, and appears to be unstable. Taken together, these observations suggest that inactivation of RNase MRP blocks cleavage at sites A0, A1, A2, and A3, which in turn, prevents precursor rRNA from entering the canonical processing pathway (35S > 20S + 27S > 18S + 25S + 5.8S rRNA). Nevertheless, at least some cleavage at the processing site in the second internal transcribed spacer takes place to form an unusual 24S intermediate, suggesting that cleavage at C2 is not blocked. Furthermore, the long form of 5.8S rRNA is made in the absence of RNase MRP activity, but only in the presence of Xrn1p (exonuclease 1), an enzyme not required for the canonical pathway. We conclude that RNase MRP is a key enzyme for initiating the canonical processing of precursor rRNA transcripts, but alternative pathway(s) might provide a backup for production of small amounts of rRNA.

  12. Metallothionein 2A core promoter region genetic polymorphism and its impact on the risk, tumor behavior, and recurrences of sinonasal inverted papilloma (Schneiderian papilloma).

    PubMed

    Starska, Katarzyna; Bryś, Magdalena; Forma, Ewa; Olszewski, Jurek; Pietkiewicz, Piotr; Lewy-Trenda, Iwona; Stasikowska-Kanicka, Olga; Danilewicz, Marian; Krześlak, Anna

    2015-11-01

    Inverted papillomas are a unique group of locally aggressive benign epithelial neoplasms in the nasal cavity and paranasal sinuses arising from the Schneiderian mucosa. Metallothioneins are sulfhydryl-rich heavy metal-binding proteins required for metal toxicity protection and regulation of biological mechanisms including proliferation and invasion. The goal of this study was to identify three SNPs at loci -5 A/G (rs28366003) and -209 A/G (rs1610216) in the core promoter region and at locus +838 C/G (rs10636) in 3'UTR region of the MT2A gene with IP risk and with tumor invasiveness according to Krouse staging. Genotyping was performed using the PCR restriction fragment length polymorphism technique in 130 genetically unrelated IP individuals, and 418 randomly selected healthy volunteers. The presence of the rs28366003 SNP was significantly related to the risk of IP within the present population-based case-control study. Compared to homozygous common allele carriers, heterozygosity and homozygosity for the G variant had a significantly increased risk of IP (adjusted odds ratio [OR] = 7.71, 95% confidence interval [CI]: 4.01-14.91, p(dominant) < 0.001). Moreover, risk allele carriers demonstrated higher Krouse stage (pT1 vs. pT2-4) (OR = 19.32; 95% CI, 2.30-173.53; p < 0.0001), diffuse tumor growth (OR = 4.58; 95% CI, 1.70-12.11; p = 0.0008), bone destruction (OR = 4.13; 95% CI, 1.50-11.60; p = 0.003), and higher incidence of tumor recurrences (OR = 5.11; 95% CI, 1.68-15.20; p = 0.001). The findings suggest that MT2A gene variation rs28366003 may be implicated in the etiology of sinonasal inverted papilloma in a Polish population.

  13. Influence of the basal core promoter and precore mutation on replication of hepatitis B virus and antiviral susceptibility of different genotypes.

    PubMed

    Cui, Xiu-Ji; Cho, Yoo-Kyung; Song, Byung-Cheol

    2015-04-01

    Mutations in the basal core promoter (BCP) and precore (PC) regions of the hepatitis B virus (HBV) are more common in genotypes B and C than in genotype A, suggesting that these mutations might affect replication competency depending on genotype. The purpose of the study was to investigate the influence of these mutations on the capacity of HBV for replication and antiviral drug susceptibility according to genotype. Genotypes A, B, and C of HBV strains with a BCP mutation, PC mutation, or BCP + PC mutation were made by site-directed mutagenesis. Replication competency of each construct and susceptibility to nucleos(t) ide analogues were tested in an Huh7 cell line. In genotype A, the BCP and BCP + PC mutations increased the viral replication around 6.5 times compared with the wild type, and the PC mutation alone similarly increased the viral replication around three times. In genotypes B and C, all three mutant types increased viral replication to a similar extent, regardless of mutation pattern. Interestingly, the BCP mutation appeared to have a greater effect on viral replication in genotype A than in genotypes B and C. This finding was unexpected because the BCP mutation is more common in HBV genotypes B and C. Moreover, the BCP, PC, and BCP + PC mutations decreased the sensitivity of HBV to antiviral agents to various degrees (2- to 10-fold) regardless of genotype. In conclusion, BCP and PC mutations increased viral replication regardless of HBV genotype and decreased in vitro antiviral susceptibility to the nucleos(t) ide analogues.

  14. Naturally occurring basal core promoter A1762T/G1764A dual mutations increase the risk of HBV-related hepatocellular carcinoma: a meta-analysis

    PubMed Central

    Lu, Yunfei; Xu, Qingnian; Tang, Bozong; Chen, Xiaorong

    2016-01-01

    Basal core promoter (BCP) A1762T/G1764A dual mutations in hepatocarcinogenesis remain controversial. Published studies up to June 1, 2015 investigating the frequency of A1762T/G1764A dual mutations from chronic hepatitis B virus (HBV) infection, including hepatocellular carcinoma (HCC), were systematically identified. A total of 10,240 patients with chronic HBV infection, including 3729 HCC cases, were included in 52 identified studies. HCC patients had a higher frequency of BCP A1762T/G1764A dual mutations compared with asymptomatic HBsAg carriers (ASC) and patients with chronic hepatitis B (CHB) and liver cirrhosis (LC) (OR = 5.59, P < 0.00001; OR = 2.87, P < 0.00001; OR = 1.55, P = 0.02, respectively). No statistically significant difference was observed in the frequency of A1762T/G1764A dual mutations in cirrhotic HCC versus non-cirrhotic HCC patients (OR = 2.06, P = 0.05). Chronic HBV-infected patients and HCC patients with genotype B had a significantly lower risk of A1762T/G1764A dual mutations compared with patients with genotype C (OR = 0.30, P < 0.0001 and OR = 0.34, P = 0.04, respectively). In HBV genotype C subjects, A1762T/G1764A dual mutations contributed to significantly higher risk for HCC developing compared with non-mutation ones (OR = 3.47, P < 0.00001). In conclusion, A1762T/G1764A dual mutations increase the risk of HBV-related hepatocellular carcinoma, particularly in an HBV genotype C population, even without progression to cirrhosis. PMID:26848866

  15. Identification of the G-protein-coupled ORL1 receptor in the mouse spinal cord by [35S]-GTPgammaS binding and immunohistochemistry.

    PubMed

    Narita, M; Mizoguchi, H; Oji, D E; Dun, N J; Hwang, B H; Nagase, H; Tseng, L F

    1999-11-01

    1 Although the ORL1 receptor is clearly located within the spinal cord, the functional signalling mechanism of the ORL1 receptor in the spinal cord has not been clearly documented. The present study was then to investigate the guanine nucleotide binding protein (G-protein) activation mediated through by the ORL1 receptor in the mouse spinal cord, measuring the modulation of guanosine-5'-o-(3-[35S]-thio) triphosphate ([35S]-GTPgammaS) binding by the putative endogenous ligand nociceptin, also referred as orphanin FQ. We also studied the anatomical distribution of nociceptin-like immunoreactivity and nociceptin-stimulated [35S]-GTPgammaS autoradiography in the spinal cord. 2 Immunohistochemical staining of mouse spinal cord sections revealed a dense plexus of nociceptin-like immunoreactive fibres in the superficial layers of the dorsal horn throughout the entire length of the spinal cord. In addition, networks of fibres were seen projecting from the lateral border of the dorsal horn to the lateral grey matter and around the central canal. 3 In vitro [35S]-GTPgammaS autoradiography showed high levels of nociceptin-stimulated [35S]-GTPgammaS binding in the superficial layers of the mouse dorsal horn and around the central canal, corresponding to the areas where nociceptin-like immunoreactive fibres were concentrated. 4 In [35S]-GTPgammaS membrane assay, nociceptin increased [35S]-GTPgammaS binding of mouse spinal cord membranes in a concentration-dependent and saturable manner, affording maximal stimulation of 64.1+/-2.4%. This effect was markedly inhibited by the specific ORL1 receptor antagonist [Phe1Psi (CH2-NH) Gly2] nociceptin (1 - 13) NH2. None of the mu-, delta-, and kappa-opioid and other G-protein-coupled receptor antagonists had a significant effect on basal or nociceptin-stimulated [35S]-GTPgammaS binding. 5 These findings suggest that nociceptin-containing fibres terminate in the superficial layers of the dorsal horn and the central canal and that

  16. Effect of chemical carcinogens and partial hepatectomy on in vivo ( sup 35 S)methionine interaction with rat liver tRNA

    SciTech Connect

    Kanduc, D.; Aresta, A.; Rossiello, M.R.; Ranieri, T.; Quagliariello, E. )

    1989-09-29

    The effect of carcinogens given by a single or multiple injections on the extent of ({sup 35}S)methionine interaction with hepatic tRNA was studied in normal and partially hepatectomized rats. Either partial hepatectomy or administration of ethionine (100 or 330 mg/kg body weight) and dimethylnitrosamine (120 mg/kg body weight) by multiple i.p. injections inhibited the ({sup 35}S)methionine-tRNA interaction, while administration of hepatocarcinogenic chemicals plus PH resulted rather in a stimulation. Methylnitrosourea enhanced the extent of interaction when administered in a single dose (100 mg per kg body weight) 18 h after partial hepatectomy.

  17. Evaluation of constitutive viral promoters in transgenic soybean roots and nodules.

    PubMed

    Govindarajulu, Manjula; Elmore, James M; Fester, Thomas; Taylor, Christopher G

    2008-08-01

    The efficiency of beta-glucuronidase (GUS) expression was evaluated with five viral promoters to identify the most suitable promoter or promoters for use in soybean hairy roots, including applications to study the symbiotic interaction with Bradyrhizobium japonicum. Levels of GUS activity were fluorimetrically and histochemically assayed when the GUS (uidA) gene was driven by the Cauliflower mosaic virus (CaMV) 35S promoter and enhanced 35S (E35S) promoter, the Cassava vein mosaic virus (CsVMV) promoter, the Figwort mosaic virus (FMV) promoter, and the Strawberry vein banding virus (SVBV2) promoter. We demonstrate that GUS activity was highest when driven by the FMV promoter and that the promoter activity of 35S and SVBV2 was significantly lower than that of the CsVMV and E35S promoters when tested in soybean hairy roots. In mature soybean root nodules, strong GUS activity was evident when the FMV, 35S, and CsVMV promoters were used. These results indicate that the FMV promoter facilitates the strong expression of target genes in soybean hairy roots and root nodules.

  18. Hepatitis B virus basal core promoter/precore mutants and association with liver cirrhosis in children with chronic hepatitis B virus infection.

    PubMed

    Zhong, Y W; Di, F L; Liu, C; Zhang, X C; Bi, J F; Li, Y L; Wu, S Q; Dong, H; Liu, L M; He, J; Shi, Y M; Zhang, H F; Zhang, M

    2016-04-01

    We investigated 168 children and analysed the virological characterization and association with disease progression in children with hepatitis B virus (HBV) basal core promoter/precore (BCP/PC) mutants. Among 168 patients with HBV infection (aged 0.5-18 years old, mean 10.1), 86 of them had HBV-related liver cirrhosis (LC) and 82 had HBV-related chronic hepatitis B (CHB). A direct sequencing method was employed to determine the HBV genotypes and the mutations in BCP/PC regions. In all, 133 of them were infected with genotype C viruses (79.17%); only 35 patients (20.83%) were infected with genotype B viruses. Both LC patients and CHB patients had significantly higher ratios of genotype C when compared with the ratios of genotype B (83.7%-16.3% versus 74.4%-25.6%). For patients with CHB, the prevalence of BCP/PC wild-type viruses was 52.4%; but this was only 4.7% in patients with LC. The C1653T, T1753C, A1762T/G1764A and G1896A mutations had a significantly higher prevalence in patients with LC. Among all the patients with genotype B viruses, those with LC had lower HBV DNA levels and higher G1899A mutation frequency than patients with CHB. Among all the patients with genotype C viruses, the patients with LC had higher prevalence of C1653T, A1762T/G1764A and G1896A mutation frequency, higher hepatitis B e antigen (HBeAg) -negative rates, lower viral load, lower elevated alanine aminotransferase and lower anti-HBe positive rates than CHB patients. The HBV BCP/PC variants were more common in HBeAg-negative LC patients than in the CHB group (BCP, 53.4% versus 15.6%; PC, 18.6% versus 3.7%, respectively, p < 0.001). Patients with HBV genotype C viruses, high viral load and C1653T, A1762T/G1764A, G1896A mutant viruses, were more susceptible to developing LC.

  19. A comparison of the effects of penicillamine, trientine, and trithiomolybdate on ( sup 35 S)-labeled metallothionein in vitro; implications for Wilson's disease therapy

    SciTech Connect

    McQuaid, A.; Mason, J. )

    1991-02-01

    The synthesis of radiolabeled metallothionein was induced in rats in vivo by the injection of CuSO{sub 4} and ({sup 35}S)-cysteine. Treatment of 'cold' rat liver cytosol 'spiked' with purified ({sup 35}S) metallothionein with Penicillamine and Trientine showed that even at relatively high concentrations (up to 50 mg/g liver, wet weight), these compounds had no effect on the copper peak or the position of the ({sup 35S}) label in the cytosol eluate after Sephadex G-75 gel filtration. By contrast, incubation of the 'spiked' liver cytosol with Trithiomolybdate, even at relatively low concentrations (0.5 mg/g liver, wet weight), resulted in a transfer of metallothionein copper to high molecular weight protein fractions; the position of the ({sup 35}S) apoprotein was unaffected. This copper 'stripping' effect on metallothionein supports clinical and other evidence that thiomolybdates have a genuine decoppering effect in vivo whereas Penicillamine and Trientine have another mode of action and indicates that thiomolybdates might provide a more rational alternate therapy for Wilson's disease patients.

  20. High affinity P2x-purinoceptor binding sites for [35S]-adenosine 5'-O-[3-thiotriphosphate] in rat vas deferens membranes.

    PubMed Central

    Michel, A. D.; Humphrey, P. P.

    1996-01-01

    1. The binding sites labelled by [35S]-adenosine 5'-O-[3-thiotriphosphate]([35S]-ATP gamma S) at 4 degrees C in rat vas deferens membranes were studied and compared to the sites labelled by [3H]-alpha,beta-methylene ATP ([3H]-alpha beta meATP) to ascertain whether [35S]-ATP gamma S can be used to label the P2x purinoceptor. 2. In the presence of 4 mM CaCl2, the binding of 0.2 nM [35S]-ATP gamma S to vas deferens membranes was increased 3.4 fold, when compared to studies performed in the absence of calcium. However, binding did not appear to be solely to P2x purinoceptors since [35S]-ATP gamma S labelled a heterogeneous population of sites and about 72% of the sites possessed high affinity (pIC50 = 7.5) for guanosine 5'-O-[3-thiotriphosphate] (GTP gamma S). Even in the presence of 1 microM GTP gamma S, to occlude the sites with high affinity for GTP gamma S, the binding of [35S]-ATP gamma S was heterogeneous and since there was also evidence of extensive metabolism of ATP in the presence of calcium, the binding of [35S]-ATP gamma S under these conditions was not studied further. 3. In the absence of calcium ions, [35S]-ATP gamma S bound to a single population of sites (pKD = 9.23; Bmax = 4270 fmol mg-1 protein). Binding reached steady state within 3 h (t1/2 = 38 min), was stable for a further 4 h and was readily reversible upon addition of 10 microM unlabelled ATP gamma S (t1/2 = 45 min). In competition studies the binding of 0.2 nM [35S]-ATP gamma S was inhibited by a number of P2x purinoceptor agonists and antagonists, but not by adenosine receptor agonists, staurosporine (1 microM) or several ATPase inhibitors. The rank order of agonist affinity estimates (pIC50 values) in competing for the [35S]-ATP gamma S binding sites was: ATP (9.01), 2-methylthio- ATP (8.79), ATP gamma S (8.73), alpha beta meATP (7.57), ADP (7.24), beta, gamma-methylene ATP (7.18), L-beta, gamma-methylene ATP (5.83), alpha, beta-methylene ADP (4.36). 4. Affinity estimates (pIC50 values) for

  1. Effects of cysteamine administration on the in vivo incorporation of (/sup 35/S)cysteine into somatostatin-14, somatostatin-28, arginine vasopressin, and oxytocin in rat hypothalamus

    SciTech Connect

    Cameron, J.L.; Fernstrom, J.D.

    1986-09-01

    The effect of cysteamine injection on the in vivo incorporation of (/sup 35/S)cysteine into somatostatin-14 (SRIF-14), SRIF-28, arginine vasopressin (AVP), and oxytocin (OXT) in rat hypothalamus was studied. (/sup 35/S)Cysteine was injected into the third ventricle 1 h, 4 h, or 1 week after cysteamine (300 mg/kg, sc) injection; animals were killed 4 h later. The drug was found to substantially reduce immunoreactive SRIF levels, but not OXT or AVP, 4 h after its injection. Cysteamine also caused large reductions in label incorporation into SRIF-14, SRIF-28, and OXT 1 and 4 h after drug injection. However, (/sup 35/S)cysteine incorporation into AVP was increased substantially at these time points, while that into acid-precipitable protein was normal. One week after cysteamine injection, label incorporation into all hypothalamic peptides was normal. Cysteine specific activity was also measured after (/sup 35/S)cysteine injection and was found to be similar in treatment and control groups. The results suggest that cysteamine inhibits the syntheses of SRIF-14, SRIF-28, and OXT and stimulates that of AVP.

  2. Ex vivo binding of t-( sup 35 S) butylbicyclophosphorothionate: A biochemical tool to study the pharmacology of ethanol at the gamma-aminobutyric acid-coupled chloride channel

    SciTech Connect

    Sanna, E.; Concas, A.; Serra, M.; Santoro, G.; Biggio, G. )

    1991-03-01

    The effects of acute administration of ethanol on t-(35S)Butylbiclophosphorothionate (35S-TBPS) binding measured ex vivo in unwashed membrane preparations of rat cerebral cortex were investigated. Ethanol, given i.g., decreased in a dose-related (0.5-4 g/kg) and time-dependent manner the binding of 35S-TBPS. This effect was similar to that induced by the administration of diazepam (0.5-4 mg/kg i.p.). Scatchard plot analysis of this radioligand binding revealed that ethanol, differently from diazepam, decreased the apparent affinity of 35S-TBPS recognition sites whereas it failed to change the density of these binding sites. The effect of ethanol on 35S-TBPS binding could not be reversed by the previous administration to rats of the benzodiazepine receptor antagonist, Ro 15-1788 (ethyl-8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H- imidazo (1,5a) (1,4) benzodiazepine-3-carboxylate). Vice versa, the benzodiazepine receptor partial inverse agonist, Ro 15-4513 (ethyl-8-azido-5,6-dihydro-5-methyl-6-oxo-4H- imidazo (1,5a) (4,4) benzodiazepine-3-carboxylate) (8 mg/kg i.p.), prevented completely ethanol-induced decrease of 35S-TBPS binding. The ability of Ro 15-4513 to prevent the action of ethanol was shared by the anxiogenic and proconvulsant beta-carboline derivatives, FG 7142 (N-methyl-beta-carboline-3-carboxamide) (12.5 mg/kg i.p.) and ethyl-beta-carboline-3-carboxylate (0.6 mg/kg i.v.), which, per se, enhanced this parameter. Moreover, ethanol (0.5-4 g/kg) was able to reverse the increase of 35S-TBPS binding elicited by the s.c. injection of isoniazid (350 mg/kg) and to clearly attenuate the severity of tonic-clonic seizures produced by this inhibitor of the GABAergic transmission.

  3. Differentiating atmospheric and mineral sources of sulfur during snowmelt using δ 34S, 35S activity, and δ 18O of sulfate and water as tracers

    NASA Astrophysics Data System (ADS)

    Shanley, J. B.; Mayer, B.; Mitchell, M. J.; Michel, R. L.; Bailey, S.; Kendall, C.

    2003-12-01

    The biogeochemical cycling of sulfur was studied during the 2000 snowmelt at Sleepers River Research Watershed in northeastern Vermont, USA using a combination of isotopic, chemical, and hydrometric measurements. The snowpack and 10 streams of varying size and land use were sampled for sulfate concentrations and isotopic analyses of 35S, δ 34S, and δ 18O of sulfate. Values of δ 18O of water were measured at one of the streams. Apportionment of atmospheric and mineral S sources based on δ 34S was possible at 7 of the 10 streams. Weathering of S-containing minerals was a major contributor to sulfate flux in streamwater, but atmospheric contributions exceeded 50% in several of the streams at peak snowmelt and averaged 41% overall. In contrast, δ 18Osulfate values of streamwater remained significantly lower than those of atmospheric sulfate throughout the melt period, indicating that atmospheric sulfate undergoes microbial redox reactions in the soil that replace the oxygen of atmospheric sulfate with isotopically lighter oxygen from soil water. Streamwater 35S activities were low relative to those of the snowpack; the youngest 35S-ages of the atmospheric S component in each of the 7 streams ranged from 184 to 320 days. Atmospheric S contributions to streamwater, as determined by δ 34S values, co-varied both with 35S activity and new water contributions as determined by δ 18Owater. However, the δ 18Osulfate and 35S ages clearly show that this new water carries very little of the atmospheric sulfate entering with the current snowmelt to the stream. Most incoming atmospheric sulfate first cycles through the organic soil S pool and ultimately reaches the stream as pedogenic sulfate.

  4. Analysis of the rolC promoter region involved in somatic embryogenesis-related activation in carrot cell cultures.

    PubMed Central

    Fujii, N; Yokoyama, R; Uchimiya, H

    1994-01-01

    In cell cultures of carrot (Daucus carota L.), somatic embryogenesis can be induced by transferring cells from a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) to one devoid of 2,4-D. Previous analysis of transgenic carrot cells containing the 5' non-coding sequence of the Ri plasmid rolC and a structural gene for bacterial beta-glucuronidase (uidA) has shown that the chimeric gene is actively expressed after induction of somatic embryogenesis. In this study, we demonstrate that activation of the rolC promoter is dependent on the process of embryo development but not on the duration of the cell culture in 2,4-D-free medium. We also analyzed the cis region of the rolC promoter that is responsible for somatic embryogenesis-related activation (SERA), namely relatively low beta-glucuronidase (GUS) activity in calli and proembryogenic masses (PEM) and high GUS activity in heart- and torpedo-stage embryos. When the -255-bp region of the rolC gene was used, SERA was retained. Internal deletions within this -255-bp region did not alter SERA by the rolC promoter. Furthermore, when a rolC promoter fragment (-848 to -94 bp) was fused to the cauliflower mosaic virus (CaMV) 35S core region (-90 to +6 bp), it conferred relatively low GUS activity in calli and PEM but high GUS activity in heart and torpedo embryos. When -848 to -255-bp or -255- to -94-bp fragments of the rolC promoter were fused to the same CaMV 35S core region, GUS activity patterns were not related to somatic embryogenesis. These results suggest that the combination of several regulatory regions in the rolC promoter may be required for SERA in carrot cell cultures. PMID:8016259

  5. Many-electron aspects of molecular promotion in ion-atom collisions - Production of core-excited states of Li in Li/+/-He collisions

    NASA Technical Reports Server (NTRS)

    Elston, S. B.; Vane, C. R.; Schumann, S.

    1979-01-01

    Production of core-excited autoionizing states of neutral Li having configurations of the form 1snln(prime)l(prime) has been observed over the impact-energy range from 10-50 keV. Although the results for production of all such states is remarkably consistent with a quasi-molecular-excitation model proposed by Stolterfoht and Leithaeuser (1976), production of individual lines in the observed spectra exhibits collision-velocity dependencies indicative of considerably more complex processes, including processes which appear to be inherently two-electron in nature. Excitation functions are presented for (1s2s/2/)/2/S, 1s(2s2p/3/P)/2/P, 1s(2s2p/1/P)/2/P, and (1s2p/2/)/2/D core-excited state of Li and for total core excitation.

  6. In vivo treatment of HCV core-positive HepG2 cells with the transfer of recombinant caspase-3 using a 2'-5' OAS promoter.

    PubMed

    Zi, Yuan; Wang, Ying; Wiegmann, Peter S; Luo, Junming; Feng, Deyun

    2012-03-01

    Hepatitis C virus (HCV) is one of the most common pathogens causing liver-related morbidity and mortality, which affect 170 million individuals worldwide. There is no vaccine available, and current therapy is only partially effective. In a previous study, we constructed a recombinant caspase-3 expression vector under the 2'-5'-oligoadenylate synthetase gene (OAS) promoter (pGL3-OAS-re-caspase-3) and demonstrated that it is an effective gene therapy for HCV core-positive liver cells in vitro. In the present study, the human hepatoma cell line HepG2 was transfected with the pcDNA3.1-HCV-core-EGFP plasmid and selected by G418. Expression of HCV core protein was confirmed by RT-PCR and immunocytochemistry. Both HepG2-expressing HCV core protein and parental HepG2 cells were inoculated subcutaneously into BALB/c mice, respectively. Tumor-bearing mice were treated with an intratumoral injection of pGL3-OAS-re-caspase-3. The mice were sacrificed after 48 h. The correlation between HCV core and caspase-3 expression in tumor tissues was analyzed by immunohistochemical staining and double-label immunofluorescence staining. The subcutaneous hepatoma in vivo mouse models stably expressing HCV core protein and co-expressing HCV core protein and pGL3-OAS-re-caspase-3 were established. Double-label immunofluorescence staining showed that the percentage of co-expression of both HCV core and caspase-3 was 76 ± 6% in the group treated with pGL3-OAS-re-caspase-3. There was a significant increase in the number of apoptotic cells in the group treated with the pGL3-OAS-re-caspase-3 system by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and transmission electron microscopy. The results suggest that the pGL3-OAS-re-caspase-3 construct can effectively induce apoptosis in HCV core-positive hepatocytes in vivo. The results presented strongly suggest that the transfer of pGL3-OAS-re-caspase-3 is an effective and promising gene therapy strategy for HCV infection.

  7. Distributions of /sup 35/S-sulfate and /sup 3/H-glucosamine in the angular region of the hamster: light and electron microscopic autoradiography

    SciTech Connect

    Ohnishi, Y.; Taniguchi, Y.

    1983-06-01

    The distribution of /sup 35/S-sulfate and /sup 3/H-glucosamine in the angular region of the hamster was studied by light and electron microscopic autoradiography following intraperitoneal injection of these compounds to hamsters. Exposed silver grains of /sup 35/S-sulfate were concentrated in the trabecular meshwork, sclera, and cornea, and grains of /sup 3/H-glucosamine were localized in the trabecular region. The radioactivity of both isotopes was observed in the Golgi apparatuses of the endothelial cells of the angular aqueous plexus and the trabecular meshwork. The grains were noted over the entire cytoplasm, except for the nucleus, and then were incorporated into the amorphous substance and collagen fibers in the region adjacent to the angular aqueous sinus. These results suggest that endothelial cells in the angular region synthesize and secrete the sulfated glycosaminoglycans and hyaluronic acid.

  8. Is there evidence for a 17 keV neutrino in the 35S β spectrum? The case of Ohi et al.

    NASA Astrophysics Data System (ADS)

    Simpson, J. J.

    1986-06-01

    It is shown that there is a threshold 17 keV below the end point of the β spectrum of 35S in the published work of Ohio et al. The distortion of the Kurie plot is consistent with that seen in the 3H β spectrum, strengthening the earlier suggestion that the distortion is due to the emission of a neutrino of mass 17 keV.

  9. In vivo biosynthesis of L-(/sup 35/S)Cys-arginine vasopressin, -oxytocin, and -somatostatin: rapid estimation using reversed phase high pressure liquid chromatography. [Rats

    SciTech Connect

    Franco-Bourland, R.E.; Fernstrom, J.D.

    1981-01-01

    L(/sup 35/S)Cys-arginine vasopressin, -oxytocin, and -somatostatin were purified from hypothalami and neurohypophyses 4 h after rats received L(/sup 35/S)Cys via the third ventricle. After acetic acid extraction, Sephadex G-25 filtration, and chemoadsorption to C18-silica (Sep-Pak cartridges), the labeled peptides were rapidly separated by gradient elution, reversed phase, high pressure liquid chromatography (HPLC). The identity and isotopic purity of the labeled peptides were determined by several reversed phase HPLC procedures in conjunction with chemical modification. The labeled peptide fractions were at least 50% radiochemically pure. Using this HPLC isolation procedure, incorporation of L-(/sup 35/S)Cys into each peptide was determined in hydrated and dehydrated rats. Label incorporation into arginine vasopressin and oxytocin in the hypothalamus and the neurohypophysis of dehydrated rats was 2-3 times greater than that in hydrated rats. Incorporation of label into hypothalamic and neurohypophyseal somatostatin was unaffected by the hydration state of the animal. This procedure thus provides a very rapid, but sensitive, set of techniques for studying the control of small peptide biosynthesis in the brain.

  10. Moving beyond Compliance: Promoting Research-Based Professional Discretion in the Implementation of the Common Core State Standards in English Language Arts

    ERIC Educational Resources Information Center

    Woodard, Rebecca; Kline, Sonia

    2015-01-01

    State- and local-level mandates are currently being implemented to ensure strict compliance to the new national Common Core State Standards for English Language Arts (CCSS for ELA) and related assessments. These standards provide many potential opportunities to improve literacy education nationally and locally. However, the CCSS for ELA will…

  11. Functional analysis of the stress-inducible soybean calmodulin isoform-4 (GmCaM-4) promoter in transgenic tobacco plants.

    PubMed

    Park, Hyeong Cheol; Kim, Man Lyang; Kang, Yun Hwan; Jeong, Jae Cheol; Cheong, Mi Sun; Choi, Wonkyun; Lee, Sang Yeol; Cho, Moo Je; Kim, Min Chul; Chung, Woo Sik; Yun, Dae-Jin

    2009-04-30

    The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

  12. Use of cosmogenic 35S for comparing ages of water from three alpine-subalpine basins in the Colorado Front Range

    USGS Publications Warehouse

    Sueker, J.K.; Turk, J.T.; Michel, R.L.

    1999-01-01

    High-elevation basins in Colorado are a major source of water for the central and western United States; however, acidic deposition may affect the quality of this water. Water that is retained in a basin for a longer period of time may be less impacted by acidic deposition. Sulfur-35 (35S), a short-lived isotope of sulfur (t( 1/2 ) = 87 days), is useful for studying short-time scale hydrologic processes in basins where biological influences and water/rock interactions are minimal. When sulfate response in a basin is conservative, the age of water may be assumed to be that of the dissolved sulfate in it. Three alpine-subalpine basins on granitic terrain in Colorado were investigated to determine the influence of basin morphology on the residence time of water in the basins. Fern and Spruce Creek basins are glaciated and accumulate deep snowpacks during the winter. These basins have hydrologic and chemical characteristics typical of systems with rapid hydrologic response times. The age of sulfate leaving these basins, determined from the activity of 35S, averages around 200 days. In contrast, Boulder Brook basin has broad, gentle slopes and an extensive cover of surficial debris. Its area above treeline, about one-half of the basin, is blown free of snow during the winter. Variations in flow and solute concentrations in Boulder Brook are quite small compared to Fern and Spruce Creeks. After peak snowmelt, sulfate in Boulder Brook is about 200 days older than sulfate in Fern and Spruce Creeks. This indicates a substantial source of older sulfate (lacking 35S) that is probably provided from water stored in pore spaces of surficial debris in Boulder Brook basin.

  13. Repeated reunions and splits feature the highly dynamic evolution of 5S and 35S ribosomal RNA genes (rDNA) in the Asteraceae family

    PubMed Central

    2010-01-01

    Background In flowering plants and animals the most common ribosomal RNA genes (rDNA) organisation is that in which 35S (encoding 18S-5.8S-26S rRNA) and 5S genes are physically separated occupying different chromosomal loci. However, recent observations established that both genes have been unified to a single 35S-5S unit in the genus Artemisia (Asteraceae), a genomic arrangement typical of primitive eukaryotes such as yeast, among others. Here we aim to reveal the origin, distribution and mechanisms leading to the linked organisation of rDNA in the Asteraceae by analysing unit structure (PCR, Southern blot, sequencing), gene copy number (quantitative PCR) and chromosomal position (FISH) of 5S and 35S rRNA genes in ~200 species representing the family diversity and other closely related groups. Results Dominant linked rDNA genotype was found within three large groups in subfamily Asteroideae: tribe Anthemideae (93% of the studied cases), tribe Gnaphalieae (100%) and in the "Heliantheae alliance" (23%). The remaining five tribes of the Asteroideae displayed canonical non linked arrangement of rDNA, as did the other groups in the Asteraceae. Nevertheless, low copy linked genes were identified among several species that amplified unlinked units. The conserved position of functional 5S insertions downstream from the 26S gene suggests a unique, perhaps retrotransposon-mediated integration event at the base of subfamily Asteroideae. Further evolution likely involved divergence of 26S-5S intergenic spacers, amplification and homogenisation of units across the chromosomes and concomitant elimination of unlinked arrays. However, the opposite trend, from linked towards unlinked arrangement was also surmised in few species indicating possible reversibility of these processes. Conclusions Our results indicate that nearly 25% of Asteraceae species may have evolved unusual linked arrangement of rRNA genes. Thus, in plants, fundamental changes in intrinsic structure of rDNA units

  14. Observation of parity violation and a left-right asymmetry in the reaction /sup 35/Cl (n, p) /sup 35/S

    SciTech Connect

    Antonov, A.; Vesna, V.A.; Gledenov, Y.M.; Lobashev, V.M.; Okunev, I.S.; Popov, Y.P.; Rigol', K.; Smotritskii, L.M.

    1984-09-10

    The P-odd and left-right asymmetry in the emission of protons by the compound nucleus in the reaction /sup 35/Cl (n, p) /sup 35/S have been measured for the first time. The coefficients are a/sub p/ = -(1.51 +- 0.34) x 10/sup -4/ and a/sup LR//sub p/ = -(2.40 +- 0.43) x 10/sup -4/. A limitation is found on the dependence of the total cross section on the neutron helicity: Vertical Bar..cap alpha../sub n/Vertical Bar<2 x 10/sup -6/ (at a 90% confidence level).

  15. Three Medicago MtFUL genes have distinct and overlapping expression patterns during vegetative and reproductive development and 35S:MtFULb accelerates flowering and causes a terminal flower phenotype in Arabidopsis.

    PubMed

    Jaudal, Mauren; Zhang, Lulu; Che, Chong; Putterill, Joanna

    2015-01-01

    The timing of the transition to flowering is carefully controlled by plants in order to optimize sexual reproduction and the ensuing production of seeds, grains, and fruits. The genetic networks that regulate floral induction are best characterized in the temperate eudicot Arabidopsis in which the florigen gene FT plays a major role in promoting the transition to flowering. Legumes are an important plant group, but less is known about the regulation of their flowering time. In the model legume Medicago truncatula (Medicago), a temperate annual plant like Arabidopsis, flowering is induced by prolonged cold (vernalization) followed by long day lengths (LD). Recent molecular-genetic experiments have revealed that a FT-like gene, MtFTa1, is a central regulator of flowering time in Medicago. Here, we characterize the three Medicago FRUITFULL (FUL) MADS transcription factors, MtFULa, MtFULb, and MtFULc using phylogenetic analyses, gene expression profiling through developmental time courses, and functional analyses in transgenic plants. MtFULa and MtFULb have similarity in sequence and expression profiles under inductive environmental conditions during both vegetative and reproductive development while MtFULc is only up regulated in the apex after flowering in LD conditions. Sustained up regulation of MtFULs requires functional MtFTa1 but their transcript levels are not affected during cold treatment. Overexpression of MtFULa and MtFULb promotes flowering in transgenic Arabidopsis plants with an additional terminal flower phenotype on some 35S:MtFULb plants. An increase in transcript levels of the MtFULs was also observed in Medicago plants overexpressing MtFTa1. Our results suggest that the MtFULs are targets of MtFTa1. Overall, this work highlights the conserved functions of FUL-like genes in promoting flowering and other roles in plant development and thus contributes to our understanding of the genetic control of the flowering process in Medicago.

  16. Incorporation of /sup 35/S-sulfate and /sup 3/H-glucosamine into heparan and chondroitin sulfates during the cell cycle of B16-F10 cells

    SciTech Connect

    Blair, O.C.; Sartorelli, A.C.

    1984-05-01

    Changes in glycosaminoglycan composition occurring during the cell cycle were determined in B16-F10 cells sorted flow cytometrically with respect to DNA content. Incorporation of /sup 35/S-sulfate into heparan sulfate and chondroitin sulfate of unsorted and G1,S, and G2 +M sorted cells was determined following chondroitinase ABC or nitrous acid treatment; the incorporation into surface material was measured as the difference between the radioactivity of control and trypsin-treated cells. Incorporation of /sup 35/S-sulfate and /sup 3/H-glucosamine into cetyl pyridinium chloride (CPC)-precipitable material was characterized before and after chondroitinase or nitrous acid treatment by Sephadex G50 chromatography. Long-term (48 h) and short-term (1 h) labeling studies demonstrate that (a) the amount of total cellular chondroitin sulfate is greater than that of heparan sulfate, with larger amounts of unsulfated heparan than chondroitin being present; (b) the rate of turnover of heparan sulfate is greater than that of chondroitin sulfate; (c) greatest short-term incorporation of 3H-glucosamine into CPC-precipitable material occurs during S phase; and (d) the rate of turnover of both heparan sulfate and chondroitin sulfate is decreased in S phase relative to G1 and G2 + M.

  17. Effects of recombinant eel growth hormone on the uptake of ( sup 35 S)sulfate by ceratobranchial cartilages of the Japanese eel, Anguilla japonica

    SciTech Connect

    Duan, C.M.; Inui, Y. )

    1990-08-01

    Effects of growth hormone (GH) on the synthesis of mucopolysaccharide by ceratobranchial cartilages of the Japanese eel, Anguilla japonica, were examined by monitoring the in vitro uptake of ({sup 35}S)sulfate. The ({sup 35}S)sulfate uptake decreased rapidly to one-third of the initial level during the first 3 days after hypophysectomy, and decreased gradually thereafter. When hypophysectomized eels were injected intramuscularly with recombinant eel GH (2 micrograms/g), the plasma GH concentrations increased maximally after 6 hr, and declined rapidly thereafter. On the other hand, the sulfate uptake increased significantly after 12 hr, and high levels were maintained until 48 hr. The stimulating effect of GH was dose dependent (0.02-2 micrograms/g). However, the addition of eel GH (0.05-5 micrograms/ml) to the culture medium did not affect the sulfate uptake by hypophysectomized eel cartilages, suggesting that the stimulative action of GH on the sulfate uptake by the cartilages is indirect.

  18. Human Promoters Are Intrinsically Directional

    PubMed Central

    Duttke, Sascha H.C.; Lacadie, Scott A.; Ibrahim, Mahmoud M.; Glass, Christopher K.; Corcoran, David L.; Benner, Christopher; Heinz, Sven; Kadonaga, James T.; Ohler, Uwe

    2015-01-01

    Divergent transcription, in which reverse-oriented transcripts occur upstream of eukaryotic promoters in regions devoid of annotated genes, has been suggested to be a general property of active promoters. Here we show that the human basal RNA polymerase II transcriptional machinery and core promoter are inherently unidirectional, and that reverse-oriented transcripts originate from their own cognate reverse-directed core promoters. In vitro transcription analysis and mapping of nascent transcripts in cells revealed that sequences at reverse start sites are similar to those of their forward counterparts. The use of DNase I accessibility to define proximal promoter borders revealed that up to half of promoters are unidirectional and that unidirectional promoters are depleted at their upstream edges of reverse core promoter sequences and their associated chromatin features. Divergent transcription is thus not an inherent property of the transcription process, but rather the consequence of the presence of both forward- and reverse-directed core promoters. PMID:25639469

  19. Recognition of a core fragment of Beauveria bassiana hydrophobin gene promoter (P hyd1) and its special use in improving fungal biocontrol potential

    PubMed Central

    Wang, Zheng-Liang; Ying, Sheng-Hua; Feng, Ming-Guang

    2013-01-01

    To identify a suitable promoter for use in engineering fungal entomopathogens to improve heterologous gene expression and fungal biocontrol potential, a 1798 bp promoter (Phyd1) upstream of Beauveria bassiana class I hydrophobin gene (hyd1) was optimized by upstream truncation and site-directed mutation. A truncated 1290 bp fragment (Phyd1-t1) drove eGFP expression in B. bassiana much more efficiently than full-length Phyd1. Further truncating Phyd1-t1 to 1179, 991 and 791 bp or mutating one of the binding domains of three transcription factors in Phyd1-t1 reduced significantly the expression of eGFP (enhanced green fluorescence protein). Under Phyd1-t1 control, eGFP was expressed more abundantly in conidiogenic cells and conidia than in mycelia. Therefore, Phyd1-t1 was used to integrate a bacterium-derived, insect midgut-specific toxin (vip3Aa1) gene into B. bassiana, yielding a transgenic strain (BbHV8) expressing 9.8-fold more toxin molecules in conidia than a counterpart strain (BbV28) expressing the toxin under the control of PgpdA, a promoter widely used for gene expression in fungi. Consequently, BbHV8 showed much higher per os virulence to Spodoptera litura larvae than BbV28 in standardized bioassays with normal conidia for both cuticle penetration and ingestion or heat-killed conidia for ingestion only. Conclusively, Phyd1-t1 is a useful tool for enhancing beneficial protein expression, such as vip3Aa1, in fungal conidia, which are the active ingredients of mycoinsecticides. PMID:22639846

  20. Recognition of a core fragment ofBeauveria bassiana hydrophobin gene promoter (P hyd1) and its special use in improving fungal biocontrol potential.

    PubMed

    Wang, Zheng-Liang; Ying, Sheng-Hua; Feng, Ming-Guang

    2013-01-01

    To identify a suitable promoter for use in engineering fungal entomopathogens to improve heterologous gene expression and fungal biocontrol potential, a 1798 bp promoter (P hyd1) upstream of Beauveria bassiana class I hydrophobin gene (hyd1) was optimized by upstream truncation and site-directed mutation. A truncated 1290 bp fragment (P hyd1-t1) drove eGFP expression in B. bassiana much more efficiently than full-length P hyd1. Further truncating P hyd1-t1 to 1179, 991 and 791 bp or mutating one of the binding domains of three transcription factors in P hyd1-t1 reduced significantly the expression of eGFP (enhanced green fluorescence protein). Under P hyd1-t1 control, eGFP was expressed more abundantly in conidiogenic cells and conidia than in mycelia. Therefore, P hyd1-t1 was used to integrate a bacterium-derived, insect midgut-specific toxin (vip3Aa1) gene into B. bassiana, yielding a transgenic strain (BbHV8) expressing 9.8-fold more toxin molecules in conidia than a counterpart strain (BbV28) expressing the toxin under the control of P gpdA, a promoter widely used for gene expression in fungi. Consequently, BbHV8 showed much higher per os virulence to Spodoptera litura larvae than BbV28 in standardized bioassays with normal conidia for both cuticle penetration and ingestion or heat-killed conidia for ingestion only. Conclusively, P hyd1-t1 is a useful tool for enhancing beneficial protein expression, such as vip3Aa1, in fungal conidia, which are the active ingredients of mycoinsecticides.

  1. Phospholipase D1 production of phosphatidic acid at the plasma membrane promotes exocytosis of large dense-core granules at a late stage.

    PubMed

    Zeniou-Meyer, Maria; Zabari, Naama; Ashery, Uri; Chasserot-Golaz, Sylvette; Haeberlé, Anne-Marie; Demais, Valérie; Bailly, Yannick; Gottfried, Irit; Nakanishi, Hideki; Neiman, Aaron M; Du, Guangwei; Frohman, Michael A; Bader, Marie-France; Vitale, Nicolas

    2007-07-27

    Substantial efforts have recently been made to demonstrate the importance of lipids and lipid-modifying enzymes in various membrane trafficking processes, including calcium-regulated exocytosis of hormones and neurotransmitters. Among bioactive lipids, phosphatidic acid (PA) is an attractive candidate to promote membrane fusion through its ability to change membrane topology. To date, however, the biosynthetic pathway, the dynamic location, and actual function of PA in secretory cells remain unknown. Using a short interference RNA strategy on chromaffin and PC12 cells, we demonstrate here that phospholipase D1 is activated in secretagogue-stimulated cells and that it produces PA at the plasma membrane at the secretory granule docking sites. We show that phospholipase D1 activation and PA production represent key events in the exocytotic progression. Membrane capacitance measurements indicate that reduction of endogenous PA impairs the formation of fusion-competent granules. Finally, we show that the PLD1 short interference RNA-mediated inhibition of exocytosis can be rescued by exogenous provision of a lipid that favors the transition of opposed bi-layer membranes to hemifused membranes having the outer leaflets fused. Our findings demonstrate that PA synthesis is required during exocytosis to facilitate a late event in the granule fusion pathway. We propose that the underlying mechanism is related to the ability of PA to alter membrane curvature and promote hemi-fusion.

  2. Screening promoters for Anthurium transformation using transient expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Different promoters and tissue types were evaluated for transient '-glucoronidase (GUS) expression in Anthurium andreanum Hort. ‘Marian Seefurth’ following microprojectile bombardment. Plasmids containing the Ubiquitin 2, Actin 1, Cytochrome C1 from rice, Ubiquitin 1 from maize and 35 S promoter fr...

  3. A Point Mutation in the N-Terminal Amphipathic Helix α0 in NS3 Promotes Hepatitis C Virus Assembly by Altering Core Localization to the Endoplasmic Reticulum and Facilitating Virus Budding.

    PubMed

    Yan, Yu; He, Ying; Boson, Bertrand; Wang, Xuesong; Cosset, François-Loïc; Zhong, Jin

    2017-03-15

    The assembly of hepatitis C virus (HCV), a complicated process in which many viral and cellular factors are involved, has not been thoroughly deciphered. NS3 is a multifunctional protein that contains an N-terminal amphipathic α helix (designated helix α0), which is crucial for the membrane association and stability of NS3 protein, followed by a serine protease domain and a C-terminal helicase/NTPase domain. NS3 participates in HCV assembly likely through its C-terminal helicase domain, in which all reported adaptive mutations enhancing virion assembly reside. In this study, we determined that the N-terminal helix α0 of NS3 may contribute to HCV assembly. We identified a single mutation from methionine to threonine at amino acid position 21 (M21T) in helix α0, which significantly promoted viral production while having no apparent effect on the membrane association and protease activity of NS3. Subsequent analyses demonstrated that the M21T mutation did not affect HCV genome replication but rather promoted virion assembly. Further study revealed a shift in the subcellular localization of core protein from lipid droplets (LD) to the endoplasmic reticulum (ER). Finally, we showed that the M21T mutation increased the colocalization of core proteins and viral envelope proteins, leading to a more efficient envelopment of viral nucleocapsids. Collectively, the results of our study revealed a new function of NS3 helix α0 and aid understanding of the role of NS3 in HCV virion morphogenesis.IMPORTANCE HCV NS3 protein possesses the protease activity in its N-terminal domain and the helicase activity in its C-terminal domain. The role of NS3 in virus assembly has been mainly attributed to its helicase domain, because all adaptive mutations enhancing progeny virus production are found to be within this domain. Our study identified, for the first time to our knowledge, an adaptive mutation within the N-terminal helix α0 domain of NS3 that significantly enhanced virus

  4. Functional Characterization of a Strong Bi-directional Constitutive Plant Promoter Isolated from Cotton Leaf Curl Burewala Virus

    PubMed Central

    Khan, Zainul A.; Abdin, Malik Z.; Khan, Jawaid A.

    2015-01-01

    Cotton leaf curl Burewala virus (CLCuBuV), belonging to the genus Begomovirus, possesses single-stranded monopartite DNA genome. The bidirectional promoters representing Rep and coat protein (CP) genes of CLCuBuV were characterized and their efficacy was assayed. Rep and CP promoters of CLCuBuV and 35S promoter of Cauliflower mosaic virus (CaMV) were fused with β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes. GUS activity in individual plant cells driven by Rep, CP and 35S promoters was estimated using real-time PCR and fluorometric GUS assay. Histochemical staining of GUS in transformed tobacco (Nicotiana tabacum cv. Xanthi) leaves showed highest expression driven by Rep promoter followed by 35S promoter and CP promoter. The expression level of GUS driven by Rep promoter in transformed tobacco plants was shown to be two to four-fold higher than that of 35S promoter, while the expression by CP promoter was slightly lower. Further, the expression of GFP was monitored in agroinfiltrated leaves of N. benthamiana, N. tabacum and cotton (Gossypium hirsutum) plants using confocal laser scanning microscopy. Rep promoter showed strong consistent transient expression in tobacco and cotton leaves as compared to 35S promoter. The strong constitutive CLCuBuV Rep promoter developed in this study could be very useful for high level expression of transgenes in a wide variety of plant cells. PMID:25799504

  5. Hepatitis B virus basal core promoter mutations A1762T/G1764A are associated with genotype C and a low serum HBsAg level in chronically-infected HBeAg-positive Chinese patients.

    PubMed

    Yan, Chun-Hui; Zhao, Cheng-Yu; Ding, Hai; Peng, Ya-Qin; Jin, Peng-Yuan; Yan, Ling; Zhuang, Hui; Li, Tong

    2012-11-01

    The present study was aimed to obtain baseline information of basal core promoter A1762T/G1764A and precore G1896A mutations of hepatitis B virus (HBV) in 192 HBeAg-positive chronically-infected Chinese patients, who were potential candidates for antiviral treatment. The detection of these mutations (including minor mutant subpopulations) was achieved by direct sequencing, whose sensitivity for minor mutant subpopulations identification was confirmed by clone sequencing. Patients enrolled were infected with either genotype B (46.35%) or C (53.65%) HBV identified by routine tests in our laboratory. The A1762T/G1764A or G1896A mutations were detected in 125specimens (125/192, 65.10%), in which 77 (77/125, 61.60%) existed as subpopulations. The A1762T/G1764A mutations were found to be more prevalent in genotype C than that in genotype B HBV [62.14% (64/103) vs. 20.22% (18/89), P<0.0001]. There is no statistically significant link between G1896A and genotypes. The emergence of A1762T/G1764A mutations was also found to be associated with an older age, an elevated ALT/AST level, and a lower HBsAg level in serum [wild-type vs. mutant: 4.57 (3.46-5.42) vs. 3.93 (2.51-5.36), P<0.0001]. In conclusion, HBV basal core promoter mutations A1762T/G1764A are associated with genotype C and a low serum HBsAg level in chronically-infected HBeAg-positive Chinese patients.

  6. Identification of a negative regulatory cis-element in the enhancer core region of the prostate-specific antigen promoter: implications for intersection of androgen receptor and nuclear factor-kappaB signalling in prostate cancer cells.

    PubMed Central

    Cinar, Bekir; Yeung, Fan; Konaka, Hiroyuki; Mayo, Marty W; Freeman, Michael R; Zhau, Haiyen E; Chung, Leland W K

    2004-01-01

    The NF-kappaB (nuclear factor-kappaB) transcription factors mediate activation of a large number of gene promoters containing diverse kappaB-site sequences. Here, PSA (prostate-specific antigen) was used as an AR (androgen receptor)-responsive gene to examine the underlying mechanism by which the NF-kappaB p65 transcription factor down-regulates the transcriptional activity of AR in cells. We observed that activation of NF-kappaB by TNFalpha (tumour necrosis factor alpha) inhibited both basal and androgen-stimulated PSA expression, and that this down-regulation occurred at the promoter level, as confirmed by the super-repressor IkappaBalpha (S32A/S36A), a dominant negative inhibitor of NF-kappaB. Using a linker-scanning mutagenesis approach, we identified a cis -element, designated XBE (X-factor-binding element), in the AREc (androgen response element enhancer core) of the PSA promoter, which negatively regulated several AR-responsive promoters, including that of PSA. When three copies of XBE in tandem were juxtaposed to GRE4 (glucocorticoid response element 4), a 4-6-fold reduction of inducible GRE4 activity was detected in three different cell lines, LNCaP, ARCaP-AR and PC3-AR. Bioinformatics and molecular biochemical studies indicated that XBE is a kappaB-like element that binds specifically to the NF-kappaB p65 subunit; consistent with these observations, only NF-kappaB p65, but not the NF-kappaB p50 subunit, was capable of inhibiting AR-mediated PSA promoter transactivation in LNCaP cells. In addition, our data also showed that AR binds to XBE, as well as to the kappaB consensus site, and that the transfection of AR inhibits the kappaB-responsive promoter in transient co-transfection assays. Collectively, these data indicate that cross-modulation between AR and NF-kappaB p65 transcription factors may occur by a novel mechanism involving binding to a common cis -DNA element. PMID:14715080

  7. Effect of x-organ sinus gland extract on [(35)S] methionine incorporation to the ovary of the red swamp crawfish Procambarus clarkii.

    PubMed

    Chaves, A R

    2000-07-01

    The presence of gonad-inhibiting hormone in the x-organ sinus gland complex was evaluated in female Procambarus clarkii. Elimination of gonad-inhibiting hormone by way of eyestalk removal resulted in a large acceleration of ovarian development. Daily injection of four sinus gland equivalents reduced ovarian growth of eyestalk-ablated females by about 50% on day 6. Use of the radiotracer [(35)S] methionine showed that gonad-inhibiting activity reached its peak effect between 12 and 24 h following sinus gland injection. Dose-response showed that at least two sinus gland equivalents were needed to significantly counter the accelerated growth induced by eyestalk ablation. The high dose of extract needed to cause significant inhibition was attributed to this delayed response, which subsequently may have required a relatively prolonged exposure to the hormone.

  8. Genome-Wide Mapping Targets of the Metazoan Chromatin Remodeling Factor NURF Reveals Nucleosome Remodeling at Enhancers, Core Promoters and Gene Insulators

    PubMed Central

    Kwon, So Yeon; Grisan, Valentina; Jang, Boyun; Herbert, John; Badenhorst, Paul

    2016-01-01

    NURF is a conserved higher eukaryotic ISWI-containing chromatin remodeling complex that catalyzes ATP-dependent nucleosome sliding. By sliding nucleosomes, NURF is able to alter chromatin dynamics to control transcription and genome organization. Previous biochemical and genetic analysis of the specificity-subunit of Drosophila NURF (Nurf301/Enhancer of Bithorax (E(bx)) has defined NURF as a critical regulator of homeotic, heat-shock and steroid-responsive gene transcription. It has been speculated that NURF controls pathway specific transcription by co-operating with sequence-specific transcription factors to remodel chromatin at dedicated enhancers. However, conclusive in vivo demonstration of this is lacking and precise regulatory elements targeted by NURF are poorly defined. To address this, we have generated a comprehensive map of in vivo NURF activity, using MNase-sequencing to determine at base pair resolution NURF target nucleosomes, and ChIP-sequencing to define sites of NURF recruitment. Our data show that, besides anticipated roles at enhancers, NURF interacts physically and functionally with the TRF2/DREF basal transcription factor to organize nucleosomes downstream of active promoters. Moreover, we detect NURF remodeling and recruitment at distal insulator sites, where NURF functionally interacts with and co-localizes with DREF and insulator proteins including CP190 to establish nucleosome-depleted domains. This insulator function of NURF is most apparent at subclasses of insulators that mark the boundaries of chromatin domains, where multiple insulator proteins co-associate. By visualizing the complete repertoire of in vivo NURF chromatin targets, our data provide new insights into how chromatin remodeling can control genome organization and regulatory interactions. PMID:27046080

  9. Uniform accumulation of recombinant miraculin protein in transgenic tomato fruit using a fruit-ripening-specific E8 promoter.

    PubMed

    Hirai, Tadayoshi; Kim, You-Wang; Kato, Kazuhisa; Hiwasa-Tanase, Kyoko; Ezura, Hiroshi

    2011-12-01

    The E8 promoter, a tomato fruit-ripening-specific promoter, and the CaMV 35S promoter, a constitutive promoter, were used to express the miraculin gene encoding the taste-modifying protein in tomato. The accumulation of miraculin protein and mRNA was compared among transgenic tomatoes expressing the miraculin gene driven by these promoters. Recombinant miraculin protein predominantly accumulated in transgenic tomato lines using the E8 promoter (E8-MIR) only at the red fruit stage. The accumulations were almost uniform among all fruit tissues. When the 35S promoter (35S-MIR) was used, miraculin accumulation in the exocarp was much higher than in other tissues, indicating that the miraculin accumulation pattern can be regulated by using different types of promoters. We also discuss the potential of the E8-MIR lines for practical use.

  10. Core bioactive components promoting blood circulation in the traditional Chinese medicine compound xueshuantong capsule (CXC) based on the relevance analysis between chemical HPLC fingerprint and in vivo biological effects.

    PubMed

    Liu, Hong; Liang, Jie-ping; Li, Pei-bo; Peng, Wei; Peng, Yao-yao; Zhang, Gao-min; Xie, Cheng-shi; Long, Chao-feng; Su, Wei-wei

    2014-01-01

    Compound xueshuantong capsule (CXC) is an oral traditional Chinese herbal formula (CHF) comprised of Panax notoginseng (PN), Radix astragali (RA), Salvia miltiorrhizae (SM), and Radix scrophulariaceae (RS). The present investigation was designed to explore the core bioactive components promoting blood circulation in CXC using high-performance liquid chromatography (HPLC) and animal studies. CXC samples were prepared with different proportions of the 4 herbs according to a four-factor, nine-level uniform design. CXC samples were assessed with HPLC, which identified 21 components. For the animal experiments, rats were soaked in ice water during the time interval between two adrenaline hydrochloride injections to reduce blood circulation. We assessed whole-blood viscosity (WBV), erythrocyte aggregation and red corpuscle electrophoresis indices (EAI and RCEI, respectively), plasma viscosity (PV), maximum platelet aggregation rate (MPAR), activated partial thromboplastin time (APTT), and prothrombin time (PT). Based on the hypothesis that CXC sample effects varied with differences in components, we performed grey relational analysis (GRA), principal component analysis (PCA), ridge regression (RR), and radial basis function (RBF) to evaluate the contribution of each identified component. Our results indicate that panaxytriol, ginsenoside Rb1, angoroside C, protocatechualdehyde, ginsenoside Rd, and calycosin-7-O-β-D-glucoside are the core bioactive components, and that they might play different roles in the alleviation of circulation dysfunction. Panaxytriol and ginsenoside Rb1 had close relevance to red blood cell (RBC) aggregation, angoroside C was related to platelet aggregation, protocatechualdehyde was involved in intrinsic clotting activity, ginsenoside Rd affected RBC deformability and plasma proteins, and calycosin-7-O-β-D-glucoside influenced extrinsic clotting activity. This study indicates that angoroside C, calycosin-7-O-β-D-glucoside, panaxytriol, and

  11. Core Bioactive Components Promoting Blood Circulation in the Traditional Chinese Medicine Compound Xueshuantong Capsule (CXC) Based on the Relevance Analysis between Chemical HPLC Fingerprint and In Vivo Biological Effects

    PubMed Central

    Liu, Hong; Liang, Jie-ping; Li, Pei-bo; Peng, Wei; Peng, Yao-yao; Zhang, Gao-min; Xie, Cheng-shi; Long, Chao-feng; Su, Wei-wei

    2014-01-01

    Compound xueshuantong capsule (CXC) is an oral traditional Chinese herbal formula (CHF) comprised of Panax notoginseng (PN), Radix astragali (RA), Salvia miltiorrhizae (SM), and Radix scrophulariaceae (RS). The present investigation was designed to explore the core bioactive components promoting blood circulation in CXC using high-performance liquid chromatography (HPLC) and animal studies. CXC samples were prepared with different proportions of the 4 herbs according to a four-factor, nine-level uniform design. CXC samples were assessed with HPLC, which identified 21 components. For the animal experiments, rats were soaked in ice water during the time interval between two adrenaline hydrochloride injections to reduce blood circulation. We assessed whole-blood viscosity (WBV), erythrocyte aggregation and red corpuscle electrophoresis indices (EAI and RCEI, respectively), plasma viscosity (PV), maximum platelet aggregation rate (MPAR), activated partial thromboplastin time (APTT), and prothrombin time (PT). Based on the hypothesis that CXC sample effects varied with differences in components, we performed grey relational analysis (GRA), principal component analysis (PCA), ridge regression (RR), and radial basis function (RBF) to evaluate the contribution of each identified component. Our results indicate that panaxytriol, ginsenoside Rb1, angoroside C, protocatechualdehyde, ginsenoside Rd, and calycosin-7-O-β-D-glucoside are the core bioactive components, and that they might play different roles in the alleviation of circulation dysfunction. Panaxytriol and ginsenoside Rb1 had close relevance to red blood cell (RBC) aggregation, angoroside C was related to platelet aggregation, protocatechualdehyde was involved in intrinsic clotting activity, ginsenoside Rd affected RBC deformability and plasma proteins, and calycosin-7-O-β-D-glucoside influenced extrinsic clotting activity. This study indicates that angoroside C, calycosin-7-O-β-D-glucoside, panaxytriol, and

  12. Differential regulation of serotonin-1A receptor-stimulated [35S]GTP gamma S binding in the dorsal raphe nucleus by citalopram and escitalopram.

    PubMed

    Rossi, Dania V; Burke, Teresa F; Hensler, Julie G

    2008-03-31

    The effect of chronic citalopram or escitalopram administration on 5-HT1A receptor function in the dorsal raphe nucleus was determined by measuring [35S]GTP gamma S binding stimulated by the 5-HT1A receptor agonist (R)-(+)-8-OH-DPAT (1nM-10 microM). Although chronic administration of citalopram or escitalopram has been shown to desensitize somatodendritic 5-HT1A autoreceptors, we found that escitalopram treatment decreased the efficacy of 5-HT1A receptors to activate G proteins, whereas citalopram treatment did not. The binding of [3H]8-OH-DPAT to the coupled, high affinity agonist state of the receptor was not altered by either treatment. Interestingly, escitalopram administration resulted in greater occupancy of serotonin transporter sites as measured by the inhibition of [3H]cyanoimipramine binding. As the binding and action of escitalopram is limited by the inactive enantiomer R-citalopram present in racemic citalopram, we propose that the regulation of 5-HT1A receptor function in the dorsal raphe nucleus at the level of receptor-G protein interaction may be a result of greater inhibition of the serotonin transporter by escitalopram.

  13. [32P]orthophosphate and [35S]methionine label separate pools of neurofilaments with markedly different axonal transport kinetics in mouse retinal ganglion cells in vivo.

    PubMed

    Nixon, R A; Lewis, S E; Mercken, M; Sihag, R K

    1994-11-01

    Newly synthesized neurofilament proteins become highly phosphorylated within axons. Within 2 days after intravitreously injecting normal adult mice with [32P]orthophosphate, we observed that neurofilaments along the entire length of optic axons were radiolabeled by a soluble 32P-carrier that was axonally transported faster than neurofilaments. 32P-incorporation into neurofilament proteins synthesized at the time of injection was comparatively low and minimally influenced the labeling pattern along axons. 32P-incorporation into axonal neurofilaments was considerably higher in the middle region of the optic axons. This characteristic non-uniform distribution of radiolabel remained nearly unchanged for at least 22 days. During this interval, less than 10% of the total 32P-labeled neurofilaments redistributed from the optic nerve to the optic tract. By contrast, newly synthesized neurofilaments were selectively pulse-labeled in ganglion cell bodies by intravitreous injection of [35S]methionine and about 60% of this pool translocated by slow axoplasmic transport to the optic tract during the same time interval. These findings indicate that the steady-state or resident pool of neurofilaments in axons is not identical to the newly synthesized neurofilament pool, the major portion of which moves at the slowest rate of axoplasmic transport. Taken together with earlier studies, these results support the idea that, depending in part on their phosphorylation state, transported neurofilaments can interact for short or very long periods with a stationary but dynamic neurofilament lattice in axons.

  14. 24. A CORE WORKER DISPLAYS THE CORE BOX AND CORES ...

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

    24. A CORE WORKER DISPLAYS THE CORE BOX AND CORES FOR A BRASS GATE VALVE BODY MADE ON A CORE BOX, CA. 1950. - Stockham Pipe & Fittings Company, 4000 Tenth Avenue North, Birmingham, Jefferson County, AL

  15. Dancing together and separate again: gymnosperms exhibit frequent changes of fundamental 5S and 35S rRNA gene (rDNA) organisation

    PubMed Central

    Garcia, S; Kovařík, A

    2013-01-01

    In higher eukaryotes, the 5S rRNA genes occur in tandem units and are arranged either separately (S-type arrangement) or linked to other repeated genes, in most cases to rDNA locus encoding 18S–5.8S–26S genes (L-type arrangement). Here we used Southern blot hybridisation, PCR and sequencing approaches to analyse genomic organisation of rRNA genes in all large gymnosperm groups, including Coniferales, Ginkgoales, Gnetales and Cycadales. The data are provided for 27 species (21 genera). The 5S units linked to the 35S rDNA units occur in some but not all Gnetales, Coniferales and in Ginkgo (∼30% of the species analysed), while the remaining exhibit separate organisation. The linked 5S rRNA genes may occur as single-copy insertions or as short tandems embedded in the 26S–18S rDNA intergenic spacer (IGS). The 5S transcript may be encoded by the same (Ginkgo, Ephedra) or opposite (Podocarpus) DNA strand as the 18S–5.8S–26S genes. In addition, pseudogenised 5S copies were also found in some IGS types. Both L- and S-type units have been largely homogenised across the genomes. Phylogenetic relationships based on the comparison of 5S coding sequences suggest that the 5S genes independently inserted IGS at least three times in the course of gymnosperm evolution. Frequent transpositions and rearrangements of basic units indicate relatively relaxed selection pressures imposed on genomic organisation of 5S genes in plants. PMID:23512008

  16. Dancing together and separate again: gymnosperms exhibit frequent changes of fundamental 5S and 35S rRNA gene (rDNA) organisation.

    PubMed

    Garcia, S; Kovařík, A

    2013-07-01

    In higher eukaryotes, the 5S rRNA genes occur in tandem units and are arranged either separately (S-type arrangement) or linked to other repeated genes, in most cases to rDNA locus encoding 18S-5.8S-26S genes (L-type arrangement). Here we used Southern blot hybridisation, PCR and sequencing approaches to analyse genomic organisation of rRNA genes in all large gymnosperm groups, including Coniferales, Ginkgoales, Gnetales and Cycadales. The data are provided for 27 species (21 genera). The 5S units linked to the 35S rDNA units occur in some but not all Gnetales, Coniferales and in Ginkgo (∼30% of the species analysed), while the remaining exhibit separate organisation. The linked 5S rRNA genes may occur as single-copy insertions or as short tandems embedded in the 26S-18S rDNA intergenic spacer (IGS). The 5S transcript may be encoded by the same (Ginkgo, Ephedra) or opposite (Podocarpus) DNA strand as the 18S-5.8S-26S genes. In addition, pseudogenised 5S copies were also found in some IGS types. Both L- and S-type units have been largely homogenised across the genomes. Phylogenetic relationships based on the comparison of 5S coding sequences suggest that the 5S genes independently inserted IGS at least three times in the course of gymnosperm evolution. Frequent transpositions and rearrangements of basic units indicate relatively relaxed selection pressures imposed on genomic organisation of 5S genes in plants.

  17. A comparison of constitutive promoters for expression of transgenes in alfalfa (Medicago sativa).

    PubMed

    Samac, Deborah A; Tesfaye, Mesfin; Dornbusch, Melinda; Saruul, Purev; Temple, Stephen J

    2004-08-01

    The activity of constitutive promoters was compared in transgenic alfalfa plants using two marker genes. Three promoters, the 35S promoter from cauliflower mosaic virus (CaMV), the cassava vein mosaic virus (CsVMV) promoter, and the sugarcane bacilliform badnavirus (ScBV) promoter were each fused to the beta-glucuronidase (gusA) gene. The highest GUS enzyme activity was obtained using the CsVMV promoter and all alfalfa cells assayed by in situ staining had high levels of enzyme activity. The 35S promoter was expressed in leaves, roots, and stems at moderate levels, but the promoter was not active in stem pith cells, root cortical cells, or in the symbiotic zones of nodules. The ScBV promoter was active primarily in vascular tissues throughout the plant. In leaves, GUS activity driven by the CsVMV promoter was approximately 24-fold greater than the activity from the 35S promoter and 38-fold greater than the activity from the ScBV promoter. Five promoters, the double 35S promoter, figwort mosaic virus (FMV) promoter, CsVMV promoter, ScBV promoter, and alfalfa small subunit Rubisco (RbcS) promoter were used to control expression of a cDNA from Trichoderma atroviride encoding an endochitinase (ech42). Highest chitinase activity in leaves, roots, and root nodules was obtained in plants containing the CsVMV:ech42 transgene. Plants expressing the endochitinase were challenged with Phoma medicaginis var. medicaginis, the causal agent of spring black stem and leaf spot of alfalfa. Although endochitinase activity in leaves of transgenic plants was 50- to 2650-fold greater than activity in control plants, none of the transgenic plants showed a consistent increase in disease resistance compared to controls. The high constitutive levels of both GUS and endochitinase activity obtained demonstrate that the CsVMV promoter is useful for high-level transgene expression in alfalfa.

  18. Students Promoted Despite Test Failure

    ERIC Educational Resources Information Center

    Wakefield, Dara V.

    2012-01-01

    "No Child Left Behind" (NCLB) dictates students in Grades 3, 5, and 8 pass state tests to be promoted. Accordingly, most state education codes require students to pass reading and math exams for promotion. The majority of those who fail, however, appear to be promoted anyway. This article addresses core questions concerning the paradigm…

  19. Core layering

    NASA Astrophysics Data System (ADS)

    Jacobson, S. A.; Rubie, D. C.; Hernlund, J. W.; Morbidelli, A.

    2015-12-01

    We have created a planetary accretion and differentiation model that self-consistently builds and evolves Earth's core. From this model, we show that the core grows stably stratified as the result of rising metal-silicate equilibration temperatures and pressures, which increases the concentrations of light element impurities into each newer core addition. This stable stratification would naturally resist convection and frustrate the onset of a geodynamo, however, late giant impacts could mechanically mix the distinct accreted core layers creating large homogenous regions. Within these regions, a geodynamo may operate. From this model, we interpret the difference between the planetary magnetic fields of Earth and Venus as a difference in giant impact histories. Our planetary accretion model is a numerical N-body integration of the Grand Tack scenario [1]—the most successful terrestrial planet formation model to date [2,3]. Then, we take the accretion histories of Earth-like and Venus-like planets from this model and post-process the growth of each terrestrial planet according to a well-tested planetary differentiation model [4,5]. This model fits Earth's mantle by modifying the oxygen content of the pre-cursor planetesimals and embryos as well as the conditions of metal-silicate equilibration. Other non-volatile major, minor and trace elements included in the model are assumed to be in CI chondrite proportions. The results from this model across many simulated terrestrial planet growth histories are robust. If the kinetic energy delivered by larger impacts is neglected, the core of each planet grows with a strong stable stratification that would significantly impede convection. However, if giant impact mixing is very efficient or if the impact history delivers large impacts late, than the stable stratification can be removed. [1] Walsh et al. Nature 475 (2011) [2] O'Brien et al. Icarus 223 (2014) [3] Jacobson & Morbidelli PTRSA 372 (2014) [4] Rubie et al. EPSL 301

  20. The − 5 A/G single-nucleotide polymorphism in the core promoter region of MT2A and its effect on allele-specific gene expression and Cd, Zn and Cu levels in laryngeal cancer

    SciTech Connect

    Starska, Katarzyna; Krześlak, Anna; Forma, Ewa; Morawiec-Sztandera, Alina; Aleksandrowicz, Paweł; Lewy-Trenda, Iwona; and others

    2014-10-15

    Metallothioneins (MTs) are low molecular weight, cysteine-rich heavy metal-binding proteins which participate in the mechanisms of Zn homeostasis, and protect against toxic metals. MTs contain metal-thiolate cluster groups and suppress metal toxicity by binding to them. The aim of this study was to determine the − 5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene and to investigate its effect on allele-specific gene expression and Cd, Zn and Cu content in squamous cell laryngeal cancer (SCC) and non-cancerous laryngeal mucosa (NCM) as a control. The MT2A promoter region − 5 A/G SNP was determined by restriction fragment length polymorphism using 323 SCC and 116 NCM. MT2A gene analysis was performed by quantitative real-time PCR. The frequency of A allele carriage was 94.2% and 91.8% in SCC and NCM, respectively, while G allele carriage was detected in 5.8% and 8.2% of SCC and NCM samples, respectively. As a result, a significant association was identified between the − 5 A/G SNP in the MT2A gene with mRNA expression in both groups. Metal levels were analyzed by flame atomic absorption spectrometry. The significant differences were identified between A/A and both the A/G and G/G genotypes, with regard to the concentration of the contaminating metal. The Spearman rank correlation results showed that the MT2A expression and Cd, Zn, Cu levels were negatively correlated. Results obtained in this study suggest that − 5 A/G SNP in MT2A gene may have an effect on allele-specific gene expression and accumulation of metal levels in laryngeal cancer. - Highlights: • MT2A gene expression and metal content in laryngeal cancer tissues • Association between SNP (rs28366003) and expression of MT2A • Significant associations between the SNP and Cd, Zn and Cu levels • Negative correlation between MT2A gene expression and Cd, Zn and Cu levels.

  1. Novel point mutations and mutational complexes in the enhancer II, core promoter and precore regions of hepatitis B virus genotype D1 associated with hepatocellular carcinoma in Saudi Arabia.

    PubMed

    Khan, Anis; Al Balwi, Mohammed A; Tanaka, Yasuhito; Hajeer, Ali; Sanai, Faisal M; Al Abdulkarim, Ibrahim; Al Ayyar, Latifah; Badri, Motasim; Saudi, Dib; Tamimi, Waleed; Mizokami, Masashi; Al Knawy, Bandar

    2013-12-15

    In this study, a cohort of 182 patients [55 hepatocellular carcinoma (HCC) and 127 non-HCC] infected with hepatitis B virus (HBV) in Saudi Arabia was investigated to study the relationship between sequence variation in the enhancer II (EnhII), basal core promoter (BCP) and precore regions of HBV genotype D (HBV/D) and the risk of HCC. HBV genotypes were determined by sequencing analysis and/or enzyme-linked immunosorbent assay. Variations in the EnhII, BCP and precore regions were compared between 107 non-HCC and 45 HCC patients infected with HBV/D, followed by age-matched analysis of 40 cases versus equal number of controls. Age and male gender were significantly associated with HCC (p = 0.0001 and p = 0.03, respectively). Serological markers such as aspartate aminotransferase, albumin and anti-HBe were significantly associated with HCC (p = 0.0001 for all), whereas HBeAg positivity was associated with non-HCC (p = 0.0001). The most prevalent HBV genotype was HBV/D (94%), followed by HBV/E (4%), HBV/A (1.6%) and HBV/C (0.5%). For HBV/D1, genomic mutations associated with HCC were T1673/G1679, G1727, C1741, C1761, A1757/T1764/G1766, T1773, T1773/G1775 and C1909. Age- and gender-adjusted stepwise logistic regression analysis indicated that mutations G1727 [odds ratio (OR) = 18.3; 95% confidence interval (CI) = 2.8-118.4; p = 0.002], A1757/T1764/G1766 (OR = 4.7; 95% CI = 1.3-17.2; p = 0.01) and T1773 (OR = 14.06; 95% CI = 2.3-84.8; p = 0.004) are independent predictors of HCC development. These results implicate novel individual and combination patterns of mutations in the X/precore region of HBV/D1 as predictors of HCC. Risk stratification based on these mutation complexes would be useful in determining high-risk patients and improving diagnostic and treatment strategies for HBV/D1.

  2. Efficient chimeric plant promoters derived from plant infecting viral promoter sequences.

    PubMed

    Acharya, Sefali; Ranjan, Rajiv; Pattanaik, Sitakanta; Maiti, Indu B; Dey, Nrisingha

    2014-02-01

    In the present study, we developed a set of three chimeric/hybrid promoters namely FSgt-PFlt, PFlt-UAS-2X and MSgt-PFlt incorporating different important domains of Figwort Mosaic Virus sub-genomic transcript promoter (FSgt, -270 to -60), Mirabilis Mosaic Virus sub-genomic transcript promoter (MSgt, -306 to -125) and Peanut Chlorotic Streak Caulimovirus full-length transcript promoter (PFlt-, -353 to +24 and PFlt-UAS, -353 to -49). We demonstrated that these chimeric/hybrid promoters can drive the expression of reporter genes in different plant species including tobacco, Arabidopsis, petunia, tomato and spinach. FSgt-PFlt, PFlt-UAS-2X and MSgt-PFlt promoters showed 4.2, 1.5 and 1.2 times stronger GUS activities compared to the activity of the CaMV35S promoter, respectively, in tobacco protoplasts. Protoplast-derived recombinant promoter driven GFP showed enhanced accumulation compared to that obtained under the CaMV35S promoter. FSgt-PFlt, PFlt-UAS-2X and MSgt-PFlt promoters showed 3.0, 1.3 and 1.0 times stronger activities than the activity of the CaMV35S² (a modified version of the CaMV35S promoter with double enhancer domain) promoter, respectively, in tobacco (Nicotiana tabacum, var. Samsun NN). Alongside, we observed a fair correlation between recombinant promoter-driven GUS accumulation with the corresponding uidA-mRNA level in transgenic tobacco. Histochemical (X-gluc) staining of whole transgenic seedlings and fluorescence images of ImaGene Green™ treated floral parts expressing the GUS under the control of recombinant promoters also support above findings. Furthermore, we confirmed that these chimeric promoters are inducible in the presence of 150 μM salicylic acid (SA) and abscisic acid (ABA). Taken altogether, we propose that SA/ABA inducible chimeric/recombinant promoters could be used for strong expression of gene(s) of interest in crop plants.

  3. Sex Difference in κ-Opioid Receptor (KOPR)-Mediated Behaviors, Brain Region KOPR Level and KOPR-Mediated Guanosine 5′-O-(3-[35S]Thiotriphosphate) Binding in the Guinea Pig

    PubMed Central

    Wang, Yu-Jun; Rasakham, Khampaseuth; Huang, Peng; Chudnovskaya, Darina; Cowan, Alan

    2011-01-01

    We examined whether sex differences in κ-opioid receptor (KOPR) pharmacology exist in guinea pigs, which are more similar to humans in the expression level and distribution of KOPR in the brain than rats and mice. The KOPR agonist trans-(±)-3,4-dichloro-N-methyl-N-(2-[1-pyrrolidinyl]-cyclohexyl)benzeneacetamide methanesulfonate (U50,488H) produced a dose-dependent increase in abnormal postures and immobility with more effects in males than females. Males also showed more U50,488H-induced antinociception in the paw pressure test than females. Pretreatment with the KOPR antagonist norbinaltorphimine blocked U50,488H-induced abnormal body postures and antinociception. In contrast, inhibition of cocaine-induced hyperambulation by U50,488H was more effective in females than males. Thus, sex differences in the effects of U50,488H are endpoint-dependent. We then examined whether sex differences in KOPR levels and KOPR-mediated G protein activation in brain regions may contribute to the observed differences using quantitative in vitro autoradiography of [3H](5a,7a,8b)-(−)-N-methyl-N-(7-(1-pyrrolidinyl)1-oxaspiro(4,5)dec-8-yl)benzeacetamide ([3H]U69,593) binding to the KOPR and U50,488H-stimulated guanosine 5′-O-(3-[35S]thiotriphosphate ([35S]GTPγS) binding. Compared with females, males exhibited more [3H]U69,593 binding in the deep layers of somatosensory and insular cortices, claustrum, endopiriform nucleus, periaqueductal gray, and substantial nigra. Concomitantly, U50,488H-stimulated [35S]GTPγS binding was greater in males than females in the superficial and deep layers of somatosensory and insular cortices, caudate putamen, claustrum, medial geniculate nucleus, and cerebellum. In contrast, compared with males, females showed more U50,488H-stimulated [35S]GTPγS binding in the dentate gyrus and a trend of higher [35S]GTPγS binding in the hypothalamus. These data demonstrate that males and females differ in KOPR expression and KOPR-mediated G protein activation

  4. The Late Developmental Pattern of Mu Transposon Excision Is Conferred by a Cauliflower Mosaic Virus 35S –Driven MURA cDNA in Transgenic Maize

    PubMed Central

    Raizada, Manish N.; Walbot, Virginia

    2000-01-01

    The MuDR element responsible for Mutator activities in maize encodes two genes, mudrA and mudrB. Each encodes multiple transcripts hypothesized to regulate, directly or indirectly, the unique late timing and switch in transposition mechanism during maize development. mudrA, which encodes the MURA transposase, is unstable in bacterial plasmids, a technical problem solved by using phage M13 as a vector to prepare DNA for biolistic transformation. In transgenic maize, a single 2.7-kb mudrA cDNA predicted to encode an 823–amino acid protein is sufficient to catalyze late somatic excisions, despite removal of the native promoter, alternative transcription start sites, known introns, polymorphic 5′ and 3′ untranslated sequences, and the mudrB gene. These results suggest that post-translational regulation confers Mu excision timing. The transgene is active in lines containing silencing MuDR elements. This suggests that endogenous MuDR transposons do not measurably immunize the host against expression of a homologous transgene. PMID:10634904

  5. Characterization of [35S]-ATP alpha S and [3H]-alpha, beta-MeATP binding sites in rat brain cortical synaptosomes: regulation of ligand binding by divalent cations.

    PubMed

    Schäfer, R; Reiser, G

    1997-07-01

    1. We made a comparative analysis of the binding characteristics of the radioligands [35S]-ATP alpha S and [3H]-alpha, beta-MeATP in order to test whether these ligands can be used to analyse P2-purinoceptors in synaptosomal membranes from rat brain cortex. 2. Synaptosomes possess sites with high affinity for [35S]-ATP alpha S (Kd = 22.2 +/- 9.1 nM, Bmax = 14.8 pmol mg-1 protein). The rank order of the competition potency of the different compounds (ATP alpha S, ATP, ATP gamma S > ADP beta S, 2-MeSATP > deoxyATP, ADP > > UTP, alpha, beta-MeATP, AMP, Reactive Blue-2, suramin, isoPPADS) is consistent with pharmacological properties of P2Y-purinoceptors. 3. Under identical conditions [35S]-ATP alpha S and [3H]-alpha, beta-MeATP bind to different binding sites at synaptosomal membranes from rat brain cortex. The affinity of the [3H]-alpha, beta-MeATP binding sites (Kd = 13.7 +/- 1.8 nM, Bmax = 6.34 +/- 0.28 pmol mg-1 protein) was 38 fold higher than the potency of alpha, beta-MeATP to displace [35S]-ATP alpha S binding (Ki = 0.52 microM). ATP and ADP beta S competed at both binding sites with different affinities, 60 fold and 175 fold, respectively. The other agonists tested (2-MeSATP, UTP, GTP) did not affect specific [35H]-alpha, beta-MeATP binding at concentrations up to 100 microM. The antagonists (suramin, isoPPADS, Evan's Blue) showed completely different affinities for both binding sites. 4. Binding of [35S]-ATP alpha S on synaptosomes was regulated by GTP, which is indicative for G-protein coupled receptors. The Kd value for the high affinity binding site was reduced in the presence of GTP about 5 fold (from 1.8 nM to 8.6 nM). In the presence of Mg2+ the affinity was increased (Kd 1.8 nM versus 22 nM in the absence of Mg2+). 5. The binding of both radioligands was regulated in an opposite manner by physiological concentrations of Ca2+ and Mg2+. Binding of [3H]-alpha, beta-MeATP to synaptosomal membranes was increased 3 fold by raising the Ca2+ concentration

  6. Progressive Curation of IODP Core Material at Kochi Core Center, Japan

    NASA Astrophysics Data System (ADS)

    Gupta, L. P.; Hisamitsu, T.; Ahagon, N.; Kuramoto, T.; Tokuyama, H.; Kinoshita, M.

    2014-12-01

    Kochi Core Center (KCC) is one of the 3 IODP core repositories in the world, and is in-charge of curating core materials collected/to be collected from most of the Indian Ocean, west Pacific Ocean and Bering Sea. Curation of IODP core material in the KCC began in 2007 as it started receiving 83 km of Legacy cores from the other IODP core repositories. Since then the KCC has not only maintained curatorial standards of the IODP, but also added many services for convenience of the IODP researchers that include curation of cuttings and deep frozen aliquots of cores, open access to logging equipment in the KCC for core measurements, virtual core library to provide quick online access to 3-D XCT images of the cores collected by the D/V Chikyu, online summary of the cores being curated in the KCC, and up-to-date online images of working half to show status of samples available for prospective users. With its existing stock of 104 km of the IODP & Legacy cores and cores to be recovered from the Indian Ocean in near future by the JOIDES Resolution, and a huge new reefer building with storage capacity of ca. 150 km core becoming part of the KCC this August, the KCC is bound to play a significant role in promoting earth and biogeo-sciences throughout the world, especially in neighboring Asian countries.

  7. Using health promotion competencies for curriculum development in higher education.

    PubMed

    Madsen, Wendy; Bell, Tanya

    2012-03-01

    Health promotion core competencies are used for a variety of reasons. Recently there have been moves to gain international consensus regarding core competencies within health promotion. One of the main reasons put forward for having core competencies is to guide curriculum development within higher education institutions. This article outlines the endeavours of one institution to develop undergraduate and postgraduate curricula around the Australian core competencies for health promotion practitioners. It argues that until core competencies have been agreed upon internationally, basing curricula on these carries a risk associated with change. However, delaying curricula until such risks are ameliorated decreases opportunities to deliver dynamic and current health promotion education within higher institutions.

  8. Study of P-even and P-odd angular correlations in /sup 35/Cl(n,p)/sup 35/S and /sup 14/N(n,p)/sup 14/C reactions

    SciTech Connect

    Antonov, A.; Vesna, V.A.; Gledenov, Y.M.; Zvarova, T.S.; Lobashev, V.M.; Okunev, I.S.; Popov, Y.P.; Rigol', K.; Smotritskii, L.M.; Shul'gina, E.V.; and others

    1988-08-01

    P-odd and left-right asymmetries have been observed in the /sup 35/Cl(n,p)/sup 35/S reaction with capture of polarized thermal neutrons. The correlation coefficients are ..cap alpha../sub n//sub p/ = -(1.51 +- 0.34)x10/sup -4/ and ..cap alpha../sup l//sup r//sub n//sub p/ = -(2.40 +- 0.43)x10/sup -4/, respectively. For the /sup 14/N(n,p)/sup 14/C reaction, and upper bound of ..cap alpha../sub n//sub p/ = (0.07 +- 0.12)x10/sup -4/ is obtained for the P-odd asymmetry, and a left-right asymmetry is found, with correlation coefficient ..cap alpha../sup l//sup r//sub n//sub p/ = (0.66 +- 0.18)x10/sup -4/. The estimated value of the weak-interaction matrix element for the /sup 35/Cl(n,p)/sup 35/S reaction is W/sub S//sub P/ = 0.06 +- 0.02 eV.

  9. Mutational analysis of Escherichia coli σ28 and its target promoters reveal recognition of a composite −10 region, comprised of an “extended −10 motif” and a core-10 element

    PubMed Central

    Koo, Byoung-Mo; Rhodius, Virgil A.; Campbell, Elizabeth A.; Gross, Carol A.

    2009-01-01

    Summary σ28 controls the expression of flagella related genes and is the most widely distributed alternative σ factor, present in motile gram-positive and gram-negative bacteria. The distinguishing feature of σ28 promoters is a long −10 region (GCCGATAA). Despite the fact that the upstream GC is highly conserved, previous studies have not indicated a functional role for this motif. Here we examine the functional relevance of the GCCG motif and determine which residues in σ28 participate in its recognition. We find that the GCCG motif is a functionally important composite element. The upstream GC constitutes an extended −10 motif and is recognized by R91, a residue in Domain 3 of σ28. The downstream CG is the upstream edge of −10 region of the promoter; two residues in Region 2.4, D81 and R84, participate in its recognition. Consistent with their role in base-specific recognition of the promoter, R91, D81 and D84 are universally conserved in σ28 orthologues. σ28 is the second Group 3 σ shown to use an extended −10 region in promoter recognition, raising the possibility that other Group 3 σs will do so as well. PMID:19400790

  10. Core-core and core-valence correlation

    NASA Technical Reports Server (NTRS)

    Bauschlicher, Charles W., Jr.; Langhoff, Stephen R.; Taylor, Peter R.

    1988-01-01

    The effect of (1s) core correlation on properties and energy separations was analyzed using full configuration-interaction (FCI) calculations. The Be 1 S - 1 P, the C 3 P - 5 S and CH+ 1 Sigma + or - 1 Pi separations, and CH+ spectroscopic constants, dipole moment and 1 Sigma + - 1 Pi transition dipole moment were studied. The results of the FCI calculations are compared to those obtained using approximate methods. In addition, the generation of atomic natural orbital (ANO) basis sets, as a method for contracting a primitive basis set for both valence and core correlation, is discussed. When both core-core and core-valence correlation are included in the calculation, no suitable truncated CI approach consistently reproduces the FCI, and contraction of the basis set is very difficult. If the (nearly constant) core-core correlation is eliminated, and only the core-valence correlation is included, CASSCF/MRCI approached reproduce the FCI results and basis set contraction is significantly easier.

  11. Functional analysis of a reproductive organ predominant expressing promoter in cotton plants.

    PubMed

    Ren, Maozhi; Chen, Quanjia; Li, Li; Zhang, Rui; Guo, Sandui

    2005-10-01

    Transgenic Bt insect-resistant cotton plants have high insect resistance in the early stage of development, but relatively low resistance in the late stage. Substituting a reproductive organ-specific promoter for the CaMV35S promoter presently being used could be an ideal solution. For the first time, the promoter sequence of ADP-ribosylation factor 1 (arf1) gene was isolated from Gossypium hirsutumY18 by means of inverse PCR. The sequencing result discovered the unique structure of the arf1 promoter, including four promoter-specific elements, the initiator, TATA box, CAAT box and GC box, and also an intron in 5'-untranslation region. Four plant expression vectors were constructed for functional analysis of the promoter. Based on the pBl121 plant expression vector, four truncated arf1 promoters took the place of the CaMV35S promoter. These vectors were different only in their promoter regions. They were introduced into cotton plants via pollen tube pathway. Histochemical GUS staining and fluorescence quantitative analyses were performed to examine the expression patterns of the GUS gene driven by the 4 arf1 truncated promoters in transgenic cotton plants respectively. The results showed that the arf1 promoter was a typical reproductive organ-specific promoter. Hopefully, the arf1 promoter can be a regulatory element for designing cotton reproductive organs with desired characteristics.

  12. Promotion of embryonic chick limb cartilage differentiation by transforming growth factor-beta.

    PubMed

    Kulyk, W M; Rodgers, B J; Greer, K; Kosher, R A

    1989-10-01

    This study represents a first step in investigating the possible involvement of transforming growth factor-beta (TGF-beta) in the regulation of embryonic chick limb cartilage differentiation. TGF-beta 1 and 2 (1-10 ng/ml) elicit a striking increase in the accumulation of Alcian blue, pH 1-positive cartilage matrix, and a corresponding twofold to threefold increase in the accumulation of 35S-sulfate- or 3H-glucosamine-labeled sulfated glycosaminoglycans (GAG) by high density micromass cultures prepared from the cells of whole stage 23/24 limb buds or the homogeneous population of chondrogenic precursor cells comprising the distal subridge mesenchyme of stage 25 wing buds. Moreover, TGF-beta causes a striking (threefold to sixfold) increase in the steady-state cytoplasmic levels of mRNAs for cartilage-characteristic type II collagen and the core protein of cartilage-specific proteoglycan. Only a brief (2 hr) exposure to TGF-beta at the initiation of culture is sufficient to stimulate chondrogenesis, indicating that the growth factor is acting at an early step in the process. Furthermore, TGF-beta promotes the formation of cartilage matrix and cartilage-specific gene expression in low density subconfluent spot cultures of limb mesenchymal cells, which are situations in which little, or no chondrogenic differentiation normally occurs. These results provide strong incentive for considering and further investigating the role of TGF-beta in the control of limb cartilage differentiation.

  13. Proximity of Radiation Desiccation Response Motif to the core promoter is essential for basal repression as well as gamma radiation-induced gyrB gene expression in Deinococcus radiodurans.

    PubMed

    Anaganti, Narasimha; Basu, Bhakti; Mukhopadhyaya, Rita; Apte, Shree Kumar

    2017-03-02

    The radioresistant D. radiodurans regulates its DNA damage regulon (DDR) through interaction between a 17bp palindromic cis-regulatory element called the Radiation Desiccation Response Motif (RDRM), the DdrO repressor and a protease IrrE. The role of RDRM in regulation of DDR was dissected by constructing RDRM sequence-, position- or deletion-variants of Deinococcal gyrB gene (DR0906) promoter and by RDRM insertion in the non-RDRM groESL gene (DR0606) promoter, and monitoring the effect of such modifications on the basal as well as gamma radiation inducible promoter activity by quantifying fluorescence of a GFP reporter. RDRM sequence-variants revealed that the conservation of sequence at the 5th and 13th position and the ends of RDRM is essential for basal repression by interaction with DdrO. RDRM position-variants showed that the sequence acts as a negative regulatory element only when located around transcription start site (TSS) and within the span of RNA polymerase (RNAP) binding region. RDRM deletion-variants indicated that the 5' sequence of RDRM possibly possesses an enhancer-like element responsible for higher expression yields upon repressor clearance post-irradiation. The results suggest that RDRM plays both a negative as well as a positive role of in the regulation of DDR in D. radiodurans.

  14. Promoter/leader deletion analysis and plant expression vectors with the figwort mosaic virus (FMV) full length transcript (FLt) promoter containing single or double enhancer domains.

    PubMed

    Maiti, I B; Gowda, S; Kiernan, J; Ghosh, S K; Shepherd, R J

    1997-03-01

    The boundaries required for maximal expression from the promoter/leader region of the full length transcript of figwort mosaic virus (FLt promoter) coupled to reporter genes were defined by 5' and 3' deletion analyses. In transient expression assays using protoplasts of Nicotiana edwardsonii, a 314 bp FLt promoter fragment sequence (-249 to +65 from the transcription start site) was sufficient for strong expression activity. Plant expression vectors developed with modified FLt promoters were tested with GUS or CAT as reporter genes in transgenic plants. The FLt promoter is a strong constitutive promoter, with strength comparable to or greater than that of the CaMV 35S promoter. The FLt promoter with its double enhancer domain linked to GUS or CAT reporter genes provides an average 4-fold greater activity than the FLt promoter with a single enhancer domain (-55 to -249 bp upstream fragment) in tests with transgenic plants and in protoplast transient expression assays.

  15. Identifying ELIXIR Core Data Resources

    PubMed Central

    Durinx, Christine; McEntyre, Jo; Appel, Ron; Apweiler, Rolf; Barlow, Mary; Blomberg, Niklas; Cook, Chuck; Gasteiger, Elisabeth; Kim, Jee-Hyub; Lopez, Rodrigo; Redaschi, Nicole; Stockinger, Heinz; Teixeira, Daniel; Valencia, Alfonso

    2017-01-01

    The core mission of ELIXIR is to build a stable and sustainable infrastructure for biological information across Europe. At the heart of this are the data resources, tools and services that ELIXIR offers to the life-sciences community, providing stable and sustainable access to biological data. ELIXIR aims to ensure that these resources are available long-term and that the life-cycles of these resources are managed such that they support the scientific needs of the life-sciences, including biological research. ELIXIR Core Data Resources are defined as a set of European data resources that are of fundamental importance to the wider life-science community and the long-term preservation of biological data. They are complete collections of generic value to life-science, are considered an authority in their field with respect to one or more characteristics, and show high levels of scientific quality and service. Thus, ELIXIR Core Data Resources are of wide applicability and usage. This paper describes the structures, governance and processes that support the identification and evaluation of ELIXIR Core Data Resources. It identifies key indicators which reflect the essence of the definition of an ELIXIR Core Data Resource and support the promotion of excellence in resource development and operation. It describes the specific indicators in more detail and explains their application within ELIXIR’s sustainability strategy and science policy actions, and in capacity building, life-cycle management and technical actions. The identification process is currently being implemented and tested for the first time. The findings and outcome will be evaluated by the ELIXIR Scientific Advisory Board in March 2017. Establishing the portfolio of ELIXIR Core Data Resources and ELIXIR Services is a key priority for ELIXIR and publicly marks the transition towards a cohesive infrastructure. PMID:27803796

  16. Identifying ELIXIR Core Data Resources.

    PubMed

    Durinx, Christine; McEntyre, Jo; Appel, Ron; Apweiler, Rolf; Barlow, Mary; Blomberg, Niklas; Cook, Chuck; Gasteiger, Elisabeth; Kim, Jee-Hyub; Lopez, Rodrigo; Redaschi, Nicole; Stockinger, Heinz; Teixeira, Daniel; Valencia, Alfonso

    2016-01-01

    The core mission of ELIXIR is to build a stable and sustainable infrastructure for biological information across Europe. At the heart of this are the data resources, tools and services that ELIXIR offers to the life-sciences community, providing stable and sustainable access to biological data. ELIXIR aims to ensure that these resources are available long-term and that the life-cycles of these resources are managed such that they support the scientific needs of the life-sciences, including biological research. ELIXIR Core Data Resources are defined as a set of European data resources that are of fundamental importance to the wider life-science community and the long-term preservation of biological data. They are complete collections of generic value to life-science, are considered an authority in their field with respect to one or more characteristics, and show high levels of scientific quality and service. Thus, ELIXIR Core Data Resources are of wide applicability and usage. This paper describes the structures, governance and processes that support the identification and evaluation of ELIXIR Core Data Resources. It identifies key indicators which reflect the essence of the definition of an ELIXIR Core Data Resource and support the promotion of excellence in resource development and operation. It describes the specific indicators in more detail and explains their application within ELIXIR's sustainability strategy and science policy actions, and in capacity building, life-cycle management and technical actions. The identification process is currently being implemented and tested for the first time. The findings and outcome will be evaluated by the ELIXIR Scientific Advisory Board in March 2017. Establishing the portfolio of ELIXIR Core Data Resources and ELIXIR Services is a key priority for ELIXIR and publicly marks the transition towards a cohesive infrastructure.

  17. Protected deoxyribonucleoside-3' aryl phosphodiesters as key intermediates in polynucleotide synthesis. Construction of an icosanucleotide analogous to the sequence at the ends of Rous sarcoma virus 35S RNA.

    PubMed Central

    Gough, G R; Singleton, C K; Weith, H L; Gilham, P T

    1979-01-01

    Several modifications have been incorporated into the phosphotriester strategy for chemical synthesis of oligodeoxyribonucleotides. These include high-yield methods of preparation and isolation of O5', N-protected deoxyribonucleoside-3' p-chlorophenyl phosphates which serve as key intermediates, and the elimination of some superfluous manipulation and purification steps commonly used in the process of synthesizing oligonucleotide blocks. In addition, two new arylsulfonyl nitroimidazole derivatives have been prepared and found to be highly effective agents for internucleotide bond formation. These techniques have been applied in construction of the iconsamer d(G-C-C-A-T-T-T-T-A-C-C-A-T-T-C-A-C-C-A)-rC, equivalent to a ribonucleotide sequence located at both the 5' and 3' ends of Rous sarcoma virus 35S RNA. Images PMID:221888

  18. Electroactive crown ester-Cu(2+) complex with in-situ modification at molecular beacon probe serving as a facile electrochemical DNA biosensor for the detection of CaMV 35s.

    PubMed

    Zhan, Fengping; Liao, Xiaolei; Gao, Feng; Qiu, Weiwei; Wang, Qingxiang

    2017-06-15

    A novel electrochemical DNA biosensor has been facilely constructed by in-situ assembly of electroactive 4'-aminobenzo-18-crown-6-copper(II) complex (AbC-Cu(2+)) on the free terminal of the hairpin-structured molecule beacon. The 3'-SH modified molecule beacon probe was first immobilized on the gold electrode (AuE) surface through self-assembly chemistry of Au-S bond. Then the crow ester of AbC was covalently coupled with 5'-COOH on the molecule beacon, and served as a platform to attach the Cu(2+) by coordination with ether bond (-O-) of the crown cycle. Thus, an electroactive molecule beacon-based biosensing interface was constructed. In comparison with conventional methods for preparation of electroactive molecule beacon, the approach presented in this work is much simpler, reagent- and labor-saving. Selectivity study shows that the in-situ fabricated electroactive molecule beacon remains excellent recognition ability of pristine molecule beacon probe to well differentiate various DNA fragments. The target DNA can be quantatively determined over the range from 0.10pM to 0.50nM. The detection limit of 0.060pM was estimated based on signal-to-noise ratio of 3. When the biosensor was applied for the detection cauliflower mosaic virus 35s (CaMV 35s) in soybean extraction samples, satisfactory results are achieved. This work opens a new strategy for facilely fabricating electrochemical sensing interface, which also shows great potential in aptasensor and immurosensor fabrication.

  19. Academic Rigor: The Core of the Core

    ERIC Educational Resources Information Center

    Brunner, Judy

    2013-01-01

    Some educators see the Common Core State Standards as reason for stress, most recognize the positive possibilities associated with them and are willing to make the professional commitment to implementing them so that academic rigor for all students will increase. But business leaders, parents, and the authors of the Common Core are not the only…

  20. Plant defense gene promoter enhances the reliability of shiva-1 gene-induced resistance to soft rot disease in potato.

    PubMed

    Yi, Jung Yoon; Seo, Hyo Won; Yang, Moon Sik; Robb, E Jane; Nazar, Ross N; Lee, Shin Woo

    2004-11-01

    PAL5, a tomato (Lycopersicon esculentum Mill.) plant defense gene that encodes phenylalanine ammonia-lyase, is known to respond to a variety of environmental stresses including pathogen infection and wounding. A shiva-1 gene recombinant that encodes a small synthetic antibacterial peptide under the PAL5 gene promoter was transformed into potato (Solanum tuberosum L.) and its ability to induce resistance to Erwinia carotovora was compared with a construct under the control of the constitutive and widely used cauliflower mosaic virus (CaMV) 35S promoter. The shiva-1 peptide, an analog of natural cecropin B, was shown previously to have high bactericidal activity in vitro, but when expressed in vivo under the control of the CaMV 35S promoter, the effects were very inconsistent. As observed previously, in the present studies a few transformants with the CaMV 35S promoter were highly resistant when assayed for susceptibility to soft rot disease. In marked contrast the majority of transformants with the PAL5 gene promoter were highly resistant. More-detailed analyses of the incorporated DNA indicated that most of the transformants with the CaMV 35S promoter contained multiple copies of the transforming DNA while all of the PAL5 recombinants contained single copies. The highly resistant CaMV 35S recombinant also was present as a single copy. The results indicate that, at least in this instance, a constitutive promoter may not be ideal for the effective expression of a foreign gene and suggest that multiple insertions may have negative consequences.

  1. Health Promotion

    DTIC Science & Technology

    1986-03-11

    Department of Defense DIRECTIVEAD-A269 638 , , AD-A29 638March 11, 1986 IIIIii!IN 111111111,11 Ii1111,111111[NUMBER 1010.10 SUBJECT: Health Promotion ...34 March 13, 1985 INC A. URPOSE SThis Directive establishes a health promotion policy within the Department of Defense to improve and maintain military...civilian employees. C. DEFINITIONS 1. Health Promotion . Any combination of health education and related organizational, social, economic or health care

  2. Analysis of cis-sequence of subgenomic transcript promoter from the Figwort mosaic virus and comparison of promoter activity with the cauliflower mosaic virus promoters in monocot and dicot cells.

    PubMed

    Bhattacharyya, Somnath; Dey, Nrisingha; Maiti, Indu B

    2002-12-01

    A sub-genomic transcript (Sgt) promoter was isolated from the Figwort mosaic virus (FMV) genomic clone. The FMV Sgt promoter was linked to heterologous coding sequences to form a chimeric gene construct. The 5'-3'-boundaries required for maximal activity and involvement of cis-sequences for optimal expression in plants were defined by 5'-, 3'-end deletion and internal deletion analysis of FMV Sgt promoter fragments coupled with a beta-glucuronidase reporter gene in both transient protoplast expression experiments and in transgenic plants. A 301 bp FMV Sgt promoter fragment (sequence -270 to +31 from the transcription start site; TSS) provided maximum promoter activity. The TSS of the FMV Sgt promoter was determined by primer extension analysis using total RNA from transgenic plants developed for FMV Sgt promoter: uidA fusion gene. An activator domain located upstream of the TATA box at -70 to -100 from TSS is absolutely required for promoter activity and its function is critically position-dependent with respect to TATA box. Two sequence motifs AGATTTTAAT (coordinates -100 to -91) and GTAAGCGC (coordinates -80 to -73) were found to be essential for promoter activity. The FMV Sgt promoter is less active in monocot cells; FMV Sgt promoter expression level was about 27.5-fold higher in tobacco cells compared to that in maize cells. Comparative expression analysis of FMV Sgt promoter with cauliflower mosaic virus (CaMV) 35S promoter showed that the FMV Sgt promoter is about 2-fold stronger than the CaMV 35S promoter. The FMV Sgt promoter is a constitutive promoter; expression level in seedlings was in the order: root>leaf>stem.

  3. Evaluating Community Health Advisor (CHA) Core Competencies: The CHA Core Competency Retrospective Pretest/Posttest (CCCRP).

    PubMed

    Story, Lachel; To, Yen M

    2016-05-01

    Health care and academic systems are increasingly collaborating with community health advisors (CHAs) to provide culturally relevant health interventions that promote sustained community transformation. Little attention has been placed on CHA training evaluation, including core competency attainment. This study identified common CHA core competencies, generated a theoretically based measure of those competencies, and explored psychometric properties of that measure. A concept synthesis revealed five CHA core competencies (leadership, translation, guidance, advocacy, and caring). The CHA Core Competency Retrospective Pretest/Posttest (CCCRP) resulted from that synthesis, which was administered using multiple approaches to individuals who previously received CHA training (N= 142). Exploratory factor analyses revealed a two-factor structure underlying the posttraining data, and Cronbach's alpha indicated high internal consistency. This study suggested some CHA core competencies might be more interrelated than previously thought, and two major competencies exist rather than five and supported the CCCRP's use to evaluate core competency attainment resulting from training.

  4. The fundamental ribosomal RNA transcription initiation factor-IB (TIF-IB, SL1, factor D) binds to the rRNA core promoter primarily by minor groove contacts.

    PubMed

    Geiss, G K; Radebaugh, C A; Paule, M R

    1997-11-14

    Acanthamoeba castellanii transcription initiation factor-IB (TIF-IB) is the TATA-binding protein-containing transcription factor that binds the rRNA promoter to form the committed complex. Minor groove-specific drugs inhibit TIF-IB binding, with higher concentrations needed to disrupt preformed complexes because of drug exclusion by bound TIF-IB. TIF-IB/DNA interactions were mapped by hydroxyl radical and uranyl nitrate footprinting. TIF-IB contacts four minor grooves in its binding site. TIF-IB and DNA wrap around each other in a right-handed superhelix of high pitch, so the upstream and downstream contacts are on opposite faces of the helix. Dimethyl sulfate protection assays revealed limited contact with a few guanines in the major groove. This detailed analysis suggests significant DNA conformation dependence of the interaction.

  5. Coring Sample Acquisition Tool

    NASA Technical Reports Server (NTRS)

    Haddad, Nicolas E.; Murray, Saben D.; Walkemeyer, Phillip E.; Badescu, Mircea; Sherrit, Stewart; Bao, Xiaoqi; Kriechbaum, Kristopher L.; Richardson, Megan; Klein, Kerry J.

    2012-01-01

    A sample acquisition tool (SAT) has been developed that can be used autonomously to sample drill and capture rock cores. The tool is designed to accommodate core transfer using a sample tube to the IMSAH (integrated Mars sample acquisition and handling) SHEC (sample handling, encapsulation, and containerization) without ever touching the pristine core sample in the transfer process.

  6. Core labeling of adenovirus with EGFP

    SciTech Connect

    Le, Long P.; Le, Helen N.; Nelson, Amy R.; Matthews, David A.; Yamamoto, Masato; Curiel, David T. . E-mail: curiel@uab.edu

    2006-08-01

    The study of adenovirus could greatly benefit from diverse methods of virus detection. Recently, it has been demonstrated that carboxy-terminal EGFP fusions of adenovirus core proteins Mu, V, and VII properly localize to the nucleus and display novel function in the cell. Based on these observations, we hypothesized that the core proteins may serve as targets for labeling the adenovirus core with fluorescent proteins. To this end, we constructed various chimeric expression vectors with fusion core genes (Mu-EGFP, V-EGFP, preVII-EGFP, and matVII-EGFP) while maintaining expression of the native proteins. Expression of the fusion core proteins was suboptimal using E1 expression vectors with both conventional CMV and modified (with adenovirus tripartite leader sequence) CMV5 promoters, resulting in non-labeled viral particles. However, robust expression equivalent to the native protein was observed when the fusion genes were placed in the deleted E3 region. The efficient Ad-wt-E3-V-EGFP and Ad-wt-E3-preVII-EGFP expression vectors were labeled allowing visualization of purified virus and tracking of the viral core during early infection. The vectors maintained their viral function, including viral DNA replication, viral DNA encapsidation, cytopathic effect, and thermostability. Core labeling offers a means to track the adenovirus core in vector targeting studies as well as basic adenovirus virology.

  7. Why the Common Core Changes Math Instruction

    ERIC Educational Resources Information Center

    Faulkner, Valerie N.

    2013-01-01

    The Common Core math standards promote several important differences in how math concepts are taught and should be talked about. These changes will make it easier for younger students to comprehend and adapt to more complex concepts in the later grades. This guide should help elementary teachers make changes and adaptations that are in line the…

  8. What Should Common Core Assessments Measure?

    ERIC Educational Resources Information Center

    Chandler, Kayla; Fortune, Nicholas; Lovett, Jennifer N.; Scherrer, Jimmy

    2016-01-01

    The Common Core State Standards for mathematics promote ideals about learning mathematics by providing specific standards focused on conceptual understanding and incorporating practices in which students must participate to develop conceptual understanding. Thus, how we define learning is pivotal because our current definition isn't aligned with…

  9. Ubiquitin promoter-terminator cassette promotes genetically stable expression of the taste-modifying protein miraculin in transgenic lettuce.

    PubMed

    Hirai, Tadayoshi; Shohael, Abdullah Mohammad; Kim, You-Wang; Yano, Megumu; Ezura, Hiroshi

    2011-12-01

    Lettuce is a commercially important leafy vegetable that is cultivated worldwide, and it is also a target crop for plant factories. In this study, lettuce was selected as an alternative platform for recombinant miraculin production because of its fast growth, agronomic value, and wide availability. The taste-modifying protein miraculin is a glycoprotein extracted from the red berries of the West African native shrub Richadella dulcifica. Because of its limited natural availability, many attempts have been made to produce this protein in suitable alternative hosts. We produced transgenic lettuce with miraculin gene driven either by the ubiquitin promoter/terminator cassette from lettuce or a 35S promoter/nos terminator cassette. Miraculin gene expression and miraculin accumulation in both cassettes were compared by quantitative real-time PCR analysis, Western blotting, and enzyme-linked immunosorbent assay. The expression level of the miraculin gene and protein in transgenic lettuce was higher and more genetically stable in the ubiquitin promoter/terminator cassette than in the 35S promoter/nos terminator cassette. These results demonstrated that the ubiquitin promoter/terminator cassette is an efficient platform for the genetically stable expression of the miraculin protein in lettuce and hence this platform is of benefit for recombinant miraculin production on a commercial scale.

  10. Banded transformer cores

    NASA Technical Reports Server (NTRS)

    Mclyman, C. W. T. (Inventor)

    1974-01-01

    A banded transformer core formed by positioning a pair of mated, similar core halves on a supporting pedestal. The core halves are encircled with a strap, selectively applying tension whereby a compressive force is applied to the core edge for reducing the innate air gap. A dc magnetic field is employed in supporting the core halves during initial phases of the banding operation, while an ac magnetic field subsequently is employed for detecting dimension changes occurring in the air gaps as tension is applied to the strap.

  11. Substrate and Lewis Acid Coordination Promote O-O Bond Cleavage of an Unreactive L2Cu(II)2(O2(2-)) Species to Form L2Cu(III)2(O)2 Cores with Enhanced Oxidative Reactivity.

    PubMed

    Garcia-Bosch, Isaac; Cowley, Ryan E; Díaz, Daniel E; Peterson, Ryan L; Solomon, Edward I; Karlin, Kenneth D

    2017-03-01

    Copper-dependent metalloenzymes are widespread throughout metabolic pathways, coupling the reduction of O2 with the oxidation of organic substrates. Small-molecule synthetic analogs are useful platforms to generate L/Cu/O2 species that reproduce the structural, spectroscopic, and reactive properties of some copper-/O2-dependent enzymes. Landmark studies have shown that the conversion between dicopper(II)-peroxo species (L2Cu(II)2(O2(2-)) either side-on peroxo, (S)P, or end-on trans-peroxo, (T)P) and dicopper(III)-bis(μ-oxo) (L2Cu(III)2(O(2-))2: O) can be controlled through ligand design, reaction conditions (temperature, solvent, and counteranion), or substrate coordination. We recently published ( J. Am. Chem. Soc. 2012 , 134 , 8513 , DOI: 10.1021/ja300674m ) the crystal structure of an unusual (S)P species [(MeAN)2Cu(II)2(O2(2-))](2+) ((S)P(MeAN), MeAN: N-methyl-N,N-bis[3-(dimethylamino)propyl]amine) that featured an elongated O-O bond but did not lead to O-O cleavage or reactivity toward external substrates. Herein, we report that (S)P(MeAN) can be activated to generate O(MeAN) and perform the oxidation of external substrates by two complementary strategies: (i) coordination of substituted sodium phenolates to form the substrate-bound O(MeAN)-RPhO(-) species that leads to ortho-hydroxylation in a tyrosinase-like fashion and (ii) addition of stoichiometric amounts (1 or 2 equiv) of Lewis acids (LA's) to form an unprecedented series of O-type species (O(MeAN)-LA) able to oxidize C-H and O-H bonds. Spectroscopic, computational, and mechanistic studies emphasize the unique plasticity of the (S)P(MeAN) core, which combines the assembly of exogenous reagents in the primary (phenolates) and secondary (Lewis acids association to the MeAN ligand) coordination spheres with O-O cleavage. These findings are reminiscent of the strategy followed by several metalloproteins and highlight the possible implication of O-type species in copper-/dioxygen-dependent enzymes such as

  12. Health Promotion

    PubMed Central

    Wilson, Ron

    1992-01-01

    How physicians address issues of disease prevention and health promotion is discussed and current standards of screening for disease and counseling practices are reviewed. Collaboration among all health professionals is necessary if preventive medicine is to be effective. PMID:21221259

  13. HYDRATE CORE DRILLING TESTS

    SciTech Connect

    John H. Cohen; Thomas E. Williams; Ali G. Kadaster; Bill V. Liddell

    2002-11-01

    The ''Methane Hydrate Production from Alaskan Permafrost'' project is a three-year endeavor being conducted by Maurer Technology Inc. (MTI), Noble, and Anadarko Petroleum, in partnership with the U.S. DOE National Energy Technology Laboratory (NETL). The project's goal is to build on previous and ongoing R&D in the area of onshore hydrate deposition. The project team plans to design and implement a program to safely and economically drill, core and produce gas from arctic hydrates. The current work scope includes drilling and coring one well on Anadarko leases in FY 2003 during the winter drilling season. A specially built on-site core analysis laboratory will be used to determine some of the physical characteristics of the hydrates and surrounding rock. Prior to going to the field, the project team designed and conducted a controlled series of coring tests for simulating coring of hydrate formations. A variety of equipment and procedures were tested and modified to develop a practical solution for this special application. This Topical Report summarizes these coring tests. A special facility was designed and installed at MTI's Drilling Research Center (DRC) in Houston and used to conduct coring tests. Equipment and procedures were tested by cutting cores from frozen mixtures of sand and water supported by casing and designed to simulate hydrate formations. Tests were conducted with chilled drilling fluids. Tests showed that frozen core can be washed out and reduced in size by the action of the drilling fluid. Washing of the core by the drilling fluid caused a reduction in core diameter, making core recovery very difficult (if not impossible). One successful solution was to drill the last 6 inches of core dry (without fluid circulation). These tests demonstrated that it will be difficult to capture core when drilling in permafrost or hydrates without implementing certain safeguards. Among the coring tests was a simulated hydrate formation comprised of coarse, large

  14. 23. CORE WORKER OPERATING A COREBLOWER THAT PNEUMATICALLY FILLED CORE ...

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

    23. CORE WORKER OPERATING A CORE-BLOWER THAT PNEUMATICALLY FILLED CORE BOXES WITH RESIGN IMPREGNATED SAND AND CREATED A CORE THAT THEN REQUIRED BAKING, CA. 1950. - Stockham Pipe & Fittings Company, 4000 Tenth Avenue North, Birmingham, Jefferson County, AL

  15. Activation of the pathogen-inducible Gst1 promoter of potato after elicitation by Venturia inaequalis and Erwinia amylovora in transgenic apple (Malus x domestica).

    PubMed

    Malnoy, M; Reynoird, J P; Borejsza-Wysocka, E E; Aldwinckle, H S

    2006-02-01

    Rather than using a constitutive promoter to drive transgenes for resistance against fungal and bacterial diseases in genetic engineering of apple (Malus x domestica) cultivars, a promoter induced only after infection was preferred. The ability of the Pgst1 promoter from potato (Solanum tuberosum L.) to drive expression of the gusA reporter gene was determined in two genotypes of apple: the fruit cultivar Royal Gala and the M.26 rootstock. beta-Glucuronidase activity in the transgenic lines grown in a growth chamber was determined quantitatively using fluorometric assays and compared to the activity in Cauliflower Mosaic Virus (CaMV) 35S promoter-driven transgenic lines. In both apple genotypes, the Pgst1 promoter exhibited a low level of expression after bacterial and fungal inoculation compared to the level obtained with the PCaMV35S promoter (15% and 8% respectively). The Pgst1 promoter was systematically activated in apple at the site of infection with a fungal pathogen. It was also activated after treatment with salicylic acid, but not after wounding. Taken together, these data show that, although the Pgst1 promoter is less active than the PCaMV35S promoter in apple, its pathogen responsiveness could be useful in driving the expression of transgenes to promote bacterial and fungal disease resistance.

  16. Core-Cutoff Tool

    NASA Technical Reports Server (NTRS)

    Gheen, Darrell

    2007-01-01

    A tool makes a cut perpendicular to the cylindrical axis of a core hole at a predetermined depth to free the core at that depth. The tool does not damage the surrounding material from which the core was cut, and it operates within the core-hole kerf. Coring usually begins with use of a hole saw or a hollow cylindrical abrasive cutting tool to make an annular hole that leaves the core (sometimes called the plug ) in place. In this approach to coring as practiced heretofore, the core is removed forcibly in a manner chosen to shear the core, preferably at or near the greatest depth of the core hole. Unfortunately, such forcible removal often damages both the core and the surrounding material (see Figure 1). In an alternative prior approach, especially applicable to toxic or fragile material, a core is formed and freed by means of milling operations that generate much material waste. In contrast, the present tool eliminates the damage associated with the hole-saw approach and reduces the extent of milling operations (and, hence, reduces the waste) associated with the milling approach. The present tool (see Figure 2) includes an inner sleeve and an outer sleeve and resembles the hollow cylindrical tool used to cut the core hole. The sleeves are thin enough that this tool fits within the kerf of the core hole. The inner sleeve is attached to a shaft that, in turn, can be attached to a drill motor or handle for turning the tool. This tool also includes a cutting wire attached to the distal ends of both sleeves. The cutting wire is long enough that with sufficient relative rotation of the inner and outer sleeves, the wire can cut all the way to the center of the core. The tool is inserted in the kerf until its distal end is seated at the full depth. The inner sleeve is then turned. During turning, frictional drag on the outer core pulls the cutting wire into contact with the core. The cutting force of the wire against the core increases with the tension in the wire and

  17. How Rhetorical Theories of Genre Address Common Core Writing Standards

    ERIC Educational Resources Information Center

    Collin, Ross

    2013-01-01

    This article begins with a review of the forms of writing promoted in the Common Core State Standards. Across content areas, Common Core encourages teachers to attune students' writing to rhetorical concerns of audience, purpose, task, and disciplinary thinking. To address these concerns, teachers might take a rhetorical approach to the study…

  18. Core Strength: Implications for Fitness and Low Back Pain.

    ERIC Educational Resources Information Center

    Liemohn, Wendell; Pariser, Gina

    2002-01-01

    Presents information to promote understanding of the concept of core strength and stability, explain why this concept is important to spine health, and evaluate trunk training activities with respect to their contribution to core strength and stability, noting implications for physical fitness and low back pain. The paper reviews the anatomy and…

  19. The Effect of Inner Core Translation on Outer Core Flow and the Geomagnetic Field

    NASA Astrophysics Data System (ADS)

    Mound, J. E.; Davies, C. J.; Silva, L.

    2015-12-01

    Bulk translation of the inner core has been proposed to explain quasi-hemispheric patterns of seismic heterogeneity. Such a translation would result in differential melting and freezing at the inner core boundary (ICB) and hence a heterogeneous pattern of buoyancy flux that could influence convection in the outer core. This heterogeneous flux at the ICB will tend to promote upwelling on the trailing hemisphere, where enhanced inner core growth results in increased latent heat and light element release, and inhibit upwelling on the leading hemisphere, where melting of the inner core occurs. If this difference in convective driving between the two hemispheres propagated across the thickness of the outer core, then flows near the surface of the core could be linked to the ICB heterogeneity and result in a hemispheric imbalance in the geomagnetic field. We have investigated the influence of such ICB boundary conditions on core flows and magnetic field structure in numerical geodynamo models and analysed the resultant hemispheric imbalance relative to the hemispheric structure in models constructed from observations of Earth's field. Inner core translation at rates consistent with estimates for the Earth produce a strong hemispheric bias in the field, one that should be readily apparent in averages of the field over tens of thousands of years. Current models of the field over the Holocene may be able to rule out the most extreme ICB forcing scenarios, but more information on the dynamic structure of the field over these time scales will be needed to adequately test all cases.

  20. Core sample extractor

    NASA Technical Reports Server (NTRS)

    Akins, James; Cobb, Billy; Hart, Steve; Leaptrotte, Jeff; Milhollin, James; Pernik, Mark

    1989-01-01

    The problem of retrieving and storing core samples from a hole drilled on the lunar surface is addressed. The total depth of the hole in question is 50 meters with a maximum diameter of 100 millimeters. The core sample itself has a diameter of 60 millimeters and will be two meters in length. It is therefore necessary to retrieve and store 25 core samples per hole. The design utilizes a control system that will stop the mechanism at a certain depth, a cam-linkage system that will fracture the core, and a storage system that will save and catalogue the cores to be extracted. The Rod Changer and Storage Design Group will provide the necessary tooling to get into the hole as well as to the core. The mechanical design for the cam-linkage system as well as the conceptual design of the storage device are described.

  1. The core paradox.

    NASA Technical Reports Server (NTRS)

    Kennedy, G. C.; Higgins, G. H.

    1973-01-01

    Rebuttal of suggestions from various critics attempting to provide an escape from the seeming paradox originated by Higgins and Kennedy's (1971) proposed possibility that the liquid in the outer core was thermally stably stratified and that this stratification might prove a powerful inhibitor to circulation of the outer core fluid of the kind postulated for the generation of the earth's magnetic field. These suggestions are examined and shown to provide no reasonable escape from the core paradox.

  2. Core Research Center

    USGS Publications Warehouse

    Hicks, Joshua; Adrian, Betty

    2009-01-01

    The Core Research Center (CRC) of the U.S. Geological Survey (USGS), located at the Denver Federal Center in Lakewood, Colo., currently houses rock core from more than 8,500 boreholes representing about 1.7 million feet of rock core from 35 States and cuttings from 54,000 boreholes representing 238 million feet of drilling in 28 States. Although most of the boreholes are located in the Rocky Mountain region, the geologic and geographic diversity of samples have helped the CRC become one of the largest and most heavily used public core repositories in the United States. Many of the boreholes represented in the collection were drilled for energy and mineral exploration, and many of the cores and cuttings were donated to the CRC by private companies in these industries. Some cores and cuttings were collected by the USGS along with other government agencies. Approximately one-half of the cores are slabbed and photographed. More than 18,000 thin sections and a large volume of analytical data from the cores and cuttings are also accessible. A growing collection of digital images of the cores are also becoming available on the CRC Web site Internet http://geology.cr.usgs.gov/crc/.

  3. Health Promotion

    PubMed Central

    Karmali-Rawji, Shameela; Kassim-Lakha, Shaheen; Taylor, Karmel

    1992-01-01

    Perceived lack or loss of control, stress, a rapidly again population and rising costs of health care necessitate effective health promotion and disease prevention in the elderly. In a collaborative health promotion effort, the private sector, public sector, and community partners have joined to increase the South Asian elders' sense of control over the decisions and circumstances that affect their everyday lives. The project was designed to help elders come to terms with the fragmentation of their extended families, cultural alienation, decreased autonomy, need for information, and greater risk of cardiovascular disease. Imagesp622-a

  4. Can Psychiatric Rehabilitation Be Core to CORE?

    ERIC Educational Resources Information Center

    Olney, Marjorie F.; Gill, Kenneth J.

    2016-01-01

    Purpose: In this article, we seek to determine whether psychiatric rehabilitation principles and practices have been more fully incorporated into the Council on Rehabilitation Education (CORE) standards, the extent to which they are covered in four rehabilitation counseling "foundations" textbooks, and how they are reflected in the…

  5. Development and Functional Analysis of Novel Genetic Promoters Using DNA Shuffling, Hybridization and a Combination Thereof

    PubMed Central

    Ranjan, Rajiv; Patro, Sunita; Pradhan, Bhubaneswar; Kumar, Alok; Maiti, Indu B.; Dey, Nrisingha

    2012-01-01

    Background Development of novel synthetic promoters with enhanced regulatory activity is of great value for a diverse range of plant biotechnology applications. Methodology Using the Figwort mosaic virus full-length transcript promoter (F) and the sub-genomic transcript promoter (FS) sequences, we generated two single shuffled promoter libraries (LssF and LssFS), two multiple shuffled promoter libraries (LmsFS-F and LmsF-FS), two hybrid promoters (FuasFScp and FSuasFcp) and two hybrid-shuffled promoter libraries (LhsFuasFScp and LhsFSuasFcp). Transient expression activities of approximately 50 shuffled promoter clones from each of these libraries were assayed in tobacco (Nicotiana tabacum cv. Xanthi) protoplasts. It was observed that most of the shuffled promoters showed reduced activity compared to the two parent promoters (F and FS) and the CaMV35S promoter. In silico studies (computer simulated analyses) revealed that the reduced promoter activities of the shuffled promoters could be due to their higher helical stability. On the contrary, the hybrid promoters FuasFScp and FSuasFcp showed enhanced activities compared to F, FS and CaMV 35S in both transient and transgenic Nicotiana tabacum and Arabidopsis plants. Northern-blot and qRT-PCR data revealed a positive correlation between transcription and enzymatic activity in transgenic tobacco plants expressing hybrid promoters. Histochemical/X-gluc staining of whole transgenic seedlings/tissue-sections and fluorescence images of ImaGene Green™ treated roots and stems expressing the GUS reporter gene under the control of the FuasFScp and FSuasFcp promoters also support the above findings. Furthermore, protein extracts made from protoplasts expressing the human defensin (HNP-1) gene driven by hybrid promoters showed enhanced antibacterial activity compared to the CaMV35S promoter. Significance/Conclusion Both shuffled and hybrid promoters developed in the present study can be used as molecular tools to study the

  6. Making an Ice Core.

    ERIC Educational Resources Information Center

    Kopaska-Merkel, David C.

    1995-01-01

    Explains an activity in which students construct a simulated ice core. Materials required include only a freezer, food coloring, a bottle, and water. This hands-on exercise demonstrates how a glacier is formed, how ice cores are studied, and the nature of precision and accuracy in measurement. Suitable for grades three through eight. (Author/PVD)

  7. Core Concepts of Kinesiology.

    ERIC Educational Resources Information Center

    Hudson, Jackie L.

    1995-01-01

    Core concepts of kinesiology are the basis of communication about movement that facilitate progression of skill levels. The article defines and exemplifies each of 10 core concepts: range of motion, speed of motion, number of segments, nature of segments, balance, coordination, compactness, extension at release/contact, path of projection, and…

  8. CORE - Performance Feedback System

    SciTech Connect

    2009-10-02

    CORE is an architecture to bridge the gaps between disparate data integration and delivery of disparate information visualization. The CORE Technology Program includes a suite of tools and user-centered staff that can facilitate rapid delivery of a deployable integrated information to users.

  9. Iowa Core Annual Report

    ERIC Educational Resources Information Center

    Iowa Department of Education, 2015

    2015-01-01

    One central component of a great school system is a clear set of expectations, or standards, that educators help all students reach. In Iowa, that effort is known as the Iowa Core. The Iowa Core represents the statewide academic standards, which describe what students should know and be able to do in math, science, English language arts, and…

  10. Ice Core Investigations

    ERIC Educational Resources Information Center

    Krim, Jessica; Brody, Michael

    2008-01-01

    What can glaciers tell us about volcanoes and atmospheric conditions? How does this information relate to our understanding of climate change? Ice Core Investigations is an original and innovative activity that explores these types of questions. It brings together popular science issues such as research, climate change, ice core drilling, and air…

  11. Modular core holder

    SciTech Connect

    Mueller, J.; Cole, C.W.; Hamid, S.; Lucas, J.K.

    1991-03-05

    This patent describes a modular core holder. It comprises: a sleeve, forming an internal cavity for receiving a core. The sleeve including segments; support means, overlying the sleeve, for supporting the sleeve; and access means, positioned between at least two of the segments of the sleeve, for allowing measurement of conditions within the internal cavity.

  12. More on the Core

    ERIC Educational Resources Information Center

    Chan, Monnica

    2013-01-01

    From a higher education perspective, new "Common Core" standards could improve student college-readiness levels, reduce institutional remediation rates, and close education gaps in and between states. As a national initiative to create common educational standards for students across multiple states, the Common Core State Standards…

  13. Mercury's core evolution

    NASA Astrophysics Data System (ADS)

    Deproost, Marie-Hélène; Rivoldini, Attilio; Van Hoolst, Tim

    2016-10-01

    Remote sensing data of Mercury's surface by MESSENGER indicate that Mercury formed under reducing conditions. As a consequence, silicon is likely the main light element in the core together with a possible small fraction of sulfur. Compared to sulfur, which does almost not partition into solid iron at Mercury's core conditions and strongly decreases the melting temperature, silicon partitions almost equally well between solid and liquid iron and is not very effective at reducing the melting temperature of iron. Silicon as the major light element constituent instead of sulfur therefore implies a significantly higher core liquidus temperature and a decrease in the vigor of compositional convection generated by the release of light elements upon inner core formation.Due to the immiscibility in liquid Fe-Si-S at low pressure (below 15 GPa), the core might also not be homogeneous and consist of an inner S-poor Fe-Si core below a thinner Si-poor Fe-S layer. Here, we study the consequences of a silicon-rich core and the effect of the blanketing Fe-S layer on the thermal evolution of Mercury's core and on the generation of a magnetic field.

  14. NFE Core Bibliographies.

    ERIC Educational Resources Information Center

    Michigan State Univ., East Lansing. Inst. for International Studies in Education.

    This collection of core bibliographies, which expands on an initial bibliography published in 1979 of the core resources housed in the Non-Formal Education Information Center at Michigan State University, comprises a basic stock of materials on nonformal education and women in development that have been contributed by development planners,…

  15. Dimethyl disulfide produced by the naturally associated bacterium bacillus sp B55 promotes Nicotiana attenuata growth by enhancing sulfur nutrition.

    PubMed

    Meldau, Dorothea G; Meldau, Stefan; Hoang, Long H; Underberg, Stefanie; Wünsche, Hendrik; Baldwin, Ian T

    2013-07-01

    Bacillus sp B55, a bacterium naturally associated with Nicotiana attenuata roots, promotes growth and survival of wild-type and, particularly, ethylene (ET)-insensitive (35)S-ethylene response1 (etr1) N. attenuata plants, which heterologously express the mutant Arabidopsis thaliana receptor ETR1-1. We found that the volatile organic compound (VOC) blend emitted by B55 promotes seedling growth, which is dominated by the S-containing compound dimethyl disulfide (DMDS). DMDS was depleted from the headspace during cocultivation with seedlings in bipartite Petri dishes, and (35)S was assimilated from the bacterial VOC bouquet and incorporated into plant proteins. In wild-type and (35)S-etr1 seedlings grown under different sulfate (SO(4)(-2)) supply conditions, exposure to synthetic DMDS led to genotype-dependent plant growth promotion effects. For the wild type, only S-starved seedlings benefited from DMDS exposure. By contrast, growth of (35)S-etr1 seedlings, which we demonstrate to have an unregulated S metabolism, increased at all SO(4)(-2) supply rates. Exposure to B55 VOCs and DMDS rescued many of the growth phenotypes exhibited by ET-insensitive plants, including the lack of root hairs, poor lateral root growth, and low chlorophyll content. DMDS supplementation significantly reduced the expression of S assimilation genes, as well as Met biosynthesis and recycling. We conclude that DMDS by B55 production is a plant growth promotion mechanism that likely enhances the availability of reduced S, which is particularly beneficial for wild-type plants growing in S-deficient soils and for (35)S-etr1 plants due to their impaired S uptake/assimilation/metabolism.

  16. Internal core tightener

    DOEpatents

    Brynsvold, Glen V.; Snyder, Jr., Harold J.

    1976-06-22

    An internal core tightener which is a linear actuated (vertical actuation motion) expanding device utilizing a minimum of moving parts to perform the lateral tightening function. The key features are: (1) large contact areas to transmit loads during reactor operation; (2) actuation cam surfaces loaded only during clamping and unclamping operation; (3) separation of the parts and internal operation involved in the holding function from those involved in the actuation function; and (4) preloaded pads with compliant travel at each face of the hexagonal assembly at the two clamping planes to accommodate thermal expansion and irradiation induced swelling. The latter feature enables use of a "fixed" outer core boundary, and thus eliminates the uncertainty in gross core dimensions, and potential for rapid core reactivity changes as a result of core dimensional change.

  17. Lunar Core and Tides

    NASA Technical Reports Server (NTRS)

    Williams, J. G.; Boggs, D. H.; Ratcliff, J. T.

    2004-01-01

    Variations in rotation and orientation of the Moon are sensitive to solid-body tidal dissipation, dissipation due to relative motion at the fluid-core/solid-mantle boundary, and tidal Love number k2 [1,2]. There is weaker sensitivity to flattening of the core-mantle boundary (CMB) [2,3,4] and fluid core moment of inertia [1]. Accurate Lunar Laser Ranging (LLR) measurements of the distance from observatories on the Earth to four retroreflector arrays on the Moon are sensitive to lunar rotation and orientation variations and tidal displacements. Past solutions using the LLR data have given results for dissipation due to solid-body tides and fluid core [1] plus Love number [1-5]. Detection of CMB flattening, which in the past has been marginal but improving [3,4,5], now seems significant. Direct detection of the core moment has not yet been achieved.

  18. Calcium ions affect the hepatitis B virus core assembly

    SciTech Connect

    Choi, Yongwook; Gyoo Park, Sung; Yoo, Jun-hi; Jung, Guhung . E-mail: drjung@snu.ac.kr

    2005-02-05

    Previous report showed that cytosolic Ca{sup 2+} induced by hepatitis B virus X protein (HBx) promotes HBV replication. In this study, in vitro experiments showed that (i) HBV core assembly in vitro was promoted by Ca{sup 2+} through the sucrose density gradient and the analytical ultracentrifuge analysis. Also (ii) transmission electron microscope analysis demonstrated these assembled HBV core particles were the capsids. Ex vivo experiments showed that the treatment of BAPTA-AM and cyclosporine A (CsA) reduced HBV capsids in the transfected HepG2 cells. In addition to that, the treatment of Thapsigargin (TG) increased HBV capsids in the transfected HepG2 cells. Furthermore, we investigated the increased HBV core assembly by HBx. The results show that the increased cytosolic calcium ions by HBx promote the HBV core assembly.

  19. Expectations for the Martian core magnetic field

    NASA Technical Reports Server (NTRS)

    Stevenson, D. J.

    1993-01-01

    In the traditional view of planetary magnetism, a planet either has a core dynamo (Earth, Jupiter, Saturn, Uranus, Neptune, maybe Mercury) or does not (Mars, Venus, Moon...) I argue that this view is simplistic in two respects. First, mantle convection in terrestrial planets is invariably ata high enough Rayleigh number that it is time variable; this leads to the intermittent arrival of mantle 'cold fingers' at the core-mantle boundary promoting at least local core convection and dynamo action even when the planetary core is stably stratified on average. Thus, I predict an intermittent dynamo regime in addition to the simple dynamo-on (Earth) and dynamo-off regimes. Second, the mantle convection-driven horizontal temperature gradients just below the core-mantle boundary can lead to unstable flows that will convert thermoelectric or electrochemical toroidal fields into externally detectable poloidal fields, even when a dynamo is not possible. It is likely that Mars possesses an interesting core magnetic field of the latter kind, complex but with a dipole that might be approximately aligned with the rotation axis and a surface field of a few to tens of gammas.

  20. Bacterial Fouling in a Model Core System

    PubMed Central

    Shaw, J. C.; Bramhill, B.; Wardlaw, N. C.; Costerton, J. W.

    1985-01-01

    We have used a sintered glass bead core to simulate the spaces and surfaces of reservoir rock in studies of the bacterial plugging phenomenon that affects waterflood oil recovery operations. The passage of pure or mixed natural populations of bacteria through this solid matrix was initially seen to promote the formation of adherent bacterial microcolonies on available surfaces. Bacteria within these microcolonies produced huge amounts of exopolysaccharides and coalesced to form a confluent plugging biofilm that eventually caused a >99% decrease in core permeability. Aerobic bacteria developed a plugging biofilm on the inlet face of the core, facultative anaerobes plugged throughout the core, and dead bacteria did not effectively plug the narrow (33-μm) spaces of this solid matrix because they neither adhered extensively to surfaces nor produced the extensive exopolysaccharides characteristic of living cells. The presence of particles in the water used in these experiments rapidly decreased the core permeability because they became trapped in the developing biofilm and accelerated the plugging of pore spaces. Once established, cells within the bacterial biofilm could be killed by treatment with a biocide (isothiazalone), but their essentially inert carbohydrate biofilm matrix persisted and continued to plug the pore spaces, whereas treatment with 5% sodium hypochlorite killed the bacteria, dissolved the exopolysaccharide biofilm matrix, and restored permeability to these plugged glass bead cores. Images PMID:16346760

  1. 34. DESPATCH CORE OVENS, GREY IRON FOUNDRY CORE ROOM, BAKES ...

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

    34. DESPATCH CORE OVENS, GREY IRON FOUNDRY CORE ROOM, BAKES CORES THAT ARE NOT MADE ON HEATED OR COLD BOX CORE MACHINES, TO SET BINDING AGENTS MIXED WITH THE SAND CREATING CORES HARD ENOUGH TO WITHSTAND THE FLOW OF MOLTEN IRON INSIDE A MOLD. - Stockham Pipe & Fittings Company, Grey Iron Foundry, 4000 Tenth Avenue North, Birmingham, Jefferson County, AL

  2. Multiple Core Galaxies

    NASA Technical Reports Server (NTRS)

    Miller, R.H.; Morrison, David (Technical Monitor)

    1994-01-01

    Nuclei of galaxies often show complicated density structures and perplexing kinematic signatures. In the past we have reported numerical experiments indicating a natural tendency for galaxies to show nuclei offset with respect to nearby isophotes and for the nucleus to have a radial velocity different from the galaxy's systemic velocity. Other experiments show normal mode oscillations in galaxies with large amplitudes. These oscillations do not damp appreciably over a Hubble time. The common thread running through all these is that galaxies often show evidence of ringing, bouncing, or sloshing around in unexpected ways, even though they have not been disturbed by any external event. Recent observational evidence shows yet another phenomenon indicating the dynamical complexity of central regions of galaxies: multiple cores (M31, Markarian 315 and 463 for example). These systems can hardly be static. We noted long-lived multiple core systems in galaxies in numerical experiments some years ago, and we have more recently followed up with a series of experiments on multiple core galaxies, starting with two cores. The relevant parameters are the energy in the orbiting clumps, their relative.masses, the (local) strength of the potential well representing the parent galaxy, and the number of cores. We have studied the dependence of the merger rates and the nature of the final merger product on these parameters. Individual cores survive much longer in stronger background potentials. Cores can survive for a substantial fraction of a Hubble time if they travel on reasonable orbits.

  3. Global Core Plasma Model

    NASA Technical Reports Server (NTRS)

    Gallagher, Dennis L.; Craven, P. D.; Comfort, R. H.

    1999-01-01

    Abstract. The Global Core Plasma Model (GCPM) provides, empirically derived, core plasma density as a function of geomagnetic and solar conditions throughout the inner magnetosphere. It is continuous in value and gradient and is composed of separate models for the ionosphere, the plasmasphere, the plasmapause, the trough, and the polar cap. The relative composition of plasmaspheric H+, He+, and O+ is included in the GCPM. A blunt plasmaspheric bulge and rotation of the bulge with changing geomagnetic conditions is included. The GCPM is an amalgam of density models, intended to serve as a framework for continued improvement as new measurements become available and are used to characterize core plasma density, composition, and temperature.

  4. Core shroud corner joints

    DOEpatents

    Gilmore, Charles B.; Forsyth, David R.

    2013-09-10

    A core shroud is provided, which includes a number of planar members, a number of unitary corners, and a number of subassemblies each comprising a combination of the planar members and the unitary corners. Each unitary corner comprises a unitary extrusion including a first planar portion and a second planar portion disposed perpendicularly with respect to the first planar portion. At least one of the subassemblies comprises a plurality of the unitary corners disposed side-by-side in an alternating opposing relationship. A plurality of the subassemblies can be combined to form a quarter perimeter segment of the core shroud. Four quarter perimeter segments join together to form the core shroud.

  5. A study on the influence of different promoter and 5'UTR (URM) cassettes from Arabidopsis thaliana on the expression level of the reporter gene β glucuronidase in tobacco and cotton.

    PubMed

    Agarwal, Parul; Garg, Varsha; Gautam, Taru; Pillai, Beena; Kanoria, Shaveta; Burma, Pradeep Kumar

    2014-04-01

    Several reports of promoters from plants, viral and artificial origin that confer high constitutive expression are known. Among these the CaMV 35S promoter is used extensively for transgene expression in plants. We identified candidate promoters from Arabidopsis based on their transcript levels (meta-analysis of available microarray control datasets) to test their activity in comparison to the CaMV 35S promoter. A set of 11 candidate genes were identified which showed high transcript levels in the aerial tissue (i.e. leaf, shoot, flower and stem). In the initial part of the study binary vectors were developed wherein the promoter and 5'UTR region of these candidate genes (Upstream Regulatory Module, URM) were cloned upstream to the reporter gene β glucuronidase (gus). The promoter strengths were tested in transformed callus of Nicotiana tabacum and Gossypium hirsutum. On the basis of the results obtained from the callus, the influence of the URM cassettes on transgene expression was tested in transgenic tobacco. The URM regions of the genes encoding a subunit of photosystem I (PHOTO) and geranyl geranyl reductase (GGR) in A. thaliana genome showed significantly high levels of GUS activity in comparison to the CaMV 35S promoter. Further, when the 5'UTRs of both the genes were placed downstream to the CaMV 35S promoter it led to a substantial increase in GUS activity in transgenic tobacco lines and cotton callus. The enhancement observed was even higher to that observed with the viral leader sequences like Ω and AMV, known translational enhancers. Our results indicate that the two URM cassettes or the 5'UTR regions of PHOTO and GGR when placed downstream to the CaMV 35S promoter can be used to drive high levels of transgene expression in dicotyledons.

  6. Hydrogen exchange, core modules, and new designed proteins.

    PubMed

    Carulla, Natàlia; Barany, George; Woodward, Clare

    2002-12-10

    A strategy for design of new proteins that mimic folding properties of native proteins is based on peptides modeled on the slow exchange cores of natural proteins. We have synthesized peptides, called core modules, that correspond to the elements of secondary structure that carry the very slowest exchanging amides in a protein. The expectation is that, if soluble in water, core modules will form conformational ensembles that favor native-like structure. Core modules modeled on natural bovine pancreatic trypsin inhibitor have been shown by NMR studies to meet this expectation. The next step toward production of a native state mimic is to further shift the conformational bias of a core module toward more ordered structure by promoting module-module interactions that are mutually stabilizing. For this, two core modules were incorporated into a single molecule by means of a long cross-link. From a panel of several two-module peptides, one very promising lead emerged; it is called BetaCore. BetaCore is monomeric in water and forms a new fold composed of a four-stranded, antiparallel beta-sheet. The single, dominant conformation of BetaCore is characterized by various NMR experiments. Here we compare the individual core module to the two-module BetaCore and discuss the progressive stabilization of intramodule structure and the formation of new intermodule interactions.

  7. Core assembly storage structure

    DOEpatents

    Jones, Jr., Charles E.; Brunings, Jay E.

    1988-01-01

    A structure for the storage of core assemblies from a liquid metal-cooled nuclear reactor. The structure comprises an enclosed housing having a substantially flat horizontal top plate, a bottom plate and substantially vertical wall members extending therebetween. A plurality of thimble members extend downwardly through the top plate. Each thimble member is closed at its bottom end and has an open end adjacent said top plate. Each thimble member has a length and diameter greater than that of the core assembly to be stored therein. The housing is provided with an inlet duct for the admission of cooling air and an exhaust duct for the discharge of air therefrom, such that when hot core assemblies are placed in the thimbles, the heat generated will by convection cause air to flow from the inlet duct around the thimbles and out the exhaust duct maintaining the core assemblies at a safe temperature without the necessity of auxiliary powered cooling equipment.

  8. Magnetorotational iron core collapse

    NASA Technical Reports Server (NTRS)

    Symbalisty, E. M. D.

    1984-01-01

    During its final evolutionary stages, a massive star, as considered in current astrophysical theory, undergoes rapid collapse, thereby triggering a sequence of a catastrophic event which results in a Type II supernova explosion. A remnant neutron star or a black hole is left after the explosion. Stellar collapse occurs, when thermonuclear fusion has consumed the lighter elements present. At this stage, the core consists of iron. Difficulties arise regarding an appropriate model with respect to the core collapse. The present investigation is concerned with the evolution of a Type II supernova core including the effects of rotation and magnetic fields. A simple neutrino model is developed which reproduced the spherically symmetric results of Bowers and Wilson (1982). Several two-dimensional computational models of stellar collapse are studied, taking into account a case in which a 15 solar masses iron core was artificially given rotational and magnetic energy.

  9. Contaminated Sediment Core Profiling

    EPA Science Inventory

    Evaluating the environmental risk of sites containing contaminated sediments often poses major challenges due in part to the absence of detailed information available for a given location. Sediment core profiling is often utilized during preliminary environmental investigations ...

  10. INTEGRAL core programme

    NASA Technical Reports Server (NTRS)

    Gehrels, N.; Schoenfelder, V.; Ubertini, P.; Winkler, C.

    1997-01-01

    The International Gamma Ray Astrophysics Laboratory (INTEGRAL) mission is described with emphasis on the INTEGRAL core program. The progress made in the planning activities for the core program is reported on. The INTEGRAL mission has a nominal lifetime of two years with a five year extension option. The observing time will be divided between the core program (between 30 and 35 percent during the first two years) and general observations. The core program consists of three main elements: the deep survey of the Galactic plane in the central radian of the Galaxy; frequent scans of the Galactic plane in the search for transient sources, and pointed observations of several selected sources. The allocation of the observation time is detailed and the sensitivities of the observations are outlined.

  11. Biospecimen Core Resource - TCGA

    Cancer.gov

    The Cancer Genome Atlas (TCGA) Biospecimen Core Resource centralized laboratory reviews and processes blood and tissue samples and their associated data using optimized standard operating procedures for the entire TCGA Research Network.

  12. Core bounce supernovae

    SciTech Connect

    Cooperstein, J.

    1987-01-01

    The gravitational collapse mechanism for Type II supernovae is considered, concentrating on the direct implosion - core bounce - hydrodynamic explosion picture. We examine the influence of the stiffness of the dense matter equation of state and discuss how the shock wave is formed. Its chances of success are determined by the equation of state, general relativistic effects, neutrino transport, and the size of presupernova iron core. 12 refs., 1 tab.

  13. Nuclear core positioning system

    DOEpatents

    Garkisch, Hans D.; Yant, Howard W.; Patterson, John F.

    1979-01-01

    A structural support system for the core of a nuclear reactor which achieves relatively restricted clearances at operating conditions and yet allows sufficient clearance between fuel assemblies at refueling temperatures. Axially displaced spacer pads having variable between pad spacing and a temperature compensated radial restraint system are utilized to maintain clearances between the fuel elements. The core support plates are constructed of metals specially chosen such that differential thermal expansion produces positive restraint at operating temperatures.

  14. Micro coring apparatus

    NASA Technical Reports Server (NTRS)

    Collins, David; Brooks, Marshall; Chen, Paul; Dwelle, Paul; Fischer, Ben

    1989-01-01

    A micro-coring apparatus for lunar exploration applications, that is compatible with the other components of the Walking Mobile Platform, was designed. The primary purpose of core sampling is to gain an understanding of the geological composition and properties of the prescribed environment. This procedure has been used extensively for Earth studies and in limited applications during lunar explorations. The corer is described and analyzed for effectiveness.

  15. MCNP LWR Core Generator

    SciTech Connect

    Fischer, Noah A.

    2012-08-14

    The reactor core input generator allows for MCNP input files to be tailored to design specifications and generated in seconds. Full reactor models can now easily be created by specifying a small set of parameters and generating an MCNP input for a full reactor core. Axial zoning of the core will allow for density variation in the fuel and moderator, with pin-by-pin fidelity, so that BWR cores can more accurately be modeled. LWR core work in progress: (1) Reflectivity option for specifying 1/4, 1/2, or full core simulation; (2) Axial zoning for moderator densities that vary with height; (3) Generating multiple types of assemblies for different fuel enrichments; and (4) Parameters for specifying BWR box walls. Fuel pin work in progress: (1) Radial and azimuthal zoning for generating further unique materials in fuel rods; (2) Options for specifying different types of fuel for MOX or multiple burn assemblies; (3) Additional options for replacing fuel rods with burnable poison rods; and (4) Control rod/blade modeling.

  16. Emergency core cooling system

    DOEpatents

    Schenewerk, William E.; Glasgow, Lyle E.

    1983-01-01

    A liquid metal cooled fast breeder reactor provided with an emergency core cooling system includes a reactor vessel which contains a reactor core comprising an array of fuel assemblies and a plurality of blanket assemblies. The reactor core is immersed in a pool of liquid metal coolant. The reactor also includes a primary coolant system comprising a pump and conduits for circulating liquid metal coolant to the reactor core and through the fuel and blanket assemblies of the core. A converging-diverging venturi nozzle with an intermediate throat section is provided in between the assemblies and the pump. The intermediate throat section of the nozzle is provided with at least one opening which is in fluid communication with the pool of liquid sodium. In normal operation, coolant flows from the pump through the nozzle to the assemblies with very little fluid flowing through the opening in the throat. However, when the pump is not running, residual heat in the core causes fluid from the pool to flow through the opening in the throat of the nozzle and outwardly through the nozzle to the assemblies, thus providing a means of removing decay heat.

  17. TERT Core Promotor Mutations in Early-Onset Bladder Cancer

    PubMed Central

    Giedl, Johannes; Rogler, Anja; Wild, Andreas; Riener, Marc-Oliver; Filbeck, Thomas; Burger, Maximilian; Rümmele, Petra; Hurst, Carolyn; Knowles, Margaret; Hartmann, Arndt; Zinnall, Ulrike; Stoehr, Robert

    2016-01-01

    Activating mutations in the core promoter of the TERT gene have been described in many different tumor entities. In vitro models showed a two- to fourfold increase in transcriptional activity of the TERT promoter through creation of a consensus binding motif for Ets/TCF transcription factors caused by these mutations. TERT core promoter mutations are the most common mutations in bladder cancer with a frequency between 55.6% and 82.8% described so far, and are independent of stage and grade. Since limited data on molecular alterations of early-onset bladder tumors exists, we assessed the frequency of TERT core promoter mutations in early-onset bladder cancer. Two cohorts of bladder tumors (early-onset patient group; n=144 (age of onset of disease ≤45 years); unselected, consecutive group; n=125) were examined for TERT core promoter mutations. After microdissection and extraction of DNA the corresponding hotspot regions in the TERT core promoter were examined by Sanger-sequencing or a SNaPshot approach. A significantly lower frequency of TERT core promoter mutations was found in tumors from the early-onset cohort compared to the consecutive cohort (57.6% vs. 84.8%, p<0.001). Among the early-onset cohort cases younger than the cohort's median age of 39 years at disease onset showed a significantly reduced number of TERT promoter mutations (31/67, 46,3%) than cases aged between 39 and 45 years (52/77, 67.5%; p=0.012). This association was not found in the consecutive cases. Mutation status was independent of tumor stage and grade. We conclude that in tumors from early-onset bladder cancer patients TERT core promoter mutations are not as frequent as in bladder tumors from consecutive cases, but seem to play an important role there as well. In patients below 39 years of age TERT core promoter mutations are a more infrequent event, suggesting different mechanisms of tumorigenesis in these young patients. PMID:27313781

  18. An engineered strong promoter for streptomycetes.

    PubMed

    Wang, Weishan; Li, Xiao; Wang, Juan; Xiang, Sihai; Feng, Xiaozhou; Yang, Keqian

    2013-07-01

    Well-characterized promoters are essential tools for metabolic engineering and synthetic biology. In Streptomyces coelicolor, the native kasOp is a temporally expressed promoter strictly controlled by two regulators, ScbR and ScbR2. In this work, first, kasOp was engineered to remove a common binding site of ScbR and ScbR2 upstream of its core region, thus generating a stronger promoter, kasOp3. Second, another ScbR binding site internal to the kasOp3 core promoter region was abolished by random mutation and screening of the mutant library to obtain the strongest promoter, kasOp* (where the asterisk is used to distinguish the engineered promoter from the native promoter). The activities of kasOp* were compared with those of two known strong promoters, ermEp* and SF14p, in three Streptomyces species. kasOp* showed the highest activity at the transcription and protein levels in all three hosts. Furthermore, relative to ermEp* and SF14p, kasOp* was shown to confer the highest actinorhodin production level when used to drive the expression of actII-ORF4 in S. coelicolor. Therefore, kasOp* is a simple and well-defined strong promoter useful for gene overexpression in streptomycetes.

  19. An Engineered Strong Promoter for Streptomycetes

    PubMed Central

    Wang, Weishan; Li, Xiao; Wang, Juan; Xiang, Sihai; Feng, Xiaozhou

    2013-01-01

    Well-characterized promoters are essential tools for metabolic engineering and synthetic biology. In Streptomyces coelicolor, the native kasOp is a temporally expressed promoter strictly controlled by two regulators, ScbR and ScbR2. In this work, first, kasOp was engineered to remove a common binding site of ScbR and ScbR2 upstream of its core region, thus generating a stronger promoter, kasOp3. Second, another ScbR binding site internal to the kasOp3 core promoter region was abolished by random mutation and screening of the mutant library to obtain the strongest promoter, kasOp* (where the asterisk is used to distinguish the engineered promoter from the native promoter). The activities of kasOp* were compared with those of two known strong promoters, ermEp* and SF14p, in three Streptomyces species. kasOp* showed the highest activity at the transcription and protein levels in all three hosts. Furthermore, relative to ermEp* and SF14p, kasOp* was shown to confer the highest actinorhodin production level when used to drive the expression of actII-ORF4 in S. coelicolor. Therefore, kasOp* is a simple and well-defined strong promoter useful for gene overexpression in streptomycetes. PMID:23686264

  20. Pineapple translation factor SUI1 and ribosomal protein L36 promoters drive constitutive transgene expression patterns in Arabidopsis thaliana.

    PubMed

    Koia, Jonni; Moyle, Richard; Hendry, Caroline; Lim, Lionel; Botella, José Ramón

    2013-03-01

    The availability of a variety of promoter sequences is necessary for the genetic engineering of plants, in basic research studies and for the development of transgenic crops. In this study, the promoter and 5' untranslated regions of the evolutionally conserved protein translation factor SUI1 gene and ribosomal protein L36 gene were isolated from pineapple and sequenced. Each promoter was translationally fused to the GUS reporter gene and transformed into the heterologous plant system Arabidopsis thaliana. Both the pineapple SUI1 and L36 promoters drove GUS expression in all tissues of Arabidopsis at levels comparable to the CaMV35S promoter. Transient assays determined that the pineapple SUI1 promoter also drove GUS expression in a variety of climacteric and non-climacteric fruit species. Thus the pineapple SUI1 and L36 promoters demonstrate the potential for using translation factor and ribosomal protein genes as a source of promoter sequences that can drive constitutive transgene expression patterns.

  1. Assessing Core Competencies

    NASA Astrophysics Data System (ADS)

    Narayanan, M.

    2004-12-01

    Catherine Palomba and Trudy Banta offer the following definition of assessment, adapted from one provided by Marches in 1987. Assessment in the systematic collection, review, and use of information about educational programs undertaken for the purpose of improving student learning and development. (Palomba and Banta 1999). It is widely recognized that sophisticated computing technologies are becoming a key element in today's classroom instructional techniques. Regardless, the Professor must be held responsible for creating an instructional environment in which the technology actually supplements learning outcomes of the students. Almost all academic disciplines have found a niche for computer-based instruction in their respective professional domain. In many cases, it is viewed as an essential and integral part of the educational process. Educational institutions are committing substantial resources to the establishment of dedicated technology-based laboratories, so that they will be able to accommodate and fulfill students' desire to master certain of these specific skills. This type of technology-based instruction may raise some fundamental questions about the core competencies of the student learner. Some of the most important questions are : 1. Is the utilization of these fast high-powered computers and user-friendly software programs creating a totally non-challenging instructional environment for the student learner ? 2. Can technology itself all too easily overshadow the learning outcomes intended ? 3. Are the educational institutions simply training students how to use technology rather than educating them in the appropriate field ? 4. Are we still teaching content-driven courses and analysis oriented subject matter ? 5. Are these sophisticated modern era technologies contributing to a decline in the Critical Thinking Capabilities of the 21st century technology-savvy students ? The author tries to focus on technology as a tool and not on the technology

  2. Core Noise - Increasing Importance

    NASA Technical Reports Server (NTRS)

    Hultgren, Lennart S.

    2011-01-01

    This presentation is a technical summary of and outlook for NASA-internal and NASA-sponsored external research on core (combustor and turbine) noise funded by the Fundamental Aeronautics Program Subsonic Fixed Wing (SFW) Project. Sections of the presentation cover: the SFW system-level noise metrics for the 2015, 2020, and 2025 timeframes; turbofan design trends and their aeroacoustic implications; the emerging importance of core noise and its relevance to the SFW Reduced-Perceived-Noise Technical Challenge; and the current research activities in the core-noise area, with additional details given about the development of a high-fidelity combustor-noise prediction capability as well as activities supporting the development of improved reduced-order, physics-based models for combustor-noise prediction. The need for benchmark data for validation of high-fidelity and modeling work and the value of a potential future diagnostic facility for testing of core-noise-reduction concepts are indicated. The NASA Fundamental Aeronautics Program has the principal objective of overcoming today's national challenges in air transportation. The SFW Reduced-Perceived-Noise Technical Challenge aims to develop concepts and technologies to dramatically reduce the perceived aircraft noise outside of airport boundaries. This reduction of aircraft noise is critical to enabling the anticipated large increase in future air traffic. Noise generated in the jet engine core, by sources such as the compressor, combustor, and turbine, can be a significant contribution to the overall noise signature at low-power conditions, typical of approach flight. At high engine power during takeoff, jet and fan noise have traditionally dominated over core noise. However, current design trends and expected technological advances in engine-cycle design as well as noise-reduction methods are likely to reduce non-core noise even at engine-power points higher than approach. In addition, future low-emission combustor

  3. Recent developments in pressure coring

    SciTech Connect

    McFall, A. L.

    1980-01-01

    The current rapid growth in the number of enhanced oil and gas recovery projects has created a strong demand for reservoir data such as true residual oil saturations. The companies providing pressure coring services have moved to fill this need. Two recent developments have emerged with the potential of significantly improving the present performance of pressure coring. Coring bits utilizing synthetic diamond cutters have demonstrated coring rates of one-foot per minute while improving core recovery. It is also apparent that cores of a near-unconsolidated nature are more easily recovered. In addition, a special low invasion fluid that is placed in the core retriever has demonstrated reduced core washing by the drilling mud and a decrease in the complexity of preparing cores for analysis. This paper describes the design, laboratory, and field testing efforts that led to these coring improvements. Also, experience in utilizing these developments while recovering over 100 cores is discussed.

  4. Pressure Core Characterization

    NASA Astrophysics Data System (ADS)

    Santamarina, J. C.

    2014-12-01

    Natural gas hydrates form under high fluid pressure and low temperature, and are found in permafrost, deep lakes or ocean sediments. Hydrate dissociation by depressurization and/or heating is accompanied by a multifold hydrate volume expansion and host sediments with low permeability experience massive destructuration. Proper characterization requires coring, recovery, manipulation and testing under P-T conditions within the stability field. Pressure core technology allows for the reliable characterization of hydrate bearing sediments within the stability field in order to address scientific and engineering needs, including the measurement of parameters used in hydro-thermo-mechanical analyses, and the monitoring of hydrate dissociation under controlled pressure, temperature, effective stress and chemical conditions. Inherent sampling effects remain and need to be addressed in test protocols and data interpretation. Pressure core technology has been deployed to study hydrate bearing sediments at several locations around the world. In addition to pressure core testing, a comprehensive characterization program should include sediment analysis, testing of reconstituted specimens (with and without synthetic hydrate), and in situ testing. Pressure core characterization technology can be used to study other gas-charged formations such as deep sea sediments, coal bed methane and gas shales.

  5. Core Noise Reduction

    NASA Technical Reports Server (NTRS)

    Hultgren, Lennart S.

    2011-01-01

    This presentation is a technical summary of and outlook for NASA-internal and NASA-sponsored external research on core (combustor and turbine) noise funded by the Fundamental Aeronautics Program Subsonic Fixed Wing (SFW) Project. Sections of the presentation cover: the SFW system-level noise metrics for the 2015, 2020, and 2025 timeframes; turbofan design trends and their aeroacoustic implications; the emerging importance of core noise and its relevance to the SFW Reduce-Perceived-Noise Technical Challenge; and the current research activities in the core noise area. Recent work1 on the turbine-transmission loss of combustor noise is briefly described, two2,3 new NRA efforts in the core-noise area are outlined, and an effort to develop CMC-based acoustic liners for broadband noise reduction suitable for turbofan-core application is delineated. The NASA Fundamental Aeronautics Program has the principal objective of overcoming today's national challenges in air transportation. The reduction of aircraft noise is critical to enabling the anticipated large increase in future air traffic. The Subsonic Fixed Wing Project's Reduce-Perceived-Noise Technical Challenge aims to develop concepts and technologies to dramatically reduce the perceived aircraft noise outside of airport boundaries.

  6. A promoter from sugarcane bacilliform badnavirus drives transgene expression in banana and other monocot and dicot plants.

    PubMed

    Schenk, P M; Sagi, L; Remans, T; Dietzgen, R G; Bernard, M J; Graham, M W; Manners, J M

    1999-04-01

    A 1369 bp DNA fragment (Sc) was isolated from a full-length clone of sugarcane bacilliform badnavirus (ScBV) and was shown to have promoter activity in transient expression assays using monocot (banana, maize, millet and sorghum) and dicot plant species (tobacco, sunflower, canola and Nicotiana benthamiana). This promoter was also tested for stable expression in transgenic banana and tobacco plants. These experiments showed that this promoter could drive high-level expression of the beta-glucuronidase (GUS) reporter gene in most plant cells. The expression level was comparable to the maize ubiquitin promoter in standardised transient assays in maize. In transgenic banana plants the expression levels were variable for different transgenic lines but was generally comparable with the activities of both the maize ubiquitin promoter and the enhanced cauliflower mosaic virus (CaMV) 35S promoter. The Sc promoter appears to express in a near-constitutive manner in transgenic banana and tobacco plants. The promoter from sugarcane bacilliform virus represents a useful tool for the high-level expression of foreign genes in both monocot and dicot transgenic plants that could be used similarly to the CaMV 35S or maize polyubiquitin promoter.

  7. Molten core retention assembly

    DOEpatents

    Lampe, Robert F.

    1976-06-22

    Molten fuel produced in a core overheating accident is caught by a molten core retention assembly consisting of a horizontal baffle plate having a plurality of openings therein, heat exchange tubes having flow holes near the top thereof mounted in the openings, and a cylindrical, imperforate baffle attached to the plate and surrounding the tubes. The baffle assembly is supported from the core support plate of the reactor by a plurality of hanger rods which are welded to radial beams passing under the baffle plate and intermittently welded thereto. Preferably the upper end of the cylindrical baffle terminates in an outwardly facing lip to which are welded a plurality of bearings having slots therein adapted to accept the hanger rods.

  8. Earth's core iron

    NASA Astrophysics Data System (ADS)

    Geophysicist J. Michael Brown of Texas A & M University noted recently at the Spring AGU Meeting in Baltimore that the structure and phase of metallic iron at pressures of the earth's inner core (approximately 3.3 Mbar) could have great significance in defining geometrical aspects of the core itself. Brown worked at the Los Alamos Scientific Laboratory with R.B. McQueen to redetermine the phase relations of metallic iron in a series of new shock-wave experiments. They found the melting point of iron at conditions equal to those at the boundary of the earth's outer (liquid) and inner (solid) cores to be 6000°±500°C (Geophysical Research Letters, 7, 533-536, 1980).

  9. Cloning and characterization of a tuberous root-specific promoter from cassava (Manihot esculenta Crantz).

    PubMed

    Koehorst-van Putten, Herma J J; Wolters, Anne-Marie A; Pereira-Bertram, Isolde M; van den Berg, Hans H J; van der Krol, Alexander R; Visser, Richard G F

    2012-12-01

    In order to obtain a tuberous root-specific promoter to be used in the transformation of cassava, a 1,728 bp sequence containing the cassava granule-bound starch synthase (GBSSI) promoter was isolated. The sequence proved to contain light- and sugar-responsive cis elements. Part of this sequence (1,167 bp) was cloned into binary vectors to drive expression of the firefly luciferase gene. Cassava cultivar Adira 4 was transformed with this construct or a control construct in which the luciferase gene was cloned behind the 35S promoter. Luciferase activity was measured in leaves, stems, roots and tuberous roots. As expected, the 35S promoter induced luciferase activity in all organs at similar levels, whereas the GBSSI promoter showed very low expression in leaves, stems and roots, but very high expression in tuberous roots. These results show that the cassava GBSSI promoter is an excellent candidate to achieve tuberous root-specific expression in cassava.

  10. CORE SATURATION BLOCKING OSCILLATOR

    DOEpatents

    Spinrad, R.J.

    1961-10-17

    A blocking oscillator which relies on core saturation regulation to control the output pulse width is described. In this arrangement an external magnetic loop is provided in which a saturable portion forms the core of a feedback transformer used with the thermionic or semi-conductor active element. A first stationary magnetic loop establishes a level of flux through the saturation portion of the loop. A second adjustable magnet moves the flux level to select a saturation point giving the desired output pulse width. (AEC)

  11. Soybean GH3 promoter contains multiple auxin-inducible elements.

    PubMed Central

    Liu, Z B; Ulmasov, T; Shi, X; Hagen, G; Guilfoyle, T J

    1994-01-01

    The soybean GH3 gene is transcriptionally induced in a wide variety of tissues and organs within minutes after auxin application. To determine the sequence elements that confer auxin inducibility to the GH3 promoter, we used gel mobility shift assays, methylation interference, deletion analysis, linker scanning, site-directed mutagenesis, and gain-of-function analysis with a minimal cauliflower mosaic virus 35S promoter. We identified at least three sequence elements within the GH3 promoter that are auxin inducible and can function independently of one another. Two of these elements are found in a 76-bp fragment, and these consist of two independent 25- and 32-bp auxin-inducible elements. Both of these 25- and 32-bp auxin-inducible elements contain the sequence TGTCTC just upstream of an AATAAG. An additional auxin-inducible element was found upstream of the 76-bp auxin-inducible fragment; this can function independently of the 76-bp fragment. Two TGA-box or Hex-like elements (TGACGTAA and TGACGTGGC) in the promoter, which are strong binding sites for proteins in plant nuclear extracts, may also elevate the level of auxin inducibility of the GH3 promoter. The multiple auxin-inducible elements within the GH3 promoter contribute incrementally to the overall level of auxin induction observed with this promoter. PMID:8038604

  12. Efficient chimeric promoters derived from full-length and sub-genomic transcript promoters of Figwort mosaic virus (FMV).

    PubMed

    Ranjan, Rajiv; Patro, Sunita; Kumari, Sangeeta; Kumar, Deepak; Dey, Nrisingha; Maiti, Indu B

    2011-03-10

    Addition of multiple repeats of the FS3 upstream activation sequence (FS3-UAS, -270 to -60) intra-molecularly to the TATA containing core-domain of the FS3 (-151 to +31) promoter resulted in 2-3-folds enhanced promoter activity. The chimeric promoter, FS3-UAS-3X with maximum activity, showed 3.31 times stronger activity in root vascular tissue compared to FS3 promoter and could be used efficiently in translational research.

  13. Promoting Retention

    PubMed Central

    Hall, LaToya N.; Ficker, Lisa J.; Chadiha, Letha A.; Green, Carmen R.; Jackson, James S.; Lichtenberg, Peter A.

    2016-01-01

    Objectives: The objectives of this study were to evaluate the capability of a research volunteer registry to retain community-dwelling African American older adults, and to explore demographic and health factors associated with retention. Method: A logistic regression model was used to determine the influence of demographics, health factors, and registry logic model activities on retention in a sample of 1,730 older African American adults. Results: Almost 80% of participants active in the volunteer research registry between January 2012 and June 2015 were retained. Employment, being referred to research studies, a higher number of medical conditions, and more follow-up contacts were associated with an increased likelihood of retention. Older age, more months in the registry, and more mobility problems decreased the likelihood of retention. Discussion: These results suggest the Michigan Center for Urban African American Aging Research logic model promotes retention through involving older African American adults in research through study referrals and intensive follow-up. The loss of participants due to age- and mobility-related issues indicate the registry may be losing its most vulnerable participants. PMID:28138501

  14. Navagating the Common Core

    ERIC Educational Resources Information Center

    McShane, Michael Q.

    2014-01-01

    This article presents a debate over the Common Core State Standards Initiative as it has rocketed to the forefront of education policy discussions around the country. The author contends that there is value in having clear cross state standards that will clarify the new online and blended learning that the growing use of technology has provided…

  15. Ultrasonic Drilling and Coring

    NASA Technical Reports Server (NTRS)

    Bar-Cohen, Yoseph

    1998-01-01

    A novel drilling and coring device, driven by a combination, of sonic and ultrasonic vibration, was developed. The device is applicable to soft and hard objects using low axial load and potentially operational under extreme conditions. The device has numerous potential planetary applications. Significant potential for commercialization in construction, demining, drilling and medical technologies.

  16. NUCLEAR REACTOR CORE DESIGN

    DOEpatents

    Mahlmeister, J.E.; Peck, W.S.; Haberer, W.V.; Williams, A.C.

    1960-03-22

    An improved core design for a sodium-cooled, graphitemoderated nuclear reactor is described. The improved reactor core comprises a number of blocks of moderator material, each block being in the shape of a regular prism. A number of channels, extending the length of each block, are disposed around the periphery. When several blocks are placed in contact to form the reactor core, the channels in adjacent blocks correspond with each other to form closed conduits extending the length of the core. Fuel element clusters are disposed in these closed conduits, and liquid coolant is forced through the annulus between the fuel cluster and the inner surface of the conduit. In a preferred embodiment of the invention, the moderator blocks are in the form of hexagonal prisms with longitudinal channels cut into the corners of the hexagon. The main advantage of an "edge-loaded" moderator block is that fewer thermal neutrons are absorbed by the moderator cladding, as compared with a conventional centrally loaded moderator block.

  17. Authentic to the Core

    ERIC Educational Resources Information Center

    Kukral, Nicole; Spector, Stacy

    2012-01-01

    When educators think about what makes learning relevant to students, often they narrow their thinking to electives or career technical education. While these provide powerful opportunities for students to make relevant connections to their learning, they can also create authentic experiences in the core curriculum. In the San Juan Unified School…

  18. Electromagnetic pump stator core

    DOEpatents

    Fanning, Alan W.; Olich, Eugene E.; Dahl, Leslie R.

    1995-01-01

    A stator core for supporting an electrical coil includes a plurality of groups of circumferentially abutting flat laminations which collectively form a bore and perimeter. A plurality of wedges are interposed between the groups, with each wedge having an inner edge and a thicker outer edge. The wedge outer edges abut adjacent ones of the groups to provide a continuous path around the perimeter.

  19. Theory of core excitons

    SciTech Connect

    Dow, J. D.; Hjalmarson, H. P.; Sankey, O. F.; Allen, R. E.; Buettner, H.

    1980-01-01

    The observation of core excitons with binding energies much larger than those of the valence excitons in the same material has posed a long-standing theoretical problem. A proposed solution to this problem is presented, and Frenkel excitons and Wannier excitons are shown to coexist naturally in a single material. (GHT)

  20. Some Core Contested Concepts

    ERIC Educational Resources Information Center

    Chomsky, Noam

    2015-01-01

    Core concepts of language are highly contested. In some cases this is legitimate: real empirical and conceptual issues arise. In other cases, it seems that controversies are based on misunderstanding. A number of crucial cases are reviewed, and an approach to language is outlined that appears to have strong conceptual and empirical motivation, and…

  1. Core Directions in HRD.

    ERIC Educational Resources Information Center

    1996

    This document consists of four papers presented at a symposium on core directions in human resource development (HRD) moderated by Verna Willis at the 1996 conference of the Academy of Human Resource Development. "Reengineering the Organizational HRD Function: Two Case Studies" (Neal Chalofsky) reports an action research study in which…

  2. Core Geometry Manual.

    ERIC Educational Resources Information Center

    Hirata, Li Ann

    Core Geometry is a course offered in the Option Y sequence of the high school mathematics program described by the Hawaii State Department of Education's guidelines. The emphasis of this course is on the general awareness and use of the relationships among points, lines, and figures in planes and space. This sample course is based on the…

  3. Life from the core

    NASA Astrophysics Data System (ADS)

    Doglioni, Carlo; Coleman, Max; Pignatti, Johannes; Glassmeier, Karl-Heinz

    2010-05-01

    Life on Earth is the result of the chaotic combination of several independent chemical and physical parameters. One of them is the shield from ionizing radiation exerted by the atmosphere and the Earth's magnetic field. We hypothesise that the first few billion years of the Earth's history, dominated by bacteria, were characterized by stronger ionizing radiation. Bacteria can survive under such conditions better than any other organism. During the Archean and early Proterozoic the shield could have been weaker, allowing the development of only a limited number of species, more resistant to the external radiation. The Cambrian explosion of life could have been enhanced by the gradual growth of the solid inner core, which was not existent possibly before 1 Ga. The cooling of the Earth generated the solidification of the iron alloy in the center of the planet. As an hypothesis, before the crystallization of the core, the turbulence in the liquid core could have resulted in a lower or different magnetic field from the one we know today, being absent the relative rotation between inner and external core.

  4. The Earth's Core.

    ERIC Educational Resources Information Center

    Jeanloz, Raymond

    1983-01-01

    The nature of the earth's core is described. Indirect evidence (such as that determined from seismological data) indicates that it is an iron alloy, solid toward its center but otherwise liquid. Evidence also suggests that it is the turbulent flow of the liquid that generates the earth's magnetic field. (JN)

  5. Modeling Core Collapse Supernovae

    NASA Astrophysics Data System (ADS)

    Mezzacappa, Anthony

    2017-01-01

    Core collapse supernovae, or the death throes of massive stars, are general relativistic, neutrino-magneto-hydrodynamic events. The core collapse supernova mechanism is still not in hand, though key components have been illuminated, and the potential for multiple mechanisms for different progenitors exists. Core collapse supernovae are the single most important source of elements in the Universe, and serve other critical roles in galactic chemical and thermal evolution, the birth of neutron stars, pulsars, and stellar mass black holes, the production of a subclass of gamma-ray bursts, and as potential cosmic laboratories for fundamental nuclear and particle physics. Given this, the so called ``supernova problem'' is one of the most important unsolved problems in astrophysics. It has been fifty years since the first numerical simulations of core collapse supernovae were performed. Progress in the past decade, and especially within the past five years, has been exponential, yet much work remains. Spherically symmetric simulations over nearly four decades laid the foundation for this progress. Two-dimensional modeling that assumes axial symmetry is maturing. And three-dimensional modeling, while in its infancy, has begun in earnest. I will present some of the recent work from the ``Oak Ridge'' group, and will discuss this work in the context of the broader work by other researchers in the field. I will then point to future requirements and challenges. Connections with other experimental, observational, and theoretical efforts will be discussed, as well.

  6. University City Core Plan.

    ERIC Educational Resources Information Center

    Philadelphia City Planning Commission, PA.

    A redevelopment plan for an urban core area of about 300 acres was warranted by--(1) unsuitable building conditions, (2) undesirable land usage, and (3) faulty traffic circulation. The plan includes expansion of two universities and creation of a regional science center, high school, and medical center. Guidelines for proposed land use and zoning…

  7. The Tom Core Complex

    PubMed Central

    Ahting, Uwe; Thun, Clemens; Hegerl, Reiner; Typke, Dieter; Nargang, Frank E.; Neupert, Walter; Nussberger, Stephan

    1999-01-01

    Translocation of nuclear-encoded preproteins across the outer membrane of mitochondria is mediated by the multicomponent transmembrane TOM complex. We have isolated the TOM core complex of Neurospora crassa by removing the receptors Tom70 and Tom20 from the isolated TOM holo complex by treatment with the detergent dodecyl maltoside. It consists of Tom40, Tom22, and the small Tom components, Tom6 and Tom7. This core complex was also purified directly from mitochondria after solubilization with dodecyl maltoside. The TOM core complex has the characteristics of the general insertion pore; it contains high-conductance channels and binds preprotein in a targeting sequence-dependent manner. It forms a double ring structure that, in contrast to the holo complex, lacks the third density seen in the latter particles. Three-dimensional reconstruction by electron tomography exhibits two open pores traversing the complex with a diameter of ∼2.1 nm and a height of ∼7 nm. Tom40 is the key structural element of the TOM core complex. PMID:10579717

  8. Nucleosome Core Particle

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Nucleosome Core Particle grown on STS-81. The fundamental structural unit of chromatin and is the basis for organization within the genome by compaction of DNA within the nucleus of the cell and by making selected regions of chromosomes available for transcription and replication. Principal Investigator's are Dr. Dan Carter and Dr. Gerard Bunick of New Century Pharmaceuticals.

  9. From Context to Core

    ERIC Educational Resources Information Center

    Campus Technology, 2008

    2008-01-01

    At Campus Technology 2008, Arizona State University Technology Officer Adrian Sannier mesmerized audiences with his mandate to become more efficient by doing only the "core" tech stuff--and getting someone else to slog through the context. This article presents an excerpt from Sannier's hour-long keynote address at Campus Technology '08. Sannier…

  10. Dimethyl Disulfide Produced by the Naturally Associated Bacterium Bacillus sp B55 Promotes Nicotiana attenuata Growth by Enhancing Sulfur Nutrition[W

    PubMed Central

    Meldau, Dorothea G.; Meldau, Stefan; Hoang, Long H.; Underberg, Stefanie; Wünsche, Hendrik; Baldwin, Ian T.

    2013-01-01

    Bacillus sp B55, a bacterium naturally associated with Nicotiana attenuata roots, promotes growth and survival of wild-type and, particularly, ethylene (ET)–insensitive 35S-ethylene response1 (etr1) N. attenuata plants, which heterologously express the mutant Arabidopsis thaliana receptor ETR1-1. We found that the volatile organic compound (VOC) blend emitted by B55 promotes seedling growth, which is dominated by the S-containing compound dimethyl disulfide (DMDS). DMDS was depleted from the headspace during cocultivation with seedlings in bipartite Petri dishes, and 35S was assimilated from the bacterial VOC bouquet and incorporated into plant proteins. In wild-type and 35S-etr1 seedlings grown under different sulfate (SO4−2) supply conditions, exposure to synthetic DMDS led to genotype-dependent plant growth promotion effects. For the wild type, only S-starved seedlings benefited from DMDS exposure. By contrast, growth of 35S-etr1 seedlings, which we demonstrate to have an unregulated S metabolism, increased at all SO4−2 supply rates. Exposure to B55 VOCs and DMDS rescued many of the growth phenotypes exhibited by ET-insensitive plants, including the lack of root hairs, poor lateral root growth, and low chlorophyll content. DMDS supplementation significantly reduced the expression of S assimilation genes, as well as Met biosynthesis and recycling. We conclude that DMDS by B55 production is a plant growth promotion mechanism that likely enhances the availability of reduced S, which is particularly beneficial for wild-type plants growing in S-deficient soils and for 35S-etr1 plants due to their impaired S uptake/assimilation/metabolism. PMID:23903320

  11. Lunar Polar Coring Lander

    NASA Technical Reports Server (NTRS)

    Angell, David; Bealmear, David; Benarroche, Patrice; Henry, Alan; Hudson, Raymond; Rivellini, Tommaso; Tolmachoff, Alex

    1990-01-01

    Plans to build a lunar base are presently being studied with a number of considerations. One of the most important considerations is qualifying the presence of water on the Moon. The existence of water on the Moon implies that future lunar settlements may be able to use this resource to produce things such as drinking water and rocket fuel. Due to the very high cost of transporting these materials to the Moon, in situ production could save billions of dollars in operating costs of the lunar base. Scientists have suggested that the polar regions of the Moon may contain some amounts of water ice in the regolith. Six possible mission scenarios are suggested which would allow lunar polar soil samples to be collected for analysis. The options presented are: remote sensing satellite, two unmanned robotic lunar coring missions (one is a sample return and one is a data return only), two combined manned and robotic polar coring missions, and one fully manned core retrieval mission. One of the combined manned and robotic missions has been singled out for detailed analysis. This mission proposes sending at least three unmanned robotic landers to the lunar pole to take core samples as deep as 15 meters. Upon successful completion of the coring operations, a manned mission would be sent to retrieve the samples and perform extensive experiments of the polar region. Man's first step in returning to the Moon is recommended to investigate the issue of lunar polar water. The potential benefits of lunar water more than warrant sending either astronauts, robots or both to the Moon before any permanent facility is constructed.

  12. The Chemistry of Curcumin, the Health Promoting Ingredient in Turmeric

    ERIC Educational Resources Information Center

    Dewprashad, Brahmadeo

    2010-01-01

    Case studies pertaining to the health benefits of foods can be particularly effective in engaging students and in teaching core concepts in science (Heidemann and Urquart 2005). This case study focuses on the chemistry of curcumin, the health-promoting ingredient in turmeric. The case was developed to review core concepts in organic chemistry and…

  13. Promoting Mental Model Building in Astronomy Education

    ERIC Educational Resources Information Center

    Taylor, Ian; Barker, Miles; Jones, Alister

    2003-01-01

    While astronomy has recently re-emerged in many science curricula, there remain unresolved teaching and learning difficulties peculiar to astronomy education. This paper argues that mental model building, the core process in astronomy itself, should be reflected in astronomy education. Also, this crucial skill may promote a better understanding of…

  14. Application of Core Dynamics Modeling to Core-Mantle Interactions

    NASA Technical Reports Server (NTRS)

    Kuang, Weijia

    2003-01-01

    Observations have demonstrated that length of day (LOD) variation on decadal time scales results from exchange of axial angular momentum between the solid mantle and the core. There are in general four core-mantle interaction mechanisms that couple the core and the mantle. Of which, three have been suggested likely the dominant coupling mechanism for the decadal core-mantle angular momentum exchange, namely, gravitational core-mantle coupling arising from density anomalies in the mantle and in the core (including the inner core), the electromagnetic coupling arising from Lorentz force in the electrically conducting lower mantle (e.g. D-layer), and the topographic coupling arising from non-hydrostatic pressure acting on the core-mantle boundary (CMB) topography. In the past decades, most effort has been on estimating the coupling torques from surface geomagnetic observations (kinematic approach), which has provided insights on the core dynamical processes. In the meantime, it also creates questions and concerns on approximations in the studies that may invalidate the corresponding conclusions. The most serious problem is perhaps the approximations that are inconsistent with dynamical processes in the core, such as inconsistencies between the core surface flow beneath the CMB and the CMB topography, and that between the D-layer electric conductivity and the approximations on toroidal field at the CMB. These inconsistencies can only be addressed with numerical core dynamics modeling. In the past few years, we applied our MoSST (Modular, Scalable, Self-consistent and Three-dimensional) core dynamics model to study core-mantle interactions together with geodynamo simulation, aiming at assessing the effect of the dynamical inconsistencies in the kinematic studies on core-mantle coupling torques. We focus on topographic and electromagnetic core-mantle couplings and find that, for the topographic coupling, the consistency between the core flow and the CMB topography is

  15. Core Outlet Temperature Study

    SciTech Connect

    Moisseytsev, A.; Hoffman, E.; Majumdar, S.

    2008-07-28

    It is a known fact that the power conversion plant efficiency increases with elevation of the heat addition temperature. The higher efficiency means better utilization of the available resources such that higher output in terms of electricity production can be achieved for the same size and power of the reactor core or, alternatively, a lower power core could be used to produce the same electrical output. Since any nuclear power plant, such as the Advanced Burner Reactor, is ultimately built to produce electricity, a higher electrical output is always desirable. However, the benefits of the higher efficiency and electricity production usually come at a price. Both the benefits and the disadvantages of higher reactor outlet temperatures are analyzed in this work.

  16. Dynamics of core accretion

    DOE PAGES

    Nelson, Andrew F.; Ruffert, Maximilian

    2012-12-21

    In this paper, we perform three-dimensional hydrodynamic simulations of gas flowing around a planetary core of mass Mpl = 10M⊕ embedded in a near Keplerian background flow, using a modified shearing box approximation. We assume an ideal gas behaviour following an equation of state with a fixed ratio of the specific heats, γ = 1.42, consistent with the conditions of a moderate-temperature background disc with solar composition. No radiative heating or cooling is included in the models. We employ a nested grid hydrodynamic code implementing the ‘Piecewise Parabolic Method’ with as many as six fixed nested grids, providing spatial resolutionmore » on the finest grid comparable to the present-day diameters of Neptune and Uranus. We find that a strongly dynamically active flow develops such that no static envelope can form. The activity is not sensitive to plausible variations in the rotation curve of the underlying disc. It is sensitive to the thermodynamic treatment of the gas, as modelled by prescribed equations of state (either ‘locally isothermal’ or ‘locally isentropic’) and the temperature of the background disc material. The activity is also sensitive to the shape and depth of the core's gravitational potential, through its mass and gravitational softening coefficient. Each of these factors influences the magnitude and character of hydrodynamic feedback of the small-scale flow on the background, and we conclude that accurate modelling of such feedback is critical to a complete understanding of the core accretion process. The varying flow pattern gives rise to large, irregular eruptions of matter from the region around the core which return matter to the background flow: mass in the envelope at one time may not be found in the envelope at any later time. No net mass accretion into the envelope is observed over the course of the simulation and none is expected, due to our neglect of cooling. Except in cases of very rapid cooling however, as

  17. Dynamics of core accretion

    SciTech Connect

    Nelson, Andrew F.; Ruffert, Maximilian

    2012-12-21

    In this paper, we perform three-dimensional hydrodynamic simulations of gas flowing around a planetary core of mass Mpl = 10M embedded in a near Keplerian background flow, using a modified shearing box approximation. We assume an ideal gas behaviour following an equation of state with a fixed ratio of the specific heats, γ = 1.42, consistent with the conditions of a moderate-temperature background disc with solar composition. No radiative heating or cooling is included in the models. We employ a nested grid hydrodynamic code implementing the ‘Piecewise Parabolic Method’ with as many as six fixed nested grids, providing spatial resolution on the finest grid comparable to the present-day diameters of Neptune and Uranus. We find that a strongly dynamically active flow develops such that no static envelope can form. The activity is not sensitive to plausible variations in the rotation curve of the underlying disc. It is sensitive to the thermodynamic treatment of the gas, as modelled by prescribed equations of state (either ‘locally isothermal’ or ‘locally isentropic’) and the temperature of the background disc material. The activity is also sensitive to the shape and depth of the core's gravitational potential, through its mass and gravitational softening coefficient. Each of these factors influences the magnitude and character of hydrodynamic feedback of the small-scale flow on the background, and we conclude that accurate modelling of such feedback is critical to a complete understanding of the core accretion process. The varying flow pattern gives rise to large, irregular eruptions of matter from the region around the core which return matter to the background flow: mass in the envelope at one time may not be found in the envelope at any later time. No net mass accretion into the envelope is observed over the course of the simulation and none is expected, due to our neglect of cooling. Except in cases of very rapid cooling

  18. Long Valley Coring Project

    USGS Publications Warehouse

    Sass, John; Finger, John; McConnel, Vicki

    1998-01-01

    In December 1997, the California Energy Commission (CEC) agreed to provide funding for Phase III continued drilling of the Long Valley Exploratory Well (LVEW) near Mammoth Lakes, CA, from its present depth. The CEC contribution of $1 million completes a funding package of $2 million from a variety of sources, which will allow the well to be cored continuously to a depth of between 11,500 and 12,500 feet. The core recovered from Phase III will be crucial to understanding the origin and history of the hydrothermal systems responsible for the filling of fractures in the basement rock. The borehole may penetrate the metamorphic roof of the large magmatic complex that has fed the volcanism responsible for the caldera and subsequent activity.

  19. Geomagnetism of earth's core

    NASA Technical Reports Server (NTRS)

    Benton, E. R.

    1983-01-01

    Instrumentation, analytical methods, and research goals for understanding the behavior and source of geophysical magnetism are reviewed. Magsat, launched in 1979, collected global magnetometer data and identified the main terrestrial magnetic fields. The data has been treated by representing the curl-free field in terms of a scalar potential which is decomposed into a truncated series of spherical harmonics. Solutions to the Laplace equation then extend the field upward or downward from the measurement level through intervening spaces with no source. Further research is necessary on the interaction between harmonics of various spatial scales. Attempts are also being made to analytically model the main field and its secular variation at the core-mantle boundary. Work is also being done on characterizing the core structure, composition, thermodynamics, energetics, and formation, as well as designing a new Magsat or a tethered satellite to be flown on the Shuttle.

  20. Banded electromagnetic stator core

    DOEpatents

    Fanning, Alan W.; Gonzales, Aaron A.; Patel, Mahadeo R.; Olich, Eugene E.

    1996-01-01

    A stator core for an electromagnetic pump includes a plurality of circumferentially adjoining groups of flat laminations disposed about a common centerline axis and collectively defining a central bore and a discontinuous outer perimeter, with adjacent groups diverging radially outwardly to form V-shaped gaps. An annular band surrounds the groups and is predeterminedly tensioned to clamp together the laminations, and has a predetermined flexibility in a radial direction to form substantially straight bridge sections between the adjacent groups.

  1. Banded electromagnetic stator core

    DOEpatents

    Fanning, A.W.; Gonzales, A.A.; Patel, M.R.; Olich, E.E.

    1994-04-05

    A stator core for an electromagnetic pump includes a plurality of circumferentially adjoining groups of flat laminations disposed about a common centerline axis and collectively defining a central bore and a discontinuous outer perimeter, with adjacent groups diverging radially outwardly to form V-shaped gaps. An annular band surrounds the groups and is predeterminedly tensioned to clamp together the laminations, and has a predetermined flexibility in a radial direction to form substantially straight bridge sections between the adjacent groups. 5 figures.

  2. Banded electromagnetic stator core

    DOEpatents

    Fanning, Alan W.; Gonzales, Aaron A.; Patel, Mahadeo R.; Olich, Eugene E.

    1994-01-01

    A stator core for an electromagnetic pump includes a plurality of circumferentially adjoining groups of flat laminations disposed about a common centerline axis and collectively defining a central bore and a discontinuous outer perimeter, with adjacent groups diverging radially outwardly to form V-shaped gaps. An annular band surrounds the groups and is predeterminedly tensioned to clamp together the laminations, and has a predetermined flexibility in a radial direction to form substantially straight bridge sections between the adjacent groups.

  3. Banded electromagnetic stator core

    DOEpatents

    Fanning, A.W.; Gonzales, A.A.; Patel, M.R.; Olich, E.E.

    1996-06-11

    A stator core for an electromagnetic pump includes a plurality of circumferentially adjoining groups of flat laminations disposed about a common centerline axis and collectively defining a central bore and a discontinuous outer perimeter, with adjacent groups diverging radially outwardly to form V-shaped gaps. An annular band surrounds the groups and is predeterminedly tensioned to clamp together the laminations, and has a predetermined flexibility in a radial direction to form substantially straight bridge sections between the adjacent groups. 5 figs.

  4. Toroidal core winder

    DOEpatents

    Potthoff, Clifford M.

    1978-01-01

    The disclosure is directed to an apparatus for placing wire windings on a toroidal body, such as a transformer core, having an orifice in its center. The apparatus comprises a wire storage spool, a wire loop holding continuous belt maintained in a C-shaped loop by a belt supporting structure and provision for turning the belt to place and tighten loops of wire on a toroidal body, which is disposed within the gap of the C-shaped belt loop.

  5. Electromagnetic pump stator core

    DOEpatents

    Fanning, A.W.; Olich, E.E.; Dahl, L.R.

    1995-01-17

    A stator core for supporting an electrical coil includes a plurality of groups of circumferentially abutting flat laminations which collectively form a bore and perimeter. A plurality of wedges are interposed between the groups, with each wedge having an inner edge and a thicker outer edge. The wedge outer edges abut adjacent ones of the groups to provide a continuous path around the perimeter. 21 figures.

  6. Fissioning Plasma Core Reactor

    NASA Technical Reports Server (NTRS)

    Albright, Dennis; Butler, Carey; West, Nicole; Cole, John W. (Technical Monitor)

    2002-01-01

    Institute for Scientific Research, Inc. (ISR) research program consist of: 1.Study core physics by adapting existing codes: MCNP4C - Monte Carlo code; COMBINE/VENTURE - diffusion theory; SCALE4 - Monte Carlo, with many utility codes. 2. Determine feasibility and study major design parameters: fuel selection, temperature and reflector sizing. 3. Study reactor kinetics: develop QCALC1 to model point kinetics; study dynamic behavior of the power release.

  7. Variable depth core sampler

    DOEpatents

    Bourgeois, Peter M.; Reger, Robert J.

    1996-01-01

    A variable depth core sampler apparatus comprising a first circular hole saw member, having longitudinal sections that collapses to form a point and capture a sample, and a second circular hole saw member residing inside said first hole saw member to support the longitudinal sections of said first hole saw member and prevent them from collapsing to form a point. The second hole saw member may be raised and lowered inside said first hole saw member.

  8. Some core contested concepts.

    PubMed

    Chomsky, Noam

    2015-02-01

    Core concepts of language are highly contested. In some cases this is legitimate: real empirical and conceptual issues arise. In other cases, it seems that controversies are based on misunderstanding. A number of crucial cases are reviewed, and an approach to language is outlined that appears to have strong conceptual and empirical motivation, and to lead to conclusions about a number of significant issues that differ from some conventional beliefs.

  9. Origin of tumor-promoter released fibronectin in fibroblasts

    SciTech Connect

    Burrous, B.A.; Wolf, G.

    1986-05-01

    Previous work from the laboratory showed that the chemical tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated release of the cell surface glycoprotein, fibronectin (FN) from human lung fibroblasts (HLF), leading to depletion of cell surface FN, while FN synthesis is not altered by TPA. To further investigate the mechanism(s) by which TPA stimulates FN release, two types of experiments were performed. In the first, HLF were pulsed with /sup 35/S-methionine-labeled medium with or without TPA. In the second, cell-surface proteins were labeled by iodination (/sup 125/I) and then incubated in unlabeled medium with or without TPA. In both cases, the fate of labeled FN was followed over 12 hr. The /sup 35/S-meth-labeled HLF showed a rapid loss of labeled FN, first into a small, highly-labeled pool of cell surface FN (1 hr), later into the medium (4 hr or longer). Specific activities showed that this small pool in the cell surface turned over rapidly. TPA treatment resulted in more rapid movement of /sup 35/S-meth pulse-labeled FN to the cell surface and into the medium than in control cells. TPA thus affected the fate of intracellular FN. TPA treatment of HLF also resulted in more rapid removal of /sup 125/I-cell surface-labeled FN into the medium than in control cells. Thus, TPA affects the fate of preexisting cell surface FN in HLF. From these results, they hypothesize that TPA has two separate effects: it stimulates depletion of preexisting intracellular FN during the first hr of treatment, and it stimulates release of preexisting cell surface FN over all treatment times.

  10. Core-collapse Supernovae

    SciTech Connect

    Hix, William Raphael; Lentz, E. J.; Baird, Mark L; Chertkow, Merek A; Lee, Ching-Tsai; Blondin, J. M.; Bruenn, S. W.; Messer, Bronson; Mezzacappa, Anthony

    2013-01-01

    Marking the inevitable death of a massive star, and the birth of a neutron star or black hole, core-collapse supernovae bring together physics at a wide range in spatial scales, from kilometer-sized hydrodynamic motions (growing to gigameter scale) down to femtometer scale nuclear reactions. Carrying 10$^{51}$ ergs of kinetic energy and a rich-mix of newly synthesized atomic nuclei, core-collapse supernovae are the preeminent foundries of the nuclear species which make up ourselves and our solar system. We will discuss our emerging understanding of the convectively unstable, neutrino-driven explosion mechanism, based on increasingly realistic neutrino-radiation hydrodynamic simulations that include progressively better nuclear and particle physics. Recent multi-dimensional models with spectral neutrino transport from several research groups, which slowly develop successful explosions for a range of progenitors, have motivated changes in our understanding of the neutrino reheating mechanism. In a similar fashion, improvements in nuclear physics, most notably explorations of weak interactions on nuclei and the nuclear equation of state, continue to refine our understanding of how supernovae explode. Recent progress on both the macroscopic and microscopic effects that affect core-collapse supernovae are discussed.

  11. GEOS-CORE

    SciTech Connect

    2014-06-24

    GEOS-CORE is a code that integrates open source Libraries for linear algebra and I/O with two main LLNL-written components: (i) a set of standard finite, discrete, and discontinuous displacement element physics solvers for resolving Darcy fluid flow, explicit mechanics, implicit mechanics, and fluid-mediated fracturing, including resolution of physical behaviors both implicitly and explicitly, and (ii) a MPI-based parallelization implementation for use on generic HPC distributed memory architectures. The resultant code can be used alone for linearly elastic and quasistatic damage problems; problems involving hydraulic fracturing, where the mesh topology is dynamically changed; and general granular materials behavior. The key application domain is for low-rate stimulation and fracture control in subsurface reservoirs (e.g., enhanced geothermal sites and unconventional shale gas stimulation). GEOS-CORE also has interfaces to call external libraries for, e.g., material models and equations fo state; however, LLNL-developed EOS and material models, beyond the aforementioned linear elastic and quasi-static damage models, will not be part of the current release. GEOS-CORE's secondary applications include granular materials behavior under different load paths.

  12. Identification of a 467 bp Promoter of Maize Phosphatidylinositol Synthase Gene (ZmPIS) Which Confers High-Level Gene Expression and Salinity or Osmotic Stress Inducibility in Transgenic Tobacco

    PubMed Central

    Zhang, Hongli; Hou, Jiajia; Jiang, Pingping; Qi, Shoumei; Xu, Changzheng; He, Qiuxia; Ding, Zhaohua; Wang, Zhiwu; Zhang, Kewei; Li, Kunpeng

    2016-01-01

    Salinity and drought often affect plant growth and crop yields. Cloning and identification of salinity and drought stress inducible promoters is of great significance for their use in the genetic improvement of crop resistance. Previous studies showed that phosphatidylinositol synthase is involved in plant salinity and drought stress responses but its promoter has not been characterized by far. In the study, the promoter (pZmPIS, 1834 bp upstream region of the translation initiation site) was isolated from maize genome. To functionally validate the promoter, eight 5′ deletion fragments of pZmPIS in different lengths were fused to GUS to produce pZmPIS::GUS constructs and transformed into tobacco, namely PZ1–PZ8. The transcription activity and expression pattern obviously changed when the promoter was truncated. Previous studies have demonstrated that NaCl and PEG treatments are usually used to simulate salinity and drought treatments. The results showed that PZ1–PZ7 can respond well upon NaCl and PEG treatments, while PZ8 not. PZ7 (467 bp) displayed the highest transcription activity in all tissues of transgenic tobacco amongst 5′ deleted promoter fragments, which corresponds to about 20 and 50% of CaMV35S under normal and NaCl or PEG treatment, respectively. This implied that PZ7 is the core region of pZmPIS which confers high-level gene expression and NaCl or PEG inducible nature. The 113 bp segment between PZ7 and PZ8 (-467 to -355 bp) was considered as the key sequence for ZmPIS responding to NaCl or PEG treatment. GUS transient assay in tobacco leaves showed that this segment was sufficient for the NaCl or PEG stress response. Bioinformatic analysis revealed that the 113 bp sequence may contain new elements that are crucial for ZmPIS response to NaCl or PEG stress. These results promote our understanding on transcriptional regulation mechanism of ZmPIS and the characterized PZ7 promoter fragment would be an ideal candidate for the overexpression of

  13. 33. BENCH CORE STATION, GREY IRON FOUNDRY CORE ROOM WHERE ...

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

    33. BENCH CORE STATION, GREY IRON FOUNDRY CORE ROOM WHERE CORE MOLDS WERE HAND FILLED AND OFTEN PNEUMATICALLY COMPRESSED WITH A HAND-HELD RAMMER BEFORE THEY WERE BAKED. - Stockham Pipe & Fittings Company, Grey Iron Foundry, 4000 Tenth Avenue North, Birmingham, Jefferson County, AL

  14. Condition for the formation of micron-sized dust grains in dense molecular cloud cores

    NASA Astrophysics Data System (ADS)

    Hirashita, Hiroyuki; Li, Zhi-Yun

    2013-07-01

    We investigate the condition for the formation of micron-sized grains in dense cores of molecular clouds. This is motivated by the detection of mid-infrared emission from deep inside a number of dense cores, the so-called `coreshine,' which is thought to come from scattering by micron (μm)-sized grains. Based on numerical calculations of coagulation starting from the typical grain-size distribution in the diffuse interstellar medium, we obtain a conservative lower limit to the time t to form μm-sized grains: t/tff > 3(5/S)(nH/105 cm-3)-1/4 (where tff is the free-fall time at hydrogen number density nH in the core and S the enhancement factor of the grain-grain collision cross-section to account for non-compact aggregates). At the typical core density nH = 105 cm-3, it takes at least a few free-fall times to form the μm-sized grains responsible for coreshine. The implication is that those dense cores observed in coreshine are relatively long-lived entities in molecular clouds, rather than dynamically transient objects that last for one free-fall time or less.

  15. Analysis of nematode-responsive promoters in sugar beet hairy roots.

    PubMed

    Van Poucke, K; Karimi, M; Gheysen, G

    2001-01-01

    One of the strategies to make crops resistant to the beet cyst nematode Heterodera schachtii is the destruction of the feeding site or syncytium. This can be achieved by local expression of the cytotoxic barnase gene under control of a nematode-inducible plant promoter that is active in the syncytium. Expression of barnase outside the feeding site has to be neutralized by its inhibitor barstar driven from a constitutive promoter that is downregulated in the syncytium. Several promoters that are upregulated in feeding structures were identified using the promoter tagging strategy in Arabidopsis thaliana (Barthels et al., 1997) or by differential cDNA screening in tomato (Van der Eycken et al., 1996). Nematode downregulated promoters in Arabidopsis were described by Goddijn et al. (1993). Five nematode-induced promoters (ARM1, 1164, 728, 25 and Lemmi9) and four downregulated promoters (CaMV35S, the nopaline synthase promoter (nos) and the rooting loci promoters RolC and RolD) fused to the beta-glucuronidase (gus) reporter gene were introduced into sugar beet hairy roots by transformation with Agrobacterium rhizogenes to evaluate their expression pattern. All upregulated promoters were found to be active at the base of lateral roots. The 728 and 25 promoter were as well active in root tips. In the 25-gus lines GUS could also be detected in the vascular tissue, while the ARM1 promoter was also active in sugar beet callus. The Lemmi9 promoter and the 4 constitutive promoters were active in the entire root. The transgenic hairy roots were inoculated with Heterodera schachtii and at different time-points (4, 8, 15, 22 days after inoculation; dpi) GUS analysis was performed on the infected roots. For the ARM1, 1164 and 728 promoter the highest gus expression level in syncytia was observed at 8 dpi. In 4 days old syncytia of the 25-gus lines the intensity of the GUS signal was of the same extent as the non-specific vascular signal. In later stages it even disappeared from

  16. Selenium semiconductor core optical fibers

    SciTech Connect

    Tang, G. W.; Qian, Q. Peng, K. L.; Wen, X.; Zhou, G. X.; Sun, M.; Chen, X. D.; Yang, Z. M.

    2015-02-15

    Phosphate glass-clad optical fibers containing selenium (Se) semiconductor core were fabricated using a molten core method. The cores were found to be amorphous as evidenced by X-ray diffraction and corroborated by Micro-Raman spectrum. Elemental analysis across the core/clad interface suggests that there is some diffusion of about 3 wt % oxygen in the core region. Phosphate glass-clad crystalline selenium core optical fibers were obtained by a postdrawing annealing process. A two-cm-long crystalline selenium semiconductor core optical fibers, electrically contacted to external circuitry through the fiber end facets, exhibit a three times change in conductivity between dark and illuminated states. Such crystalline selenium semiconductor core optical fibers have promising utility in optical switch and photoconductivity of optical fiber array.

  17. Hollow-Core Fiber Lamp

    NASA Technical Reports Server (NTRS)

    Yi, Lin (Inventor); Tjoelker, Robert L. (Inventor); Burt, Eric A. (Inventor); Huang, Shouhua (Inventor)

    2016-01-01

    Hollow-core capillary discharge lamps on the millimeter or sub-millimeter scale are provided. The hollow-core capillary discharge lamps achieve an increased light intensity ratio between 194 millimeters (useful) and 254 millimeters (useless) light than conventional lamps. The capillary discharge lamps may include a cone to increase light output. Hollow-core photonic crystal fiber (HCPCF) may also be used.

  18. Sneak in Some Core Subjects

    ERIC Educational Resources Information Center

    Clarke, Lynne

    2011-01-01

    Even if students don't have an aversion to core subjects, they may not see the relationship between the core subjects and their career path. In this article, the author outlines a career path project that can be adapted to work in any career and technical education (CTE) class to highlight the relationship between core subjects and the real world.…

  19. Faculty Supports Communication Core Courses.

    ERIC Educational Resources Information Center

    Kopenhaver, Lillian Lodge; And Others

    1989-01-01

    Asks public relations educators what they think about core classes required for students in their field. Finds they generally support the idea that their students should take core mass communications courses, even if such core courses are developed from a traditional journalism/news-editorial standpoint. (MS)

  20. Mercury's inner core size and core-crystallization regime

    NASA Astrophysics Data System (ADS)

    Dumberry, Mathieu; Rivoldini, Attilio

    2015-03-01

    Earth-based radar observation of Mercury's rotation vector combined with gravity observation by the MESSENGER spacecraft yield a measure of Mercury's moment of inertia and the amplitude of the 88-day libration of its silicate shell. These two geodetic constraints provide information on Mercury's interior structure, including the presence of a fluid core, the radius of the core-mantle boundary and the bulk densities of the core and mantle. In this work, we show how they further provide information on the size of the solid inner core and on the crystallization regime of the fluid core. If Mercury's fluid core is a Fe-FeS alloy with a sulfur concentration on the Fe-rich side of the eutectic, the largest inner core compatible with geodetic observations at the 1σ level is 1325 ± 250 km. Our results further suggest that the crystallization scenario that best fits the geodetic observations involves the formation of Fe-snow within the fluid core, and that this scenario is preferred for models with an iron-poor mantle composition. Consequently, Mercury's dynamo most likely operates in concert with snow formation. For an inner core larger than ∼650 km, snow formation extends to the inner core boundary. If a dynamo cannot be maintained by the dynamics of snow formation, or if such dynamo produces a magnetic field incompatible with observation, Mercury's inner core must then be smaller than 650 km.

  1. Integrative Biology: A Capstone Course for an Introductory Biology Core

    ERIC Educational Resources Information Center

    Chaplin, Susan B.; Hartung, Nancy Z.

    2012-01-01

    A capstone to the biology introductory curriculum was developed with the specific goals of enhancing integration of course content, promoting development of oral presentation skills and critical reading and thinking skills, and introducing ecological principles omitted from the rest of the core. Classes of 12 to 16 students were team taught by…

  2. Shakespeare and the Common Core: An Opportunity to Reboot

    ERIC Educational Resources Information Center

    Turchi, Laura; Thompson, Ayanna

    2013-01-01

    The Common Core generally eschews mandating texts in favor of promoting critical analysis and rigor. So it's significant that Shakespeare is the only author invoked in imperatives. His explicit inclusion offers a significant opportunity for educators to rethink how we approach Shakespearean instruction. Rather than the traditional learning of…

  3. Connecting the "Missing Words" to the Common Core

    ERIC Educational Resources Information Center

    Wilburne, Jane M.; Kulbacki, Ashley

    2014-01-01

    This article describes how a sixth-grade teacher's "missing word" task uncovered higher-level thinking and engaged her students in the Standards for Mathematical Practice. The role and selection of the task promotes higher-level thinking and connects to the Common Core State Standards for Mathematics. The task required students to apply…

  4. 85+ Ways to Implement Home Economics I, Basic Core.

    ERIC Educational Resources Information Center

    Wilson, Wanda; Sawatzky, Joyce

    Designed to help build basic homemaking skills, this core curriculum instructional package, which was teacher-developed in Oklahoma, is a collection of teaching ideas to promote motivation and learning in the classroom. Activities are included for the following subjects: (1) orientation to Future Homemakers of America; (2) occupational…

  5. CANOPEN Controller IP Core

    NASA Astrophysics Data System (ADS)

    Caramia, Maurizio; Montagna, Mario; Furano, Gianluca; Winton, Alistair

    2010-08-01

    This paper will describe the activities performed by Thales Alenia Space Italia supported by the European Space Agency in the definition of a CAN bus interface to be used on Exomars. The final goal of this activity is the development of an IP core, to be used in a slave node, able to manage both the CAN bus Data Link and Application Layer totally in hardware. The activity has been focused on the needs of the EXOMARS mission where devices with different computational performances are all managed by the onboard computer through the CAN bus.

  6. Automated Core Design

    SciTech Connect

    Kobayashi, Yoko; Aiyoshi, Eitaro

    2005-07-15

    Multistate searching methods are a subfield of distributed artificial intelligence that aims to provide both principles for construction of complex systems involving multiple states and mechanisms for coordination of independent agents' actions. This paper proposes a multistate searching algorithm with reinforcement learning for the automatic core design of a boiling water reactor. The characteristics of this algorithm are that the coupling structure and the coupling operation suitable for the assigned problem are assumed and an optimal solution is obtained by mutual interference in multistate transitions using multiagents. Calculations in an actual plant confirmed that the proposed algorithm increased the convergence ability of the optimization process.

  7. The development and characterization of synthetic minimal yeast promoters

    PubMed Central

    Redden, Heidi; Alper, Hal S.

    2015-01-01

    Synthetic promoters, especially minimally sized, are critical for advancing fungal synthetic biology. Fungal promoters often span hundreds of base pairs, nearly ten times the amount of bacterial counterparts. This size limits large-scale synthetic biology efforts in yeasts. Here we address this shortcoming by establishing a methodical workflow necessary to identify robust minimal core elements that can be linked with minimal upstream activating sequences to develop short, yet strong yeast promoters. Through a series of library-based synthesis, analysis and robustness tests, we create a set of non-homologous, purely synthetic, minimal promoters for yeast. These promoters are comprised of short core elements that are generic and interoperable and 10 bp UAS elements that impart strong, constitutive function. Through this methodology, we are able to generate the shortest fungal promoters to date, which can achieve high levels of both inducible and constitutive expression with up to an 80% reduction in size. PMID:26183606

  8. Characterization of a Maize Wip1 Promoter in Transgenic Plants

    PubMed Central

    Zhang, Shengxue; Lian, Yun; Liu, Yan; Wang, Xiaoqing; Liu, Yunjun; Wang, Guoying

    2013-01-01

    The Maize Wip1 gene encodes a wound-induced Bowman-Birk inhibitor (BBI) protein which is a type of serine protease inhibitor, and its expression is induced by wounding or infection, conferring resistance against pathogens and pests. In this study, the maize Wip1 promoter was isolated and its function was analyzed. Different truncated Wip1 promoters were fused upstream of the GUS reporter gene and transformed into Arabidopsis, tobacco and rice plants. We found that (1) several truncated maize Wip1 promoters led to strong GUS activities in both transgenic Arabidopsis and tobacco leaves, whereas low GUS activity was detected in transgenic rice leaves; (2) the Wip1 promoter was not wound-induced in transgenic tobacco leaves, but was induced by wounding in transgenic rice leaves; (3) the truncated Wip1 promoter had different activity in different organs of transgenic tobacco plants; (4) the transgenic plant leaves containing different truncated Wip1 promoters had low GUS transcripts, even though high GUS protein level and GUS activities were observed; (5) there was one transcription start site of Wip1 gene in maize and two transcription start sites of GUS in Wip1::GUS transgenic lines; (6) the adjacent 35S promoter which is present in the transformation vectors enhanced the activity of the truncated Wip1 promoters in transgenic tobacco leaves, but did not influence the disability of truncated Wip1231 promoter to respond to wounding signals. We speculate that an ACAAAA hexamer, several CAA trimers and several elements similar to ACAATTAC octamer in the 5′-untranslated region might contribute to the strong GUS activity in Wip1231 transgenic lines, meanwhile, compared to the 5′-untranslated region from Wip1231 transgenic lines, the additional upstream open reading frames (uORFs) in the 5′-untranslated region from Wip1737 transgenic lines might contribute to the lower level of GUS transcript and GUS activity. PMID:24322445

  9. Metrics for Success: Strategies for Enabling Core Facility Performance and Assessing Outcomes.

    PubMed

    Turpen, Paula B; Hockberger, Philip E; Meyn, Susan M; Nicklin, Connie; Tabarini, Diane; Auger, Julie A

    2016-04-01

    Core Facilities are key elements in the research portfolio of academic and private research institutions. Administrators overseeing core facilities (core administrators) require assessment tools for evaluating the need and effectiveness of these facilities at their institutions. This article discusses ways to promote best practices in core facilities as well as ways to evaluate their performance across 8 of the following categories: general management, research and technical staff, financial management, customer base and satisfaction, resource management, communications, institutional impact, and strategic planning. For each category, we provide lessons learned that we believe contribute to the effective and efficient overall management of core facilities. If done well, we believe that encouraging best practices and evaluating performance in core facilities will demonstrate and reinforce the importance of core facilities in the research and educational mission of institutions. It will also increase job satisfaction of those working in core facilities and improve the likelihood of sustainability of both facilities and personnel.

  10. Metrics for Success: Strategies for Enabling Core Facility Performance and Assessing Outcomes

    PubMed Central

    Hockberger, Philip E.; Meyn, Susan M.; Nicklin, Connie; Tabarini, Diane; Auger, Julie A.

    2016-01-01

    Core Facilities are key elements in the research portfolio of academic and private research institutions. Administrators overseeing core facilities (core administrators) require assessment tools for evaluating the need and effectiveness of these facilities at their institutions. This article discusses ways to promote best practices in core facilities as well as ways to evaluate their performance across 8 of the following categories: general management, research and technical staff, financial management, customer base and satisfaction, resource management, communications, institutional impact, and strategic planning. For each category, we provide lessons learned that we believe contribute to the effective and efficient overall management of core facilities. If done well, we believe that encouraging best practices and evaluating performance in core facilities will demonstrate and reinforce the importance of core facilities in the research and educational mission of institutions. It will also increase job satisfaction of those working in core facilities and improve the likelihood of sustainability of both facilities and personnel. PMID:26848284

  11. PROCESS FOR JACKETING A CORE

    DOEpatents

    Last, G.A.

    1960-07-19

    A process is given for enclosing the uranium core of a nuclear fuel element by placing the core in an aluminum cup and closing the open end of the cup over the core. As the metal of the cup is brought together in a weld over the center of the end of the core, it is extruded inwardly as internal projection into a central recess in the core and outwardly as an external projection. Thus oxide inclusions in the weld of the cup are spread out into the internal and external projections and do not interfere with the integrity of the weld.

  12. An alternative method of promoter assessment by confocal laser scanning microscopy.

    PubMed

    Sahoo, Dipak K; Ranjan, Rajiv; Kumar, Deepak; Kumar, Alok; Sahoo, Bhabani S; Raha, Sumita; Maiti, Indu B; Dey, Nrisingha

    2009-10-01

    A rapid and useful method of promoter activity analysis using techniques of confocal laser scanning microscopy (CLSM) is described in the present study. The activities of some pararetroviral promoters such as CaMV35S (Cauliflower mosaic virus), FMVSgt3 (Figwort mosaic virus sub-genomic transcript) and MMVFLt12 (Mirabilis mosaic virus full-length transcript) coupled to GFP (green fluorescent protein) and GUS (beta-glucuronidase) reporter genes were determined simultaneously by the CLSM technique and other available conventional methods for reporter gene assay based on relevant biochemical and molecular approaches. Consistent and comparable results obtained by CLSM as well as by other conventional assay methods confirm the effectiveness of the CLSM approach for assessment of promoter activity. Hence the CLSM method can be suggested as an alternative way for promoter analysis on the basis of high throughput.

  13. High level transgenic expression of soybean (Glycine max) GmERF and Gmubi gene promoters isolated by a novel promoter analysis pipeline

    PubMed Central

    2010-01-01

    Background Although numerous factors can influence gene expression, promoters are perhaps the most important component of the regulatory control process. Promoter regions are often defined as a region upstream of the transcriptional start. They contain regulatory elements that interact with regulatory proteins to modulate gene expression. Most genes possess their own unique promoter and large numbers of promoters are therefore available for study. Unfortunately, relatively few promoters have been isolated and characterized; particularly from soybean (Glycine max). Results In this research, a bioinformatics approach was first performed to identify members of the Gmubi (G.max ubiquitin) and the GmERF (G. max Ethylene Response Factor) gene families of soybean. Ten Gmubi and ten GmERF promoters from selected genes were cloned upstream of the gfp gene and successfully characterized using rapid validation tools developed for both transient and stable expression. Quantification of promoter strength using transient expression in lima bean (Phaseolus lunatus) cotyledonary tissue and stable expression in soybean hairy roots showed that the intensity of gfp gene expression was mostly conserved across the two expression systems. Seven of the ten Gmubi promoters yielded from 2- to 7-fold higher expression than a standard CaMV35S promoter while four of the ten GmERF promoters showed from 1.5- to 2.2-times higher GFP levels compared to the CaMV35S promoter. Quantification of GFP expression in stably-transformed hairy roots of soybean was variable among roots derived from different transformation events but consistent among secondary roots, derived from the same primary transformation events. Molecular analysis of hairy root events revealed a direct relationship between copy number and expression intensity; higher copy number events displayed higher GFP expression. Conclusion In this study, we present expression intensity data on 20 novel soybean promoters from two different gene

  14. Functional characterization of the pollen-specific SBgLR promoter from potato (Solanum tuberosum L.).

    PubMed

    Lang, Zhihong; Zhou, Peng; Yu, Jingjuan; Ao, Guangming; Zhao, Qian

    2008-01-01

    SBgLR (Solanum tuberosum genomic lysine-rich) gene was isolated from a potato genomic library using SB401 (S. berthaultii 401) cDNA as probe. RT-PCR analysis of SBgLR gene expression profile and microscopic analysis of green fluorescent protein (GFP) expression in tobacco plants transformed with SBgLR promoter-GFP reporters indicate that SBgLR is a pollen-specific gene. A series of 5'deletions of SBgLR promoter were fused to the beta-glucuronidase (GUS) gene and stably introduced into tobacco plants. Histochemical and quantitative assays of GUS expression in transgenic plants allowed us to localize an enhancer of SBgLR promoter to the region -345 to -269 relative to the translation start site. This 76 bp (-345 to -269) fragment enhanced GUS expression in leaves, stems and roots when fused to -90/+6 CaMV 35S minimal promoter. Deletion analysis showed that a cis-element, which can repress gene expression in root hairs, was located in the region -345 to -311. Further study indicated that the -269 to -9 region was sufficient to confer pollen-specific expression of GFP when fused to CaMV 35S enhancer.

  15. The Potential of Digital Technologies to Support Literacy Instruction Relevant to the Common Core State Standards

    ERIC Educational Resources Information Center

    Hutchison, Amy C.; Colwell, Jamie

    2014-01-01

    Digital tools have the potential to transform instruction and promote literacies outlined in the Common Core State Standards. Empirical research is examined to illustrate this potential in grades 6-12 instruction.

  16. Plasma core reactor applications

    NASA Technical Reports Server (NTRS)

    Latham, T. S.; Rodgers, R. J.

    1976-01-01

    Analytical and experimental investigations were conducted to demonstrate the feasibility of fissioning uranium plasma core reactors and to characterize space and terrestrial applications for such reactors. Uranium hexafluoride fuel is injected into core cavities and confined away from the surface by argon buffer gas injected tangentially from the peripheral walls. Radiant heat transfer calculations were performed for a six-cavity reactor configuration. Axial working fluid channels are located along a fraction of each cavity peripheral wall. Results of calculations for outward-directed radiant energy fluxes corresponding to radiating temperatures of 2000 to 5000 K indicate total operating pressures from 80 to 650 atm, centerline temperatures from 6900 to 30,000 K, and total radiated powers from 25 to 2500 MW, respectively. Applications are described for this type of reactor such as (1) high-thrust, high specific impulse space propulsion, (2) highly efficient systems for generation of electricity, and (3) hydrogen or synthetic fuel production systems using the intense radiant energy fluxes.

  17. NEUTRONIC REACTOR CORE

    DOEpatents

    Thomson, W.B.; Corbin, A. Jr.

    1961-07-18

    An improved core for a gas-cooled power reactor which admits gas coolant at high temperatures while affording strong integral supporting structure and efficient moderation of neutrons is described. The multiplicities of fuel elements constituting the critical amassment of fissionable material are supported and confined by a matrix of metallic structure which is interspersed therebetween. Thermal insulation is interposed between substantially all of the metallic matrix and the fuel elements; the insulation then defines the principal conduit system for conducting the coolant gas in heat-transfer relationship with the fuel elements. The metallic matrix itseif comprises a system of ducts through which an externally-cooled hydrogeneous liquid, such as water, is circulated to serve as the principal neutron moderant for the core and conjointly as the principal coolant for the insulated metallic structure. In this way, use of substantially neutron transparent metals, such as aluminum, becomes possible for the supporting structure, despite the high temperatures of the proximate gas. The Aircraft Nuclear Propulsion program's "R-1" reactor design is a preferred embodiment.

  18. Core-tube data logger

    SciTech Connect

    Henfling, J.A.; Normann, R.A.; Knudsen, S.; Drumheller, D.

    1997-01-01

    Wireline core drilling, increasingly used for geothermal exploration, employs a core-tube to capture a rock core sample during drilling. Three types of core-tube data loggers (CTDL) have been built and tested to date by Sandia national Laboratories. They are: (1) temperature-only logger, (2) temperature/inclinometer logger and (3) heat-shielded temperature/inclinometer logger. All were tested during core drilling operations using standard wireline diamond core drilling equipment. While these tools are designed for core-tube deployment, the tool lends itself to be adapted to other drilling modes and equipment. Topics covered in this paper include: (1) description on how the CTDLs are implemented, (2) the components of the system, (3) the type of data one can expect from this type of tool, (4) lessons learned, (5) comparison to its counterpart and (6) future work.

  19. Models of the earth's core

    NASA Technical Reports Server (NTRS)

    Stevenson, D. J.

    1981-01-01

    Combined inferences from seismology, high-pressure experiment and theory, geomagnetism, fluid dynamics, and current views of terrestrial planetary evolution lead to models of the earth's core with five basic properties. These are that core formation was contemporaneous with earth accretion; the core is not in chemical equilibrium with the mantle; the outer core is a fluid iron alloy containing significant quantities of lighter elements and is probably almost adiabatic and compositionally uniform; the more iron-rich inner solid core is a consequence of partial freezing of the outer core, and the energy release from this process sustains the earth's magnetic field; and the thermodynamic properties of the core are well constrained by the application of liquid-state theory to seismic and labroatory data.

  20. Models of the Earth's Core.

    PubMed

    Stevenson, D J

    1981-11-06

    Combined inferences from seismology, high-pressure experiment and theory, geomagnetism, fluid dynamics, and current views of terrestrial planetary evolution lead to models of the earth's core with the following properties. Core formation was contemporaneous with earth accretion; the core is not in chemical equilibrium with the mantle; the outer core is a fluid iron alloy containing significant quantities of lighter elements and is probably almost adiabatic and compositionally uniform; the more iron-rich inner solid core is a consequence of partial freezing of the outer core, and the energy release from this process sustains the earth's magnetic field; and the thermodynamic properties of the core are well constrained by the application of liquid-state theory to seismic and laboratory data.

  1. Heterologous activation of the Porphyra tenera HSP70 promoter in Bangiophycean algal cells.

    PubMed

    Hirata, Ryo; Jeong, Won-Joong; Saga, Naotsune; Mikami, Koji

    2011-01-01

    Porphyra has attracted great attention for its biological and industrial importance. However, establishment of a stable nuclear transformation has not yet been achieved in these organisms, which impedes the molecular biological study and the development of a molecular breeding method for them. Toward establishing the stable transformation, we have recently developed an efficient transient gene expression system in Bangiophycean algae, in which the HSP70 promoter from P. tenera (PtHSP70 promoter) was activated heterologously in P. yezoensis cells. Since heterologous promoters are required for homologous recombination-based stable transformation, the identification of heterologously activated promoters is important in establishing a stable transformation system in individual Bangiophycean alga. We here examined the activation of the PtHSP70 promoter using the GC-rich PyGUS reporter system in additional Porphyra and Bangia species. The results indicated that this promoter drove expression of the PyGUS gene efficiently in all examined algae, whereas there was quite low expression of PyGUS by the cauliflower mosaic virus 35S promoter that is widely used as a heterologous promoter in the transformation of green land plants. Therefore, heterologous activation of the PtHSP70 promoter could promote the establishment of the stable transformation system in various kinds of Bangiophycean algae.

  2. Expression of wheat expansin driven by the RD29 promoter in tobacco confers water-stress tolerance without impacting growth and development.

    PubMed

    Li, Feng; Han, Yangyang; Feng, Yanan; Xing, Shichao; Zhao, Meirong; Chen, Yanhui; Wang, Wei

    2013-02-10

    Expansins are the key regulators of cell wall extension during plant growth. Previously, we produced transgenic tobacco plants with increased tolerance to water stress by overexpressing the wheat expansin gene TaEXPB23 driven by the constitutive 35S cauliflower mosaic virus (CaMV) promoter. However, the growth and development of 35S::TaEXPB23 transgenic tobacco plants were altered under normal growth conditions, with a faster growth rate at the seedling stage, earlier flowering and maturation, and a shorter plant height compared to WT. In the current study, we determined that cellular characteristics and carbohydrate metabolism were altered in 35S::TaEXPB23 transgenic tobacco plants. We also generated transgenic Arabidopsis plants using the same vector. The transgenic Arabidopsis plants had the same phenotype as the transgenic tobacco plants, which may have resulted from the altered expression of several flowering-related genes. We then produced TaEXPB23 transgenic tobacco plants using the stress-inducible RD29A promoter. The use of this promoter reduced the negative effects of TaEXPB23 on plant growth and development. The RD29A::TaEXPB23 transgenic tobacco plants had greater tolerance to water stress than WT, as determined by examining physiological and biochemical parameters. Therefore, the use of stress-inducible promoters, such as RD29A, may minimize the negative effects of constitutive transgene expression and improve the water-stress tolerance of plants.

  3. A negative element in the downstream region of the Rice tungro bacilliform virus promoter is orientation- and position-independent and is active with heterologous promoters.

    PubMed

    Purkayastha, Arunima; Sharma, Shweta; Dasgupta, Indranil

    2010-10-01

    The promoter of an Indian isolate of the pararetrovirus Rice tungro bacilliform virus (RTBV-WB) contains a negative element downstream of the transcription start site (TSS), between nucleotide residues +58 and +195 (Mathur and Dasgupta, 2007). To further characterize the element, we show, by using transient gus reporter gene assays in the cells of onion peel, rice calli and Arabidopsis leaves, that it down-regulates heterologous promoters CaMV35S and Maize ubiquitin. Quantitative measurements of transient GUS activity indicated more than 90% inhibition of reporter gene expression by the negative element. We also show, by reversing the orientation of the element downstream and by placing it in a position upstream to a constitutively expressing RTBV promoter, that the negative element is orientation- and position-independent, pointing towards its activity at the transcriptional and not post-transcriptional level.

  4. NEUTRONIC REACTOR CORE INSTRUMENT

    DOEpatents

    Mims, L.S.

    1961-08-22

    A multi-purpose instrument for measuring neutron flux, coolant flow rate, and coolant temperature in a nuclear reactor is described. The device consists essentially of a hollow thimble containing a heat conducting element protruding from the inner wall, the element containing on its innermost end an amount of fissionsble materinl to function as a heat source when subjected to neutron flux irradiation. Thermocouple type temperature sensing means are placed on the heat conducting element adjacent the fissionable material and at a point spaced therefrom, and at a point on the thimble which is in contact with the coolant fluid. The temperature differentials measured between the thermocouples are determinative of the neutron flux, coolant flow, and temperature being measured. The device may be utilized as a probe or may be incorporated in a reactor core. (AE C)

  5. HTTF Core Stress Analysis

    SciTech Connect

    Brian D. Hawkes; Richard Schultz

    2012-07-01

    In accordance with the need to determine whether cracking of the ceramic core disks which will be constructed and used in the High Temperature Test Facility (HTTF) for heatup and cooldown experiments, a set of calculation were performed using Abaqus to investigate the thermal stresses levels and likelihood for cracking. The calculations showed that using the material properties provided for the Greencast 94F ceramic, cracking is predicted to occur. However, this modeling does not predict the size or length of the actual cracks. It is quite likely that cracks will be narrow with rough walls which would impede the flow of coolant gases entering the cracks. Based on data recorded at Oregon State University using Greencast 94F samples that were heated and cooled at prescribed rates, it was concluded that the likelihood that the cracks would be detrimental to the experimental objectives is small.

  6. NEW SOIL VOC SAMPLERS: EN CORE AND ACCU CORE SAMPLING/STORAGE DEVICES FOR VOC ANALYSIS

    SciTech Connect

    Susan S. Sorini; John F. Schabron; Joseph F. Rovani Jr

    2006-06-01

    Soil sampling and storage practices for volatile organic analysis must be designed to minimize loss of volatile organic compounds (VOCs) from samples. The En Core{reg_sign} sampler is designed to collect and store soil samples in a manner that minimizes loss of contaminants due to volatilization and/or biodegradation. An ASTM International (ASTM) standard practice, D 6418, Standard Practice for Using the Disposable En Core Sampler for Sampling and Storing Soil for Volatile Organic Analysis, describes use of the En Core sampler to collect and store a soil sample of approximately 5 grams or 25 grams for volatile organic analysis and specifies sample storage in the En Core sampler at 4 {+-} 2 C for up to 48 hours; -7 to -21 C for up to 14 days; or 4 {+-} 2 C for up to 48 hours followed by storage at -7 to -21 C for up to five days. This report discusses activities performed during the past year to promote and continue acceptance of the En Core samplers based on their performance to store soil samples for VOC analysis. The En Core sampler is designed to collect soil samples for VOC analysis at the soil surface. To date, a sampling tool for collecting and storing subsurface soil samples for VOC analysis is not available. Development of a subsurface VOC sampling/storage device was initiated in 1999. This device, which is called the Accu Core{trademark} sampler, is designed so that a soil sample can be collected below the surface using a dual-tube penetrometer and transported to the laboratory for analysis in the same container. Laboratory testing of the current Accu Core design shows that the device holds low-level concentrations of VOCs in soil samples during 48-hour storage at 4 {+-} 2 C and that the device is ready for field evaluation to generate additional performance data. This report discusses a field validation exercise that was attempted in Pennsylvania in 2004 and activities being performed to plan and conduct a field validation study in 2006. A draft ASTM

  7. Growth outside the core.

    PubMed

    Zook, Chris; Allen, James

    2003-12-01

    Growth in an adjacent market is tougher than it looks; three-quarters of the time, the effort fails. But companies can change those odds dramatically. Results from a five-year study of corporate growth conducted by Bain & Company reveal that adjacency expansion succeeds only when built around strong core businesses that have the potential to become market leaders. And the best place to look for adjacency opportunities is inside a company's strongest customers. The study also found that the most successful companies were able to consistently, profitably outgrow their rivals by developing a formula for pushing out the boundaries of their core businesses in predictable, repeatable ways. Companies use their repeatability formulas to expand into any number of adjacencies. Some companies make repeated geographic moves, as Vodafone has done in expanding from one geographic market to another over the past 13 years, building revenues from $1 billion in 1990 to $48 billion in 2003. Others apply a superior business model to new segments. Dell, for example, has repeatedly adapted its direct-to-customer model to new customer segments and new product categories. In other cases, companies develop hybrid approaches. Nike executed a series of different types of adjacency moves: it expanded into adjacent customer segments, introduced new products, developed new distribution channels, and then moved into adjacent geographic markets. The successful repeaters in the study had two common characteristics. First, they were extraordinarily disciplined, applying rigorous screens before they made an adjacency move. This discipline paid off in the form of learning curve benefits, increased speed, and lower complexity. And second, in almost all cases, they developed their repeatable formulas by studying their customers and their customers' economics very, very carefully.

  8. Glass-clad semiconductor core optical fibers

    NASA Astrophysics Data System (ADS)

    Morris, Stephanie Lynn

    ; experimental results show a decrease in fiber core oxygen content in the fibers drawn with the tailored glass composition. In a further attempt to reduce the presence of oxide species in the core, a reactive molten core approach to semiconductor optical fibers are developed. Specifically, the addition of silicon carbide (SiC) into a silicon (Si) core provides an in-situ reactive getter of oxygen during the draw process to achieve oxygen-free silicon optical fibers. Elemental analysis and x-ray diffraction of fibers drawn using this reactive chemistry approach show negligible oxygen concentration in the highly crystalline silicon core, a significant departure from the nearly 18 atom percent oxygen in previous fibers. Scattering of light out of the core is shown qualitatively to have been reduced in the process. The role of the cross-sectional geometry on the resultant core crystallography with respect to the fiber axis is explored in a continued effort to better understand the nature of the crystal formation and structural properties in these semiconductor core optical fibers. A square cross-sectional geometry was explored to determine if core non-circularity can enhance or promote single crystallinity, as the semiconductors studied have a preference to form cubic crystals. Resultant crystallography of the non-circular core showed a significant improvement in maintaining a preferred crystallographic orientation, with the square core fibers exhibiting a 90% preference for the < 1 1 0 > family of directions occurring closest to the longitudinal direction of the fiber. The ability to orient the crystallography with respect to the fiber axis could be of great value to future nonlinear optical fiber-based devices. In summary, this dissertation begins to elucidate some of the microstructural features, not present in conventional glass optical fibers, which could be important for future low-loss single crystalline semiconductor optical fibers. Additionally, this

  9. Genome-wide analysis of promoter architecture in Drosophila melanogaster

    SciTech Connect

    Hoskins, Roger A.; Landolin, Jane M.; Brown, James B.; Sandler, Jeremy E.; Takahashi, Hazuki; Lassmann, Timo; Yu, Charles; Booth, Benjamin W.; Zhang, Dayu; Wan, Kenneth H.; Yang, Li; Boley, Nathan; Andrews, Justen; Kaufman, Thomas C.; Graveley, Brenton R.; Bickel, Peter J.; Carninci, Piero; Carlson, Joseph W.; Celniker, Susan E.

    2010-10-20

    Core promoters are critical regions for gene regulation in higher eukaryotes. However, the boundaries of promoter regions, the relative rates of initiation at the transcription start sites (TSSs) distributed within them, and the functional significance of promoter architecture remain poorly understood. We produced a high-resolution map of promoters active in the Drosophila melanogaster embryo by integrating data from three independent and complementary methods: 21 million cap analysis of gene expression (CAGE) tags, 1.2 million RNA ligase mediated rapid amplification of cDNA ends (RLMRACE) reads, and 50,000 cap-trapped expressed sequence tags (ESTs). We defined 12,454 promoters of 8037 genes. Our analysis indicates that, due to non-promoter-associated RNA background signal, previous studies have likely overestimated the number of promoter-associated CAGE clusters by fivefold. We show that TSS distributions form a complex continuum of shapes, and that promoters active in the embryo and adult have highly similar shapes in 95% of cases. This suggests that these distributions are generally determined by static elements such as local DNA sequence and are not modulated by dynamic signals such as histone modifications. Transcription factor binding motifs are differentially enriched as a function of promoter shape, and peaked promoter shape is correlated with both temporal and spatial regulation of gene expression. Our results contribute to the emerging view that core promoters are functionally diverse and control patterning of gene expression in Drosophila and mammals.

  10. Cestrum yellow leaf curling virus (CmYLCV) promoter: a new strong constitutive promoter for heterologous gene expression in a wide variety of crops.

    PubMed

    Stavolone, Livia; Kononova, Maria; Pauli, Sandra; Ragozzino, Antonio; de Haan, Peter; Milligan, Steve; Lawton, Kay; Hohn, Thomas

    2003-11-01

    Appropriately regulated gene expression requires a suitable promoter. A number of promoters have been isolated and shown to be functional in plants, but only a few of them activate transcription of transgenes at high levels constitutively. We report here the cloning and characterization of a novel, constitutively expressed promoter isolated from Cestrum yellow leaf curling virus (CmYLCV), a double-stranded DNA plant pararetrovirus belonging to the Caulimoviridae family. The CmYLCV promoter is highly active in callus, meristems and vegetative and reproductive tissues in Arabidopsis thaliana, Nicotiana tabacum, Lycopersicon esculentum, Zea mays and Oryza sativa. Furthermore, the level of expression is comparable to, or higher than, that from the CaMV 35S, the 'super-promoter' or the maize ubiquitin 1 promoters, three frequently used promoters in agricultural biotechnology. The heritable, strong and constitutive activity in both monocotyledonous and dicotyledonous plants, combined with the extremely narrow CmYLCV host range, makes the CmYLCV promoter an attractive tool for regulating transgene expression in a wide variety of plant species.

  11. GPM Core Observatory Launch Animation

    NASA Video Gallery

    This animation depicts the launch of the Global Precipitation Measurement (GPM) Core Observatory satellite from Tanegashima Space Center, Japan. The launch is currently scheduled for Feb. 27, 2014....

  12. IN-CORE FUEL MANAGEMENT: PWR Core Calculations Using MCRAC

    NASA Astrophysics Data System (ADS)

    PetroviĆ, B. G.

    1991-01-01

    The following sections are included: * INTRODUCTION * IN-CORE FUEL MANAGEMENT CALCULATIONS * In-Core Fuel Management * Methodological Problems of In-Core Fuel Management * In-Core Fuel Management Analytical Tools * PENN STATE FUEL MANAGEMENT PACKAGE * Penn State Fuel Management Package (PFMP) * Assembly Data Description (ADD) * Linking PSU-LEOPARD and MCRAC: An Example * MULTICYCLE REACTOR ANALYSIS CODE (MCRAC) * Main Features and Options of MCRAC code * Core geometry * Diffusion equations * 1.5-group model * Multicycle neutronic analysis * Multicycle cost analysis * Criticality search * Power-dependent xenon feedback calculations * Control rod and burnable absorber simulation * Search for LP with flat BOC power distribution * Artificial ADD option * Variable dimensioning technique * RBI version of MCRAC code * Programming changes in PC version * Fuel interchange option * MCRAC Input/Output * General input description * Sample input * Sample output * EXPERIENCE WITH MCRAC CODE * CONCLUSIONS * REFERENCES

  13. Relativistic frozen core potential scheme with relaxation of core electrons

    NASA Astrophysics Data System (ADS)

    Nakajima, Yuya; Seino, Junji; Hayami, Masao; Nakai, Hiromi

    2016-10-01

    This letter proposes a relaxation scheme for core electrons based on the frozen core potential method at the infinite-order Douglas-Kroll-Hess level, called FCP-CR. The core electrons are self-consistently relaxed using frozen molecular valence potentials after the valence SCF calculation is performed. The efficiency of FCP-CR is confirmed by calculations of gold clusters. Furthermore, FCP-CR reproduces the results of the all-electron method for the energies of coinage metal dimers and the core ionization energies and core level shifts of vinyl acetate and three tungsten complexes at the Hartree-Fock and/or symmetry-adapted cluster configuration interaction levels.

  14. Induction and maintenance of DNA methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing

    PubMed Central

    Kon, Tatsuya; Yoshikawa, Nobuyuki

    2014-01-01

    Apple latent spherical virus (ALSV) is an efficient virus-induced gene silencing vector in functional genomics analyses of a broad range of plant species. Here, an Agrobacterium-mediated inoculation (agroinoculation) system was developed for the ALSV vector, and virus-induced transcriptional gene silencing (VITGS) is described in plants infected with the ALSV vector. The cDNAs of ALSV RNA1 and RNA2 were inserted between the cauliflower mosaic virus 35S promoter and the NOS-T sequences in a binary vector pCAMBIA1300 to produce pCALSR1 and pCALSR2-XSB or pCALSR2-XSB/MN. When these vector constructs were agroinoculated into Nicotiana benthamiana plants with a construct expressing a viral silencing suppressor, the infection efficiency of the vectors was 100%. A recombinant ALSV vector carrying part of the 35S promoter sequence induced transcriptional gene silencing of the green fluorescent protein gene in a line of N. benthamiana plants, resulting in the disappearance of green fluorescence of infected plants. Bisulfite sequencing showed that cytosine residues at CG and CHG sites of the 35S promoter sequence were highly methylated in the silenced generation zero plants infected with the ALSV carrying the promoter sequence as well as in progeny. The ALSV-mediated VITGS state was inherited by progeny for multiple generations. In addition, induction of VITGS of an endogenous gene (chalcone synthase-A) was demonstrated in petunia plants infected with an ALSV vector carrying the native promoter sequence. These results suggest that ALSV-based vectors can be applied to study DNA methylation in plant genomes, and provide a useful tool for plant breeding via epigenetic modification. PMID:25426109

  15. Business Planning Core Facilities

    PubMed Central

    Itzkowitz, G.N.

    2014-01-01

    Thoughtful business planning is pivotal to the success of any business/operational venture. When planned in a thoughtful and detailed manner there are very few operational or financial surprises for an institution or facility (service center) to contend with. At Stony Brook Medicine we include SWOT analysis and a detailed Market Analysis as part of the process. This is bolstered by an initiative to ensure institutional policies are met so that facilities remain in compliance throughout their lifecycle. As we operate 14 facilities we have had the opportunity to become creative in our approach to coordinate activities, virtualize services, integrate new software business-to-business partners, and finally coordinate plans for phased consolidation instead of outright termination of services when required. As the Associate Dean for Scientific Operations and Research Facilities, the shared research facilities (cores) of the Medical School are in my direct line of sight. We understand their value to the meeting our overall research mission. We have found that an active process of monitoring to predict trouble as much as possible is the best approach for facilities. Some case analysis of this type of interaction will be presented as well.

  16. Adult educators' core competences

    NASA Astrophysics Data System (ADS)

    Wahlgren, Bjarne

    2016-06-01

    Which competences do professional adult educators need? This research note discusses the topic from a comparative perspective, finding that adult educators' required competences are wide-ranging, heterogeneous and complex. They are subject to context in terms of national and cultural environment as well as the kind of adult education concerned (e.g. basic education, work-related education etc.). However, it seems that it is possible to identify certain competence requirements which transcend national, cultural and functional boundaries. This research note summarises these common or "core" requirements, organising them into four thematic subcategories: (1) communicating subject knowledge; (2) taking students' prior learning into account; (3) supporting a learning environment; and (4) the adult educator's reflection on his or her own performance. At the end of his analysis of different competence profiles, the author notes that adult educators' ability to train adult learners in a way which then enables them to apply and use what they have learned in practice (thus performing knowledge transfer) still seems to be overlooked.

  17. Variable depth core sampler

    SciTech Connect

    Bourgeois, P.M.; Reger, R.J.

    1994-12-31

    This invention relates to a sampling means, more particularly to a device to sample hard surfaces at varying depths. Often it is desirable to take samples of a hard surface wherein the samples are of the same diameter but of varying depths. Current practice requires that a full top-to-bottom sample of the material be taken, using a hole saw, and boring a hole from one end of the material to the other. The sample thus taken is removed from the hole saw and the middle of said sample is then subjected to further investigation. This paper describes a variable depth core sampler comprimising a circular hole saw member, having longitudinal sections that collapse to form a point and capture a sample, and a second saw member residing inside the first hole saw member to support the longitudinal sections of the first member and prevent them from collapsing to form a point. The second hole saw member may be raised and lowered inside the the first hole saw member.

  18. The complete sequence of soybean chlorotic mottle virus DNA and the identification of a novel promoter.

    PubMed

    Hasegawa, A; Verver, J; Shimada, A; Saito, M; Goldbach, R; Van Kammen, A; Miki, K; Kameya-Iwaki, M; Hibi, T

    1989-12-11

    The complete nucleotide sequence of an infectious clone of soybean chlorotic mottle virus (SoyCMV) DNA was determined and compared with those of three other caulimoviruses, cauliflower mosaic virus (CaMV), carnation etched ring virus and figwort mosaic virus. The double-stranded DNA genome of SoyCMV (8,175 bp) contained nine open reading frames (ORFs) and one large intergenic region. The primer binding sites, gene organization and size of ORFs were similar to those of the other caulimoviruses, except for ORF I, which was split into ORF Ia and Ib. The amino acid sequences deduced from each ORF showed only short, highly homologous regions in several of the corresponding ORFs of the three other caulimoviruses. A promoter fragment of 378 bp in SoyCMV ORF III showed a strong expression activity, comparable to that of the CaMV 35S promoter, in tobacco mesophyll protoplasts as determined by a beta-glucuronidase assay using electrotransfection. The fragment contained CAAT and TATA boxes but no transcriptional enhancer signal as reported for the CaMV 35S promoter. Instead, it had sequences homologous to a part of the translational enhancer signal reported for the 5'-leader sequence of tobacco mosaic virus RNA.

  19. Public/Private Partnerships in Health Promotion: A Guide for the Aging Network.

    ERIC Educational Resources Information Center

    Atlanta Regional Commission, GA.

    The aging network has been slow to embrace health promotion activities as core services to the aging population. Therefore, advocates for business and aging partnerships in health promotion activities must first educate their colleagues about the value of health promotion programs before looking for partners in the business community. The goal of…

  20. Core-to-core uniformity improvement in multi-core fiber Bragg gratings

    NASA Astrophysics Data System (ADS)

    Lindley, Emma; Min, Seong-Sik; Leon-Saval, Sergio; Cvetojevic, Nick; Jovanovic, Nemanja; Bland-Hawthorn, Joss; Lawrence, Jon; Gris-Sanchez, Itandehui; Birks, Tim; Haynes, Roger; Haynes, Dionne

    2014-07-01

    Multi-core fiber Bragg gratings (MCFBGs) will be a valuable tool not only in communications but also various astronomical, sensing and industry applications. In this paper we address some of the technical challenges of fabricating effective multi-core gratings by simulating improvements to the writing method. These methods allow a system designed for inscribing single-core fibers to cope with MCFBG fabrication with only minor, passive changes to the writing process. Using a capillary tube that was polished on one side, the field entering the fiber was flattened which improved the coverage and uniformity of all cores.

  1. Combination pipe rupture mitigator and in-vessel core catcher

    DOEpatents

    Tilbrook, Roger W.; Markowski, Franz J.

    1983-01-01

    A device which mitigates against the effects of a failed coolant loop in a nuclear reactor by restricting the outflow of coolant from the reactor through the failed loop and by retaining any particulated debris from a molten core which may result from coolant loss or other cause. The device reduces the reverse pressure drop through the failed loop by limiting the access of coolant in the reactor to the inlet of the failed loop. The device also spreads any particulated core debris over a large area to promote cooling.

  2. COVERING A CORE BY EXTRUSION

    DOEpatents

    Karnie, A.J.

    1963-07-16

    A method of covering a cylindrical fuel core with a cladding metal ms described. The metal is forced between dies around the core from both ends in two opposing skirts, and as these meet the ends turn outward into an annular recess in the dics. By cutting off the raised portion formed by the recess, oxide impurities are eliminated. (AEC)

  3. Common Core: Victory Is Yours!

    ERIC Educational Resources Information Center

    Fink, Jennifer L. W.

    2012-01-01

    In this article, the author discusses how to implement the Common Core State Standards in the classroom. She presents examples and activities that will leave teachers feeling "rosy" about tackling the new standards. She breaks down important benchmarks and shows how other teachers are doing the Core--and loving it!

  4. Complicated Politics to the Core

    ERIC Educational Resources Information Center

    McGuinn, Patrick

    2015-01-01

    People dislike the Common Core for several different reasons, and so it is important to disaggregate the sources of opposition and to assess and then to dispel some of the myths that have built up around it. It also is important to understand the unusual political alliances that have emerged in opposition to Common Core implementation and how they…

  5. Data interchange across cores of multi-core optical fibers

    NASA Astrophysics Data System (ADS)

    Awad, Ehab S.

    2015-12-01

    A novel device for data interchange among space-division multiplexed cores inside MCF is demonstrated using numerical simulations. The device allows complete exchange of all WDM data channels between MCF cores in propagation direction whether the channels have the same or different sets of wavelengths. This is crucial in future MCF optical networks where in-fiber data interchange over space-division multiplexed cores can allow for a simple and fast data swapping among cores without a need for space-division demultiplexing to single-mode single-core fibers. The data core-interchange (DCI) device consists of a graded refractive-index rectangular waveguide enclosing the two interchanged cores in addition to the cladding region in between them. Both finite-difference-time-domain (FDTD) and eigenmode expansion (EME) simulations are performed to verify the device operation and characterize its performance. The simulations demonstrate that the DCI has a very short-length with polarization independent operation, and high performance over the broadband wavelength range S, C, L, and U bands. Moreover, the device shows a high coupling-factor of -0.13 dB with small cross-talk, back-reflection, and return-loss of -26.3, -46.1, and -48.8 dB, respectively.

  6. The EPOS Integrated Core Services

    NASA Astrophysics Data System (ADS)

    Jeffery, Keith; Michelini, Alberto; Bailo, Daniele

    2013-04-01

    The European Plate Observing System (EPOS) is integrating the diverse, but advanced Research Infrastructures in Europe for solid Earth Science, and will build on new e-science opportunities to monitor and understand the dynamic and complex solid-Earth System. This integration requires a significant coordination between, among others, disciplinary (thematic) communities, national RIs policies and initiatives, and geo- and IT-scientists. The RIs that EPOS will coordinate include at least, but not only: regionally-distributed geophysical observing systems (seismological and geodetic networks), local observatories (including geomagnetic, permanent in-situ and volcano observatories), experimental & analogue laboratories in Europe, integrated satellite data and geological information. EPOS is promoting open access to geophysical and geological data as well as modelling/processing tools, enabling a step change in multidisciplinary scientific research for Earth Sciences The EPOS e-infrastructure is developed through strawman (initial design / architecture), woodman (refined design/architecture) and ironman (final design/architecture) phases. Midway in the project we are in the woodman phase based on extensive primary requirements from users and secondary requirements for interoperation with other geoscience systems, other European environmental research infrastructure projects and e-infrastructure projects (e.g. EUDAT). The EPOS e-infrastructure is being developed along 3 parallel tracks: (a) an inventory of assets offered by organisations within the EPOS community. The RIDE (Research Infrastructure Database for EPOS) system from the strawman phase is being extended in the woodman phase to the metadata catalog describing computing and scientific resources, data, services (software), and users which will drive the EPOS e-infrastructure; (b) refining an architecture to meet the requirements. This is an iterative process with the working groups (organised thematically) within

  7. Crystallization in Earth's Core after High-Temperature Core Formation

    NASA Astrophysics Data System (ADS)

    Hirose, K.; Morard, G.; Hernlund, J. W.; Helffrich, G. R.; Ozawa, H.

    2015-12-01

    Recent core formation models based on the metal-silicate partitioning of siderophile elements suggest that the Earth's core was formed by metal segregation at high pressure and high temperature in a deep magma ocean. It is also thought that the simultaneous solubility of silicon and oxygen in liquid iron are strongly enhanced at high pressure and high temperature, such that at the end of accretion the core was rich in both silicon and oxygen. Here we performed crystallization experiments on the Fe-Si binary and Fe-Si-O ternary systems up to core pressure in a laser-heated diamond-anvil cell. The starting material for the latter was a homogeneous mixture of fine-grain Fe-Si and SiO2 (<1 µm). We prepared cross sections of samples recovered from the DAC using a focused ion beam (FIB) and subsequently performed textural and chemical characterization with field-emission-type electron microprobe (FE-EPMA). Quenched liquid alloy was found at the hottest part coexisting with a solid phase (liquidus phase) at the periphery. These results combined with literature data on the melting phase relations in the Fe-FeO binary system demonstrate that the liquidus field of SiO2 is very wide at the Fe-rich portion of the Fe-Si-O ternary system at the core pressure range. It indicates that the original Fe-Si-O core liquid should have crystallized a large amount SiO2 until it lost either silicon or oxygen. The recent finding of high thermal conductivity of the core suggests that core thermal convection is difficult to sustain without extreme degrees of secular cooling. However, even for modest degrees of joint Si-O incorporation into the early core, the buoyancy released by crystallization of SiO2 is sufficient to overcome thermal stratification and sustain the geodynamo.

  8. Production of 35S for a Liquid Semiconductor Betavoltaic

    SciTech Connect

    Meier, David E.; Garnov, A. Y.; Robertson, J. D.; Kwon, J. W.; Wacharasindhu, T.

    2009-10-01

    The specific energy density from radioactive decay is five to six orders of magnitude greater than the specific energy density in conventional chemical battery and fuel cell technologies. We are currently investigating the use of liquid semiconductor based betavoltaics as a way to directly convert the energy of radioactive decay into electrical power and potentially avoid the radiation damage that occurs in solid state semiconductor devices due to non-ionizing energy loss. Sulfur-35 was selected as the isotope for the liquid semiconductor demonstrations because it can be produced in high specific activity and it is chemically compatible with known liquid semiconductor media.

  9. Developing a Promotional Video

    ERIC Educational Resources Information Center

    Epley, Hannah K.

    2014-01-01

    There is a need for Extension professionals to show clientele the benefits of their program. This article shares how promotional videos are one way of reaching audiences online. An example is given on how a promotional video has been used and developed using iMovie software. Tips are offered for how professionals can create a promotional video and…

  10. Isolation and functional characterization of two novel seed-specific promoters from sunflower (Helianthus annuus L.).

    PubMed

    Zavallo, Diego; Lopez Bilbao, Marisa; Hopp, H Esteban; Heinz, Ruth

    2010-03-01

    The promoter region of two sunflower (Helianthus annuus L. HA89 genotype) seed specifically expressed genes, coding for an oleate desaturase (HaFAD2-1) and a lipid transfer protein (HaAP10), were cloned and in silico characterized. The isolated fragments are 867 and 964 bp long, respectively, and contain several seed-specific motifs, such as AACA motif, ACGT element, E-Boxes, SEF binding sites and GCN4 motif. Functional analysis of these promoters in transgenic Arabidopsis plants was investigated after fusing them with the beta-glucuronidase (GUS) reporter gene. None of the promoters triggered GUS activity in any vegetative tissue, with the exception of early seedling cotyledons. HaFAD2-1 and HaAP10 promoters were tested along seed development from globular stage to mature seeds. GUS staining was restricted to embryonic tissue and quantitative fluorometric assays showed high activity values at the later stages of development. In this work we demonstrate that HaFAD2-1 promoter is as strong as 35S promoter even though it is a tissue-specific promoter and its activity derived just from the embryo, thus confirming that it can be considered a strong highly specific seed promoter useful for biotechnology applications.

  11. Usefulness of a Competency-Based Reflective Portfolio for Student Learning on a Masters Health Promotion Programme

    ERIC Educational Resources Information Center

    McKenna, Verna; Connolly, Claire; Hodgins, Margaret

    2011-01-01

    Background: Efforts to identify core competencies within health promotion and health education have been on-going for a number of years. These efforts include work carried out by the International Union for Health Promotion and Education (IUHPE) in drawing up a draft list of 11 core competencies which were incorporated into the practice module on…

  12. Radiation Effects: Core Project

    NASA Technical Reports Server (NTRS)

    Dicello, John F.

    1999-01-01

    The risks to personnel in space from the naturally occurring radiations are generally considered to be one of the most serious limitations to human space missions, as noted in two recent reports of the National Research Council/National Academy of Sciences. The Core Project of the Radiation Effects Team for the National Space Biomedical Research Institute is the consequences of radiations in space in order to develop countermeasure, both physical and pharmaceutical, to reduce the risks of cancer and other diseases associated with such exposures. During interplanetary missions, personnel in space will be exposed to galactic cosmic rays, including high-energy protons and energetic ions with atomic masses of iron or higher. In addition, solar events will produce radiation fields of high intensity for short but irregular durations. The level of intensity of these radiations is considerably higher than that on Earth's surface, and the biological risks to astronauts is consequently increased, including increased risks of carcinogenesis and other diseases. This group is examining the risk of cancers resulting from low-dose, low-dose rate exposures of model systems to photons, protons, and iron by using ground-based accelerators which are capable of producing beams of protons, iron, and other heavy ions at energies comparable to those encountered in space. They have begun the first series of experiments using a 1-GeV iron beam at the Brookhaven National Laboratory and 250-MeV protons at Loma Linda University Medical Center's proton synchrotron facility. As part of these studies, this group will be investigating the potential for the pharmaceutical, Tamoxifen, to reduce the risk of breast cancer in astronauts exposed to the level of doses and particle types expected in space. Theoretical studies are being carried out in a collaboration between scientists at NASA's Johnson Space Center and Johns Hopkins University in parallel with the experimental program have provided

  13. HOW STARLESS ARE STARLESS CORES?

    SciTech Connect

    Schnee, Scott; Friesen, Rachel; Di Francesco, James; Johnstone, Doug; Enoch, Melissa; Sadavoy, Sarah

    2012-01-20

    In this paper, we present the results of Combined Array for Research in Millimeter-wave Astronomy continuum and spectral line observations of the dense core Per-Bolo 45. Although this core has previously been classified as starless, we find evidence for an outflow and conclude that Per-Bolo 45 is actually an embedded, low-luminosity protostar. We discuss the impact of newly discovered, low-luminosity, embedded objects in the Perseus molecular cloud on starless core and protostar lifetimes. We estimate that the starless core lifetime has been overestimated by 4%-18% and the Class 0/I protostellar lifetime has been underestimated by 5%-20%. Given the relatively large systematic uncertainties involved in these calculations, variations on the order of 10% do not significantly change either core lifetimes or the expected protostellar luminosity function. Finally, we suggest that high-resolution (sub)millimeter surveys of known cores lacking near-infrared and mid-infrared emission are necessary to make an accurate census of starless cores.

  14. Dynamics of the core, geodynamo

    NASA Astrophysics Data System (ADS)

    Roberts, Paul H.

    1995-07-01

    "The mechanism for generating the geomagnetic field remains one of the central unsolved problems in geoscience." So states the report on the National Geomagnetic Initiative (NGI) prepared by the U.S. Geodynamics Committee, et al [1993], with advice from the NGI Workshop held in Washington D.C. in March 1992. All analyses of the geomagnetic data point to the core as containing the source of the field and "The basic premise that virtually everyone accepts is that the Earth's magnetism is created by a self-sustaining dynamo driven by fluid motions in Earth's core" (NGI, p.135). Dynamical questions at once arise, such as "What is the energy source driving those motions?" Jacobs [1953] proposed that the solid inner core (SIC) is the result of the freezing of the fluid outer core (FOC). Verhoogen [1961] noticed that the release of latent heat at the inner core boundary (ICB) during freezing would help drive thermal convection in the FOC, and Braginsky [1963] pointed out that the release of the light alloying elements during fractionation at the ICB would provide compositional buoyancy. These two sources suffice to supply the geodynamo with energy throughout geological time, even in the absence of dissolved radioactivity in the core [Braginsky and Roberts, 1994a; Kuang et al, 1994]. Stevenson [1991] argues that potential differences on the core-mantle boundary (CMB) of electrochemical origin may be partially responsible for the geomagnetic field.

  15. Simplified cut core inductor design

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1974-01-01

    Although filter inductor designers have routinely tended to specify molypermalloy powder cores for use in high frequency power converters and pulse-width modulated switching regulators, there are sigificant advantages in specifying C cores and cut toroids fabricated from grain oriented silicon steels which should not be overlooked. Such steel cores can develop flux densities of 1.6 tesla, with useful linearity to 1.2 tesla, whereas molypermalloy cores carrying d.c. current have useful flux density capabilities only to about 0.3 tesla. The use of silicon steel cores thus makes it possible to design more compact cores, and therefore inductors of reduced volume, or conversely to provide greater load capacity in inductors of a given volume. Information is available which makes it possible to obtain quick and close approximations of significant parameters such as size, weight and temperature rise for silicon steel cores for breadboarding. Graphs, nomographs and tables are presented for this purpose, but more complete mathematical derivations of some of the important parameters are also included for a more rigorous treatment.

  16. Core physics analysis of 100% MOX Core in IRIS

    SciTech Connect

    Franceschini, F.; Petrovic, B.

    2006-07-01

    International Reactor Innovative and Secure (IRIS) is an advanced small-to-medium-size (1000 MWt) Pressurized Water Reactor (PWR), targeting deployment around 2015. Its reference core design is based on the current Westinghouse UO{sub 2} fuel with less than 5% {sup 235}U, and the analysis has been previously completed confirming good performance. The full MOX fuel core is currently under evaluation as one of the alternatives for the second wave of IRIS reactors. A full 3-D neutronic analysis has been performed to examine main core performance parameters, such as critical boron concentration, peaking factors, discharge burnup, etc. The enhanced moderation of the IRIS fuel lattice facilitates MOX core design, and all the obtained results are within the requirements, confirming viability of this option from the reactor physics standpoint. (authors)

  17. Core Exercises: Why You Should Strengthen Your Core Muscles

    MedlinePlus

    ... Read on to find out why. Core exercises train the muscles in your pelvis, lower back, hips ... manner that involves maintaining a stable trunk can train and strengthen several of your muscles, including your ...

  18. Core principles in treating suicidal patients.

    PubMed

    Fowler, James Christopher

    2013-09-01

    The treatment of suicidal individuals requires special attention to therapist interventions that promote a viable treatment alliance in the context of shared responsibilities for patient safety. Three core principles in the treatment process (alliance building, enhancing curiosity about the function of suicidal thoughts and urges, as well as enhancing experience and expression of intense emotions) are articulated and brief case vignettes are used to illuminate the principles. Results from open trails and randomized control trials involving suicidal patients are examined to support the evidence-based practice of these principles. The overarching principle undergirding the utility of the principles is a collaborative joining with the patient to decrease isolation and alienation when facing intense and overwhelming emotions.

  19. [Seven core principles for treatment of hypertension].

    PubMed

    Hu, Chun-song; Gao, Run-lin; Liu, Li-sheng

    2006-04-01

    The seven core principles (SeCP) for treatment of hypertension were (1) early identification, early diagnosis, early and life-long treatment; (2) application of long-acting and slow-released anti-hypertension drugs to control blood pressure smoothly; (3) use low dosage and combined therapy; (4) individual and racial therapy; (5) integrated traditional Chinese and Western medicine; (6) life style improvement; (7) enhancing compliance. Being more comprehensive and detailed than the Seventh Report of the Joint National Committee (JNC-7), the 2003' European Society of Hypertension/European Society of Cardiology (ESH/ESC2003), the report of the fourth working party of the British Hypertension Society (2004-BHS IV), and the 2004' Chinese Guideline of Hypertension (CGH2004), the programmatic SeCP should be promoted in clinical practice for hypertension patients and doctors to follow and apply.

  20. Uranium droplet core nuclear rocket

    NASA Technical Reports Server (NTRS)

    Anghaie, Samim

    1991-01-01

    Uranium droplet nuclear rocket is conceptually designed to utilize the broad temperature range ofthe liquid phase of metallic uranium in droplet configuration which maximizes the energy transfer area per unit fuel volume. In a baseline system dissociated hydrogen at 100 bar is heated to 6000 K, providing 2000 second of Isp. Fission fragments and intense radian field enhance the dissociation of molecular hydrogen beyond the equilibrium thermodynamic level. Uranium droplets in the core are confined and separated by an axisymmetric vortex flow generated by high velocity tangential injection of hydrogen in the mid-core regions. Droplet uranium flow to the core is controlled and adjusted by a twin flow nozzle injection system.

  1. Surface-core fiber gratings

    NASA Astrophysics Data System (ADS)

    Osório, Jonas H.; Oliveira, Ricardo; Mosquera, L.; Franco, Marcos A. R.; Heidarialamdarloo, Jamshid; Bilro, Lúcia; Nogueira, Rogério N.; Cordeiro, Cristiano M. B.

    2015-09-01

    In this paper, we report, to our knowledge, the first demonstration of the induction of long-period and Bragg gratings on surface-core optical fibers. Surface-core fibers described herein were fabricated from commercial silica tubes and germanium-doped silica rods by employing a very simple procedure. Being the core on the fiber surface, it can be sensitive to refractive index variations in the environment in which the fiber is immersed. Thus, results concerning the sensitivity of these gratings to environmental refractive index variations are presented. Besides, simulation data are presented for comparison to the experimental behavior and for projecting future steps in this research.

  2. Selective Promoter Recognition by Chlamydial σ28 Holoenzyme▿ †

    PubMed Central

    Shen, Li; Feng, Xiaogeng; Yuan, Yuan; Luo, Xudong; Hatch, Thomas P.; Hughes, Kelly T.; Liu, Jun S.; Zhang, You-xun

    2006-01-01

    The σ transcription factor confers the promoter recognition specificity of RNA polymerase (RNAP) in eubacteria. Chlamydia trachomatis has three known sigma factors, σ66, σ54, and σ28. We developed two methods to facilitate the characterization of promoter sequences recognized by C. trachomatis σ28 (σ28Ct). One involved the arabinose-induced expression of plasmid-encoded σ28Ct in a strain of Escherichia coli defective in the σ28 structural gene, fliA. The second was an analysis of transcription in vitro with a hybrid holoenzyme reconstituted with E. coli RNAP core and recombinant σ28Ct. These approaches were used to investigate the interactions of σ28Ct with the σ28Ct-dependent hctB promoter and selected E. coli σ28 (σ28Ec)-dependent promoters, in parallel, compared with the promoter recognition properties of σ28EC. Our results indicate that RNAP containing σ28Ct has at least three characteristics: (i) it is capable of recognizing some but not all σ28EC-dependent promoters; (ii) it can distinguish different promoter structures, preferentially activating promoters with upstream AT-rich sequences; and (iii) it possesses a greater flexibility than σ28EC in recognizing variants with different spacing lengths separating the −35 and −10 elements of the core promoter. PMID:16936033

  3. Characterizing Facesheet/Core Disbonding in Honeycomb Core Sandwich Structure

    NASA Technical Reports Server (NTRS)

    Rinker, Martin; Ratcliffe, James G.; Adams, Daniel O.; Krueger, Ronald

    2013-01-01

    Results are presented from an experimental investigation into facesheet core disbonding in carbon fiber reinforced plastic/Nomex honeycomb sandwich structures using a Single Cantilever Beam test. Specimens with three, six and twelve-ply facesheets were tested. Specimens with different honeycomb cores consisting of four different cell sizes were also tested, in addition to specimens with three different widths. Three different data reduction methods were employed for computing apparent fracture toughness values from the test data, namely an area method, a compliance calibration technique and a modified beam theory method. The compliance calibration and modified beam theory approaches yielded comparable apparent fracture toughness values, which were generally lower than those computed using the area method. Disbonding in the three-ply facesheet specimens took place at the facesheet/core interface and yielded the lowest apparent fracture toughness values. Disbonding in the six and twelve-ply facesheet specimens took place within the core, near to the facesheet/core interface. Specimen width was not found to have a significant effect on apparent fracture toughness. The amount of scatter in the apparent fracture toughness data was found to increase with honeycomb core cell size.

  4. Powder metallurgy process for manufacturing core projectile

    NASA Astrophysics Data System (ADS)

    Akbar, Taufik; Setyowati, Vuri Ayu; Widyastuti

    2013-09-01

    Bullets are part of the defense equipment which the development is very rapid. There are a variety of forms but the bullet Lead is a metal that has always been used for applications projectiles. Lead core constituent materials are combined with antimony. In this research will be conducted by making the material for the core projectile with Tin Lead. The addition of Tin will increase the stiffness of Lead which is soft in nature. The Lead Tin composition variation was given in 10% weight of Sn. The manufacturing process using powder metallurgy using temperature and holding time variations of sintering at 100, 150, and 200°C for 1,2, and 3 hours. XRD samples will be tested to determine the form and phase morphology was observed using SEM-EDX. These results revealed that Pb-10%wtSn Composite which is sintered in temperature 200°C for 3 hours has the greatest density, 10.695 g/cm3 as well as the smallest porosity, 2.2%. In agreement with theoretical analysis that increasing higher temperature and longer holding time give decrease in porosity level due to activation energy which further promotes grain growth. Moreover, there is no intermetallic phase formation as well as no oxide found on composites.

  5. Best Practices for Core Facilities: Handling External Customers

    PubMed Central

    Hockberger, Philip; Meyn, Susan; Nicklin, Connie; Tabarini, Diane; Turpen, Paula; Auger, Julie

    2013-01-01

    This article addresses the growing interest among U.S. scientific organizations and federal funding agencies in strengthening research partnerships between American universities and the private sector. It outlines how core facilities at universities can contribute to this partnership by offering services and access to high-end instrumentation to both nonprofit organizations and commercial organizations. We describe institutional policies (best practices) and procedures (terms and conditions) that are essential for facilitating and enabling such partnerships. In addition, we provide an overview of the relevant federal regulations that apply to external use of academic core facilities and offer a set of guidelines for handling them. We conclude by encouraging directors and managers of core facilities to work with the relevant organizational offices to promote and nurture such partnerships. If handled appropriately, we believe such partnerships can be a win-win situation for both organizations that will support research and bolster the American economy. PMID:23814500

  6. Best practices for core facilities: handling external customers.

    PubMed

    Hockberger, Philip; Meyn, Susan; Nicklin, Connie; Tabarini, Diane; Turpen, Paula; Auger, Julie

    2013-07-01

    This article addresses the growing interest among U.S. scientific organizations and federal funding agencies in strengthening research partnerships between American universities and the private sector. It outlines how core facilities at universities can contribute to this partnership by offering services and access to high-end instrumentation to both nonprofit organizations and commercial organizations. We describe institutional policies (best practices) and procedures (terms and conditions) that are essential for facilitating and enabling such partnerships. In addition, we provide an overview of the relevant federal regulations that apply to external use of academic core facilities and offer a set of guidelines for handling them. We conclude by encouraging directors and managers of core facilities to work with the relevant organizational offices to promote and nurture such partnerships. If handled appropriately, we believe such partnerships can be a win-win situation for both organizations that will support research and bolster the American economy.

  7. The ADNI PET Core: 2015

    PubMed Central

    Jagust, William J.; Landau, Susan M.; Koeppe, Robert A.; Reiman, Eric M.; Chen, Kewei; Mathis, Chester A.; Price, Julie C.; Foster, Norman L.; Wang, Angela Y.

    2015-01-01

    INTRODUCTION This paper reviews the work done in the ADNI PET core over the past 5 years, largely concerning techniques, methods, and results related to amyloid imaging in ADNI. METHODS The PET Core has utilized [18F]florbetapir routinely on ADNI participants, with over 1600 scans available for download. Four different laboratories are involved in data analysis, and have examined factors such as longitudinal florbetapir analysis, use of FDG-PET in clinical trials, and relationships between different biomarkers and cognition. RESULTS Converging evidence from the PET Core has indicated that cross-sectional and longitudinal florbetapir analyses require different reference regions. Studies have also examined the relationship between florbetapir data obtained immediately after injection, which reflects perfusion, and FDG-PET results. Finally, standardization has included the translation of florbetapir PET data to a centiloid scale. CONCLUSION The PET Core has demonstrated a variety of methods for standardization of biomarkers such as florbetapir PET in a multicenter setting. PMID:26194311

  8. Convection, nucleosynthesis, and core collapse

    NASA Technical Reports Server (NTRS)

    Bazan, Grant; Arnett, David

    1994-01-01

    We use a piecewise parabolic method hydrodynamics code (PROMETHEUS) to study convective burning in two dimensions in an oxygen shell prior to core collapse. Significant mixing beyond convective boundaries determined by mixing-length theory brings fuel (C-12) into the convective regon, causing hot spots of nuclear burning. Plumes dominate the velocity structure. Finite perturbations arise in a region in which O-16 will be explosively burned to Ni-56 when the star explodes; the resulting instabilities and mixing are likely to distribute Ni-56 throughout the supernova envelope. Inhomogeneities in Y(sub e) may be large enough to affect core collapse and will affect explosive nucleosynthesis. The nature of convective burning is dramatically different from that assumed in one-dimensional simulations; quantitative estimates of nucleosynthetic yields, core masses, and the approach to core collapse will be affected.

  9. Simplifier cut core inductor design

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1976-01-01

    Advantages of specifying C cores and cut toroids fabricated from grain oriented silicon steels for use in high frequency power converters and pulse width modulated switching regulators are discussed. A method for rating cores assigns to each core a number which is the product of its window and core cross section area, called 'Area Product A sub p.' A correlation between the A sub p numbers and current density for a given temperature rise was developed. Also, straight line relationships were developed for A sub p and volume, A sub p and surface area, and A sub p and weight. These relationships can be used to simplify and standardize the process of inductor design. They also make it possible to design inductors of small bulk and volume or to optimize efficiency.

  10. Core Recursive Hierarchical Image Segmentation

    NASA Technical Reports Server (NTRS)

    Tilton, James

    2011-01-01

    The Recursive Hierarchical Image Segmentation (RHSEG) software has been repackaged to provide a version of the RHSEG software that is not subject to patent restrictions and that can be released to the general public through NASA GSFC's Open Source release process. Like the Core HSEG Software Package, this Core RHSEG Software Package also includes a visualization program called HSEGViewer along with a utility program HSEGReader. It also includes an additional utility program called HSEGExtract. The unique feature of the Core RHSEG package is that it is a repackaging of the RHSEG technology designed to specifically avoid the inclusion of the certain software technology. Unlike the Core HSEG package, it includes the recursive portions of the technology, but does not include processing window artifact elimination technology.

  11. Viscosity of the earth's core.

    NASA Technical Reports Server (NTRS)

    Gans, R. F.

    1972-01-01

    Calculation of the viscosity of the core at the boundary of the inner and outer core. It is assumed that this boundary is a melting transition and the viscosity limits of the Andrade (1934,1952) hypothesis (3.7 to 18.5 cp) are adopted. The corresponding kinematic viscosities are such that the precessional system explored by Malkus (1968) would be unstable. Whether it would be sufficiently unstable to overcome a severely subadiabatic temperature gradient cannot be determined.

  12. Lunar magnetism. [primordial core model

    NASA Technical Reports Server (NTRS)

    Goldstein, M. L.

    1975-01-01

    It is shown, for a very simple model of the moon, that the existence of a primordial core magnetic field would give rise to a present day nonzero dipole external field. In the investigation a uniformly magnetized core embedded in a permeable mantle is considered. The significance of the obtained results for the conclusions reported by Runcorn (1975) is discussed. Comments provided by Runcorn to the discussion are also presented.

  13. Precession of the lunar core

    NASA Astrophysics Data System (ADS)

    Meyer, Jennifer; Wisdom, Jack

    2011-01-01

    Goldreich (Goldreich, P. [1967]. J. Geophys. Res. 72, 3135) showed that a lunar core of low viscosity would not precess with the mantle. We show that this is also the case for much of lunar history. But when the Moon was close to the Earth, the Moon's core was forced to follow closely the precessing mantle, in that the rotation axis of the core remained nearly aligned with the symmetry axis of the mantle. The transition from locked to unlocked core precession occurred between 26.0 and 29.0 Earth radii, thus it is likely that the lunar core did not follow the mantle during the Cassini transition. Dwyer and Stevenson (Dwyer, C.A., Stevenson, D.J. [2005]. An Early Nutation-Driven Lunar Dynamo. AGU Fall Meeting Abstracts GP42A-06) suggested that the lunar dynamo needs mechanical stirring to power it. The stirring is caused by the lack of locked precession of the lunar core. So, we do not expect a lunar dynamo powered by mechanical stirring when the Moon was closer to the Earth than 26.0-29.0 Earth radii. A lunar dynamo powered by mechanical stirring might have been strongest near the Cassini transition.

  14. Precession of the Lunar Core

    NASA Astrophysics Data System (ADS)

    Meyer, J.; Wisdom, J.

    2011-10-01

    Goldreich [3] showed that a lunar core of low viscosity would not precess with the mantle. We show that this is also the case for much of lunar history. But when the Moon was close to the Earth the Moon's core was forced to follow closely the precessing mantle, in that the rotation axis of the core remained nearly aligned with the symmetry axis of the mantle. The transition from locked to unlocked core precession occurred between 26.0 and 29.0 Earth radii, thus it is likely that the lunar core did not follow the mantle during the Cassini transition. Dwyer and Stevenson [1] suggested that the lunar dynamo needs mechanical stirring to power it. The stirring is caused by the lack of locked precession of the lunar core. So, we do not expect a lunar dynamo powered by mechanical stirring when the Moon was closer to the Earth than 26.0 to 29.0 Earth radii. A lunar dynamo powered by mechanical stirring might have been strongest near the Cassini transition.

  15. The sweet potato ADP-glucose pyrophosphorylase gene (ibAGP1) promoter confers high-level expression of the GUS reporter gene in the potato tuber.

    PubMed

    Kim, Tae-Won; Goo, Young-Min; Lee, Cheol-Ho; Lee, Byung-Hyun; Bae, Jung-Myung; Lee, Shin-Woo

    2009-10-01

    Molecular farming refers to the process of creating bioengineered plants with the capability of producing potentially valuable products, such as drugs, vaccines, and chemicals. We have investigated the potential of the sweet potato ADP-glucose pyrophosphorylase gene (ibAGP1) promoter and its transit peptide (TP) as an expression system for the mass production of foreign proteins in potato. The ibAGP1 promoter and its TP sequence were transformed into potato along with beta-glucuronidase (GUS) as a reporter gene, and GUS activity was subsequently analyzed in the transgenic potato plants. In tuber tissues, GUS activity in transgenic plants carrying only the ibAGP1 promoter (ibAGP1::GUS) increased up to 15.6-fold compared with that of transgenic plants carrying only the CaMV35S promoter (CaMV35S::GUS). GUS activity in transgenic plants was further enhanced by the addition of the sweetpotato TP to the recombinant vector (ibAGP1::TP::GUS), with tuber tissues showing a 26-fold increase in activity compared with that in the CaMV35S::GUS-transgenic lines. In leaf tissues, the levels of GUS activity found in ibAGP1::GUS-transgenic lines were similar to those in CaMV35S::GUS-lines, but they were significantly enhanced in ibAGP1::TP::GUS-lines. GUS activity gradually increased with increasing tuber diameter in ibAGP1::GUS-transgenic plants, reaching a maximum level when the tuber was 35 mm in diameter. In contrast, extremely elevated levels of GUS activity - up to about 10-fold higher than that found in CaMV35S::GUS-lines - were found in ibAGP1::TP::GUS-transgenic lines at a much earlier stage of tuber development (diameter 4 mm), and these higher levels were maintained throughout the entire tuber developmental stage. These results suggest that the sweetpotato ibAGP1 promoter and its TP are a potentially strong foreign gene expression system that can be used for molecular farming in potato plants.

  16. Bonding core mating surfaces improves transformer

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1978-01-01

    Modifications to assembly procedures for C-core transformers virtually eliminates changes in core end gaps due to temperature cycling during impregnation and potting stages, thus stabilizing magnetization properties of core.

  17. Residential Utility Core Wall System - ResCore

    SciTech Connect

    Boyd, G.; Lundell, C.; Wendt, R.

    1999-06-01

    This paper describes activities associated with the RESidential utility CORE wall system (ResCore) developed by students and faculty in the Department of Industrial Design at Auburn University between 1996 and 1998. These activities analyize three operational prototype units installed in Habitat for Humanity Houses. The paper contains two Parts: 1) analysis of the three operational prototype units, 2) exploration of alternative design solutions. ResCore is a manufactured construction component designed to expedite home building by decreasing the need for skilled labor at the work site. The unit concentrates untility elements into a wall unit(s), which is shipped to the construction site and installed in minimum time. The ResCore unit is intended to be built off-site in a manufacturing environment where the impact of vagaries of weather and work-crew coordination and scheduling are minimized. The controlled environment of the factory enhances efficient production of building components through material and labor throughput controls, enabling the production of components at a substantially reduced per-unit cost. The ResCore unit when compared to traditional "stick-built" utility wall components is in may ways analogous to the factory built roof truss compared to on-site "stick-Built" roof framing.

  18. Dynamic Cores in Hydrostatic Disguise

    NASA Astrophysics Data System (ADS)

    Ballesteros-Paredes, Javier; Klessen, Ralf S.; Vázquez-Semadeni, Enrique

    2003-07-01

    We discuss the column density profiles of ``cores'' in three-dimensional smoothed particle hydrodynamics (SPH) numerical simulations of turbulent molecular clouds. The SPH scheme allows us to perform a high spatial resolution analysis of the density maxima (cores) at scales between ~0.003 and 0.3 pc. We analyze simulations in three different physical conditions: large-scale driving (LSD), small-scale driving (SSD), and random Gaussian initial conditions without driving (GC), each one at two different time steps: just before self-gravity is turned on (t0) and when gravity has been operating such that 5% of the total mass in the box has been accreted into cores (t1). For this data set, we perform Bonnor-Ebert fits to the column density profiles of cores found by a clump-finding algorithm. We find that, for the particular fitting procedure we use, 65% of the cores can be matched to Bonnor-Ebert (BE) profiles, and of these, 47% correspond to stable equilibrium configurations with ξmax<6.5, even though the cores analyzed in the simulations are not in equilibrium but instead are dynamically evolving. The temperatures obtained with the fitting procedure vary between 5 and 60 K (in spite of the simulations being isothermal, with T=11.3 K), with the peak of the distribution being at T=11 K and most clumps having fitted temperatures between 5 and 30 K. Central densities obtained with the BE fit tend to be smaller than the actual central densities of the cores. We also find that for the LSD and GC cases, there are more BE-like cores at t0 than at t1 with ξmax<=20, while in the case of SSD, there are more such cores at t1 than at t0. We interpret this as a consequence of the stronger turbulence present in the cores of run SSD, which prevents good BE fits in the absence of gravity, and delays collapse in its presence. Finally, in some cases we find substantial superposition effects when we analyze the projection of the density structures, even though the scales over which we

  19. 38 CFR 0.601 - Core Values.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Core Values. VA's Core Values define VA employees. They describe the organization's culture and... their individual responsibilities and organizational responsibilities. (c) Advocacy. VA employees...

  20. 38 CFR 0.601 - Core Values.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Core Values. VA's Core Values define VA employees. They describe the organization's culture and... their individual responsibilities and organizational responsibilities. (c) Advocacy. VA employees...

  1. Counterrotating cores in elliptical galaxies

    NASA Astrophysics Data System (ADS)

    Balcella, Marc Comas

    The dynamics of the merger between a high- and a low-elliptical galaxy was studied to understand how kinematically peculiar cores in elliptical galaxies might form. Numerical simulations of mergers provide rotation curves, surface density profiles, surface density contour plots and velocity maps of the merger remnants, as well as diagnostics on the dynamics such as phase-space diagrams. This type of merger can create counterrotating cores. The core of the smaller galaxy, of higher density, is not disrupted by the primary tidal field and sinks to the center of the primary as an independent dynamical subsystem. Core counterrotation occurs only when the initial merger orbit is retrograde with respect to the pin of the primary. The remnant has higher effective radius and lower mean central surface density than the primary galaxy, but a smaller core radius. The adsorption of orbital energy and angular momentum by the primary particles greatly modifies the kinematic structure of the larger galaxy. Twisted rotation axes and isophote twists appear over the whole body of the remnant. These diagnostics may be used to determine whether observed peculiar cores might have formed via an elliptical-elliptical merger. Galaxies with counterrotating cores should show a complex velocity field, isophotal irregularities, and, in general, a slow rotation in the main body of the galaxy. The present experiments are the first galaxy-satellite merger experiments involving an active, rotating secondary. They show that part of the orbital angular momentum is absorbed by the secondary, thus the secondary contributes to its own sinking: the sinking rate depends on the orientation of the secondary spin. Long-slit spectroscopic observations of NGC 3656 are reported.

  2. Isotopic composition of ice cores and meltwater from upper fremont glacier and Galena Creek rock glacier, Wyoming

    USGS Publications Warehouse

    DeWayne, Cecil L.; Green, J.R.; Vogt, S.; Michel, R.; Cottrell, G.

    1998-01-01

    Meltwater runoff from glaciers can result from various sources, including recent precipitation and melted glacial ice. Determining the origin of the meltwater from glaciers through isotopic analysis can provide information about such things as the character and distribution of ablation on glaciers. A 9.4 m ice core and meltwater were collected in 1995 and 1996 at the glacigenic Galena Creek rock glacier in Wyoming's Absaroka Mountains. Measurements of chlorine-36 (36Cl), tritium (3H), sulphur-35 (35S), and delta oxygen-18 (??18O) were compared to similar measurements from an ice core taken from the Upper Fremont Glacier in the Wind River Range of Wyoming collected in 1991-95. Meltwater samples from three sites on the rock glacier yielded 36Cl concentrations that ranged from 2.1 ?? 1.0 X 106 to 5.8??0.3 X 106 atoms/l. The ice-core 36Cl concentrations from Galena Creek ranged from 3.4??0.3 X 105 to 1.0??0.1 X 106 atoms/l. Analysis of an ice core from the Upper Fremont Glacier yielded 36Cl concentrations of 1.2??0.2 X 106 and 5.2??0.2 X 106 atoms/l for pre- 1940 ice and between 2 X 106 and 3 X 106 atoms/l for post-1980 ice. Purdue's PRIME Lab analyzed the ice from the Upper Fremont Glacier. The highest concentration of 36Cl in the ice was 77 ?? 2 X 106 atoms/l and was deposited during the peak of atmospheric nuclear weapons testing in the late 1950s. This is an order of magnitude greater than the largest measured concentration from both the Upper Fremont Glacier ice core that was not affected by weapons testing fallout and the ice core collected from the Galena Creek rock glacier. Tritium concentrations from the rock glacier ranged from 9.2??0.6 to 13.2??0.8 tritium units (TU) in the meltwater to -1.3??1.3 TU in the ice core. Concentrations of 3H in the Upper Fremont Glacier ice core ranged from 0 TU in the ice older than 50 years to 6-12 TU in the ice deposited in the last 10 years. The maximum 3H concentration in ice from the Upper Fremont Glacier deposited in the

  3. Health promotion in Brazil.

    PubMed

    Buss, Paulo Marchiori; de Carvalho, Antonio Ivo

    2007-01-01

    The evolution of health promotion within the Brazilian health system is examined, including an assessment of the intersectoral and development policies that have influenced the process. Particular attention is paid to the legal characteristics of the Unified Health System. Human resources formation and research initiatives in health promotion are outlined, with a summary of the obstacles that need to be overcome in order to ensure the effective implementation of health promotion in the future. Up to the end of the 20th Century health promotion was not used as a term in the Brazilian public heath context. Health promoting activities were concentrated in the area of health education, although targeting the social determinants of health and the principle of intersectoral action were part of the rhetoric. The situation has changed during the last decade, with the publication of a national policy of health promotion, issued by the Ministry of Health and jointly implemented with the States and Municipals Health Secretaries. More recently there has been a re-emergence of the discourse on the social determinants of health and the formation of intersectoral public policies as the basis of a comprehensive health promotion. Health promotion infrastructure, particularly around human resources and financing, requires strengthening in order to ensure capacity and sustainability in health promotion practice.

  4. Advances in health promotion in Asia-Pacific: promoting health through hospitals.

    PubMed

    Huang, Nicole; Chien, Li-Yin; Chiou, Shu-Ti

    2016-03-01

    Since 1990, the WHO Health Promoting Hospital (HPH) movement has tried to facilitate and support hospitals to assume a core responsibility in health promotion. The Taiwan HPH Network was established in December 2006, and became the largest HPH network in the world in 2013. Compared to Europe where the HPH has been more established, the pace of HPH development has been much more rapid. This rapid development provides an inspiring example for research and health promotion practice. Systematic data and empirical information have been collected about HPH in Taiwan, allowing for research to be published about the achievements of the HPH movement. This paper provides an overview of the existing literature on current progress of the HPH project according to the four main perspectives of the WHO-HPH movement: promoting the health of patients, promoting the health of staff, changing the organization to a health-promoting setting, and promoting the health of the community in the catchment area of the hospital. The assessment can serve as a stepping stone in understanding current HPH development in Taiwan and as a reference for future research.

  5. Sensitivity of tropical cyclone intensification to inner-core structure

    NASA Astrophysics Data System (ADS)

    Ge, Xuyang; Xu, Wei; Zhou, Shunwu

    2015-10-01

    In this study, the dependence of tropical cyclone (TC) development on the inner-core structure of the parent vortex is examined using a pair of idealized numerical simulations. It is found that the radial profile of inner-core relative vorticity may have a great impact on its subsequent development. For a system with a larger inner-core relative vorticity/inertial stability, the conversion ratio of the diabatic heating to kinetic energy is greater. Furthermore, the behavior of the convective vorticity eddies is likely modulated by the system-scale circulation. For a parent vortex with a relatively higher inner-core vorticity and larger negative radial vorticity gradient, convective eddy formation and radially inward propagation is promoted through vorticity segregation. This provides a greater potential for these small-scale convective cells to self-organize into a mesoscale inner-core structure in the TC. In turn, convectively induced diabatic heating that is close to the center, along with higher inertial stability, efficiently enhances system-scale secondary circulation. This study provides a solid basis for further research into how the initial structure of a TC influences storm dynamics and thermodynamics.

  6. Core and Off-Core Processes in Systems Engineering

    NASA Technical Reports Server (NTRS)

    Breidenthal, Julian; Forsberg, Kevin

    2010-01-01

    An emerging methodology of organizing systems-engineering plans is based on a concept of core and off-core processes or activities. This concept has emerged as a result of recognition of a risk in the traditional representation of systems-engineering plans by a Vee model alone, according to which a large system is decomposed into levels of smaller subsystems, then integrated through levels of increasing scope until the full system is constructed. Actual systems-engineering activity is more complicated, raising the possibility that the staff will become confused in the absence of plans which explain the nature and ordering of work beyond the traditional Vee model.

  7. Counterrotating Cores in Elliptical Galaxies.

    NASA Astrophysics Data System (ADS)

    Balcells, Marc Comas

    The dynamics of the merger between a high- and a low-luminosity elliptical galaxy has been studied to understand how kinematically peculiar cores in elliptical galaxies might form. Numerical simulations of mergers provide rotation curves, surface density profiles, surface density contour plots and velocity maps of the merger remnants, as well as diagnostics on the dynamics such as phase-space diagrams. This type of merger can create counterrotating cores. The core of the smaller galaxy, of higher density, is not disrupted by the primary tidal field and sinks to the center of the primary as an independent dynamical subsystem. Core counterrotation occurs only when the initial merger orbit is retrograde with respect to the spin of the primary. The remnant has higher effective radius and lower mean central surface density than the primary galaxy, but a smaller core radius. The adsorption of orbital energy and angular momentum by the primary particles greatly modifies the kinematic structure of the larger galaxy. Twisted rotation axes and isophote twists appear over the whole body of the remnant. These diagnostics may be used to determine whether observed peculiar cores might have formed via an elliptical-elliptical merger. Galaxies with counterrotating cores should show a complex velocity field, isophotal irregularities, and, in general, a slow rotation in the main body of the galaxy. The present experiments are the first galaxy-satellite merger experiments involving an active, rotating secondary. They show that part of the orbital angular momentum is absorbed by the secondary, thus the secondary contributes to its own sinking: the sinking rate depends on the orientation of the secondary spin. Long-slit spectroscopic observations of NGC 3656 are reported. Rotation curves indicate that NGC 3656 contains a core spinning in a direction perpendicular to the rotation in the main body of the galaxy. Velocity reversals at intermediate radii are also observed. These features

  8. Single-domain intrabodies against hepatitis C virus core inhibit viral propagation and core-induced NFκB activation.

    PubMed

    Suzuki, Ryosuke; Saito, Kenji; Matsuda, Mami; Sato, Mitsuru; Kanegae, Yumi; Shi, Guoli; Watashi, Koichi; Aizaki, Hideki; Chiba, Joe; Saito, Izumu; Wakita, Takaji; Suzuki, Tetsuro

    2016-04-01

    Hepatitis C virus (HCV) core plays a key role in viral particle formation and is involved in viral pathogenesis. Here, constructs for single-domain intrabodies consisting of variable regions derived from mouse mAbs against HCV core were established. Expressed single-domain intrabodies were shown to bind to HCV core, and inhibit the growth of cell culture-produced HCV derived from JFH-1 (genotype 2a) and a TH (genotype 1b)/JFH-1 chimera. Adenovirus vectors expressing intrabodies were also capable of reducing HCV propagation. Intrabody expression did not affect viral entry or genome replication of single-round infectious trans-complemented HCV particles. However, intrabody expression reduced intracellular and extracellular infectious titres in CD81-defective Huh7-25 cells transfected with the HCV genome, suggesting that these intrabodies impair HCV assembly. Furthermore, intrabody expression suppressed HCV core-induced NFκB promoter activity. These intrabodies may therefore serve as tools for elucidating the role of core in HCV pathogenesis.

  9. Reactor core isolation cooling system

    DOEpatents

    Cooke, F.E.

    1992-12-08

    A reactor core isolation cooling system includes a reactor pressure vessel containing a reactor core, a drywell vessel, a containment vessel, and an isolation pool containing an isolation condenser. A turbine is operatively joined to the pressure vessel outlet steamline and powers a pump operatively joined to the pressure vessel feedwater line. In operation, steam from the pressure vessel powers the turbine which in turn powers the pump to pump makeup water from a pool to the feedwater line into the pressure vessel for maintaining water level over the reactor core. Steam discharged from the turbine is channeled to the isolation condenser and is condensed therein. The resulting heat is discharged into the isolation pool and vented to the atmosphere outside the containment vessel for removing heat therefrom. 1 figure.

  10. Reactor core isolation cooling system

    DOEpatents

    Cooke, Franklin E.

    1992-01-01

    A reactor core isolation cooling system includes a reactor pressure vessel containing a reactor core, a drywell vessel, a containment vessel, and an isolation pool containing an isolation condenser. A turbine is operatively joined to the pressure vessel outlet steamline and powers a pump operatively joined to the pressure vessel feedwater line. In operation, steam from the pressure vessel powers the turbine which in turn powers the pump to pump makeup water from a pool to the feedwater line into the pressure vessel for maintaining water level over the reactor core. Steam discharged from the turbine is channeled to the isolation condenser and is condensed therein. The resulting heat is discharged into the isolation pool and vented to the atmosphere outside the containment vessel for removing heat therefrom.

  11. High expression Zymomonas promoters

    DOEpatents

    Viitanen, Paul V.; Tao, Luan; Zhang, Yuying; Caimi, Perry G.; McCole, Laura : Zhang, Min; Chou, Yat-Chen; McCutchen, Carol M.; Franden, Mary Ann

    2011-08-02

    Identified are mutants of the promoter of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase gene, which direct improved expression levels of operably linked heterologous nucleic acids. These are high expression promoters useful for expression of chimeric genes in Zymomonas, Zymobacter, and other related bacteria.

  12. Health Promotion Program.

    ERIC Educational Resources Information Center

    McClary, Cheryl

    The Health Promotion Program began with establishment of a one-credit course in health promotion and wellness and the training of family practice residents at the Mountain Area Health Education Center to serve as lab leaders in the course. The course later became part of the university's general education requirements. In addition, a health…

  13. Promoting Resilience in Children.

    ERIC Educational Resources Information Center

    Rolfe, Sharne A.

    2002-01-01

    This booklet invites reflection on ways in which childhood resilience can be promoted, thereby helping children to adapt effectively in the face of adversity. The attributes of resilient children are described, as is the importance of protective factors in building or promoting resilience. The booklet discusses the complex interplay between risk…

  14. Assessing and Promoting Cultural Relativism in Students of Counseling

    ERIC Educational Resources Information Center

    Mcauliffe, Garrett John; Grothaus, Tim; Jensen, Margaret; Michel, Rebecca

    2012-01-01

    Multicultural counseling is often promoted as a core element in counselor development. As such, educational efforts aim to increase counselors' cultural relativism, or their ability to recognize their own enculturation and to appreciate the value of other cultural norms. This mixed qualitative-quantitative study explored the relationship between…

  15. Magnet Schools: Promoting Equal Opportunity and Quality Education.

    ERIC Educational Resources Information Center

    Office for Civil Rights (ED), Washington, DC.

    While adhering to state standards for the core curriculum required for graduation, magnet schools offer innovative, specialized instructional approaches to attract students of all socioeconomic, ethnic, and racial backgrounds. While the magnet programs vary in design, scope, and outcome, they share important objectives: (1) to promote educational…

  16. Ice Core Dating Software for Interactive Dating of Ice Cores

    NASA Astrophysics Data System (ADS)

    Kurbatov, A. V.; Mayewski, P. A.; Abdul Jawad, B. S.

    2005-12-01

    Scientists involved in ice core dating are well familiar with the problem of identification and recording the depth of annual signals using stable isotopes, glaciochemistry, ECM (electrical conductivity), DEP (dielectric properties) and particle counter data. Traditionally all parameters used for ice core dating were plotted as a function of depth, printed and after years were marked on the paper, converted to depth vs. age time scale. To expedite this tedious and manual process we developed interactive computer software, Ice core Dating (ICD) program. ICD is written in Java programming language, and uses GPL and GPL site licensed graphic libraries. The same 3.5 Mb in size pre-compiled single jar file, that includes all libraries and application code, was successfully tested on WinOS, Mac OSX, Linux, and Solaris operating systems running Java VM version 1.4. We have followed the modular design philosophy in our source code so potential integration with other software modules, data bases and server side distributed computer environments can be easily implemented. We expect to continue development of new suites of tools for easy integration of ice core data with other available time proxies. ICD is thoroughly documented and comes with a technical reference and cookbook that explains the purpose of the software and its many features, and provides examples to help new users quickly become familiar with the operation and philosophy of the software. ICD is available as a free download from the Climate Change Institute web site ( under the terms of GNU GPL public license.

  17. Core Leadership: Teacher Leaders and Common Core Implementation in Tennessee

    ERIC Educational Resources Information Center

    Aspen Institute, 2014

    2014-01-01

    In the summer of 2012, thousands of teachers across the United States attended several days of professional development workshops. The workshops, which focused on the Common Core State Standards, were part of a Tennessee Department of Education initiative in teacher leadership. The department recruited and trained 200 highly-effective teachers to…

  18. Earth's core and the geodynamo

    PubMed

    Buffett

    2000-06-16

    Earth's magnetic field is generated by fluid motion in the liquid iron core. Details of how this occurs are now emerging from numerical simulations that achieve a self-sustaining magnetic field. Early results predict a dominant dipole field outside the core, and some models even reproduce magnetic reversals. The simulations also show how different patterns of flow can produce similar external fields. Efforts to distinguish between the various possibilities appeal to observations of the time-dependent behavior of the field. Important constraints will come from geological records of the magnetic field in the past.

  19. Magnetic Probing of Core Geodynamics

    NASA Technical Reports Server (NTRS)

    Voorhies, Coerte V.

    2004-01-01

    To better understand geomagnetic theory and observation, we can use spatial magnetic spectra for the main field and secular variation to test core dynamical hypotheses against seismology. The hypotheses lead to theoretical spectra which are fitted to observational spectra. Each fit yields an estimate of the radius of Earth's core and uncertainty. If this agrees with the seismologic value, then the hypothesis passes the test. A new way to obtain theoretical spectra extends the hydromagnetic scale analysis of Benton to scale-variant field and flow. For narrow scale flow and a dynamically weak field by the top of Earth's core, this yields a generalized Stevenson-McLeod spectrum for the core-source field, and a secular variation spectrum modulated by a cubic polynomial in spherical harmonic degree n. The former passes the tests. The latter passes many tests, but does not describe rapid dipole decline and quadrupole rebound; some tests suggest it is a bit hard, or rich in narrow scale range. In a core geodynamo, motion of the fluid conductor does work against the Lorentz force. This converts kinetic into magnetic energy which, in turn, is lost to heat via Ohmic dissipation. In the analysis at length-scale 1/k, if one presumes kinetic energy is converted in either eddy-overturning or magnetic free-decay time-scales, then Kolmogorov or other spectra in conflict with observational spectra can result. Instead, the rate work is done roughly balances the dissipation rate, which is consistent with small-scale flow. The conversion time-scale depends on dynamical constraints. These are summarized by the magnetogeostrophic vertical vorticity balance by the top of the core, which includes anisotropic effects of rotation, the magnetic field, and the core-mantle boundary. The resulting theoretical spectra for the core-source field and its SV are far more compatible with observation. The conversion time-scale of order 120 years is pseudo-scale-invariant. Magnetic spectra of other

  20. Magnetic Probing of Core Geodynamics

    NASA Technical Reports Server (NTRS)

    Voorhies, Coerte V.

    2004-01-01

    To better understand geomagnetic theory and observation, we can use spatial magnetic spectra for the main field and secular variation to test core dynmcal hypotheses against seismology. The hypotheses lead to theoretical spectra which are fitted to observational spectra. Each fit yields an estimate of the radius of Earth's core and uncertainty. If this agrees with the seismologic value, then the hypothes pass the test. A new way to obtain theoretical spectra extends the hydromagnetic scale analysis of Benton to scale-variant field and flow. For narrow scale flow and a dynamically weak field by the top of Earth's core, this yields a generalized Stevenson-McLeod spectrum for the core-source field, and a secular variation spectrum modulated by a cubic polynomial in spherical harmonic degree n. The former passes the tests. The latter passes many tests, but does not describe rapid dipole decline and quadrupole rebound; some tests suggest it is a bit hard, or rich in narrow scale change. In a core geodynamo, motion of the fluid conductor does work against the Lorentz force. This converts kinetic into magnetic energy which, in turn, is lost to heat via Ohmic dissipation. In the analysis at lentgh-scale l/k, if one presumes kinetic energy is converted in either eddy- overturning or magnetic free-decay time-scales, then Kolmogorov or other spectra in conflict with observational spectra can result. Instead, the rate work is done roughly balances the dissipation rate, which is consistent with small scale flow. The conversion time-scale depends on dynamical constraints. These are summarized by the magneto-geostrophic vertical vorticity balance by the top of the core, which includes anisotropic effects of rotation, the magnetic field, and the core- mantle boundary. The resulting theoretical spectra for the core-source field and its SV are far more compatible with observation. The conversion time-scale of order l20 years is pseudo-scale-invarient. Magnetic spectra of other

  1. Core competencies in internal medicine.

    PubMed

    Porcel, José Manuel; Casademont, Jordi; Conthe, Pedro; Pinilla, Blanca; Pujol, Ramón; García-Alegría, Javier

    2012-06-01

    The working group on Competencies of Internal Medicine from the Spanish Society of Internal Medicine (SEMI) proposes a series of core competencies that we consider should be common to all European internal medicine specialists. The competencies include aspects related to patient care, clinical knowledge, technical skills, communication skills, professionalism, cost-awareness in medical care and academic activities. The proposal could be used as a working document for the Internal Medicine core curriculum in the context of the educational framework of medical specialties in Europe.

  2. Magnetic Probing of Core Geodynamics

    NASA Astrophysics Data System (ADS)

    Voorhies, C. V.

    2004-05-01

    To better understand geomagnetic theory and observation, we can use spatial magnetic spectra for the main field and secular variation to test core dynamical hypotheses against seismology. The hypotheses lead to theoretical spectra which are fitted to observational spectra. Each fit yields an estimate of the radius of Earth's core and uncertainty. If this agrees with the seismologic value, then the hypotheses pass the test. A new way to obtain theoretical spectra extends the hydromagnetic scale analysis of Benton [1992; GAFD] to scale-variant field and flow [Voorhies, 2004; JGR-SE, in press]. For narrow scale flow and a dynamically weak field by the top of Earth's core, this yields a generalized Stevenson-McLeod spectrum for the core-source field [Voorhies, Sabaka and Purucker, 2002; JGR-P], and a secular variation spectrum modulated by a cubic polynomial in spherical harmonic degree n. The former passes the tests. The latter passes many tests, but does not describe rapid dipole decline and quadrupole rebound; some tests suggest it is a bit hard, or rich in narrow scale change. In a core geodynamo, motion of the fluid conductor does work against the Lorentz force. This converts kinetic into magnetic energy which, in turn, is lost to heat via Ohmic dissipation. In the analysis at length-scale 1/k, if one presumes kinetic energy is converted in either eddy-overturning or magnetic free-decay time-scales, then Kolmogorov or other spectra in conflict with observational spectra can result. Instead, the rate work is done roughly balances the dissipation rate, which is consistent with small scale flow. The conversion time-scale depends on dynamical constraints. These are summarized by the magneto-geostrophic vertical vorticity balance by the top of the core, which includes anisotropic effects of rotation, the magnetic field, and the core-mantle boundary. The resulting theoretical spectra for the core-source field and its SV are far more compatible with observation. The

  3. Use of a stress inducible promoter to drive ectopic AtCBF expression improves potato freezing tolerance while minimizing negative effects on tuber yield.

    PubMed

    Pino, María-Teresa; Skinner, Jeffrey S; Park, Eung-Jun; Jeknić, Zoran; Hayes, Patrick M; Thomashow, Michael F; Chen, Tony H H

    2007-09-01

    Solanum tuberosum is a frost-sensitive species incapable of cold acclimation. A brief exposure to frost can significantly reduce its yields, while hard frosts can completely destroy entire crops. Thus, gains in freezing tolerance of even a few degrees would be of considerable benefit relative to frost damage. The S. tuberosum cv. Umatilla was transformed with three Arabidopsis CBF genes (AtCBF1-3) driven by either a constitutive CaMV35S or a stress-inducible Arabidopsis rd29A promoter. AtCBF1 and AtCBF3 over-expression via the 35S promoter increased freezing tolerance about 2 degrees C, whereas AtCBF2 over-expression failed to increase freezing tolerance. Transgenic plants of AtCBF1 and AtCBF3 driven by the rd29A promoter reached the same level of freezing tolerance as the 35S versions within a few hours of exposure to low but non-freezing temperatures. Constitutive expression of AtCBF genes was associated with negative phenotypes, including smaller leaves, stunted plants, delayed flowering, and reduction or lack of tuber production. While imparting the same degree of freezing tolerance, control of AtCBF expression via the stress-inducible promoter ameliorated these negative phenotypic effects and restored tuber production to levels similar to wild-type plants. These results suggest that use of a stress-inducible promoter to direct CBF transgene expression can yield significant gains in freezing tolerance without negatively impacting agronomically important traits in potato.

  4. City Core - detecting the anthropocene in urban lake cores

    NASA Astrophysics Data System (ADS)

    Kjaer, K. H.; Ilsøe, P.; Andresen, C. S.; Rasmussen, P.; Andersen, T. J.; Frei, R.; Schreiber, N.; Odgaard, B.; Funder, S.; Holm, J. M.; Andersen, K.

    2011-12-01

    Here, we presents the preliminary results from lake cores taken in ditches associated with the historical fortifications enclosing the oldest - central Copenhagen to achieve new knowledge from sediment deposits related to anthropogenic activities. We have examined sediment cores with X-ray fluorescence (XRF) analyzers to correlate element patterns from urban and industrial emissions. Thus, we aim to track these patterns back in time - long before regular routines of recording of atmospheric environment began around 1978. Furthermore, we compare our data to alternative sources of information in order to constrain and expand the temporal dating limits (approximately 1890) achieved from 210Pb activity. From custom reports and statistic sources, information on imported volumes from coal, metal and oil was obtained and related contaminants from these substances to the sediment archives. Intriguingly, we find a steep increase in import of coal and metals matching the exponential increase of lead and zinc counts from XRF-recordings of the sediment cores. In this finding, we claim to have constrain the initiation of urban industrialization. In order to confirm the age resolution of the lake cores, DNA was extracted from sediments, sedaDNA. Thus we attempt to trace plantation of well documented exotic plants to, for instance, the Botanical Garden. Through extraction and sampling of sedaDNA from these floral and arboreal specimens we intend to locate their strataigraphic horizons in the sediment core. These findings may correlate data back to 1872, when the garden was established on the area of the former fortification. In this line of research, we hope to achieve important supplementary knowledge of sedaDNA-leaching frequencies within freshwater sediments.

  5. Producing gapped-ferrite transformer cores

    NASA Technical Reports Server (NTRS)

    Mclyman, W. T.

    1980-01-01

    Improved manufacturing techniques make reproducible gaps and minimize cracking. Molded, unfired transformer cores are cut with thin saw and then fired. Hardened semicircular core sections are bonded together, placed in aluminum core box, and fluidized-coated. After winding is run over box, core is potted. Economical method significantly reduces number of rejects.

  6. Gelcasting Alumina Cores for Investment Casting

    SciTech Connect

    Janney, M A; Klug, F J

    2001-01-01

    General Electric currently uses silica investment casting cores for making superalloy turbine blades. The silica core technology does not provide the degree of dimensional control needed for advanced turbine system manufacture. The sum of the various process variables in silica core manufacturing produces cores that have more variability than is allowed for in advanced, power-generation gas turbine airfoils.

  7. Both core and F proteins of hepatitis C virus could enhance cell proliferation in transgenic mice

    SciTech Connect

    Hu, Wen-Ta; Li, Hui-Chun; Lee, Shen-Kao; Ma, Hsin-Chieh; Yang, Chee-Hing; Chen, Hung-Ling; Lo, Shih-Yen

    2013-05-24

    Highlights: •HCV core and F proteins could induce hepatocyte proliferation in the transgenic mice. •β-Catenin signaling pathway was activated by core protein in the transgenic mice. •β-Catenin signaling pathway was activated by myc-F protein in the transgenic mice. •Expression of SMA protein was enhanced by core but not myc-F protein. -- Abstract: The role of the protein encoded by the alternative open reading frame (ARF/F/core+1) of the Hepatitis C virus (HCV) genome in viral pathogenesis remains unknown. The different forms of ARF/F/core+1 protein were labile in cultured cells, a myc-tag fused at the N-terminus of the F protein made it more stable. To determine the role of core and F proteins in HCV pathogenesis, transgenic mice with either protein expression under the control of Albumin promoter were generated. Expression of core protein and F protein with myc tag (myc-F) could be detected by Western blotting analysis in the livers of these mice. The ratio of liver to body weight is increased for both core and myc-F transgenic mice compared to that of wild type mice. Indeed, the proliferating cell nuclear antigen protein, a proliferation marker, was up-regulated in the transgenic mice with core or myc-F protein. Further analyses by microarray and Western blotting suggested that β-catenin signaling pathway was activated by either core or myc-F protein in the transgenic mice. These transgenic mice were further treated with either Diethynitrosamine (a tumor initiator) or Phenobarbital (a tumor promoter). Phenobarbital but not Diethynitrosamine treatment could increase the liver/body weight ratio of these mice. However, no tumor formation was observed in these mice. In conclusion, HCV core and myc-F proteins could induce hepatocyte proliferation in the transgenic mice possibly through β-catenin signaling pathway.

  8. Visual Feedback for Rover-based Coring

    NASA Technical Reports Server (NTRS)

    Backes, Paul; Helmick, Daniel; Bajracharya, Max

    2008-01-01

    Technology for coring from a low-mass rover has been developed to enable core sample acquisition where a planetary rover experiences moderate slip during the coring operation. A new stereo vision technique, Absolute Motion Visual Odometry, is used to measure rover slip during coring and the slip is accommodated through corresponding arm pose updating. Coring rate is controlled by feedback of themeasured force of the coring tool against the environment. Test results in the JPL Marsyard show for the first time that coring from a low-mass rover with slip is feasible.

  9. Core-melt source reduction system

    DOEpatents

    Forsberg, Charles W.; Beahm, Edward C.; Parker, George W.

    1995-01-01

    A core-melt source reduction system for ending the progression of a molten core during a core-melt accident and resulting in a stable solid cool matrix. The system includes alternating layers of a core debris absorbing material and a barrier material. The core debris absorbing material serves to react with and absorb the molten core such that containment overpressurization and/or failure does not occur. The barrier material slows the progression of the molten core debris through the system such that the molten core has sufficient time to react with the core absorbing material. The system includes a provision for cooling the glass/molten core mass after the reaction such that a stable solid cool matrix results.

  10. Core-melt source reduction system

    DOEpatents

    Forsberg, C.W.; Beahm, E.C.; Parker, G.W.

    1995-04-25

    A core-melt source reduction system for ending the progression of a molten core during a core-melt accident and resulting in a stable solid cool matrix. The system includes alternating layers of a core debris absorbing material and a barrier material. The core debris absorbing material serves to react with and absorb the molten core such that containment overpressurization and/or failure does not occur. The barrier material slows the progression of the molten core debris through the system such that the molten core has sufficient time to react with the core absorbing material. The system includes a provision for cooling the glass/molten core mass after the reaction such that a stable solid cool matrix results. 4 figs.

  11. Core 1- and core 3-derived O-glycans collectively maintain the colonic mucus barrier and protect against spontaneous colitis in mice

    PubMed Central

    Bergstrom, Kirk; Fu, Jianxin; Johansson, Malin EV; Liu, Xiaowei; Gao, Nan; Wu, Qian; Song, Jianhua; McDaniel, J Michael; McGee, Samuel; Chen, Weichang; Braun, Jonathan; Hansson, Gunnar C; Xia, Lijun

    2016-01-01

    Core 1- and core 3-derived mucin-type O-glycans are primary components of the mucus layer in the colon. Reduced mucus thickness and impaired O-glycosylation are observed in human ulcerative colitis. However, how both types of O-glycans maintain mucus barrier function in the colon is unclear. We found that C1galt1 expression, which synthesizes core 1 O-glycans, was detected throughout the colon, whereas C3GnT, which controls core 3 O-glycan formation, was most highly expressed in the proximal colon. Consistent with this, mice lacking intestinal core 1-derived O-glycans (IEC C1galt1−/−) developed spontaneous colitis primarily in the distal colon, whereas mice lacking both intestinal core 1- and core 3-derived O-glycans (DKO) developed spontaneous colitis in both distal and proximal colon. DKO mice showed an early onset and more severe colitis than IEC C1galt1−/− mice. Antibiotic treatment restored the mucus layer and attenuated colitis in DKO mice. Mucins from DKO mice were more susceptible to proteolysis than WT mucins. This study indicates that core 1- and 3-derived O-glycans collectively contribute to the mucus barrier by protecting it from bacterial protease degradation and suggests new therapeutic targets to promote mucus barrier function in colitis patients. PMID:27143302

  12. Experience with the BEACON core monitoring system

    SciTech Connect

    Beard, C.L. ); Icide, C.A. )

    1992-01-01

    The BEACON operational core support system was developed for use in pressurized water reactors to provide an integrated system to perform reactor core monitoring, core measurement reduction, core analysis and follow, and core predictions. It is based on the very fast and accurate three-dimensional SPNOVA nodal program. The experience to date has shown the importance of an accurate integrated system. The benefits accrued are greater for the total system than the benefits that are possible separately.

  13. Laminated grid and web magnetic cores

    DOEpatents

    Sefko, John; Pavlik, Norman M.

    1984-01-01

    A laminated magnetic core characterized by an electromagnetic core having core legs which comprise elongated apertures and edge notches disposed transversely to the longitudinal axis of the legs, such as high reluctance cores with linear magnetization characteristics for high voltage shunt reactors. In one embodiment the apertures include compact bodies of microlaminations for more flexibility and control in adjusting permeability and/or core reluctance.

  14. Stellar core collapse and supernova

    SciTech Connect

    Wilson, J.R.; Mayle, R.; Woosley, S.E.; Weaver, T.

    1985-04-01

    Massive stars that end their stable evolution as their iron cores collapse to a neutron star or black hole long been considered good candidates for producing Type II supernovae. For many years the outward propagation of the shock wave produced by the bounce of these iron cores has been studied as a possible mechanism for the explosion. For the most part, the results of these studies have not been particularly encouraging, except, perhaps, in the case of very low mass iron cores or very soft nuclear equations of state. The shock stalls, overwhelmed by photodisintegration and neutrino losses, and the star does not explode. More recently, slow late time heating of the envelope of the incipient neutron star has been found to be capable of rejuvenating the stalled shock and producing an explosion after all. The present paper discusses this late time heating and presents results from numerical calculations of the evolution, core collapse, and subsequent explosion of a number of recent stellar models. For the first time they all, except perhaps the most massive, explode with reasonable choices of input physics. 39 refs., 17 figs., 1 tab.

  15. Common Core State Standards 101

    ERIC Educational Resources Information Center

    Rothman, Robert

    2013-01-01

    The Common Core State Standards (CCSS) represent the first time that nearly every state has set common expectations for what students should know and be able to do. In the past, each state set its own standards, and the results varied widely. And while states collectively developed these common standards, decisions about the curriculum and…

  16. Large core fiber optic cleaver

    DOEpatents

    Halpin, John M.

    1996-01-01

    The present invention relates to a device and method for cleaving optical fibers which yields cleaved optical fiber ends possessing high damage threshold surfaces. The device can be used to cleave optical fibers with core diameters greater than 400 .mu.m.

  17. Droplet Core Nuclear Rocket (DCNR)

    NASA Technical Reports Server (NTRS)

    Anghaie, Samim

    1991-01-01

    The most basic design feature of the droplet core nuclear reactor is to spray liquid uranium into the core in the form of droplets on the order of five to ten microns in size, to bring the reactor to critical conditions. The liquid uranium fuel ejector is driven by hydrogen, and more hydrogen is injected from the side of the reactor to about one and a half meters from the top. High temperature hydrogen is expanded through a nozzle to produce thrust. The hydrogen pressure in the system can be somewhere between 50 and 500 atmospheres; the higher pressure is more desirable. In the lower core region, hydrogen is tangentially injected to serve two purposes: (1) to provide a swirling flow to protect the wall from impingement of hot uranium droplets: (2) to generate a vortex flow that can be used for fuel separation. The reactor is designed to maximize the energy generation in the upper region of the core. The system can result in and Isp of 2000 per second, and a thrust-to-weight ratio of 1.6 for the shielded reactor. The nuclear engine system can reduce the Mars mission duration to less than 200 days. It can reduce the hydrogen consumption by a factor of 2 to 3, which reduces the hydrogen load by about 130 to 150 metric tons.

  18. 10 Core External Environmental Trends.

    ERIC Educational Resources Information Center

    El Camino Coll., Torrance, CA.

    This is an institutional report summarizing 10 core external environmental trends and their implications for El Camino College and the surrounding community. The report offers a brief description for the following trends: (1) there is more emphasis on colleges becoming learning institutions rather than teaching institutions; (2) the current and…

  19. Stability of Molten Core Materials

    SciTech Connect

    Layne Pincock; Wendell Hintze

    2013-01-01

    The purpose of this report is to document a literature and data search for data and information pertaining to the stability of nuclear reactor molten core materials. This includes data and analysis from TMI-2 fuel and INL’s LOFT (Loss of Fluid Test) reactor project and other sources.

  20. The fluffy core of Enceladus

    NASA Astrophysics Data System (ADS)

    Roberts, James H.

    2015-09-01

    Enceladus is well known for its young south polar terrain, observed by Cassini to emit several GW of heat as well as plumes of vapor and ice. The source of this energy is believed to be tidal dissipation. However, the observed south polar heat flux cannot be sustained over the age of the Solar System. Furthermore, thermal evolution models suggest that any global subsurface ocean should freeze on a timescale of tens to hundreds of My, sharply reducing future tidal heating, unless large amounts of antifreeze are present in the ocean. Here I propose an alternative internal structure for Enceladus, in which the silicate core is fragmented, and that the tidal deformation of the core may be partially controlled by interstitial ice. I find that fragmentation of the core increases tidal dissipation by a factor of 20, consistent with the long-term dynamically sustainable level, even when the interior is completely frozen, but only if the interior starts out warm and tidal heating is strong from the beginning. If this is not the case, radioactive heating will be insufficient to prevent the interior from cooling. Although an ocean need not be present in order for the interior to experience significant tidal heating, all models that dissipate enough heat to prevent runaway cooling are also warm enough to have an ocean. Tidal dissipation in the weak core provides an additional source of heat that may prevent a global subsurface ocean from freezing.

  1. Earth rotation and core topography

    NASA Technical Reports Server (NTRS)

    Hager, Bradford H.; Clayton, Robert W.; Spieth, Mary Ann

    1988-01-01

    The NASA Geodynamics program has as one of its missions highly accurate monitoring of polar motion, including changes in length of day (LOD). These observations place fundamental constraints on processes occurring in the atmosphere, in the mantle, and in the core of the planet. Short-timescale (t less than or approx 1 yr) variations in LOD are mainly the result of interaction between the atmosphere and the solid earth, while variations in LOD on decade timescales result from the exchange of angular momentum between the mantle and the fluid core. One mechanism for this exchange of angular momentum is through topographic coupling between pressure variations associated with flow in the core interacting with topography at the core-mantel boundary (CMB). Work done under another NASA grant addressing the origin of long-wavelength geoid anomalies as well as evidence from seismology, resulted in several models of CMB topography. The purpose of work supported by NAG5-819 was to study further the problem of CMB topography, using geodesy, fluid mechanics, geomagnetics, and seismology. This is a final report.

  2. Common Core: Solve Math Problems

    ERIC Educational Resources Information Center

    Strom, Erich

    2012-01-01

    The new common core standards for mathematics demand that students (and teachers!) exhibit deeper conceptual understanding. That's music to the ears of education professor John Tapper, who says teachers have overemphasized teaching procedures--and getting right answers. In his new book, "Solving for Why," he makes a powerful case for moving beyond…

  3. Large core fiber optic cleaver

    DOEpatents

    Halpin, J.M.

    1996-03-26

    The present invention relates to a device and method for cleaving optical fibers which yields cleaved optical fiber ends possessing high damage threshold surfaces. The device can be used to cleave optical fibers with core diameters greater than 400 {micro}m. 30 figs.

  4. [Core competencies in internal medicine].

    PubMed

    Porcel, J M; Casademont, J; Conthe, P; Pinilla, B; Pujol, R; García-Alegría, J

    2011-06-01

    The working group of the Spanish Society of Internal Medicine (SEMI) on "Competencies of the Internist" has defined the basic medical knowledge, skills and attitudes that all internists in Spain should have. This list of competencies represents the Internal Medicine core curriculum within the context of the future educational framework of medical specialties in Health Sciences.

  5. Common Core: Fact vs. Fiction

    ERIC Educational Resources Information Center

    Greene, Kim

    2012-01-01

    Despite students' interest in informational text, it has played second fiddle in literacy instruction for years. Now, though, nonfiction is getting its turn in the spotlight. The Common Core State Standards require that students become thoughtful consumers of complex, informative texts--taking them beyond the realm of dry textbooks and…

  6. "Common Core Implementation Best Practices"

    ERIC Educational Resources Information Center

    Martin, Carmel

    2014-01-01

    This document presents the testimony of Carmel Martin, Executive Vice President for Policy at the Center for American Progress, delivered at the New York State Office of the Governor Common Core Implementation Panel on Wednesday, February 19, 2014. In this statement, Martin began by saying that The Center for American Progress believes that this…

  7. One Health Core Competency Domains

    PubMed Central

    Frankson, Rebekah; Hueston, William; Christian, Kira; Olson, Debra; Lee, Mary; Valeri, Linda; Hyatt, Raymond; Annelli, Joseph; Rubin, Carol

    2016-01-01

    The emergence of complex global challenges at the convergence of human, animal, and environmental health has catalyzed a movement supporting “One Health” approaches. Despite recognition of the importance of One Health approaches to address these complex challenges, little effort has been directed at identifying the seminal knowledge, skills, and attitudes necessary for individuals to successfully contribute to One Health efforts. Between 2008 and 2011, three groups independently embarked on separate initiatives to identify core competencies for professionals involved with One Health approaches. Core competencies were considered critically important for guiding curriculum development and continuing professional education, as they describe the knowledge, skills, and attitudes required to be effective. A workshop was convened in 2012 to synthesize the various strands of work on One Health competencies. Despite having different mandates, participants, and approaches, all of these initiatives identified similar core competency domains: management; communication and informatics; values and ethics; leadership; teams and collaboration; roles and responsibilities; and systems thinking. These core competency domains have been used to develop new continuing professional education programs for One Health professionals and help university curricula prepare new graduates to be able to contribute more effectively to One Health approaches. PMID:27679794

  8. Common Core: Rx for Change

    ERIC Educational Resources Information Center

    Jaeger, Paige

    2012-01-01

    When David Coleman, one of the authors of the Common Core State Standards (CCSS), spoke to New York educators, he stated that over the last forty years 8th grade reading scores have been flat. Despite doubling expenditures on classroom instruction, there has been little growth. Most educators are aware that what worked for the students of the…

  9. CopperCore Service Integration

    ERIC Educational Resources Information Center

    Vogten, Hubert; Martens, Harrie; Nadolski, Rob; Tattersall, Colin; van Rosmalen, Peter; Koper, Rob

    2007-01-01

    In an e-learning environment there is a need to integrate various e-learning services like assessment services, collaboration services, learning design services and communication services. In this article we present the design and implementation of a generic integrative service framework, called CopperCore Service Integration (CCSI). We will…

  10. Wasting Away: Chicago's Declining Core

    ERIC Educational Resources Information Center

    Kissel, Adam

    2009-01-01

    The University of Chicago met widespread national opposition ten years ago after it instituted a new, less demanding core curriculum to make way for more electives. It was part of a plan to make the curriculum significantly less demanding in order to attract more students and improve the school's bottom line in a time of putative budget deficits.…

  11. A Core Program in JIAFS

    NASA Technical Reports Server (NTRS)

    Whitesides, John L.

    2000-01-01

    This paper presents a final report on A Core Program in JIAFS (Joint Institute for Advancement of Flight Sciences). The objectives of the program were to conduct high-risk innovative research, administer and direct the on-going programs, and appoint additional Graduate Research Scholar Assistants depending on availability of applicants and funds.

  12. Magnectic Probing of Core Geodynamics

    NASA Technical Reports Server (NTRS)

    Voorhies, Coerte

    2004-01-01

    To better understand geomagnetic theory and observation, we can use spatial magnetic spectra for the main field and secular variation to test core dynamical hypotheses against seismology. The hypotheses lead to theoretical spectra which are fitted to observational spectra. Each fit yields an estimate of the radius of Earth s core and uncertainty. If this agrees with the seismologic value, then the hypotheses pass the test. A new way to obtain theoretical spectra extends the hydromagnetic scale analysis of Benton to scale-variant field and flow. For narrow scale flow and a dynamically weak field by the top of Earth s core, this yields a JGR-PI, and a secular variation spectrum modulated by a cubic polynomial in spherical harmonic degree n. The former passes the tests. The latter passes many tests, but does not describe rapid dipole decline and quadrupole rebound; some tests suggest it is a bit hard, or rich in narrow scale change.In a core geodynamo, motion of the fluid conductor does work against the Lorentz force. This converts kinetic into magnetic energy which, in turn, is lost to heat via Ohmic dissipation. In the analysis at length- scale l/k, if one presumes kinetic energy is converted in either eddy- overturning or magnetic free-decay time-scales, then Kolmogorov or other spectra in conflict with observational spectra can result. Instead, the rate work is done roughly balances the dissipation rate, which is consistent with small scale flow. The conversion time-scale depends on dynamical constraints. These are summarized by the magneto- geostrophic vertical vorticity balance by the top of the core, which includes anisotropic effects of rotation, the magnetic field, and the core-mantle boundary. The resulting theoretical spectra for the core- source field and its SV are far more compatible with observation. The conversion time-scale of order 120 years is pseudo-scale-invariant. Magnetic spectra of other planets may differ; however, if a transition to non

  13. 77 FR 47820 - Invention Promoters/Promotion Firms Complaints

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-10

    ... United States Patent and Trademark Office Invention Promoters/Promotion Firms Complaints ACTION: Proposed... participate in any legal proceedings against invention promoters or promotion firms. Complaints submitted to... promotion firm, explain the basis for the complaint, and include the signature of the complainant....

  14. Translating social ecological theory into guidelines for community health promotion.

    PubMed

    Stokols, D

    1996-01-01

    Health promotion programs often lack a clearly specified theoretical foundation or are based on narrowly conceived conceptual models. For example, lifestyle modification programs typically emphasize individually focused behavior change strategies, while neglecting the environmental underpinnings of health and illness. This article compares three distinct, yet complementary, theoretical perspectives on health promotion: behavioral change, environmental enhancement, and social ecological models. Key strengths and limitations of each perspective are examined, and core principles of social ecological theory are used to derive practical guidelines for designing and evaluating community health promotion programs. Directions for future health promotion research are discussed, including studies examining the role of intermediaries (e.g., corporate decision-makers, legislators) in promoting the well-being of others, and those evaluating the duration and scope of intervention outcomes.

  15. Health promotion by social cognitive means.

    PubMed

    Bandura, Albert

    2004-04-01

    This article examines health promotion and disease prevention from the perspective of social cognitive theory. This theory posits a multifaceted causal structure in which self-efficacy beliefs operate together with goals, outcome expectations, and perceived environmental impediments and facilitators in the regulation of human motivation, behavior, and well-being. Belief in one's efficacy to exercise control is a common pathway through which psychosocial influences affect health functioning. This core belief affects each of the basic processes of personal change--whether people even consider changing their health habits, whether they mobilize the motivation and perseverance needed to succeed should they do so, their ability to recover from setbacks and relapses, and how well they maintain the habit changes they have achieved. Human health is a social matter, not just an individual one. A comprehensive approach to health promotion also requires changing the practices of social systems that have widespread effects on human health.

  16. 43 CFR 3593.1 - Core or test hole cores, samples, cuttings.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 43 Public Lands: Interior 2 2014-10-01 2014-10-01 false Core or test hole cores, samples, cuttings...) EXPLORATION AND MINING OPERATIONS Bore Holes and Samples § 3593.1 Core or test hole cores, samples, cuttings... all core or test holes made on the lands covered by the lease, license or permit. The records shall...

  17. 43 CFR 3593.1 - Core or test hole cores, samples, cuttings.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 2 2012-10-01 2012-10-01 false Core or test hole cores, samples, cuttings...) EXPLORATION AND MINING OPERATIONS Bore Holes and Samples § 3593.1 Core or test hole cores, samples, cuttings... all core or test holes made on the lands covered by the lease, license or permit. The records shall...

  18. 43 CFR 3593.1 - Core or test hole cores, samples, cuttings.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 2 2013-10-01 2013-10-01 false Core or test hole cores, samples, cuttings...) EXPLORATION AND MINING OPERATIONS Bore Holes and Samples § 3593.1 Core or test hole cores, samples, cuttings... all core or test holes made on the lands covered by the lease, license or permit. The records shall...

  19. 43 CFR 3593.1 - Core or test hole cores, samples, cuttings.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Core or test hole cores, samples, cuttings...) EXPLORATION AND MINING OPERATIONS Bore Holes and Samples § 3593.1 Core or test hole cores, samples, cuttings... officer. The authorized officer may cut such cores and receive samples as appropriate. Upon the request...

  20. Novelty and Promotional Items

    EPA Pesticide Factsheets

    Small novelty or promotional products, primarily used for outreach and educational purposes, must effectively convey a message, and their purchase will only be allowed if the item will contribute to the accomplishment of the Agency's mission.