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  1. The protein network surrounding the human telomere repeat binding factors TRF1, TRF2, and POT1.

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    Richard J Giannone

    2010-08-01

    Full Text Available Telomere integrity (including telomere length and capping is critical in overall genomic stability. Telomere repeat binding factors and their associated proteins play vital roles in telomere length regulation and end protection. In this study, we explore the protein network surrounding telomere repeat binding factors, TRF1, TRF2, and POT1 using dual-tag affinity purification in combination with multidimensional protein identification technology liquid chromatography--tandem mass spectrometry (MudPIT LC-MS/MS. After control subtraction and data filtering, we found that TRF2 and POT1 co-purified all six members of the telomere protein complex, while TRF1 identified five of six components at frequencies that lend evidence towards the currently accepted telomere architecture. Many of the known TRF1 or TRF2 interacting proteins were also identified. Moreover, putative associating partners identified for each of the three core components fell into functional categories such as DNA damage repair, ubiquitination, chromosome cohesion, chromatin modification/remodeling, DNA replication, cell cycle and transcription regulation, nucleotide metabolism, RNA processing, and nuclear transport. These putative protein-protein associations may participate in different biological processes at telomeres or, intriguingly, outside telomeres.

  2. Development and characterization of polyclonal antibodies against the linker region of the telomere-binding protein TRF2

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    Nadya V. Ilicheva

    2018-03-01

    Full Text Available Background: TRF2 (telomeric repeat binding factor 2 is an essential component of the telomere-binding protein complex shelterin. TRF2 induces the formation of a special structure of telomeric DNA and counteracts activation of DNA damage-response pathways telomeres. TRF2 has a poorly characterized linker region (udTRF2 between its homodimerization and DNA-binding domains. Some lines of evidence have shown that this region could be involved in TRF2 interaction with nuclear lamina. Results: In this study, the fragment of the TERF2 gene encoding udTRF2 domain of telomere-binding protein TRF2 was produced by PCR and cloned into the pET32a vector. The resulting plasmid pET32a-udTRF2 was used for the expression of the recombinant udTRF2 in E. coli RosettaBlue (DE3. The protein was isolated and purified using ammonium sulfate precipitation followed by ion-exchange chromatography. The purified recombinant protein udTRF2 was injected into guinea pigs to generate polyclonal antibodies. The ability of anti-udTRF2 antibodies to bind endogenous TRF2 in human skin fibroblasts was tested by western blotting and immunofluorescent staining. Conclusions: In this study, the recombinant protein udTRF2 and antibodies to it were generated. Both protein and antibodies will provide a useful tool for investigation of the functions of the udTRF2 domain and its role in the interaction between TRF2 and nuclear lamina. Keywords: Chromosomes, Molecular cloning, Nuclear lamina, Nucleoprotein complexes, Polyclonal antibodies, Recombinant polypeptide, Shelterin, Telomere-binding protein TRF2, Telomeres, Telomeric DNA, TTAGGG repeats

  3. Molecular recognition in complexes of TRF proteins with telomeric DNA.

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    Miłosz Wieczór

    Full Text Available Telomeres are specialized nucleoprotein assemblies that protect the ends of linear chromosomes. In humans and many other species, telomeres consist of tandem TTAGGG repeats bound by a protein complex known as shelterin that remodels telomeric DNA into a protective loop structure and regulates telomere homeostasis. Shelterin recognizes telomeric repeats through its two major components known as Telomere Repeat-Binding Factors, TRF1 and TRF2. These two homologous proteins are therefore essential for the formation and normal function of telomeres. Indeed, TRF1 and TRF2 are implicated in a plethora of different cellular functions and their depletion leads to telomere dysfunction with chromosomal fusions, followed by apoptotic cell death. More specifically, it was found that TRF1 acts as a negative regulator of telomere length, and TRF2 is involved in stabilizing the loop structure. Consequently, these proteins are of great interest, not only because of their key role in telomere maintenance and stability, but also as potential drug targets. In the current study, we investigated the molecular basis of telomeric sequence recognition by TRF1 and TRF2 and their DNA binding mechanism. We used molecular dynamics (MD to calculate the free energy profiles for binding of TRFs to telomeric DNA. We found that the predicted binding free energies were in good agreement with experimental data. Further, different molecular determinants of binding, such as binding enthalpies and entropies, the hydrogen bonding pattern and changes in surface area, were analyzed to decompose and examine the overall binding free energies at the structural level. With this approach, we were able to draw conclusions regarding the consecutive stages of sequence-specific association, and propose a novel aspartate-dependent mechanism of sequence recognition. Finally, our work demonstrates the applicability of computational MD-based methods to studying protein-DNA interactions.

  4. Telomere Restriction Fragment (TRF) Analysis.

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    Mender, Ilgen; Shay, Jerry W

    2015-11-20

    While telomerase is expressed in ~90% of primary human tumors, most somatic tissue cells except transiently proliferating stem-like cells do not have detectable telomerase activity (Shay and Wright, 1996; Shay and Wright, 2001). Telomeres progressively shorten with each cell division in normal cells, including proliferating stem-like cells, due to the end replication (lagging strand synthesis) problem and other causes such as oxidative damage, therefore all somatic cells have limited cell proliferation capacity (Hayflick limit) (Hayflick and Moorhead, 1961; Olovnikov, 1973). The progressive telomere shortening eventually leads to growth arrest in normal cells, which is known as replicative senescence (Shay et al. , 1991). Once telomerase is activated in cancer cells, telomere length is stabilized by the addition of TTAGGG repeats to the end of chromosomes, thus enabling the limitless continuation of cell division (Shay and Wright, 1996; Shay and Wright, 2001). Therefore, the link between aging and cancer can be partially explained by telomere biology. There are many rapid and convenient methods to study telomere biology such as Telomere Restriction Fragment (TRF), Telomere Repeat Amplification Protocol (TRAP) (Mender and Shay, 2015b) and Telomere dysfunction Induced Foci (TIF) analysis (Mender and Shay, 2015a). In this protocol paper we describe Telomere Restriction Fragment (TRF) analysis to determine average telomeric length of cells. Telomeric length can be indirectly measured by a technique called Telomere Restriction Fragment analysis (TRF). This technique is a modified Southern blot, which measures the heterogeneous range of telomere lengths in a cell population using the length distribution of the terminal restriction fragments (Harley et al. , 1990; Ouellette et al. , 2000). This method can be used in eukaryotic cells. The description below focuses on the measurement of human cancer cells telomere length. The principle of this method relies on the lack of

  5. Identification of TTAGGG-binding proteins in Neurospora crassa, a fungus with vertebrate-like telomere repeats.

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    Casas-Vila, Núria; Scheibe, Marion; Freiwald, Anja; Kappei, Dennis; Butter, Falk

    2015-11-17

    To date, telomere research in fungi has mainly focused on Saccharomyces cerevisiae and Schizosaccharomyces pombe, despite the fact that both yeasts have degenerated telomeric repeats in contrast to the canonical TTAGGG motif found in vertebrates and also several other fungi. Using label-free quantitative proteomics, we here investigate the telosome of Neurospora crassa, a fungus with canonical telomeric repeats. We show that at least six of the candidates detected in our screen are direct TTAGGG-repeat binding proteins. While three of the direct interactors (NCU03416 [ncTbf1], NCU01991 [ncTbf2] and NCU02182 [ncTay1]) feature the known myb/homeobox DNA interaction domain also found in the vertebrate telomeric factors, we additionally show that a zinc-finger protein (NCU07846) and two proteins without any annotated DNA-binding domain (NCU02644 and NCU05718) are also direct double-strand TTAGGG binders. We further find two single-strand binders (NCU02404 [ncGbp2] and NCU07735 [ncTcg1]). By quantitative label-free interactomics we identify TTAGGG-binding proteins in Neurospora crassa, suggesting candidates for telomeric factors that are supported by phylogenomic comparison with yeast species. Intriguingly, homologs in yeast species with degenerated telomeric repeats are also TTAGGG-binding proteins, e.g. in S. cerevisiae Tbf1 recognizes the TTAGGG motif found in its subtelomeres. However, there is also a subset of proteins that is not conserved. While a rudimentary core TTAGGG-recognition machinery may be conserved across yeast species, our data suggests Neurospora as an emerging model organism with unique features.

  6. TRF1 and TRF2 use different mechanisms to find telomeric DNA but share a novel mechanism to search for protein partners at telomeres.

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    Lin, Jiangguo; Countryman, Preston; Buncher, Noah; Kaur, Parminder; E, Longjiang; Zhang, Yiyun; Gibson, Greg; You, Changjiang; Watkins, Simon C; Piehler, Jacob; Opresko, Patricia L; Kad, Neil M; Wang, Hong

    2014-02-01

    Human telomeres are maintained by the shelterin protein complex in which TRF1 and TRF2 bind directly to duplex telomeric DNA. How these proteins find telomeric sequences among a genome of billions of base pairs and how they find protein partners to form the shelterin complex remains uncertain. Using single-molecule fluorescence imaging of quantum dot-labeled TRF1 and TRF2, we study how these proteins locate TTAGGG repeats on DNA tightropes. By virtue of its basic domain TRF2 performs an extensive 1D search on nontelomeric DNA, whereas TRF1's 1D search is limited. Unlike the stable and static associations observed for other proteins at specific binding sites, TRF proteins possess reduced binding stability marked by transient binding (∼ 9-17 s) and slow 1D diffusion on specific telomeric regions. These slow diffusion constants yield activation energy barriers to sliding ∼ 2.8-3.6 κ(B)T greater than those for nontelomeric DNA. We propose that the TRF proteins use 1D sliding to find protein partners and assemble the shelterin complex, which in turn stabilizes the interaction with specific telomeric DNA. This 'tag-team proofreading' represents a more general mechanism to ensure a specific set of proteins interact with each other on long repetitive specific DNA sequences without requiring external energy sources.

  7. Resolution of telomere associations by TRF1 cleavage in mouse embryonic stem cells

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    Lisaingo, Kathleen; Uringa, Evert-Jan; Lansdorp, Peter M.

    2014-01-01

    Telomere associations have been observed during key cellular processes such as mitosis, meiosis, and carcinogenesis and must be resolved before cell division to prevent genome instability. Here we establish that telomeric repeat-binding factor 1 (TRF1), a core component of the telomere protein

  8. Nucleolar TRF2 attenuated nucleolus stress-induced HCC cell-cycle arrest by altering rRNA synthesis.

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    Yuan, Fuwen; Xu, Chenzhong; Li, Guodong; Tong, Tanjun

    2018-05-03

    The nucleolus is an important organelle that is responsible for the biogenesis of ribosome RNA (rRNA) and ribosomal subunits assembly. It is also deemed to be the center of metabolic control, considering the critical role of ribosomes in protein translation. Perturbations of rRNA synthesis are closely related to cell proliferation and tumor progression. Telomeric repeat-binding factor 2 (TRF2) is a member of shelterin complex that is responsible for telomere DNA protection. Interestingly, it was recently reported to localize in the nucleolus of human cells in a cell-cycle-dependent manner, while the underlying mechanism and its role on the nucleolus remained unclear. In this study, we found that nucleolar and coiled-body phosphoprotein 1 (NOLC1), a nucleolar protein that is responsible for the nucleolus construction and rRNA synthesis, interacted with TRF2 and mediated the shuttle of TRF2 between the nucleolus and nucleus. Abating the expression of NOLC1 decreased the nucleolar-resident TRF2. Besides, the nucleolar TRF2 could bind rDNA and promoted rRNA transcription. Furthermore, in hepatocellular carcinoma (HCC) cell lines HepG2 and SMMC7721, TRF2 overexpression participated in the nucleolus stress-induced rRNA inhibition and cell-cycle arrest.

  9. Telomere shortening is associated to TRF1 and PARP1 overexpression in Duchenne muscular dystrophy.

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    Aguennouz, M'Hammed; Vita, Gian Luca; Messina, Sonia; Cama, Annamaria; Lanzano, Natalia; Ciranni, Annamaria; Rodolico, Carmelo; Di Giorgio, Rosa Maria; Vita, Giuseppe

    2011-12-01

    Telomere shortening is thought to contribute to premature senescence of satellite cells in Duchenne muscular dystrophy (DMD) muscle. Telomeric repeat binding factor-1 (TRF1) and poly (ADP-ribose) polymerase-1 (PARP1) are proteins known to modulate telomerase reverse transcriptase (TERT) activity, which controls telomere elongation. Here we show that an age-dependent telomere shortening occurs in DMD muscles and is associated to overexpression of mRNA and protein levels of TRF1 and PARP1. TERT expression and activity are detectable in normal control muscles and they slightly increase in DMD. This is the first demonstration of TRF1 and PARP1 overexpression in DMD muscles. They can be directly involved in replicative senescence of satellite cells and/or in the pathogenetic cascade through a cross-talk with oxidative stress and inflammatory response. Modulation of these events by TRF1 or PARP1 inhibition might represent a novel strategy for treatment of DMD and other muscular dystrophies. Copyright © 2010 Elsevier Inc. All rights reserved.

  10. Caspase-Dependent Apoptosis Induced by Telomere Cleavage and TRF2 Loss

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    Asha S. Multani

    2000-07-01

    Full Text Available Chromosomal abnormalities involving telomeric associations (TAs often precede replicative senescence and abnormal chromosome configurations. We report here that telomere cleavage following exposure to proapoptotic agents is an early event in apoptosis. Exposure of human and murine cancer cells to a variety of pro-apoptotic stimuli (staurosporine, thapsigargin, anti-Fas antibody, cancer chemotherapeutic agents resulted in telomere cleavage and aggregation, finally their extrusion from the nuclei. Telomere loss was associated with arrest of cells in G2/M phase and preceded DNA fragmentation. Telomere erosion and subsequent large-scale chromatin cleavage were inhibited by overexpression of the anti -apoptotic protein, bcl-2, two peptide caspase inhibitors (BACMK and zVADfmk, indicating that both events are regulated by caspase activation. The results demonstrate that telomere cleavage is an early chromatin alteration detected in various cancer cell lines leading to drug-induced apoptosis, suggest that this event contributes to mitotic catastrophe and induction of cell death. Results also suggest that the decrease of telomeric-repeat binding factor 2 (TRF2 may be the earliest event in the ara-C-induced telomere shortening, induction of endoreduplication and chromosomal fragmentation leading to cell death.

  11. Analysis of the nuclear localization signal of TRF1 in non-small cell lung cancer

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    HU JIAN

    2009-01-01

    Full Text Available Several studies revealed a similar down-regulation of telomeric repeat binding factor 1 (TRF1 in tumors. We have previously reported the TRFl expression levels were down-regulation in non-small cell lung cancer (NSCLC. The regulation of TRFl localization is proposed to be important for the function and expression. The nuclear localization signal (NLS and nuclear export signal (NES are often important clues to localization of protein. The objective of the present study was to investigate the NLS and NES of TRFl in NSCLC patients. Thirty (30 patients with NSCLCs had undergone radical operations in The First Affiliated Hospital, College of Medicine, Zhejiang University. DNA sequences of NLSs and NESs were amplified by PCR. The PCR products were analyzed by DNA sequencing. There were four NLSs of the TRFl protein, including two monopartite and two bipartite NLSs. The NLSs sequences were included in 337KKERRVGTPQSTKKKKESRR356. The exon 8 and exon 9 of TRFl DNA were covered the NLS sequences. The sequences of predicted NESs were 11WMLDFLCLSL86 and 174NLLKLQALAV183, respectively. The exon 1, exon 3 and exon 4 of TRFl were covered the NES sequences. In NSCLCs, there was no a mutation, deletion, or substitution in NLS and NES of TRFl. We conclude that the NLS and NES sequences in NSCLCs patients did not have mutations. Down-expression of TRFl does not indicate gene mutation of NLS and NES in NSCLCs.

  12. Solution properties of the archaeal CRISPR DNA repeat-binding homeodomain protein Cbp2

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    Kenchappa, Chandra; Heiðarsson, Pétur Orri; Kragelund, Birthe

    2013-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) form the basis of diverse adaptive immune systems directed primarily against invading genetic elements of archaea and bacteria. Cbp1 of the crenarchaeal thermoacidophilic order Sulfolobales, carrying three imperfect repeats, binds...... specifically to CRISPR DNA repeats and has been implicated in facilitating production of long transcripts from CRISPR loci. Here, a second related class of CRISPR DNA repeat-binding protein, denoted Cbp2, is characterized that contains two imperfect repeats and is found amongst members of the crenarchaeal...... in facilitating high affinity DNA binding of Cbp2 by tethering the two domains. Structural studies on mutant proteins provide support for Cys(7) and Cys(28) enhancing high thermal stability of Cbp2(Hb) through disulphide bridge formation. Consistent with their proposed CRISPR transcriptional regulatory role, Cbp2...

  13. Suppressive oligodeoxynucleotides containing TTAGGG motifs inhibit cGAS activation in human monocytes.

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    Steinhagen, Folkert; Zillinger, Thomas; Peukert, Konrad; Fox, Mario; Thudium, Marcus; Barchet, Winfried; Putensen, Christian; Klinman, Dennis; Latz, Eicke; Bode, Christian

    2018-04-01

    Type I interferon (IFN) is a critical mediator of autoimmune diseases such as systemic lupus erythematosus (SLE) and Aicardi-Goutières Syndrome (AGS). The recently discovered cyclic-GMP-AMP (cGAMP) synthase (cGAS) induces the production of type I IFN in response to cytosolic DNA and is potentially linked to SLE and AGS. Suppressive oligodeoxynucleotides (ODN) containing repetitive TTAGGG motifs present in mammalian telomeres have proven useful in the treatment of autoimmune diseases including SLE. In this study, we demonstrate that the suppressive ODN A151 effectively inhibits activation of cGAS in response to cytosolic DNA, thereby inhibiting type I IFN production by human monocytes. In addition, A151 abrogated cGAS activation in response to endogenous accumulation of DNA using TREX1-deficient monocytes. We demonstrate that A151 prevents cGAS activation in a manner that is competitive with DNA. This suppressive activity of A151 was dependent on both telomeric sequence and phosphorothioate backbone. To our knowledge this report presents the first cGAS inhibitor capable of blocking self-DNA. Collectively, these findings might lead to the development of new therapeutics against IFN-driven pathologies due to cGAS activation. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. TRF2 and the evolution of the bilateria.

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    Duttke, Sascha H C; Doolittle, Russell F; Wang, Yuan-Liang; Kadonaga, James T

    2014-10-01

    The development of a complex body plan requires a diversity of regulatory networks. Here we consider the concept of TATA-box-binding protein (TBP) family proteins as "system factors" that each supports a distinct set of transcriptional programs. For instance, TBP activates TATA-box-dependent core promoters, whereas TBP-related factor 2 (TRF2) activates TATA-less core promoters that are dependent on a TCT or downstream core promoter element (DPE) motif. These findings led us to investigate the evolution of TRF2. TBP occurs in Archaea and eukaryotes, but TRF2 evolved prior to the emergence of the bilateria and subsequent to the evolutionary split between bilaterians and nonbilaterian animals. Unlike TBP, TRF2 does not bind to the TATA box and could thus function as a new system factor that is largely independent of TBP. We postulate that this TRF2-based system served as the foundation for new transcriptional programs, such as those involved in triploblasty and body plan development, that facilitated the evolution of bilateria. © 2014 Duttke et al.; Published by Cold Spring Harbor Laboratory Press.

  15. Modulation of CRISPR locus transcription by the repeat-binding protein Cbp1 in Sulfolobus

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    Deng, Ling; Kenchappa, Chandra Shekar; Peng, Xu

    2012-01-01

    CRISPR loci are essential components of the adaptive immune system of archaea and bacteria. They consist of long arrays of repeats separated by DNA spacers encoding guide RNAs (crRNA), which target foreign genetic elements. Cbp1 (CRISPR DNA repeat binding protein) binds specifically to the multiple...... direct repeats of CRISPR loci of members of the acidothermophilic, crenarchaeal order Sulfolobales. cbp1 gene deletion from Sulfolobus islandicus REY15A produced a strong reduction in pre-crRNA yields from CRISPR loci but did not inhibit the foreign DNA targeting capacity of the CRISPR/Cas system....... Conversely, overexpression of Cbp1 in S. islandicus generated an increase in pre-crRNA yields while the level of reverse strand transcripts from CRISPR loci remained unchanged. It is proposed that Cbp1 modulates production of longer pre-crRNA transcripts from CRISPR loci. A possible mechanism...

  16. Drosophila TRF2 and TAF9 regulate lipid droplet size and phospholipid fatty acid composition.

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    Fan, Wei; Lam, Sin Man; Xin, Jingxue; Yang, Xiao; Liu, Zhonghua; Liu, Yuan; Wang, Yong; Shui, Guanghou; Huang, Xun

    2017-03-01

    The general transcription factor TBP (TATA-box binding protein) and its associated factors (TAFs) together form the TFIID complex, which directs transcription initiation. Through RNAi and mutant analysis, we identified a specific TBP family protein, TRF2, and a set of TAFs that regulate lipid droplet (LD) size in the Drosophila larval fat body. Among the three Drosophila TBP genes, trf2, tbp and trf1, only loss of function of trf2 results in increased LD size. Moreover, TRF2 and TAF9 regulate fatty acid composition of several classes of phospholipids. Through RNA profiling, we found that TRF2 and TAF9 affects the transcription of a common set of genes, including peroxisomal fatty acid β-oxidation-related genes that affect phospholipid fatty acid composition. We also found that knockdown of several TRF2 and TAF9 target genes results in large LDs, a phenotype which is similar to that of trf2 mutants. Together, these findings provide new insights into the specific role of the general transcription machinery in lipid homeostasis.

  17. A generic Transcriptomics Reporting Framework (TRF) for 'omics data processing and analysis.

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    Gant, Timothy W; Sauer, Ursula G; Zhang, Shu-Dong; Chorley, Brian N; Hackermüller, Jörg; Perdichizzi, Stefania; Tollefsen, Knut E; van Ravenzwaay, Ben; Yauk, Carole; Tong, Weida; Poole, Alan

    2017-12-01

    A generic Transcriptomics Reporting Framework (TRF) is presented that lists parameters that should be reported in 'omics studies used in a regulatory context. The TRF encompasses the processes from transcriptome profiling from data generation to a processed list of differentially expressed genes (DEGs) ready for interpretation. Included within the TRF is a reference baseline analysis (RBA) that encompasses raw data selection; data normalisation; recognition of outliers; and statistical analysis. The TRF itself does not dictate the methodology for data processing, but deals with what should be reported. Its principles are also applicable to sequencing data and other 'omics. In contrast, the RBA specifies a simple data processing and analysis methodology that is designed to provide a comparison point for other approaches and is exemplified here by a case study. By providing transparency on the steps applied during 'omics data processing and analysis, the TRF will increase confidence processing of 'omics data, and regulatory use. Applicability of the TRF is ensured by its simplicity and generality. The TRF can be applied to all types of regulatory 'omics studies, and it can be executed using different commonly available software tools. Crown Copyright © 2017. Published by Elsevier Inc. All rights reserved.

  18. TRF2 Protein Interacts with Core Histones to Stabilize Chromosome Ends*

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    Izumi, Takashi; Shimizu, Shigeomi

    2016-01-01

    Mammalian chromosome ends are protected by a specialized nucleoprotein complex called telomeres. Both shelterin, a telomere-specific multi-protein complex, and higher order telomeric chromatin structures combine to stabilize the chromosome ends. Here, we showed that TRF2, a component of shelterin, binds to core histones to protect chromosome ends from inappropriate DNA damage response and loss of telomeric DNA. The N-terminal Gly/Arg-rich domain (GAR domain) of TRF2 directly binds to the globular domain of core histones. The conserved arginine residues in the GAR domain of TRF2 are required for this interaction. A TRF2 mutant with these arginine residues substituted by alanine lost the ability to protect telomeres and induced rapid telomere shortening caused by the cleavage of a loop structure of the telomeric chromatin. These findings showed a previously unnoticed interaction between the shelterin complex and nucleosomal histones to stabilize the chromosome ends. PMID:27514743

  19. Cloning of Ammopiptanthus mongolicus C-repeat-binding factor gene and its cold-induced tolerance in transgenic tobacco

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    Lijiang Gu

    2013-12-01

    Full Text Available C-repeat-binding factors (CBFs are a type of important regulon in stress-related signal transduction pathways that control plant tolerance of abiotic stress. Ammopiptanthus mongolicus is the only evergreen broadleaf shrub in the northwest desert of China. The species shows strong resistance to environmental stress, especially to cold stress. An A. mongolicus CBF1 gene (AmCBF1 was cloned and transformed into tobacco. Expression of AmCBF1 could be detected in A. mongolicus shortly after exposure to low temperature of 4°C. Analysis on ratio of electrolytic leakage, soluble sugar content, free proline content, malondialdehyde (MDA content and peroxidase (POD activity before and after cold treatment (4°C for 24 h indicated AmCBF1 conferred higher cold tolerance to AmCBF1 transgenic tobacco compared with the wild type and empty vector transformed tobacco.

  20. TRF2 recruits RTEL1 to telomeres in S phase to promote t-loop unwinding.

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    Sarek, Grzegorz; Vannier, Jean-Baptiste; Panier, Stephanie; Petrini, John H J; Boulton, Simon J

    2015-02-19

    The helicase RTEL1 promotes t-loop unwinding and suppresses telomere fragility to maintain the integrity of vertebrate telomeres. An interaction between RTEL1 and PCNA is important to prevent telomere fragility, but how RTEL1 engages with the telomere to promote t-loop unwinding is unclear. Here, we establish that the shelterin protein TRF2 recruits RTEL1 to telomeres in S phase, which is required to prevent catastrophic t-loop processing by structure-specific nucleases. We show that the TRF2-RTEL1 interaction is mediated by a metal-coordinating C4C4 motif in RTEL1, which is compromised by the Hoyeraal-Hreidarsson syndrome (HHS) mutation, RTEL1(R1264H). Conversely, we define a TRF2(I124D) substitution mutation within the TRFH domain of TRF2, which eliminates RTEL1 binding and phenocopies the RTEL1(R1264H) mutation, giving rise to aberrant t-loop excision, telomere length heterogeneity, and loss of the telomere as a circle. These results implicate TRF2 in the recruitment of RTEL1 to facilitate t-loop disassembly at telomeres in S phase. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  1. Genome-wide Control of Heterochromatin Replication by the Telomere Capping Protein TRF2.

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    Mendez-Bermudez, Aaron; Lototska, Liudmyla; Bauwens, Serge; Giraud-Panis, Marie-Josèphe; Croce, Olivier; Jamet, Karine; Irizar, Agurtzane; Mowinckel, Macarena; Koundrioukoff, Stephane; Nottet, Nicolas; Almouzni, Genevieve; Teulade-Fichou, Mare-Paule; Schertzer, Michael; Perderiset, Mylène; Londoño-Vallejo, Arturo; Debatisse, Michelle; Gilson, Eric; Ye, Jing

    2018-05-03

    Hard-to-replicate regions of chromosomes (e.g., pericentromeres, centromeres, and telomeres) impede replication fork progression, eventually leading, in the event of replication stress, to chromosome fragility, aging, and cancer. Our knowledge of the mechanisms controlling the stability of these regions is essentially limited to telomeres, where fragility is counteracted by the shelterin proteins. Here we show that the shelterin subunit TRF2 ensures progression of the replication fork through pericentromeric heterochromatin, but not centromeric chromatin. In a process involving its N-terminal basic domain, TRF2 binds to pericentromeric Satellite III sequences during S phase, allowing the recruitment of the G-quadruplex-resolving helicase RTEL1 to facilitate fork progression. We also show that TRF2 is required for the stability of other heterochromatic regions localized throughout the genome, paving the way for future research on heterochromatic replication and its relationship with aging and cancer. Copyright © 2018 Elsevier Inc. All rights reserved.

  2. The telomeric protein TRF2 binds the ATM kinase and can inhibit the ATM-dependent DNA damage response.

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    Jan Karlseder

    2004-08-01

    Full Text Available The telomeric protein TRF2 is required to prevent mammalian telomeres from activating DNA damage checkpoints. Here we show that overexpression of TRF2 affects the response of the ATM kinase to DNA damage. Overexpression of TRF2 abrogated the cell cycle arrest after ionizing radiation and diminished several other readouts of the DNA damage response, including phosphorylation of Nbs1, induction of p53, and upregulation of p53 targets. TRF2 inhibited autophosphorylation of ATM on S1981, an early step in the activation of this kinase. A region of ATM containing S1981 was found to directly interact with TRF2 in vitro, and ATM immunoprecipitates contained TRF2. We propose that TRF2 has the ability to inhibit ATM activation at telomeres. Because TRF2 is abundant at chromosome ends but not elsewhere in the nucleus, this mechanism of checkpoint control could specifically block a DNA damage response at telomeres without affecting the surveillance of chromosome internal damage.

  3. TRF2 Protein Interacts with Core Histones to Stabilize Chromosome Ends.

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    Konishi, Akimitsu; Izumi, Takashi; Shimizu, Shigeomi

    2016-09-23

    Mammalian chromosome ends are protected by a specialized nucleoprotein complex called telomeres. Both shelterin, a telomere-specific multi-protein complex, and higher order telomeric chromatin structures combine to stabilize the chromosome ends. Here, we showed that TRF2, a component of shelterin, binds to core histones to protect chromosome ends from inappropriate DNA damage response and loss of telomeric DNA. The N-terminal Gly/Arg-rich domain (GAR domain) of TRF2 directly binds to the globular domain of core histones. The conserved arginine residues in the GAR domain of TRF2 are required for this interaction. A TRF2 mutant with these arginine residues substituted by alanine lost the ability to protect telomeres and induced rapid telomere shortening caused by the cleavage of a loop structure of the telomeric chromatin. These findings showed a previously unnoticed interaction between the shelterin complex and nucleosomal histones to stabilize the chromosome ends. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Strand exchange of telomeric DNA catalyzed by the Werner syndrome protein (WRN) is specifically stimulated by TRF2

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    Edwards, Deanna N.; Orren, David K.; Machwe, Amrita

    2014-01-01

    Werner syndrome (WS), caused by loss of function of the RecQ helicase WRN, is a hereditary disease characterized by premature aging and elevated cancer incidence. WRN has DNA binding, exonuclease, ATPase, helicase and strand annealing activities, suggesting possible roles in recombination-related processes. Evidence indicates that WRN deficiency causes telomeric abnormalities that likely underlie early onset of aging phenotypes in WS. Furthermore, TRF2, a protein essential for telomere protection, interacts with WRN and influences its basic helicase and exonuclease activities. However, these studies provided little insight into WRN's specific function at telomeres. Here, we explored the possibility that WRN and TRF2 cooperate during telomeric recombination processes. Our results indicate that TRF2, through its interactions with both WRN and telomeric DNA, stimulates WRN-mediated strand exchange specifically between telomeric substrates; TRF2's basic domain is particularly important for this stimulation. Although TRF1 binds telomeric DNA with similar affinity, it has minimal effects on WRN-mediated strand exchange of telomeric DNA. Moreover, TRF2 is displaced from telomeric DNA by WRN, independent of its ATPase and helicase activities. Together, these results suggest that TRF2 and WRN act coordinately during telomeric recombination processes, consistent with certain telomeric abnormalities associated with alteration of WRN function. PMID:24880691

  5. TRF2 Recruits RTEL1 to Telomeres in S Phase to Promote T-Loop Unwinding

    OpenAIRE

    Sarek, Grzegorz; Vannier, Jean-Baptiste; Panier, Stephanie; Petrini, John?H.J.; Boulton, Simon?J.

    2015-01-01

    Summary The helicase RTEL1 promotes t-loop unwinding and suppresses telomere fragility to maintain the integrity of vertebrate telomeres. An interaction between RTEL1 and PCNA is important to prevent telomere fragility, but how RTEL1 engages with the telomere to promote t-loop unwinding is unclear. Here, we establish that the shelterin protein TRF2 recruits RTEL1 to telomeres in S phase, which is required to?prevent catastrophic t-loop processing by structure-specific nucleases. We show that ...

  6. Accumulation and altered localization of telomere-associated protein TRF2 in immortally transformed and tumor-derived human breast cells

    Energy Technology Data Exchange (ETDEWEB)

    Nijjar, Tarlochan; Bassett, Ekaterina; Garbe, James; Takenaka, Yasuhiro; Stampfer, Martha R.; Gilley, David; Yaswen, Paul

    2004-12-23

    We have used cultured human mammary epithelial cells (HMEC) and breast tumor-derived lines to gain information on defects that occur during breast cancer progression. HMEC immortalized by a variety of agents (the chemical carcinogen benzo(a)pyrene, oncogenes c-myc and ZNF217, and/or dominant negative p53 genetic suppressor element GSE22) displayed marked up regulation (10-15 fold) of the telomere binding protein, TRF2. Up-regulation of TRF2 protein was apparently due to differences in post-transcriptional regulation, as mRNA levels remained comparable in finite life span and immortal HMEC. TRF2 protein was not up-regulated by the oncogenic agents alone in the absence of immortalization, nor by expression of exogenously introduced hTERT genes. We found TRF2 levels to be at least 2-fold higher than in control cells in 11/15 breast tumor cell lines, suggesting that elevated TRF2 levels are a frequent occurrence during the transformation of breast tumor cells in vivo. The dispersed distribution of TRF2 throughout the nuclei in some immortalized and tumor-derived cells indicated that not all the TRF2 was associated with telomeres in these cells. The process responsible for accumulation of TRF2 in immortalized HMEC and breast tumor-derived cell lines may promote tumorigenesis by contributing to the cells ability to maintain an indefinite life span.

  7. Dual roles of TRF1 in tethering telomeres to the nuclear envelope and protecting them from fusion during meiosis.

    Science.gov (United States)

    Wang, Lina; Tu, Zhaowei; Liu, Chao; Liu, Hongbin; Kaldis, Philipp; Chen, Zijiang; Li, Wei

    2018-01-08

    Telomeres integrity is indispensable for chromosomal stability by preventing chromosome erosion and end-to-end fusions. During meiosis, telomeres attach to the inner nuclear envelope and cluster into a highly crowded microenvironment at the bouquet stage, which requires specific mechanisms to protect the telomeres from fusion. Here, we demonstrate that germ cell-specific knockout of a shelterin complex subunit, Trf1, results in arrest of spermatocytes at two different stages. The obliterated telomere-nuclear envelope attachment in Trf1-deficient spermatocytes impairs homologue synapsis and recombination, resulting in a pachytene-like arrest, while the meiotic division arrest might stem from chromosome end-to-end fusion due to the failure of recruiting meiosis specific telomere associated proteins. Further investigations uncovered that TRF1 could directly interact with Speedy A, and Speedy A might work as a scaffold protein to further recruit Cdk2, thus protecting telomeres from fusion at this stage. Together, our results reveal a novel mechanism of TRF1, Speedy A, and Cdk2 in protecting telomere from fusion in a highly crowded microenvironment during meiosis.

  8. Transcriptome Profiling Reveals the Negative Regulation of Multiple Plant Hormone Signaling Pathways Elicited by Overexpression of C-Repeat Binding Factors

    Directory of Open Access Journals (Sweden)

    Aixin Li

    2017-09-01

    Full Text Available C-repeat binding factors (CBF are a subfamily of AP2 transcription factors that play critical roles in the regulation of plant cold tolerance and growth in low temperature. In the present work, we sought to perform a detailed investigation into global transcriptional regulation of plant hormone signaling associated genes in transgenic plants engineered with CBF genes. RNA samples from Arabidopsis thaliana plants overexpressing two CBF genes, CBF2 and CBF3, were subjected to Illumina HiSeq 2000 RNA sequencing (RNA-Seq. Our results showed that more than half of the hormone associated genes that were differentially expressed in CBF2 or CBF3 transgenic plants were related to auxin signal transduction and metabolism. Most of these alterations in gene expression could lead to repression of auxin signaling. Accordingly, the IAA content was significantly decreased in young tissues of plants overexpressing CBF2 and CBF3 compared with wild type. In addition, genes associated with the biosynthesis of Jasmonate (JA and Salicylic acid (SA, as well as the signal sensing of Brassinolide (BR and SA, were down-regulated, while genes associated with Gibberellin (GA deactivation were up-regulated. In general, overexpression of CBF2 and CBF3 negatively affects multiple plant hormone signaling pathways in Arabidopsis. The transcriptome analysis using CBF2 and CBF3 transgenic plants provides novel and integrated insights into the interaction between CBFs and plant hormones, particularly the modulation of auxin signaling, which may contribute to the improvement of crop yields under abiotic stress via molecular engineering using CBF genes.

  9. Synthetic Oligodeoxynucleotides Containing Multiple Telemeric TTAGGG Motifs Suppress Inflammasome Activity in Macrophages Subjected to Oxygen and Glucose Deprivation and Reduce Ischemic Brain Injury in Stroke-Prone Spontaneously Hypertensive Rats.

    Directory of Open Access Journals (Sweden)

    Jing Zhao

    Full Text Available The immune system plays a fundamental role in both the development and pathobiology of stroke. Inflammasomes are multiprotein complexes that have come to be recognized as critical players in the inflammation that ultimately contributes to stroke severity. Inflammasomes recognize microbial and host-derived danger signals and activate caspase-1, which in turn controls the production of the pro-inflammatory cytokine IL-1β. We have shown that A151, a synthetic oligodeoxynucleotide containing multiple telemeric TTAGGG motifs, reduces IL-1β production by activated bone marrow derived macrophages that have been subjected to oxygen-glucose deprivation and LPS stimulation. Further, we demonstrate that A151 reduces the maturation of caspase-1 and IL-1β, the levels of both the iNOS and NLRP3 proteins, and the depolarization of mitochondrial membrane potential within such cells. In addition, we have demonstrated that A151 reduces ischemic brain damage and NLRP3 mRNA levels in SHR-SP rats that have undergone permanent middle cerebral artery occlusion. These findings clearly suggest that the modulation of inflammasome activity via A151 may contribute to a reduction in pro-inflammatory cytokine production by macrophages subjected to conditions that model brain ischemia and modulate ischemic brain damage in an animal model of stroke. Therefore, modulation of ischemic pathobiology by A151 may have a role in the development of novel stroke prevention and therapeutic strategies.

  10. The C-terminal extension unique to the long isoform of the shelterin component TIN2 enhances its interaction with TRF2 in a phosphorylation- and dyskeratosis congenita-cluster-dependent fashion.

    Science.gov (United States)

    Nelson, Nya D; Dodson, Lois M; Escudero, Laura; Sukumar, Ann T; Williams, Christopher L; Mihalek, Ivana; Baldan, Alessandro; Baird, Duncan M; Bertuch, Alison A

    2018-03-26

    TIN2 is central to the shelterin complex, linking the telomeric proteins TRF1 and TRF2 with TPP1/POT1. Mutations in TINF2 , which encodes TIN2, that are found in dyskeratosis congenita (DC) result in very short telomeres and cluster in a region shared by the two TIN2 isoforms, TIN2S (short) and TIN2L (long). Here we show that TIN2L, but not TIN2S, is phosphorylated. TRF2 interacts more with TIN2L than TIN2S, and both the DC-cluster and phosphorylation promote this enhanced interaction. The binding of TIN2L, but not TIN2S, is affected by TRF2-F120, which is also required for TRF2's interaction with end processing factors such as Apollo. Conversely, TRF1 interacts more with TIN2S than with TIN2L. A DC-associated mutation further reduces TIN2L-TRF1, but not TIN2S-TRF1, interaction. Cells overexpressing TIN2L or phosphomimetic-TIN2L are permissive to telomere elongation, whereas cells overexpressing TIN2S or phosphodead-TIN2L are not. Telomere lengths are unchanged in cell lines in which TIN2L expression has been eliminated by CRISPR/Cas9-mediated mutation. These results indicate that TIN2 isoforms are biochemically and functionally distinguishable, and that shelterin composition could be fundamentally altered in patients with TINF2 mutations. Copyright © 2018 Nelson et al.

  11. Simultaneous pyrosequencing of the 16S rRNA, IncP-1 trfA, and merA genes

    DEFF Research Database (Denmark)

    Holmsgaard, Peter Nikolai; Sørensen, Søren Johannes; Hansen, Lars H.

    2013-01-01

    The use of amplicon pyrosequencing makes it possible to produce thousands of sequences of the same gene at relatively low costs. Here we show that it is possible to simultaneously sequence the 16S rRNA gene, IncP-1 trfA gene and mercury reductase gene (merA) as a way for screening the diversity...

  12. Synaptic vesicles contain small ribonucleic acids (sRNAs) including transfer RNA fragments (trfRNA) and microRNAs (miRNA).

    Science.gov (United States)

    Li, Huinan; Wu, Cheng; Aramayo, Rodolfo; Sachs, Matthew S; Harlow, Mark L

    2015-10-08

    Synaptic vesicles (SVs) are neuronal presynaptic organelles that load and release neurotransmitter at chemical synapses. In addition to classic neurotransmitters, we have found that synaptic vesicles isolated from the electric organ of Torpedo californica, a model cholinergic synapse, contain small ribonucleic acids (sRNAs), primarily the 5' ends of transfer RNAs (tRNAs) termed tRNA fragments (trfRNAs). To test the evolutionary conservation of SV sRNAs we examined isolated SVs from the mouse central nervous system (CNS). We found abundant levels of sRNAs in mouse SVs, including trfRNAs and micro RNAs (miRNAs) known to be involved in transcriptional and translational regulation. This discovery suggests that, in addition to inducing changes in local dendritic excitability through the release of neurotransmitters, SVs may, through the release of specific trfRNAs and miRNAs, directly regulate local protein synthesis. We believe these findings have broad implications for the study of chemical synaptic transmission.

  13. Low-temperature-induced expression of rice ureidoglycolate amidohydrolase is mediated by a C-repeat/dehydration-responsive element that specifically interacts with rice C-repeat-binding factor 3

    Directory of Open Access Journals (Sweden)

    Juan eLi

    2015-11-01

    Full Text Available Nitrogen recycling and redistribution are important for the environmental stress response of plants. In non nitrogen-fixing plants, ureide metabolism is crucial to nitrogen recycling from organic sources. Various studies have suggested that the rate-limiting components of ureide metabolism respond to environmental stresses. However, the underlying regulation mechanism is not well understood. In this report, rice ureidoglycolate amidohydrolase (OsUAH, which is a recently identified enzyme catalyzing the final step of ureide degradation, was identified as low-temperature- (LT but not abscisic acid- (ABA regulated. To elucidate the LT regulatory mechanism at the transcriptional level, we isolated and characterized the promoter region of OsUAH (POsUAH. Series deletions revealed that a minimal region between -522 and -420 relative to the transcriptional start site was sufficient for the cold induction of POsUAH. Detailed analyses of this 103-bp fragment indicated that a C-repeat/dehydration-responsive (CRT/DRE element localized at position -434 was essential for LT-responsive expression. A rice C-repeat-binding factors/DRE-binding proteins 1 (CBFs/DREB1s subfamily member, OsCBF3, was screened to specifically bind to the CRT/DRE element in the minimal region both in yeast one-hybrid assays and in in vitro gel-shift analysis. Moreover, the promoter could be exclusively trans-activated by the interaction between the CRT/DRE element and OsCBF3 in vivo. These findings may help to elucidate the regulation mechanism of stress-responsive ureide metabolism genes and provide an example of the member-specific manipulation of the CBF/DREB1 subfamily.

  14. High expression of TRF2, SOX10, and CD10 in circulating tumor microemboli detected in metastatic melanoma patients. A potential impact for the assessment of disease aggressiveness.

    Science.gov (United States)

    Long, Elodie; Ilie, Marius; Bence, Coraline; Butori, Catherine; Selva, Eric; Lalvée, Salomé; Bonnetaud, Christelle; Poissonnet, Gilles; Lacour, Jean-Philippe; Bahadoran, Philippe; Brest, Patrick; Gilson, Eric; Ballotti, Robert; Hofman, Véronique; Hofman, Paul

    2016-06-01

    Circulating tumors cells (CTCs) can be detected in the blood of metastatic melanoma patients (MMPs) both as isolated circulating tumor cells (iCTCs) and circulating tumor microemboli (CTMs), but their clinical significance remains unknown. The aim of this work was to evaluate the prognostic impact in metastatic cutaneous melanoma of CTMs and iCTCs identified by a cytomorphological approach using the isolation by size of tumor cell (ISET) method. We characterized the phenotype of CTCs using anti-PS100, anti-SOX10, anti-CD10, and anti-TRF2 antibodies. 128 MMPs and 37 control healthy individuals with benign nevi were included in this study. Results were compared to the follow-up of patients. 109/128 (85%) MMPs showed CTCs, 44/128 (34%) with 2 to 6 CTMs and 65/128 (51%) with 4 to 9 iCTCs. PS100 expression was homogeneous in iCTCs and heterogeneous in CTMs. SOX10, CD10, and TRF2 were mainly expressed in CTMs. None of the control subjects demonstrated circulating malignant tumor cells. Overall survival was significantly decreased in patients with CTMs, independently of the therapeutic strategies. In conclusion, the presence of CTMs is an independent predictor of shorter survival from the time of diagnosis of MMPs. © 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  15. Child Behavior Checklist (CBCL,Youth Self-Report (YSR and Teacher's Report Form(TRF: an overview of the development of the original and Brazilian versions Child Behavior Checklist (CBCL, Youth Self- Report (YSR y Teacher's Report Form (TRF: una visión general sobre el desarrollo de las versiones originales y brasileñas Child Behavior Checklist (CBCL, Youth Self-Report (YSR e Teacher's Report Form (TRF: uma visão geral sobre o desenvolvimento das versões originais e brasileiras

    Directory of Open Access Journals (Sweden)

    Isabel A. Bordin

    2013-01-01

    Full Text Available The Achenbach System of Empirically Based Assessment (ASEBA for school-age children includes three instruments for assessing emotional and/or behavioral problems: Child Behavior Checklist (CBCL, completed by parents, Youth Self-Report (YSR, completed by adolescents and Teacher's Report Form (TRF, completed by teachers. This review article gives detailed information on the development of these forms in the United States and Brazil, describing the main changes to the items, scales and score cut-off points in original versions between 1991 and 2001, as well as the process involved in the translation, back-translation and cultural adaptation of the original questionnaires to develop the current official Brazilian versions of the CBCL, YSR and TRF. The utility of these tools for research and clinical practice is highlighted, mentioning epidemiological studies and evaluation of interventions conducted in Brazil. Researchers' and clinicians' doubts regarding the correct use of the current official Brazilian versions are answered, giving examples of frequently asked questions relevant to the Brazilian context.El sistema de evaluación de base empírica de Achenbach para niños/adolescentes en edad escolar incluye tres instrumentos para evaluar problemas emocionales y/o de comportamiento: Child Behavior Checklist (CBCL [padres], Youth Self-Report (YSR [adolescentes] y Teacher's Report Form (TRF [profesores]. Este artículo de revisión proporciona información detallada sobre el desarrollo de estos instrumentos en los Estados Unidos y en Brasil, describiendo las principales alteraciones en ítems, escalas y puntos de corte en la puntuación, que se realizaron en las versiones originales de 1991 a 2001, y el proceso de traducción, retrotraducción y adaptación cultural de los cuestionarios originales, con el fin de desarrollar las actuales versiones brasileñas oficiales del CBCL, YSR y TRF. La utilidad de estos instrumentos en investigación y en la pr

  16. Ia-restricted B-B cell interaction. I. The MHC haplotype of bone marrow cells present during B cell ontogeny dictates the self-recognition specificity of B cells in the polyclonal B cell activation by a B cell differentiation factor, B151-TRF2

    International Nuclear Information System (INIS)

    Ono, S.; Takahama, Y.; Hamaoka, T.

    1987-01-01

    We have demonstrated that B cell recognition of Ia molecules is involved in polyclonal B cell differentiation by B151-TRF2. The present study was undertaken to examine the Ia recognition specificity of B151-TRF2-responsive B cells in fully major histocompatibility complex (MHC)-allogeneic P1----P2, semiallogeneic P1----(P1 x P2)F1, and double donor (P1 + P2)----(P1 x P2)F1 and (P1 + P2)----P1 radiation bone marrow chimeras. The B cells from both P1----P2 and P1----(P1 x P2)F1 chimeras could give rise to in vitro immunoglobulin M-producing cells upon stimulation with B151-TRF2 comparable in magnitude to that of normal P1 B cells, and their responses were inhibited by anti-I-AP1 but not by anti-I-AP2 monoclonal antibody even in the presence of mitomycin C-treated T cell-depleted P2 spleen cells as auxiliary cells. In contrast, the B151-TRF2 responses of P1 B cells isolated from both (P1 + P2)----(P1 x P2)F1 and (P1 + P2)----P1 double bone marrow chimeras became sensitive to the inhibition of not only anti-I-AP1 but also anti-I-AP2 monoclonal antibody only when the culture was conducted in the presence of P2 auxiliary cells, demonstrating that they adaptively differentiate to recognize as self-structures allogeneic as well as syngeneic Ia molecules. Moreover, the experiments utilizing B cells from H-2-congenic mice and B cell hybridoma clones as auxiliary cells revealed that B151-TRF2-responsive B cells recognize Ia molecules expressed on B cells. Taken together, these results demonstrate that B151-TRF2-responsive B cells recognize Ia molecules expressed by B cells as self-structures and that their self-recognition specificity is dictated by the MHC haplotype of bone marrow cells present during the B cell ontogeny but not by the MHC haplotype of a radiation-resistant host environment

  17. Disruption and functional analysis of six ORFs on chromosome XV: YOL117w, YOL115w ( TRF4), YOL114c, YOL112w ( MSB4), YOL111c and YOL072w.

    Science.gov (United States)

    Iwanejko, L; Smith, K N; Loeillet, S; Nicolas, A; Fabre, F

    1999-10-01

    We have carried out the systematic disruption of six ORFs on chromosome XV, of Saccharomyces cerevisiae using the long flanking homology technique to replace each with the KanMX cassette; we have also constructed plasmids containing replacement cassettes and cognate clones for each ORF. Disruption of three of the ORFs-YOL117w, YOL114c, and YOL112w (also known as MSB4)-does not result in any noteworthy phenotype with respect to temperature or nutritional requirements, but yol112w mutants with an additional disruption of YNL293w, which encodes a protein similar to Yol112w, exhibit a slow growth phenotype. The protein specified by YOL114c shares similarity with the human DS-1 protein. Disruption of YOL115w confers slow growth, cold sensitivity and poor sporulation; this ORF has been described elsewhere as TRF4, which encodes a topoisomerase I-related protein. Cells with disruptions of YOL111c, whose product is weakly similar to the human ubiquitin-like protein GdX, are slightly impaired in mating. Mutants disrupted for YOL072w, the predicted product of which is unrelated to any protein of known function, grow slowly, are cold-sensitive and sporulate with reduced efficiency. Copyright 1999 John Wiley & Sons, Ltd.

  18. VLBI TRF Combination Using GNSS Software

    Directory of Open Access Journals (Sweden)

    Younghee Kwak

    2013-12-01

    Full Text Available Space geodetic techniques can be used to obtain precise shape and rotation information of the Earth. To achieve this, the representative combination solution of each space geodetic technique has to be produced, and then those solutions need to be combined. In this study, the representative combination solution of very long baseline interferometry (VLBI, which is one of the space geodetic techniques, was produced, and the variations in the position coordinate of each station during 7 years were analyzed. Products from five analysis centers of the International VLBI Service for Geodesy and Astrometry (IVS were used as the input data, and Bernese 5.0, which is the global navigation satellite system (GNSS data processing software, was used. The analysis of the coordinate time series for the 43 VLBI stations indicated that the latitude component error was about 15.6 mm, the longitude component error was about 37.7 mm, and the height component error was about 30.9 mm, with respect to the reference frame, International Terrestrial Reference Frame 2008 (ITRF2008. The velocity vector of the 42 stations excluding the YEBES station showed a magnitude difference of 7.3 mm/yr (30.2% and a direction difference of 13.8° (3.8%, with respect to ITRF2008. Among these, the 10 stations in Europe showed a magnitude difference of 7.8 mm/yr (30.3% and a direction difference of 3.7° (1.0%, while the 14 stations in North America showed a magnitude difference of 2.7 mm/yr (15.8% and a direction difference of 10.3° (2.9%.

  19. HOT1 is a mammalian direct telomere repeat-binding protein contributing to telomerase recruitment

    NARCIS (Netherlands)

    Kappei, D.; Butter, F.; Benda, C.; Scheibe, M.; Draskovic, Irena; Stevense, M.; Novo, C.L.; Basquin, C.; Araki, M.; Araki, K.; Krastev, D.B.; Kittler, R.; Jessberger, R.; Londono-Vallejo, J.A.; Mann, M.; Buchholz, F.

    2013-01-01

    Telomeres are repetitive DNA structures that, together with the shelterin and the CST complex, protect the ends of chromosomes. Telomere shortening is mitigated in stem and cancer cells through the de novo addition of telomeric repeats by telomerase. Telomere elongation requires the delivery of the

  20. A simulation of the Wolsong TRF LPCE process

    International Nuclear Information System (INIS)

    Kim, K. S.; Lee, S. K.; Song, K. M.; Son, S. H.

    2000-01-01

    The concentric tubular type hydrogen isotope liquid phase catalytic exchange(LPCE) column for the front end process of the Wolsong Tritium Removal Facility(WTRF) was simulated. Both D/T and H/D systems were simulated for the WTRE and the pilot plant for the WTRE respectively. As the difference of latent heats among the hydrogen isotopes and the amount of tritium in the feed heavy water are very small as well as the column is thermally insulated, energy balance was ignored in the model. Pressure drop, the efficiencies of the catalytic bed and the hydrophilic bed, and the recycle of the condensed vapour to the top of the column were considered in the model. The column is simulated from the bottom progressively so that no iteration was required. The performance of the column was enhanced as the increase of the column temperature and the decrease of the pressure. To process the feed heavy water of 100 kg/h and tritium content of 6.3x10 -6 in mole fraction with the detriation factor of 35 using a 55 stage column the column temperature would be 70 .deg. C and the efficiency of the hydrophilic bed should be more than 0.69 when the efficiency of the catalytic bed was 0.8

  1. Poly-dipeptides encoded by the C9orf72 repeats bind nucleoli, impede RNA biogenesis, and kill cells.

    Science.gov (United States)

    Kwon, Ilmin; Xiang, Siheng; Kato, Masato; Wu, Leeju; Theodoropoulos, Pano; Wang, Tao; Kim, Jiwoong; Yun, Jonghyun; Xie, Yang; McKnight, Steven L

    2014-09-05

    Many RNA regulatory proteins controlling pre-messenger RNA splicing contain serine:arginine (SR) repeats. Here, we found that these SR domains bound hydrogel droplets composed of fibrous polymers of the low-complexity domain of heterogeneous ribonucleoprotein A2 (hnRNPA2). Hydrogel binding was reversed upon phosphorylation of the SR domain by CDC2-like kinases 1 and 2 (CLK1/2). Mutated variants of the SR domains changing serine to glycine (SR-to-GR variants) also bound to hnRNPA2 hydrogels but were not affected by CLK1/2. When expressed in mammalian cells, these variants bound nucleoli. The translation products of the sense and antisense transcripts of the expansion repeats associated with the C9orf72 gene altered in neurodegenerative disease encode GRn and PRn repeat polypeptides. Both peptides bound to hnRNPA2 hydrogels independent of CLK1/2 activity. When applied to cultured cells, both peptides entered cells, migrated to the nucleus, bound nucleoli, and poisoned RNA biogenesis, which caused cell death. Copyright © 2014, American Association for the Advancement of Science.

  2. Telomere length modulation in human astroglial brain tumors.

    Directory of Open Access Journals (Sweden)

    Domenico La Torre

    Full Text Available BACKGROUND: Telomeres alteration during carcinogenesis and tumor progression has been described in several cancer types. Telomeres length is stabilized by telomerase (h-TERT and controlled by several proteins that protect telomere integrity, such as the Telomere Repeat-binding Factor (TRF 1 and 2 and the tankyrase-poli-ADP-ribose polymerase (TANKs-PARP complex. OBJECTIVE: To investigate telomere dysfunction in astroglial brain tumors we analyzed telomeres length, telomerase activity and the expression of a panel of genes controlling the length and structure of telomeres in tissue samples obtained in vivo from astroglial brain tumors with different grade of malignancy. MATERIALS AND METHODS: Eight Low Grade Astrocytomas (LGA, 11 Anaplastic Astrocytomas (AA and 11 Glioblastoma Multiforme (GBM samples were analyzed. Three samples of normal brain tissue (NBT were used as controls. Telomeres length was assessed through Southern Blotting. Telomerase activity was evaluated by a telomere repeat amplification protocol (TRAP assay. The expression levels of TRF1, TRF2, h-TERT and TANKs-PARP complex were determined through Immunoblotting and RT-PCR. RESULTS: LGA were featured by an up-regulation of TRF1 and 2 and by shorter telomeres. Conversely, AA and GBM were featured by a down-regulation of TRF1 and 2 and an up-regulation of both telomerase and TANKs-PARP complex. CONCLUSIONS: In human astroglial brain tumours, up-regulation of TRF1 and TRF2 occurs in the early stages of carcinogenesis determining telomeres shortening and genomic instability. In a later stage, up-regulation of PARP-TANKs and telomerase activation may occur together with an ADP-ribosylation of TRF1, causing a reduced ability to bind telomeric DNA, telomeres elongation and tumor malignant progression.

  3. AKTIP/Ft1, a New Shelterin-Interacting Factor Required for Telomere Maintenance.

    KAUST Repository

    Burla, Romina; Carcuro, Mariateresa; Raffa, Grazia D; Galati, Alessandra; Raimondo, Domenico; Rizzo, Angela; La Torre, Mattia; Micheli, Emanuela; Ciapponi, Laura; Cenci, Giovanni; Cundari, Enrico; Musio, Antonio; Biroccio, Annamaria; Cacchione, Stefano; Gatti, Maurizio; Saggio, Isabella

    2015-01-01

    Telomeres are nucleoprotein complexes that protect the ends of linear chromosomes from incomplete replication, degradation and detection as DNA breaks. Mammalian telomeres are protected by shelterin, a multiprotein complex that binds the TTAGGG telomeric repeats and recruits a series of additional factors that are essential for telomere function. Although many shelterin-associated proteins have been so far identified, the inventory of shelterin-interacting factors required for telomere maintenance is still largely incomplete. Here, we characterize AKTIP/Ft1 (human AKTIP and mouse Ft1 are orthologous), a novel mammalian shelterin-bound factor identified on the basis of its homology with the Drosophila telomere protein Pendolino. AKTIP/Ft1 shares homology with the E2 variant ubiquitin-conjugating (UEV) enzymes and has been previously implicated in the control of apoptosis and in vesicle trafficking. RNAi-mediated depletion of AKTIP results in formation of telomere dysfunction foci (TIFs). Consistent with these results, AKTIP interacts with telomeric DNA and binds the shelterin components TRF1 and TRF2 both in vivo and in vitro. Analysis of AKTIP- depleted human primary fibroblasts showed that they are defective in PCNA recruiting and arrest in the S phase due to the activation of the intra S checkpoint. Accordingly, AKTIP physically interacts with PCNA and the RPA70 DNA replication factor. Ft1-depleted p53-/- MEFs did not arrest in the S phase but displayed significant increases in multiple telomeric signals (MTS) and sister telomere associations (STAs), two hallmarks of defective telomere replication. In addition, we found an epistatic relation for MST formation between Ft1 and TRF1, which has been previously shown to be required for replication fork progression through telomeric DNA. Ch-IP experiments further suggested that in AKTIP-depleted cells undergoing the S phase, TRF1 is less tightly bound to telomeric DNA than in controls. Thus, our results collectively

  4. AKTIP/Ft1, a New Shelterin-Interacting Factor Required for Telomere Maintenance.

    KAUST Repository

    Burla, Romina

    2015-06-25

    Telomeres are nucleoprotein complexes that protect the ends of linear chromosomes from incomplete replication, degradation and detection as DNA breaks. Mammalian telomeres are protected by shelterin, a multiprotein complex that binds the TTAGGG telomeric repeats and recruits a series of additional factors that are essential for telomere function. Although many shelterin-associated proteins have been so far identified, the inventory of shelterin-interacting factors required for telomere maintenance is still largely incomplete. Here, we characterize AKTIP/Ft1 (human AKTIP and mouse Ft1 are orthologous), a novel mammalian shelterin-bound factor identified on the basis of its homology with the Drosophila telomere protein Pendolino. AKTIP/Ft1 shares homology with the E2 variant ubiquitin-conjugating (UEV) enzymes and has been previously implicated in the control of apoptosis and in vesicle trafficking. RNAi-mediated depletion of AKTIP results in formation of telomere dysfunction foci (TIFs). Consistent with these results, AKTIP interacts with telomeric DNA and binds the shelterin components TRF1 and TRF2 both in vivo and in vitro. Analysis of AKTIP- depleted human primary fibroblasts showed that they are defective in PCNA recruiting and arrest in the S phase due to the activation of the intra S checkpoint. Accordingly, AKTIP physically interacts with PCNA and the RPA70 DNA replication factor. Ft1-depleted p53-/- MEFs did not arrest in the S phase but displayed significant increases in multiple telomeric signals (MTS) and sister telomere associations (STAs), two hallmarks of defective telomere replication. In addition, we found an epistatic relation for MST formation between Ft1 and TRF1, which has been previously shown to be required for replication fork progression through telomeric DNA. Ch-IP experiments further suggested that in AKTIP-depleted cells undergoing the S phase, TRF1 is less tightly bound to telomeric DNA than in controls. Thus, our results collectively

  5. Cassava C-repeat binding factor 1 gene responds to low temperature and enhances cold tolerance when overexpressed in Arabidopsis and cassava.

    Science.gov (United States)

    An, Dong; Ma, Qiuxiang; Wang, Hongxia; Yang, Jun; Zhou, Wenzhi; Zhang, Peng

    2017-05-01

    Cassava MeCBF1 is a typical CBF transcription factor mediating cold responses but its low expression in apical buds along with a retarded response cause inefficient upregulation of downstream cold-related genes, rendering cassava chilling-sensitive. Low temperature is a major abiotic stress factor affecting survival, productivity and geographic distribution of important crops worldwide. The C-repeat/dehydration-responsive element binding transcription factors (CBF/DREB) are important regulators of abiotic stress response in plants. In this study, MeCBF1, a CBF-like gene, was identified in the tropical root crop cassava (Manihot esculenta Crantz). The MeCBF1 encodes a protein that shares strong homology with DREB1As/CBFs from Arabidopsis as well as other species. The MeCBF1 was localized to the nucleus and is mainly expressed in stem and mature leaves, but not in apical buds or stem cambium. MeCBF1 expression was not only highly responsive to cold, but also significantly induced by salt, PEG and ABA treatment. Several stress-associated cis-elements were found in its promoter region, e.g., ABRE-related, MYC recognition sites, and MYB responsive element. Compared with AtCBF1, the MeCBF1 expression induced by cold in cassava was retarded and upregulated only after 4 h, which was also confirmed by its promoter activity. Overexpression of MeCBF1 in transgenic Arabidopsis and cassava plants conferred enhanced crytolerance. The CBF regulon was smaller and not entirely co-regulated with MeCBF1 expression in overexpressed cassava. The retarded MeCBF1 expression in response to cold and attenuated CBF-regulon might lead cassava to chilling sensitivity.

  6. Role for a region of helically unstable DNA within the Epstein-Barr virus latent cycle origin of DNA replication oriP in origin function

    International Nuclear Information System (INIS)

    Polonskaya, Zhanna; Benham, Craig J.; Hearing, Janet

    2004-01-01

    The minimal replicator of the Epstein-Barr virus (EBV) latent cycle origin of DNA replication oriP is composed of two binding sites for the Epstein-Barr virus nuclear antigen-1 (EBNA-1) and flanking inverted repeats that bind the telomere repeat binding factor TRF2. Although not required for minimal replicator activity, additional binding sites for EBNA-1 and TRF2 and one or more auxiliary elements located to the right of the EBNA-1/TRF2 sites are required for the efficient replication of oriP plasmids. Another region of oriP that is predicted to be destabilized by DNA supercoiling is shown here to be an important functional component of oriP. The ability of DNA fragments of unrelated sequence and possessing supercoiled-induced DNA duplex destabilized (SIDD) structures, but not fragments characterized by helically stable DNA, to substitute for this component of oriP demonstrates a role for the SIDD region in the initiation of oriP-plasmid DNA replication

  7. The Monitoring and Affinity Purification of Proteins Using Dual Tags with Tetracysteine Motifs

    Science.gov (United States)

    Giannone, Richard J.; Liu, Yie; Wang, Yisong

    Identification and characterization of protein-protein interaction networks is essential for the elucidation of biochemical mechanisms and cellular function. Affinity purification in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as a very powerful tactic for the identification of specific protein-protein interactions. In this chapter, we describe a comprehensive methodology that uses our recently developed dual-tag affinity purification system for the enrichment and identification of mammalian protein complexes. The protocol covers a series of separate but sequentially related techniques focused on the facile monitoring and purification of a dual-tagged protein of interest and its interacting partners via a system built with tetracysteine motifs and various combinations of affinity tags. Using human telomeric repeat binding factor 2 (TRF2) as an example, we demonstrate the power of the system in terms of bait protein recovery after dual-tag affinity purification, detection of bait protein subcellular localization and expression, and successful identification of known and potentially novel TRF2 interacting proteins. Although the protocol described here has been optimized for the identification and characterization of TRF2-associated proteins, it is, in principle, applicable to the study of any other mammalian protein complexes that may be of interest to the research community.

  8. DNA damage response and role of shelterin complex in human peripheral blood mononuclear cells exposed to gamma radiation

    International Nuclear Information System (INIS)

    Saini, Divyalakshmi; Das, Birajalaxmi

    2013-01-01

    Telomeres are the DNA protein structures that cap the ends of linear DNA. It consists of short repetitive DNA sequences (TTAGGG)n and specialized telomere binding proteins. There are six telomeric proteins (TRF1, TRF2, TIN2, TERF2, PTOP and POT1) called as shelterin complex/telosome which maintains telomere integrity. The function of this 'telosome' is to protect the natural ends of the chromosomes from being recognized as artificial DNA breaks, thereby preventing chromosome end-to-end fusions. DNA Damage Response (DDR) induced by radiation and its interaction with telomeric protein complex is poorly understood in human PBMCs at G 0 stage. Alterations in either telomeric DNA or telomere binding proteins can impair the function of the telosome, which may lead to senescence or apoptosis. Ionizing radiation which induces a plethora of DNA lesions in human cell may also alter the expression of telomere associated proteins. In the present study, we have made an attempt to study the DNA damage response of telomere proteins in human peripheral blood mononuclear cells exposed to gamma radiation. Venous blood samples were collected from eight random healthy volunteers and PBMCs were separated. Dose response as well as time point kinetics study was carried out at transcription as well as protein level. PBMCs were irradiated at various doses between 10 cGy to 2.0 Gy at a dose rate of 1.0 Gy/min. Total RNA was isolated for gene expression analysis at 0 hour and 4 hours respectively. cDNA was prepared and transcriptional pattern as studied using real time q-PCR where Taqman probes were used. Time point kinetics of transcriptional pattern of TRF1, TRF2, TIN2, TERF2, PTOP and POT1 was carried out at 0 min, 15 min, 30 min, 60 min, and 120 min for two different doses (1.0 Gy and 2.0 Gy). Dose response and time point kinetics of TRF2 was studied at similar doses using confocal microscopy. Our results revealed that at 2.0 Gy there was a two fold increase at the level of transcription

  9. Inherited mutations in the helicase RTEL1 cause telomere dysfunction and Hoyeraal-Hreidarsson syndrome.

    Science.gov (United States)

    Deng, Zhong; Glousker, Galina; Molczan, Aliah; Fox, Alan J; Lamm, Noa; Dheekollu, Jayaraju; Weizman, Orr-El; Schertzer, Michael; Wang, Zhuo; Vladimirova, Olga; Schug, Jonathan; Aker, Memet; Londoño-Vallejo, Arturo; Kaestner, Klaus H; Lieberman, Paul M; Tzfati, Yehuda

    2013-09-03

    Telomeres repress the DNA damage response at the natural chromosome ends to prevent cell-cycle arrest and maintain genome stability. Telomeres are elongated by telomerase in a tightly regulated manner to ensure a sufficient number of cell divisions throughout life, yet prevent unlimited cell division and cancer development. Hoyeraal-Hreidarsson syndrome (HHS) is characterized by accelerated telomere shortening and a broad range of pathologies, including bone marrow failure, immunodeficiency, and developmental defects. HHS-causing mutations have previously been found in telomerase and the shelterin component telomeric repeat binding factor 1 (TRF1)-interacting nuclear factor 2 (TIN2). We identified by whole-genome exome sequencing compound heterozygous mutations in four siblings affected with HHS, in the gene encoding the regulator of telomere elongation helicase 1 (RTEL1). Rtel1 was identified in mouse by its genetic association with telomere length. However, its mechanism of action and whether it regulates telomere length in human remained unknown. Lymphoblastoid cell lines obtained from a patient and from the healthy parents carrying heterozygous RTEL1 mutations displayed telomere shortening, fragility and fusion, and growth defects in culture. Ectopic expression of WT RTEL1 suppressed the telomere shortening and growth defect, confirming the causal role of the RTEL1 mutations in HHS and demonstrating the essential function of human RTEL1 in telomere protection and elongation. Finally, we show that human RTEL1 interacts with the shelterin protein TRF1, providing a potential recruitment mechanism of RTEL1 to telomeres.

  10. Inherited mutations in the helicase RTEL1 cause telomere dysfunction and Hoyeraal–Hreidarsson syndrome

    Science.gov (United States)

    Deng, Zhong; Glousker, Galina; Molczan, Aliah; Fox, Alan J.; Lamm, Noa; Dheekollu, Jayaraju; Weizman, Orr-El; Schertzer, Michael; Wang, Zhuo; Vladimirova, Olga; Schug, Jonathan; Aker, Memet; Londoño-Vallejo, Arturo; Kaestner, Klaus H.; Lieberman, Paul M.; Tzfati, Yehuda

    2013-01-01

    Telomeres repress the DNA damage response at the natural chromosome ends to prevent cell-cycle arrest and maintain genome stability. Telomeres are elongated by telomerase in a tightly regulated manner to ensure a sufficient number of cell divisions throughout life, yet prevent unlimited cell division and cancer development. Hoyeraal–Hreidarsson syndrome (HHS) is characterized by accelerated telomere shortening and a broad range of pathologies, including bone marrow failure, immunodeficiency, and developmental defects. HHS-causing mutations have previously been found in telomerase and the shelterin component telomeric repeat binding factor 1 (TRF1)-interacting nuclear factor 2 (TIN2). We identified by whole-genome exome sequencing compound heterozygous mutations in four siblings affected with HHS, in the gene encoding the regulator of telomere elongation helicase 1 (RTEL1). Rtel1 was identified in mouse by its genetic association with telomere length. However, its mechanism of action and whether it regulates telomere length in human remained unknown. Lymphoblastoid cell lines obtained from a patient and from the healthy parents carrying heterozygous RTEL1 mutations displayed telomere shortening, fragility and fusion, and growth defects in culture. Ectopic expression of WT RTEL1 suppressed the telomere shortening and growth defect, confirming the causal role of the RTEL1 mutations in HHS and demonstrating the essential function of human RTEL1 in telomere protection and elongation. Finally, we show that human RTEL1 interacts with the shelterin protein TRF1, providing a potential recruitment mechanism of RTEL1 to telomeres. PMID:23959892

  11. Measurement of Telomere Length in Colorectal Cancers for Improved Molecular Diagnosis

    Directory of Open Access Journals (Sweden)

    Eric Le Balc’h

    2017-08-01

    Full Text Available All tumors have in common to reactivate a telomere maintenance mechanism to allow for unlimited proliferation. On the other hand, genetic instability found in some tumors can result from the loss of telomeres. Here, we measured telomere length in colorectal cancers (CRCs using TRF (Telomere Restriction Fragment analysis. Telomeric DNA content was also quantified as the ratio of total telomeric (TTAGGG sequences over that of the invariable Alu sequences. In most of the 125 CRCs analyzed, there was a significant diminution in telomere length compared with that in control healthy tissue. Only 34 tumors exhibited no telomere erosion and, in some cases, a slight telomere lengthening. Telomere length did not correlate with age, gender, tumor stage, tumor localization or stage of tumor differentiation. In addition, while telomere length did not correlate with the presence of a mutation in BRAF (V-raf murine sarcoma viral oncogene homolog B, PIK3CA (phosphatidylinositol 3-kinase catalytic subunit, or MSI status, it was significantly associated with the occurrence of a mutation in KRAS. Interestingly, we found that the shorter the telomeres in healthy tissue of a patient, the larger an increase in telomere length in the tumor. Our study points to the existence of two types of CRCs based on telomere length and reveals that telomere length in healthy tissue might influence telomere maintenance mechanisms in the tumor.

  12. Étude structurale de l'assemblage du complexe télomérique humain TRF2/RAP1

    OpenAIRE

    Gaullier , Guillaume

    2015-01-01

    Telomeres are the ends of eukaryotic linear chromosomes. They are made oftandem repeats of a short guanine-rich motif and bound by specific proteins.In vertebrates, these proteins form a complex called shelterin, theintegrity of which is critical to ensure proper replication of chromosomeends and to protect them against illicit targeting by DNA double-strandbreak repair pathways. Telomere dysfunctions lead to genome instability,which can ultimately cause senescence or cancer. Telomeres are a ...

  13. Contribution to the study of TRH (thyrotropin-releasing hormone) conformation using circular dichroism. Physico-chemical studies, radioactive labelling and biological applications

    International Nuclear Information System (INIS)

    Pradelles, Philippe.

    1977-01-01

    In an attempt to reach a better understanding at the molecular level of phenomena connected with the action of TRF the conformation and radioactive labelling of this hormone were investigated. The specific detection of a hormone at its action site is only possible if labelled substances of very high specific activity are used. TRF was tritium labelled by three methods: direct catalytic exchange; catalytic dehalogenation of mono- and di-iodo TRF; catalytic denitrogenation of mono-azo-TRF. Whatever the method used the tritiated TRF has a very high specific activity and keeps all its biological properties. Biological activity measurements carried out on labelled TRF, in vivo in rats and in vitro on a TRF-sensitive prolactine cell clone, are described. TRF tritiated by the above methods is shown to have the same biological activity as standard TRF. Some results are given concerning the application of labelled TRF to research on the hormone action mechanism. The tritiated TRF distribution kinetics were examined in vivo and in vitro. The kinetics of hormone fixation on the antehypophysary tissue match those of in vivo release of the plasma thyreotropic hormone, confirming the relationships between the hormone fixation on its target tissue and its biological effect. Finally an outline is given of work on the interaction of tritiated TRF with prolactine cell receptors and on the penetration of intact tritiated TRF into these cells. In addition the radioimmunological analysis of TRF was developed by the use of 125 I-mono-iodo-TRF at high specific activity (above 2000 Ci/mmole) [fr

  14. Human liver cell trafficking mutants: characterization and whole exome sequencing.

    Directory of Open Access Journals (Sweden)

    Fei Yuan

    Full Text Available The HuH7 liver cell mutant Trf1 is defective in membrane trafficking and is complemented by the casein kinase 2α subunit CK2α''. Here we identify characteristic morphologies, trafficking and mutational changes in six additional HuH7 mutants Trf2-Trf7. Trf1 cells were previously shown to be severely defective in gap junction functions. Using a Lucifer yellow transfer assay, remarkable attenuation of gap junction communication was revealed in each of the mutants Trf2-Trf7. Electron microscopy and light microscopy of thiamine pyrophosphatase showed that several mutants exhibited fragmented Golgi apparatus cisternae compared to parental HuH7 cells. Intracellular trafficking was investigated using assays of transferrin endocytosis and recycling and VSV G secretion. Surface binding of transferrin was reduced in all six Trf2-Trf7 mutants, which generally correlated with the degree of reduced expression of the transferrin receptor at the cell surface. The mutants displayed the same transferrin influx rates as HuH7, and for efflux rate, only Trf6 differed, having a slower transferrin efflux rate than HuH7. The kinetics of VSV G transport along the exocytic pathway were altered in Trf2 and Trf5 mutants. Genetic changes unique to particular Trf mutants were identified by exome sequencing, and one was investigated in depth. The novel mutation Ile34Phe in the GTPase RAB22A was identified in Trf4. RNA interference knockdown of RAB22A or overexpression of RAB22AI34F in HuH7 cells caused phenotypic changes characteristic of the Trf4 mutant. In addition, the Ile34Phe mutation reduced both guanine nucleotide binding and hydrolysis activities of RAB22A. Thus, the RAB22A Ile34Phe mutation appears to contribute to the Trf4 mutant phenotype.

  15. Tocotrienols are good adjuvants for developing cancer vaccines

    International Nuclear Information System (INIS)

    Abdul Hafid, Sitti Rahma; Radhakrishnan, Ammu Kutty; Nesaretnam, Kalanithi

    2010-01-01

    Dendritic cells (DCs) have the potential for cancer immunotherapy due to their ability to process and present antigens to T-cells and also in stimulating immune responses. However, DC-based vaccines have only exhibited minimal effectiveness against established tumours in mice and humans. The use of appropriate adjuvant enhances the efficacy of DC based cancer vaccines in treating tumours. In this study we have used tocotrienol-rich fraction (TRF), a non-toxic natural compound, as an adjuvant to enhance the effectiveness of DC vaccines in treating mouse mammary cancers. In the mouse model, six-week-old female BALB/c mice were injected subcutaneously with DC and supplemented with oral TRF daily (DC+TRF) and DC pulsed with tumour lysate from 4T1 cells (DC+TL). Experimental mice were also injected with DC pulsed with tumour lysate and supplemented daily with oral TRF (DC+TL+TRF) while two groups of animal which were supplemented daily with carrier oil (control) and with TRF (TRF). After three times vaccination, mice were inoculated with 4T1 cells in the mammary breast pad to induce tumour. Our study showed that TRF in combination with DC pulsed with tumour lysate (DC+TL+TRF) injected subcutaneously significantly inhibited the growth of 4T1 mammary tumour cells as compared to control group. Analysis of cytokines production from murine splenocytes showed significant increased productions of IFN-γ and IL-12 in experimental mice (DC+TL+TRF) compared to control, mice injected with DC without TRF, mice injected with DC pulsed with tumour lysate and mice supplemented with TRF alone. Higher numbers of cytotoxic T cells (CD8) and natural killer cells (NK) were observed in the peripheral blood of TRF adjuvanted DC pulsed tumour lysate mice. Our study show that TRF has the potential to be an adjuvant to augment DC based immunotherapy

  16. Tocotrienols are good adjuvants for developing cancer vaccines

    Directory of Open Access Journals (Sweden)

    Radhakrishnan Ammu

    2010-01-01

    Full Text Available Abstract Background Dendritic cells (DCs have the potential for cancer immunotherapy due to their ability to process and present antigens to T-cells and also in stimulating immune responses. However, DC-based vaccines have only exhibited minimal effectiveness against established tumours in mice and humans. The use of appropriate adjuvant enhances the efficacy of DC based cancer vaccines in treating tumours. Methods In this study we have used tocotrienol-rich fraction (TRF, a non-toxic natural compound, as an adjuvant to enhance the effectiveness of DC vaccines in treating mouse mammary cancers. In the mouse model, six-week-old female BALB/c mice were injected subcutaneously with DC and supplemented with oral TRF daily (DC+TRF and DC pulsed with tumour lysate from 4T1 cells (DC+TL. Experimental mice were also injected with DC pulsed with tumour lysate and supplemented daily with oral TRF (DC+TL+TRF while two groups of animal which were supplemented daily with carrier oil (control and with TRF (TRF. After three times vaccination, mice were inoculated with 4T1 cells in the mammary breast pad to induce tumour. Results Our study showed that TRF in combination with DC pulsed with tumour lysate (DC+TL+TRF injected subcutaneously significantly inhibited the growth of 4T1 mammary tumour cells as compared to control group. Analysis of cytokines production from murine splenocytes showed significant increased productions of IFN-γ and IL-12 in experimental mice (DC+TL+TRF compared to control, mice injected with DC without TRF, mice injected with DC pulsed with tumour lysate and mice supplemented with TRF alone. Higher numbers of cytotoxic T cells (CD8 and natural killer cells (NK were observed in the peripheral blood of TRF adjuvanted DC pulsed tumour lysate mice. Conclusion Our study show that TRF has the potential to be an adjuvant to augment DC based immunotherapy.

  17. Radioactive labelling of peptidic hormones

    International Nuclear Information System (INIS)

    Fromageot, P.; Pradelles, P.; Morgat, J.L.; Levine, H.

    1976-01-01

    The labelling of peptidic hormones requires stability, specificity and sensitivity of the label. Introduction of a radioactive atome is one way to satisfy these criteria. Several processes have been described to prepare radioactive TRF: synthesis of the peptide with labelled aminoacids or introduction of the label into the hormone. In that approach, tritium can be substituted in the imidazole ring, via precursors activating the proper carbon. Monoiodo TRF leads essentially to tritium labelling of the 5 positions whereas monoazo TRF allows the preparation of 3 H TRF labelled in the 2 positions. Di-substituted TRF leads to labelling into the 2 and 5 carbons. Labelled analogs of TRF can be prepared with labelled iodine; further developments of peptide labelling, will be presented. In particular, the homolytic scission of the C-iodine, bond by photochemical activation. The nascent carbon radical can be stabilized by a tritiated scavenger. This approach eliminates the use of heavy metal catalysts

  18. Telomeric repeat factor 1 protein levels correlates with telomere length in colorectal cancer Los niveles proteicos del factor de repetición telomérico 1 se correlacionan con la longitud del telómero en el cáncer colorrectal

    Directory of Open Access Journals (Sweden)

    Cristina Valls-Bautista

    2012-11-01

    Full Text Available Background: colorectal cancer is the third cancer cause of death in Spain. It is important to investigate new tumoral markers for early diagnosis, disease monitoring and prevention strategies. Telomeres protect the chromosome from degradation by nucleases and end-to-end fusion. The progressive loss of the telomeric ends of chromosomes is an important mechanism in the timing of human cellular aging. Telomeric Repeat Factor 1 (TRF1 is a protein that binds at telomere ends. Purpose: to measure the concentrations of TRF1 and the relationships among telomere length, telomerase activity, and TRF1 levels in tumor and normal colorectal mucosa. Method: from normal and tumoral samples of 83 patients who underwent surgery for colorectal cancer we analyzed TRF1 protein concentration by Western Blot, telomerase activity, by the fluorescent-telomeric repeat amplification protocol assay and telomere length by Southern Blot. Results: high levels of TRF1 were observed in 68.7% of tumor samples, while the majority of normal samples (59% showed negative or weak TRF1 concentrations. Among the tumor samples, telomere length was significantly associated with TRF1 protein levels (p = 0.023. Conclusions: a relationship was found between telomere length and TRF1 abundance protein in tumor samples, which means that TRF1 is an important factor in the tumor progression and maybe a diagnostic factor.

  19. Working Memory, Attention, Inhibition, and Their Relation to Adaptive Functioning and Behavioral/Emotional Symptoms in School-Aged Children

    Science.gov (United States)

    Vuontela, Virve; Carlson, Synnove; Troberg, Anna-Maria; Fontell, Tuija; Simola, Petteri; Saarinen, Suvi; Aronen, Eeva T.

    2013-01-01

    The present study investigated the development of executive functions (EFs) and their associations with performance and behavior at school in 8-12-year-old children. The EFs were measured by computer-based n-back, Continuous Performance and Go/Nogo tasks. School performance was evaluated by Teacher Report Form (TRF) and behavior by TRF and Child…

  20. Plant phylogeny as a window on the evolution of hyperdiversity in the tropical rainforest biome.

    Science.gov (United States)

    Eiserhardt, Wolf L; Couvreur, Thomas L P; Baker, William J

    2017-06-01

    I. II. III. IV. V. VI. VII. VIII. IX. References SUMMARY: Tropical rainforest (TRF) is the most species-rich terrestrial biome on Earth, harbouring just under half of the world's plant species in c. 7% of the land surface. Phylogenetic trees provide important insights into mechanisms underpinning TRF hyperdiversity that are complementary to those obtained from the fossil record. Phylogenetic studies of TRF plant diversity have mainly focused on whether this biome is an evolutionary 'cradle' or 'museum', emphasizing speciation and extinction rates. However, other explanations, such as biome age, immigration and ecological limits, must also be considered. We present a conceptual framework for addressing the drivers of TRF diversity, and review plant studies that have tested them with phylogenetic data. Although surprisingly few in number, these studies point to old age of TRF, low extinction and high speciation rates as credible drivers of TRF hyperdiversity. There is less evidence for immigration and ecological limits, but these cannot be dismissed owing to the limited number of studies. Rapid methodological developments in DNA sequencing, macroevolutionary analysis and the integration of phylogenetics with other disciplines may improve our grasp of TRF hyperdiversity in the future. However, such advances are critically dependent on fundamental systematic research, yielding numerous, additional, well-sampled phylogenies of TRF lineages. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  1. T-cell mean telomere lengths changes in treatment naïve HIV-infected patients randomized to G-CSF or placebo simultaneously with initiation of HAART

    DEFF Research Database (Denmark)

    Aladdin, H; Von Essen, M; Schjerling, P

    2001-01-01

    The effect of highly active antiretroviral therapy (HAART) and granulocyte colony stimulating factor (G-CSF) on mean telomere restriction fragment (TRF) length of peripheral blood mononuclear cells (PBMC) was examined in 11 treatment naïve human immunodeficiency virus (HIV)-infected individuals...... with a CD4+ T-cell count CSF thrice weekly for 12 weeks (n = 6) or placebo (n = 5). An increase in the mean TRF lengths was observed in PBMC of patients on HAART after 24 weeks of treatment mainly owing to increased mean CD8+ T-cell TRF...... lengths. However, in the group of patients on HAART combined with G-CSF no changes of PBMC mean TRF length was observed during treatment or during 12 weeks of follow-up. The mean CD4+ T-cell TRF length did not change in any of the two groups. These results confirm that HAART induces mainly the lengthening...

  2. Dissecting mechanisms of nuclear mRNA surveillance in THO/sub2 complex mutants

    DEFF Research Database (Denmark)

    Rougemaille, Mathieu; Gudipati, Rajani K; Olesen, Jens Raabjerg

    2007-01-01

    The nuclear exosome is involved in numerous RNA metabolic processes. Exosome degradation of rRNA, snoRNA, snRNA and tRNA in Saccharomyces cerevisiae is activated by TRAMP complexes, containing either the Trf4p or Trf5p poly(A) polymerase. These enzymes are presumed to facilitate exosome access...... is required for both retention and degradation of nuclear restricted mRNAs. We show here that Trf4p, in the context of TRAMP, is an mRNA surveillance factor. However, unlike Rrp6p, Trf4p only partakes in RNA degradation and not in transcript retention. Surprisingly, a polyadenylation-defective Trf4p protein...

  3. The Influence of the Terrestrial Reference Frame on Studies of Sea Level Change

    Science.gov (United States)

    Nerem, R. S.; Bar-Sever, Y. E.; Haines, B. J.; Desai, S.; Heflin, M. B.

    2015-12-01

    The terrestrial reference frame (TRF) provides the foundation for the accurate monitoring of sea level using both ground-based (tide gauges) and space-based (satellite altimetry) techniques. For the latter, tide gauges are also used to monitor drifts in the satellite instruments over time. The accuracy of the terrestrial reference frame (TRF) is thus a critical component for both types of sea level measurements. The TRF is central to the formation of geocentric sea-surface height (SSH) measurements from satellite altimeter data. The computed satellite orbits are linked to a particular TRF via the assumed locations of the ground-based tracking systems. The manner in which TRF errors are expressed in the orbit solution (and thus SSH) is not straightforward, and depends on the models of the forces underlying the satellite's motion. We discuss this relationship, and provide examples of the systematic TRF-induced errors in the altimeter derived sea-level record. The TRF is also crucial to the interpretation of tide-gauge measurements, as it enables the separation of vertical land motion from volumetric changes in the water level. TRF errors affect tide gauge measurements through GNSS estimates of the vertical land motion at each tide gauge. This talk will discuss the current accuracy of the TRF and how errors in the TRF impact both satellite altimeter and tide gauge sea level measurements. We will also discuss simulations of how the proposed Geodetic Reference Antenna in SPace (GRASP) satellite mission could reduce these errors and revolutionize how reference frames are computed in general.

  4. Common carotid artery intima-media thickness is as good as carotid intima-media thickness of all carotid artery segments in improving prediction of coronary heart disease risk in the Atherosclerosis Risk in Communities (ARIC) study.

    Science.gov (United States)

    Nambi, Vijay; Chambless, Lloyd; He, Max; Folsom, Aaron R; Mosley, Tom; Boerwinkle, Eric; Ballantyne, Christie M

    2012-01-01

    Carotid intima-media thickness (CIMT) and plaque information can improve coronary heart disease (CHD) risk prediction when added to traditional risk factors (TRF). However, obtaining adequate images of all carotid artery segments (A-CIMT) may be difficult. Of A-CIMT, the common carotid artery intima-media thickness (CCA-IMT) is relatively more reliable and easier to measure. We evaluated whether CCA-IMT is comparable to A-CIMT when added to TRF and plaque information in improving CHD risk prediction in the Atherosclerosis Risk in Communities (ARIC) study. Ten-year CHD risk prediction models using TRF alone, TRF + A-CIMT + plaque, and TRF + CCA-IMT + plaque were developed for the overall cohort, men, and women. The area under the receiver operator characteristic curve (AUC), per cent individuals reclassified, net reclassification index (NRI), and model calibration by the Grønnesby-Borgan test were estimated. There were 1722 incident CHD events in 12 576 individuals over a mean follow-up of 15.2 years. The AUC for TRF only, TRF + A-CIMT + plaque, and TRF + CCA-IMT + plaque models were 0.741, 0.754, and 0.753, respectively. Although there was some discordance when the CCA-IMT + plaque- and A-CIMT + plaque-based risk estimation was compared, the NRI and clinical NRI (NRI in the intermediate-risk group) when comparing the CIMT models with TRF-only model, per cent reclassified, and test for model calibration were not significantly different. Coronary heart disease risk prediction can be improved by adding A-CIMT + plaque or CCA-IMT + plaque information to TRF. Therefore, evaluating the carotid artery for plaque presence and measuring CCA-IMT, which is easier and more reliable than measuring A-CIMT, provide a good alternative to measuring A-CIMT for CHD risk prediction.

  5. Protein composition of catalytically active human telomerase from immortal cells

    DEFF Research Database (Denmark)

    Cohen, Scott B; Graham, Mark E; Lovrecz, George O

    2007-01-01

    Telomerase is a ribonucleoprotein enzyme complex that adds 5'-TTAGGG-3' repeats onto the ends of human chromosomes, providing a telomere maintenance mechanism for approximately 90% of human cancers. We have purified human telomerase approximately 10(8)-fold, with the final elution dependent on th...

  6. Deciphering the Role of Alternative Non-Homologous End Joining (Alt-NHEJ) DNA Repair in Breast Cancer

    Science.gov (United States)

    2017-12-01

    consecutive TTAGGG repeats. To detect rare reads containing fusion junctions, we exploited the novel sequence arrangement created by the ligation of the 39G...journal.pgen.1001005 (2010). 14 van Kregten, M. et al. T-DNA integration in plants results from polymerase-theta-mediated DNA repair. Nat Plants 2

  7. Protective effects of tocotrienols against lipid-induced nephropathy in experimental type-2 diabetic rats by modulation in TGF-β expression

    Energy Technology Data Exchange (ETDEWEB)

    Siddiqui, Shabeena [Department of Biochemistry, Lipid Metabolism Laboratory, Jamia Hamdard (Hamdard University), New Delhi 110062 (India); Ahsan, Haseeb [Department of Biochemistry, Faculty of Dentistry, Jamia Millia Islamia, New Delhi 110025 (India); Khan, Mohammad Rashid [Department of Biochemistry, Lipid Metabolism Laboratory, Jamia Hamdard (Hamdard University), New Delhi 110062 (India); Siddiqui, Waseem A., E-mail: wasiddiqui01@gmail.com [Department of Biochemistry, Lipid Metabolism Laboratory, Jamia Hamdard (Hamdard University), New Delhi 110062 (India)

    2013-12-01

    Dyslipidemia is common in patients with diabetes mellitus (DM) and is considered a risk factor for the progression of diabetic nephropathy (DN). Hyperlipidemia and hyperglycemia act synergistically to induce renal injury. The present study was designed to investigate the protective effects of tocotrienols as tocotrienol-rich fraction (TRF) extracted from palm (PO) and rice bran oils (RBO) against lipid induced nephropathy in type-2 diabetic rats and its probable molecular mechanism. Male Wistar rats (175–200 g) were divided into four groups. The first group served as diabetic control, while the second and third groups received PO-TRF and RBO-TRF, respectively by gavage over a period of sixteen weeks post-induction of diabetes. The fourth group comprised of age-matched rats that served as normal control. The effects of TRF on serum lipid profile, oxidative stress markers, expression of TGF-β, fibronectin and collagen type IV were analyzed in the kidney of diabetic rats. Treatment with PO-TRF and RBO-TRF significantly improved glycemic status, serum lipid profile and renal function in type-2 diabetic rats. In addition, TRF supplementation down-regulated the expression of TGF-β, fibronectin and collagen type IV in the kidney of diabetic rats. Transforming growth factor-β (TGF-β) plays a critical role in progression of DN, but its modulation by tocotrienols in DN remains unexplored. TRF ameliorated lipid induced nephropathy in type-2 diabetes by its hypoglycemic, hypolipidemic and antioxidant activities as well as by modulation of TGF-β to prevent increased expression of collagen type IV and fibrinogen. We finally propose a mechanism for the expression of molecular markers that are significant in the events leading to diabetic nephropathy and its modulation by tocotrienols/TRF. - Highlights: • The nephroprotective effect of TRF in type-2 diabetic rats was investigated. • Treatment with TRF improved glycemic status, lipid profile and renal functions in rats

  8. Protective effects of tocotrienols against lipid-induced nephropathy in experimental type-2 diabetic rats by modulation in TGF-β expression

    International Nuclear Information System (INIS)

    Siddiqui, Shabeena; Ahsan, Haseeb; Khan, Mohammad Rashid; Siddiqui, Waseem A.

    2013-01-01

    Dyslipidemia is common in patients with diabetes mellitus (DM) and is considered a risk factor for the progression of diabetic nephropathy (DN). Hyperlipidemia and hyperglycemia act synergistically to induce renal injury. The present study was designed to investigate the protective effects of tocotrienols as tocotrienol-rich fraction (TRF) extracted from palm (PO) and rice bran oils (RBO) against lipid induced nephropathy in type-2 diabetic rats and its probable molecular mechanism. Male Wistar rats (175–200 g) were divided into four groups. The first group served as diabetic control, while the second and third groups received PO-TRF and RBO-TRF, respectively by gavage over a period of sixteen weeks post-induction of diabetes. The fourth group comprised of age-matched rats that served as normal control. The effects of TRF on serum lipid profile, oxidative stress markers, expression of TGF-β, fibronectin and collagen type IV were analyzed in the kidney of diabetic rats. Treatment with PO-TRF and RBO-TRF significantly improved glycemic status, serum lipid profile and renal function in type-2 diabetic rats. In addition, TRF supplementation down-regulated the expression of TGF-β, fibronectin and collagen type IV in the kidney of diabetic rats. Transforming growth factor-β (TGF-β) plays a critical role in progression of DN, but its modulation by tocotrienols in DN remains unexplored. TRF ameliorated lipid induced nephropathy in type-2 diabetes by its hypoglycemic, hypolipidemic and antioxidant activities as well as by modulation of TGF-β to prevent increased expression of collagen type IV and fibrinogen. We finally propose a mechanism for the expression of molecular markers that are significant in the events leading to diabetic nephropathy and its modulation by tocotrienols/TRF. - Highlights: • The nephroprotective effect of TRF in type-2 diabetic rats was investigated. • Treatment with TRF improved glycemic status, lipid profile and renal functions in rats

  9. Protective effects of tocotrienols against lipid-induced nephropathy in experimental type-2 diabetic rats by modulation in TGF-β expression

    Energy Technology Data Exchange (ETDEWEB)

    Siddiqui, Shabeena [Department of Biochemistry, Lipid Metabolism Laboratory, Jamia Hamdard (Hamdard University), New Delhi 110062 (India); Ahsan, Haseeb [Department of Biochemistry, Faculty of Dentistry, Jamia Millia Islamia, New Delhi 110025 (India); Khan, Mohammad Rashid [Department of Biochemistry, Lipid Metabolism Laboratory, Jamia Hamdard (Hamdard University), New Delhi 110062 (India); Siddiqui, Waseem A., E-mail: wasiddiqui01@gmail.com [Department of Biochemistry, Lipid Metabolism Laboratory, Jamia Hamdard (Hamdard University), New Delhi 110062 (India)

    2013-12-01

    Dyslipidemia is common in patients with diabetes mellitus (DM) and is considered a risk factor for the progression of diabetic nephropathy (DN). Hyperlipidemia and hyperglycemia act synergistically to induce renal injury. The present study was designed to investigate the protective effects of tocotrienols as tocotrienol-rich fraction (TRF) extracted from palm (PO) and rice bran oils (RBO) against lipid induced nephropathy in type-2 diabetic rats and its probable molecular mechanism. Male Wistar rats (175–200 g) were divided into four groups. The first group served as diabetic control, while the second and third groups received PO-TRF and RBO-TRF, respectively by gavage over a period of sixteen weeks post-induction of diabetes. The fourth group comprised of age-matched rats that served as normal control. The effects of TRF on serum lipid profile, oxidative stress markers, expression of TGF-β, fibronectin and collagen type IV were analyzed in the kidney of diabetic rats. Treatment with PO-TRF and RBO-TRF significantly improved glycemic status, serum lipid profile and renal function in type-2 diabetic rats. In addition, TRF supplementation down-regulated the expression of TGF-β, fibronectin and collagen type IV in the kidney of diabetic rats. Transforming growth factor-β (TGF-β) plays a critical role in progression of DN, but its modulation by tocotrienols in DN remains unexplored. TRF ameliorated lipid induced nephropathy in type-2 diabetes by its hypoglycemic, hypolipidemic and antioxidant activities as well as by modulation of TGF-β to prevent increased expression of collagen type IV and fibrinogen. We finally propose a mechanism for the expression of molecular markers that are significant in the events leading to diabetic nephropathy and its modulation by tocotrienols/TRF. - Highlights: • The nephroprotective effect of TRF in type-2 diabetic rats was investigated. • Treatment with TRF improved glycemic status, lipid profile and renal functions in rats

  10. Development of a Conceptual Etiological Model of Treatment Regimen Fatigue Among Patients Engaged in HIV Care: A Qualitative Study.

    Science.gov (United States)

    Claborn, Kasey; Miller, Mary Beth; Meier, Ellen; Carbone, Sofia

    Treatment regimen fatigue (TRF) is a decreased desire and motivation to maintain vigilance in adhering to treatment, and little is known about TRF in people living with HIV. We aimed to develop a conceptual framework of TRF. Five focus groups were conducted in 2014. Eligible participants were (a) HIV infected, (b) at least 18 years of age, (c) prescribed antiretroviral therapy, and (d) fluent in English. Data were analyzed using thematic analysis. Analyses revealed these themes: patient experiences of TRF; etiological factors at the systems, provider, and patient levels; strategies to manage TRF; consequences of TRF; and protective factors that prevent the occurrence of TRF. The results provided a conceptual framework for future investigations to build on in an effort to improve adherence and retention in HIV care. Study results indicate avenues for intervention at multiple levels (systems, provider, and patient) to reduce treatment burden and improve patient resources and capacity. Copyright © 2017 Association of Nurses in AIDS Care. Published by Elsevier Inc. All rights reserved.

  11. The neuroprotective effects of tocotrienol rich fraction and alpha tocopherol against glutamate injury in astrocytes

    Directory of Open Access Journals (Sweden)

    Thilaga Rati Selvaraju

    2014-11-01

    Full Text Available Tocotrienol rich fraction (TRF is an extract of palm oil, which consists of 25% alpha tocopherol (α-TCP and 75% tocotrienols. TRF has been shown to possess potent antioxidant, anti-inflammatory, anticancer, neuroprotection, and cholesterol lowering activities. Glutamate is the main excitatory amino acid neurotransmitter in the central nervous system of mammalian, which can be excitotoxic, and it has been suggested to play a key role in neurodegenerative disorders like Parkinson’s and Alzheimer’s diseases. In this present study, the effects of vitamin E (TRF and α-TCP in protecting astrocytes against glutamate injury were elucidated. Astrocytes induced with 180 mM of glutamate lead to significant cell death. However, glutamate mediated cytotoxicity was diminished via pre and post supplementation of TRF and α-TCP. Hence, vitamin E acted as a potent antioxidant agent in recovering mitochondrial injury due to elevated oxidative stress, and enhanced better survivability upon glutamate toxicity.  

  12. Treatment-related fluctuation in Guillain-Barre syndrome

    Directory of Open Access Journals (Sweden)

    Thirunavukkarasu Thivakaran

    2011-01-01

    Full Text Available Guillain-Barre syndrome (GBS is usually a monophasic illness but relapses occur. A 55-year-old female with hypertension and vitiligo presented with acute inflammatory demyelinating polyradiculoneuropathy. She improved with immunoglobulin treatment started on day 6 of illness, but relapsed on day 14 warranting repeat immunoglobulin therapy. Thereafter recovery was complete. Her relapse was due to treatment-related fluctuation (TRF. TRF is improvement in the GBS disability scale of at least one grade after completion of immunotherapy followed by worsening of the disability scale of at least one grade within the first 2 months after disease onset. Recurrent GBS and chronic inflammatory demyelinating polyradiculoneuropathy were excluded. During the peak of the illness ANA titres were transiently high. The presence of other medical conditions, predominant proximal weakness and the absence of preceding diarrhea are predictors for TRF seen in this patient. Early treatment and evidence of ongoing immune activation have contributed toward TRF.

  13. Photophysical characterization and time-resolved spectroscopy of a anthradithiophene dimer: exploring the role of conformation in singlet fission

    KAUST Repository

    Dean, Jacob C.; Zhang, Ruomeng; Hallani, Rawad K.; Pensack, Ryan D.; Sanders, Samuel N.; Oblinsky, Daniel G.; Parkin, Sean R.; Campos, Luis M.; Anthony, John E.; Scholes, Gregory D.

    2017-01-01

    carried out in toluene and acetone solution via absorption and fluorescence spectroscopy, and their photo-initiated dynamics were investigated with time-resolved fluorescence (TRF) and transient absorption (TA) spectroscopy. In accordance

  14. Effects of Aging and Tocotrienol-Rich Fraction Supplementation on Brain Arginine Metabolism in Rats

    Directory of Open Access Journals (Sweden)

    Musalmah Mazlan

    2017-01-01

    Full Text Available Accumulating evidence suggests that altered arginine metabolism is involved in the aging and neurodegenerative processes. This study sought to determine the effects of age and vitamin E supplementation in the form of tocotrienol-rich fraction (TRF on brain arginine metabolism. Male Wistar rats at ages of 3 and 21 months were supplemented with TRF orally for 3 months prior to the dissection of tissue from five brain regions. The tissue concentrations of L-arginine and its nine downstream metabolites were quantified using high-performance liquid chromatography and liquid chromatography tandem mass spectrometry. We found age-related alterations in L-arginine metabolites in the chemical- and region-specific manners. Moreover, TRF supplementation reversed age-associated changes in arginine metabolites in the entorhinal cortex and cerebellum. Multiple regression analysis revealed a number of significant neurochemical-behavioral correlations, indicating the beneficial effects of TRF supplementation on memory and motor function.

  15. Origin and global diversification patterns of tropical rain forests: inferences from a complete genus-level phylogeny of palms

    Directory of Open Access Journals (Sweden)

    Couvreur Thomas LP

    2011-06-01

    Full Text Available Abstract Background Understanding how biodiversity is shaped through time is a fundamental question in biology. Even though tropical rain forests (TRF represent the most diverse terrestrial biomes on the planet, the timing, location and mechanisms of their diversification remain poorly understood. Molecular phylogenies are valuable tools for exploring these issues, but to date most studies have focused only on recent time scales, which minimises their explanatory potential. In order to provide a long-term view of TRF diversification, we constructed the first complete genus-level dated phylogeny of a largely TRF-restricted plant family with a known history dating back to the Cretaceous. Palms (Arecaceae/Palmae are one of the most characteristic and ecologically important components of TRF worldwide, and represent a model group for the investigation of TRF evolution. Results We provide evidence that diversification of extant lineages of palms started during the mid-Cretaceous period about 100 million years ago. Ancestral biome and area reconstructions for the whole family strongly support the hypothesis that palms diversified in a TRF-like environment at northern latitudes. Finally, our results suggest that palms conform to a constant diversification model (the 'museum' model or Yule process, at least until the Neogene, with no evidence for any change in diversification rates even through the Cretaceous/Paleogene mass extinction event. Conclusions Because palms are restricted to TRF and assuming biome conservatism over time, our results suggest the presence of a TRF-like biome in the mid-Cretaceous period of Laurasia, consistent with controversial fossil evidence of the earliest TRF. Throughout its history, the TRF biome is thought to have been highly dynamic and to have fluctuated greatly in extent, but it has persisted even during climatically unfavourable periods. This may have allowed old lineages to survive and contribute to the steady

  16. Non-erythroid alpha spectrin prevents telomere dysfunction after DNA interstrand cross-link damage

    OpenAIRE

    Zhang, Pan; Herbig, Utz; Coffman, Frederick; Lambert, Muriel W.

    2013-01-01

    Telomere integrity is critical for telomere function and genomic stability. We previously demonstrated that non-erythroid ?-spectrin (?IISp) is present in mammalian cell nuclei where it is important in repair of DNA interstrand cross-links (ICLs) and chromosome stability. We now demonstrate that ?IISp is also important for telomere maintenance after ICL damage. It localizes to telomeres in S phase after ICL damage where it has enhanced association with TRF1 and TRF2 and is required for recrui...

  17. Interpreting ecological diversity indices applied to terminal restriction fragment length polymorphism data: insights from simulated microbial communities.

    Science.gov (United States)

    Blackwood, Christopher B; Hudleston, Deborah; Zak, Donald R; Buyer, Jeffrey S

    2007-08-01

    Ecological diversity indices are frequently applied to molecular profiling methods, such as terminal restriction fragment length polymorphism (T-RFLP), in order to compare diversity among microbial communities. We performed simulations to determine whether diversity indices calculated from T-RFLP profiles could reflect the true diversity of the underlying communities despite potential analytical artifacts. These include multiple taxa generating the same terminal restriction fragment (TRF) and rare TRFs being excluded by a relative abundance (fluorescence) threshold. True community diversity was simulated using the lognormal species abundance distribution. Simulated T-RFLP profiles were generated by assigning each species a TRF size based on an empirical or modeled TRF size distribution. With a typical threshold (1%), the only consistently useful relationship was between Smith and Wilson evenness applied to T-RFLP data (TRF-E(var)) and true Shannon diversity (H'), with correlations between 0.71 and 0.81. TRF-H' and true H' were well correlated in the simulations using the lowest number of species, but this correlation declined substantially in simulations using greater numbers of species, to the point where TRF-H' cannot be considered a useful statistic. The relationships between TRF diversity indices and true indices were sensitive to the relative abundance threshold, with greatly improved correlations observed using a 0.1% threshold, which was investigated for comparative purposes but is not possible to consistently achieve with current technology. In general, the use of diversity indices on T-RFLP data provides inaccurate estimates of true diversity in microbial communities (with the possible exception of TRF-E(var)). We suggest that, where significant differences in T-RFLP diversity indices were found in previous work, these should be reinterpreted as a reflection of differences in community composition rather than a true difference in community diversity.

  18. The neutrino mass from beta spectrum (ITEP-85)

    International Nuclear Information System (INIS)

    Lubimov, V.

    1986-01-01

    The new cycle of tritium beta spectrum measurements in valine with the ITEP spectrometer is discussed (ITEP-85). The detailed investigation of the total response function (TRF) has been performed. The special run of beta spectrum measurements carried out in a wide energy interval (3.4 KeV) has turned out to be a sensitive mode for the TRF experimental test. The results confirm the indication of the neutrino nonzero mass. 11 refs., 10 figs

  19. Interaction Profiling Identifies the Human Nuclear Exosome Targeting Complex

    DEFF Research Database (Denmark)

    Lubas, Michal Szymon; Christensen, Marianne Skovgaard; Kristiansen, Maiken Søndergaard

    2011-01-01

    from nucleoli, and consistently NEXT is specifically required for the exosomal degradation of promoter upstream transcripts (PROMPTs). We also detect putative homolog TRAMP subunits hTRF4-2 (Trf4p) and ZCCHC7 (Air2p) in hRRP6 and hMTR4 precipitates. However, at least ZCCHC7 function is restricted...... to nucleoli. Our results suggest that human nuclear exosome degradation pathways comprise modules of spatially organized cofactors that diverge from the yeast model....

  20. Time-restricted feeding improves insulin resistance and hepatic steatosis in a mouse model of postmenopausal obesity.

    Science.gov (United States)

    Chung, Heekyung; Chou, Winjet; Sears, Dorothy D; Patterson, Ruth E; Webster, Nicholas J G; Ellies, Lesley G

    2016-12-01

    Menopause is associated with significant hormonal changes that result in increased total body fat and abdominal fat, amplifying the risk for metabolic syndrome and diseases such as diabetes, cardiovascular disease and cancer in postmenopausal women. Intermittent fasting regimens hold significant health benefit promise for obese humans, however, regimens that include extreme daytime calorie restriction or daytime fasting are generally associated with hunger and irritability, hampering long-term compliance and adoption in the clinical setting. Time-restricted feeding (TRF), a regimen allowing eating only during a specific period in the normal circadian feeding cycle, without calorie restriction, may increase compliance and provide a more clinically viable method for reducing the detrimental metabolic consequences associated with obesity. We tested TRF as an intervention in a mouse model of postmenopausal obesity. Metabolic parameters were measured using Clinical Laboratory Animal Monitoring System (CLAMS) and we carried out glucose tolerance tests. We also stained liver sections with oil red O to examine steatosis and measured gene expression related to gluconeogenesis. Preexisting metabolic disease was significantly attenuated during 7 weeks of TRF. Despite having access to the same high fat diet (HFD) as ad libitum fed (ALF) mice, TRF mice experienced rapid weight loss followed by a delayed improvement in insulin resistance and a reduced severity of hepatic steatosis by having access to the HFD for only 8h during their normal nocturnal feeding period. The lower respiratory exchange ratio in the TRF group compared with the ALF group early in the dark phase suggested that fat was the predominant fuel source in the TRF group and correlated with gene expression analyses that suggested a switch from gluconeogenesis to ketogenesis. In addition, TRF mice were more physically active than ALF fed mice. Our data support further analysis of TRF as a clinically viable form of

  1. Minimizing the total harmonic distortion for a 3 kW, 20 kHz ac to dc converter using SPICE

    Science.gov (United States)

    Lollar, Louis F.; Kapustka, Robert E.

    1988-01-01

    This paper describes the SPICE model of a transformer-rectified-filter (TRF) circuit and the Micro-CAP (Microcomputer Circuit Analysis Program) model and their application. The models were used to develop an actual circuit with reduced input current THD. The SPICE analysis consistently predicted the THD improvements in actual circuits as various designs were attempted. In an effort to predict and verify load regulation, the incorporation of saturable inductor models significantly improved the fidelity of the TRF circuit output voltage.

  2. Telomere dysfunction and cell survival: Roles for distinct TIN2-containing complexes

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sahn-ho; Davalos, Albert R.; Heo, Seok-Jin; Rodier, Francis; Zou, Ying; Beausejour, Christian; Kaminker, Patrick; Yannone, Steven M.; Campisi, Judith

    2007-10-02

    Telomeres are maintained by three DNA binding proteins (TRF1, TRF2 and POT1), and several associated factors. One factor, TIN2, binds TRF1 and TRF2 directly and POT1 indirectly. Along with two other proteins, TPP1 and hRap1, these form a soluble complex that may be the core telomere maintenance complex. It is not clear whether sub-complexes also exist in vivo. We provide evidence for two TIN2 sub-complexes with distinct functions in human cells. We isolated these two TIN2 sub-complexes from nuclear lysates of unperturbed cells and cells expressing TIN2 mutants TIN2-13, TIN2-15C, which cannot bind TRF2 or TRF1, respectively. In cells with wild-type p53 function, TIN2-15C was more potent than TIN2-13 in causing telomere uncapping and eventual growth arrest. In cells lacking p53 function, TIN2-15C was more potent than TIN2-13 in causing telomere dysfunction and cell death. Our findings suggest that distinct TIN2 complexes exist, and that TIN2-15C-sensitive subcomplexes are particularly important for cell survival in the absence of functional p53.

  3. Palm Tocotrienol-Rich Fraction Improves Vascular Proatherosclerotic Changes in Hyperhomocysteinemic Rats

    Directory of Open Access Journals (Sweden)

    Ku-Zaifah Norsidah

    2013-01-01

    Full Text Available This study investigated the effects of palm tocotrienol-rich fraction (TRF on aortic proatherosclerotic changes in rats fed with a high methionine diet. Forty-two male Wistar rats were divided into six groups. The first group was the control (fed with a basal diet. Another five groups were fed with 1% methionine diet for 10 weeks. From week 6 onward, folate (8 mg/kg diet or palm TRF (30, 60, and 150 mg/kg diets was added into the diet of the last four rat groups, respectively. The high methionine diet raised the plasma total homocysteine and aortic lipid peroxidation, which were reduced by the palm TRF and folate supplementations. Plasma nitric oxide was reduced in the high methionine group compared to the control (3.72±0.57 versus 6.65±0.53 μmol/L, P<0.05, which reduction was reversed by the palm TRF (60 and 150 mg/kg and folate supplementations. The increased aortic vascular cell adhesion molecule-1 expression in the methionine group (2.58±0.29 was significantly reduced by the folate (1.38±0.18 and palm TRF at 150 mg/kg (1.19±0.23. Palm TRF was comparable to folate in reducing high methionine diet-induced plasma hyperhomocysteinemia, aortic oxidative stress, and inflammatory changes in rats.

  4. Comparative Effects of Biodynes, Tocotrienol-Rich Fraction, and Tocopherol in Enhancing Collagen Synthesis and Inhibiting Collagen Degradation in Stress-Induced Premature Senescence Model of Human Diploid Fibroblasts

    Science.gov (United States)

    Jam, Faidruz Azura; Ismail, Zahariah; Wan Ngah, Wan Zurinah

    2013-01-01

    Biodynes, tocotrienol-rich fraction (TRF), and tocopherol have shown antiaging properties. However, the combined effects of these compounds on skin aging are yet to be investigated. This study aimed to elucidate the skin aging effects of biodynes, TRF, and tocopherol on stress-induced premature senescence (SIPS) model of human diploid fibroblasts (HDFs) by determining the expression of collagen and MMPs at gene and protein levels. Primary HDFs were treated with biodynes, TRF, and tocopherol prior to hydrogen peroxide (H2O2) exposure. The expression of COL1A1, COL3A1, MMP1, MMP2, MMP3, and MMP9 genes was determined by qRT-PCR. Type I and type III procollagen proteins were measured by Western blotting while the activities of MMPs were quantified by fluorometric Sensolyte MMP Kit. Our results showed that biodynes, TRF, and tocopherol upregulated collagen genes and downregulated MMP genes (P < 0.05). Type I procollagen and type III procollagen protein levels were significantly increased in response to biodynes, TRF, and tocopherol treatment (P < 0.05) with reduction in MMP-1, MMP-2, MMP-3, and MMP-9 activities (P < 0.05). These findings indicated that biodynes, TRF, and tocopherol effectively enhanced collagen synthesis and inhibited collagen degradation and therefore may protect the skin from aging. PMID:24396567

  5. Comparative Effects of Biodynes, Tocotrienol-Rich Fraction, and Tocopherol in Enhancing Collagen Synthesis and Inhibiting Collagen Degradation in Stress-Induced Premature Senescence Model of Human Diploid Fibroblasts

    Directory of Open Access Journals (Sweden)

    Suzana Makpol

    2013-01-01

    Full Text Available Biodynes, tocotrienol-rich fraction (TRF, and tocopherol have shown antiaging properties. However, the combined effects of these compounds on skin aging are yet to be investigated. This study aimed to elucidate the skin aging effects of biodynes, TRF, and tocopherol on stress-induced premature senescence (SIPS model of human diploid fibroblasts (HDFs by determining the expression of collagen and MMPs at gene and protein levels. Primary HDFs were treated with biodynes, TRF, and tocopherol prior to hydrogen peroxide (H2O2 exposure. The expression of COL1A1, COL3A1, MMP1, MMP2, MMP3, and MMP9 genes was determined by qRT-PCR. Type I and type III procollagen proteins were measured by Western blotting while the activities of MMPs were quantified by fluorometric Sensolyte MMP Kit. Our results showed that biodynes, TRF, and tocopherol upregulated collagen genes and downregulated MMP genes (P<0.05. Type I procollagen and type III procollagen protein levels were significantly increased in response to biodynes, TRF, and tocopherol treatment (P<0.05 with reduction in MMP-1, MMP-2, MMP-3, and MMP-9 activities (P<0.05. These findings indicated that biodynes, TRF, and tocopherol effectively enhanced collagen synthesis and inhibited collagen degradation and therefore may protect the skin from aging.

  6. Ultrasensitive Sensing Material Based on Opal Photonic Crystal for Label-Free Monitoring of Transferrin.

    Science.gov (United States)

    Wu, Enqi; Peng, Yuan; Zhang, Xihao; Bai, Jialei; Song, Yanqiu; He, Houluo; Fan, Longxing; Qu, Xiaochen; Gao, Zhixian; Liu, Ying; Ning, Baoan

    2017-02-22

    A new opal photonic crystal (PC) sensing material, allowing label-free detection of transferrin (TRF), is proposed in the current study. This photonic crystal was prepared via a vertical convective self-assembly method with monodisperse microspheres polymerized by methyl methacrylate (MMA) and 3-acrylamidophenylboronic acid (AAPBA). FTIR, TG, and DLS were used to characterize the components and particle size of the monodisperse microspheres. SEM was used to observe the morphology of the PC. The diffraction peak intensity decreases as the TRF concentration increase. This was due to the combination of TRF to the boronic acid group of the photonic crystal. After condition optimization, a standard curve was obtained and the linear range of TRF concentration was from 2 × 10 -3 ng/mL to 200 ng/mL. Measurement of TRF concentration in simulated urine sample was also investigated using the sensing material. The results indicated that the PC provided a cheap, label-free, and easy-to-use alternative for TRF determination in clinical diagnostics.

  7. Endogenous and ectopic expression of telomere regulating genes in chicken embryonic fibroblasts

    International Nuclear Information System (INIS)

    Michailidis, Georgios; Saretzki, Gabriele; Hall, Judith

    2005-01-01

    In this study, we compared the endogenous expression of genes encoding telomere regulating proteins in cultured chicken embryonic fibroblasts (CEFs) and 10-day-old chicken embryos. CEFs maintained in vitro senesced and senescence was accompanied by reduced telomere length, telomerase activity, and expression of the chicken (c) TRF1 gene. There was no change in TRF2 gene expression although the major TRF2 transcript identified in 10-day-old chicken embryos encoded a truncated TRF2 protein (TRF2'), containing an N-terminal dimerisation domain but lacking a myb-related DNA binding domain and nuclear localisation signal. Senescence of the CEFs in vitro was associated with the loss of the TRF2' transcript, indicative of a novel function for the encoded protein. Senescence was also coupled with decreased expression of RAD51, but increased RAD52 expression. These data support that RAD51 independent recombination mechanisms do not function in vitro to maintain chicken telomeres. To attempt to rescue the CEFs from replicative senescence, we stably transfected passage 3 CEFs with the human telomerase reverse transcriptase (hTERT) catalytic subunit. While hTERT expression was detected in the stable transfectants neither telomerase activity nor the stabilisation of telomere length was observed, and the transfectant cells senesced at the same passage number as the untransfected cells. These data indicate that the human TERT is incompatible with the avian telomere maintenance apparatus and suggest the functioning of a species specific telomere system in the avian

  8. Disordering of human telomeric G-quadruplex with novel antiproliferative anthrathiophenedione.

    Directory of Open Access Journals (Sweden)

    Dmitry Kaluzhny

    Full Text Available Linear heteroareneanthracenediones have been shown to interfere with DNA functions, thereby causing death of human tumor cells and their drug resistant counterparts. Here we report the interaction of our novel antiproliferative agent 4,11-bis[(2-{[acetimido]amino}ethylamino]anthra[2,3-b]thiophene-5,10-dione with telomeric DNA structures studied by isothermal titration calorimetry, circular dichroism and UV absorption spectroscopy. New compound demonstrated a high affinity (K(ass∼10⁶ M⁻¹ for human telomeric antiparallel quadruplex d(TTAGGG₄ and duplex d(TTAGGG₄∶d(CCCTAA₄. Importantly, a ∼100-fold higher affinity was determined for the ligand binding to an unordered oligonucleotide d(TTAGGG TTAGAG TTAGGG TTAGGG unable to form quadruplex structures. Moreover, in the presence of Na+ the compound caused dramatic conformational perturbation of the telomeric G-quadruplex, namely, almost complete disordering of G-quartets. Disorganization of a portion of G-quartets in the presence of K+ was also detected. Molecular dynamics simulations were performed to illustrate how the binding of one molecule of the ligand might disrupt the G-quartet adjacent to the diagonal loop of telomeric G-quadruplex. Our results provide evidence for a non-trivial mode of alteration of G-quadruplex structure by tentative antiproliferative drugs.

  9. The SpTransformer Gene Family (Formerly Sp185/333) in the Purple Sea Urchin and the Functional Diversity of the Anti-Pathogen rSpTransformer-E1 Protein

    Science.gov (United States)

    Smith, L. Courtney; Lun, Cheng Man

    2017-01-01

    The complex innate immune system of sea urchins is underpinned by several multigene families including the SpTransformer family (SpTrf; formerly Sp185/333) with estimates of ~50 members, although the family size is likely variable among individuals of Strongylocentrotus purpuratus. The genes are small with similar structure, are tightly clustered, and have several types of repeats in the second of two exons and that surround each gene. The density of repeats suggests that the genes are positioned within regions of genomic instability, which may be required to drive sequence diversification. The second exon encodes the mature protein and is composed of blocks of sequence called elements that are present in mosaics of defined element patterns and are the major source of sequence diversity. The SpTrf genes respond swiftly to immune challenge, but only a single gene is expressed per phagocyte. Many of the mRNAs appear to be edited and encode proteins with altered and/or missense sequence that are often truncated, of which some may be functional. The standard SpTrf protein structure is an N-terminal glycine-rich region, a central RGD motif, a histidine-rich region, and a C-terminal region. Function is predicted from a recombinant protein, rSpTransformer-E1 (rSpTrf-E1), which binds to Vibrio and Saccharomyces, but not to Bacillus, and binds tightly to lipopolysaccharide, β-1,3-glucan, and flagellin, but not to peptidoglycan. rSpTrf-E1 is intrinsically disordered but transforms to α helical structure in the presence of binding targets including lipopolysaccharide, which may underpin the characteristics of binding to multiple targets. SpTrf proteins associate with coelomocyte membranes, and rSpTrf-E1 binds specifically to phosphatidic acid (PA). When rSpTrf-E1 is bound to PA in liposome membranes, it induces morphological changes in liposomes that correlate with PA clustering and leakage of luminal contents, and it extracts or removes PA from the bilayer. The

  10. Association of Circulating Transfer RNA fragments with antibody response to Mycoplasma bovis in beef cattle.

    Science.gov (United States)

    Casas, Eduardo; Cai, Guohong; Kuehn, Larry A; Register, Karen B; McDaneld, Tara G; Neill, John D

    2018-03-13

    High throughput sequencing allows identification of small non-coding RNAs. Transfer RNA Fragments are a class of small non-coding RNAs, and have been identified as being involved in inhibition of gene expression. Given their role, it is possible they may be involved in mediating the infection-induced defense response in the host. Therefore, the objective of this study was to identify 5' transfer RNA fragments (tRF5s) associated with a serum antibody response to M. bovis in beef cattle. The tRF5s encoding alanine, glutamic acid, glycine, lysine, proline, selenocysteine, threonine, and valine were associated (P < 0.05) with antibody response against M. bovis. tRF5s encoding alanine, glutamine, glutamic acid, glycine, histidine, lysine, proline, selenocysteine, threonine, and valine were associated (P < 0.05) with season, which could be attributed to calf growth. There were interactions (P < 0.05) between antibody response to M. bovis and season for tRF5 encoding selenocysteine (anticodon UGA), proline (anticodon CGG), and glutamine (anticodon TTG). Selenocysteine is a rarely used amino acid that is incorporated into proteins by the opal stop codon (UGA), and its function is not well understood. Differential expression of tRF5s was identified between ELISA-positive and negative animals. Production of tRF5s may be associated with a host defense mechanism triggered by bacterial infection, or it may provide some advantage to a pathogen during infection of a host. Further studies are needed to establish if tRF5s could be used as a diagnostic marker of chronic exposure.

  11. Alterations of telomerase activity and terminal restriction fragment in gastric cancer and its premalignant lesions.

    Science.gov (United States)

    Yang, S M; Fang, D C; Luo, Y H; Lu, R; Battle, P D; Liu, W W

    2001-08-01

    In order to explore the role of alterations of telomerase activity and terminal restriction fragment (TRF) length in the development and progression of gastric cancer. Telomerase activity was detected in 176 specimens of gastric mucosa obtained through an operation or endoscopical biopsy by using the telomeric repeat amplification protocol (TRAP) assay. Meanwhile, the mean length of TRF was measured with the use of a Southern blot in part of those samples. Telomerase activity was detected in 14 of 57 (24.6%) chronic atrophy gastritis patients, six of 18 (33.3%) intestinal metaplasia patients, three of eight (37.5%) dysplasia patients and 60 of 65 (92.3%) gastric cancer patients, respectively. Normal gastric mucosa revealed no telomerase activity. No association was found between telomerase activity and any clinicopathological parameters. The mean TRF length was decreased gradually with age in normal mucosa and in gastric cancer tissue. Regression analysis demonstrated that the reduction rate in these tissues was 41 +/- 12 base pairs/year. Among 35 gastric cancers, TRF length was shown to be shorter in 20 cases (57.1%), similar in 12 cases (34.3%) and elongated in three cases (7.6%), compared to the corresponding adjacent tissues. The mean TRF length tended to decrease as the mucosa underwent chronic atrophy gastritis, intestinal metaplasia, dysplasia and into gastric cancer. The mean TRF length in gastric cancer was not statistically correlated with clinicopathological parameters and telomerase activity. Our results suggest that telomerase is expressed during the early stage of gastric carcinogenesis, and that the clinical significance of TRF length appears to be limited in gastric cancer.

  12. Thujone-Rich Fraction of Thuja occidentalis Demonstrates Major Anti-Cancer Potentials: Evidences from In Vitro Studies on A375 Cells

    Directory of Open Access Journals (Sweden)

    Raktim Biswas

    2011-01-01

    Full Text Available Crude ethanolic extract of Thuja occidentalis (Fam: Cupressaceae is used as homeopathic mother tincture (TOΦ to treat various ailments, particularly moles and tumors, and also used in various other systems of traditional medicine. Anti-proliferative and apoptosis-inducing properties of TOΦ and the thujone-rich fraction (TRF separated from it have been evaluated for their possible anti-cancer potentials in the malignant melanoma cell line A375. On initial trial by S-diphenyltetrazolium bromide assay, both TOΦ and TRF showed maximum cytotoxic effect on A375 cell line while the other three principal fractions separated by chromatography had negligible or no such effect, because of which only TRF was further characterized and subjected to certain other assays for determining its precise anti-proliferative and apoptotic potentials. TRF was reported to have a molecular formula of C10H16O with a molecular weight of 152. Exposure of TRF of Thuja occidentalis to A375 cells in vitro showed more cytotoxic, anti-proliferative and apoptotic effects as compared with TOΦ, but had minimal growth inhibitory responses when exposed to normal cells (peripheral blood mononuclear cell. Furthermore, both TOΦ and TRF also caused a significant decrease in cell viability, induced inter-nucleosomal DNA fragmentation, mitochondrial transmembrane potential collapse, increase in ROS generation, and release of cytochrome c and caspase-3 activation, all of which are closely related to the induction of apoptosis in A375 cells. Thus, TRF showed and matched all the anti-cancer responses of TOΦ and could be the main bio-active fraction. The use of TOΦ in traditional medicines against tumors has, therefore, a scientific basis.

  13. Manganese Exposure from Drinking Water and Children’s Classroom Behavior in Bangladesh

    Science.gov (United States)

    Khan, Khalid; Factor-Litvak, Pam; Wasserman, Gail A.; Liu, Xinhua; Ahmed, Ershad; Parvez, Faruque; Slavkovich, Vesna; Levy, Diane; Mey, Jacob; van Geen, Alexander

    2011-01-01

    Background: Evidence of neurological, cognitive, and neuropsychological effects of manganese (Mn) exposure from drinking water (WMn) in children has generated widespread public health concern. At elevated exposures, Mn has been associated with increased levels of externalizing behaviors, including irritability, aggression, and impulsivity. Little is known about potential effects at lower exposures, especially in children. Moreover, little is known regarding potential interactions between exposure to Mn and other metals, especially arsenic (As). Objectives: We conducted a cross-sectional study of 201 children to investigate associations of Mn and As in tube well water with classroom behavior among elementary school children, 8–11 years of age, in Araihazar, Bangladesh. Methods: Data on exposures and behavioral outcomes were collected from the participants at the baseline of an ongoing longitudinal study of child intelligence. Study children were rated by their school teachers on externalizing and internalizing items of classroom behavior using the standardized Child Behavior Checklist-Teacher’s Report Form (CBCL-TRF). Results: Log-transformed WMn was positively and significantly associated with TRF internalizing [estimated β = 0.82; 95% confidence interval (CI), 0.08–1.56; p = 0.03], TRF externalizing (estimated β = 2.59; 95% CI, 0.81–4.37; p =0.004), and TRF total scores (estimated β = 3.35; 95% CI, 0.86–5.83; p = 0.008) in models that adjusted for log-transformed water arsenic (WAs) and sociodemographic covariates. We also observed a positive monotonic dose–response relationship between WMn and TRF externalizing and TRF total scores among the participants of the study. We did not find any significant associations between WAs and various scales of TRF scores. Conclusion: These observations reinforce the growing concern regarding the neurotoxicologic effects of WMn in children. PMID:21493178

  14. Comparative effect of Piper betle, Chlorella vulgaris and tocotrienol-rich fraction on antioxidant enzymes activity in cellular ageing of human diploid fibroblasts.

    Science.gov (United States)

    Makpol, Suzana; Yeoh, Thong Wei; Ruslam, Farah Adilah Che; Arifin, Khaizurin Tajul; Yusof, Yasmin Anum Mohd

    2013-08-16

    Human diploid fibroblasts (HDFs) undergo a limited number of cellular divisions in culture and progressively reach a state of irreversible growth arrest, a process termed cellular ageing. Even though beneficial effects of Piper betle, Chlorella vulgaris and tocotrienol-rich fraction (TRF) have been reported, ongoing studies in relation to ageing is of interest to determine possible protective effects that may reverse the effect of ageing. The aim of this study was to evaluate the effect of P. betle, C. vulgaris and TRF in preventing cellular ageing of HDFs by determining the activity of antioxidant enzymes viz.; catalase, superoxide dismutase (SOD) and glutathione peroxidase. Different passages of HDFs were treated with P. betle, C. vulgaris and TRF for 24 h prior to enzymes activity determination. Senescence-associated beta-galactosidase (SA β-gal) expression was assayed to validate cellular ageing. In cellular ageing of HDFs, catalase and glutathione peroxidase activities were reduced, but SOD activity was heightened during pre-senescence. P. betle exhibited the strongest antioxidant activity by reducing SA β-gal expression, catalase activities in all age groups, and SOD activity. TRF exhibited a strong antioxidant activity by reducing SA β-gal expression, and SOD activity in senescent HDFs. C. vulgaris extract managed to reduce SOD activity in senescent HDFs. P. betle, C. vulgaris, and TRF have the potential as anti-ageing entities which compensated the role of antioxidant enzymes in cellular ageing of HDFs.

  15. Comparative effect of Piper betle, Chlorella vulgaris and tocotrienol-rich fraction on antioxidant enzymes activity in cellular ageing of human diploid fibroblasts

    Science.gov (United States)

    2013-01-01

    Background Human diploid fibroblasts (HDFs) undergo a limited number of cellular divisions in culture and progressively reach a state of irreversible growth arrest, a process termed cellular ageing. Even though beneficial effects of Piper betle, Chlorella vulgaris and tocotrienol-rich fraction (TRF) have been reported, ongoing studies in relation to ageing is of interest to determine possible protective effects that may reverse the effect of ageing. The aim of this study was to evaluate the effect of P. betle, C. vulgaris and TRF in preventing cellular ageing of HDFs by determining the activity of antioxidant enzymes viz.; catalase, superoxide dismutase (SOD) and glutathione peroxidase. Methods Different passages of HDFs were treated with P. betle, C. vulgaris and TRF for 24 h prior to enzymes activity determination. Senescence-associated beta-galactosidase (SA β-gal) expression was assayed to validate cellular ageing. Results In cellular ageing of HDFs, catalase and glutathione peroxidase activities were reduced, but SOD activity was heightened during pre-senescence. P. betle exhibited the strongest antioxidant activity by reducing SA β-gal expression, catalase activities in all age groups, and SOD activity. TRF exhibited a strong antioxidant activity by reducing SA β-gal expression, and SOD activity in senescent HDFs. C. vulgaris extract managed to reduce SOD activity in senescent HDFs. Conclusion P. betle, C. vulgaris, and TRF have the potential as anti-ageing entities which compensated the role of antioxidant enzymes in cellular ageing of HDFs. PMID:23948056

  16. Treatment-related fluctuations in guillain barre syndrome and the conundrum of additional cycles of plasmapheresis

    Directory of Open Access Journals (Sweden)

    Subasree Ramakrishnan

    2017-01-01

    Full Text Available Introduction: In Guillain Barre syndrome (GBS, worsening of weakness or disability after initial period of recovery or stabilization is described as treatment-related fluctuations (TRF. Aim: This study aims to describe the clinical characteristics and outcome of six patients with GBS and TRF. Patients and Methods: Six patients with GBS fulfilling NINCDS criteria, evaluated at a tertiary care university hospital during 2008–2017, were diagnosed to have TRF. They form the basis of this report. Results: All patients were men and their mean age was 40 years. At presentation, mean duration of illness was 15 days; the illness had plateaued in three and progressive in other three patients. Two of the four patients had variant GBS. Initially, five patients were treated with large volume plasmapheresis (LVPP and one patient with methyl prednisolone. At 17–28 days after disease onset, three patients developed new neurologic deficits (bilateral facial paresis in two; paralytic ileus in one. Other three patients with worsening of limb weakness (medical research council sum score of >5 and disability (Hughes disability grade by ≥1 fulfilled Kleyweg's criteria for TRF. All the six patients were treated with the completion of five cycles or additional cycles of LVPP. Conclusion: Awareness about TRF is essential for correct diagnosis and management of patients with GBS.

  17. Radioisotopic evaluation of portal circulation

    International Nuclear Information System (INIS)

    Maliska, C.; Rosenthal, D.

    1986-01-01

    The use of a radio-tracer of portal circulation through the intestine, should prevent cruel punctures in the portal-vein or spleen as it is usually the case with traditional methods in the study of portal-system. The absorption of I-131 and Tc-99m, previously cheked in rabbits presented similar results in dogs. The time of circulation between terminal large-intestine and the liver (t-RF) was determined by external counting at hepatic level by recording radioactivity variation-time. In healthy animals the t-RF was from 20to 60 seconds, with average time of 42 seconds. In 2 animals with partial binding of portal-vein the t-RF went up to 110 and 120 seconds. (Author) [pt

  18. The Child Behavior Checklist for Ages 1.5-5 (CBCL/1(1/2)-5): Assessment and analysis of parent- and caregiver-reported problems in a population-based sample of Danish preschool children

    DEFF Research Database (Denmark)

    Kristensen, Solvejg; Henriksen, Tine Brink; Bilenberg, Niels

    2010-01-01

    Background: Psychometric instruments are used increasingly within research and clinical settings, and therefore standardization has become an important prerequisite, even for investigating very young children. Currently, there are no standardized psychometric instruments available for assessment...... of preschool children in Denmark. Aims: The aim was to achieve Danish national norm scores for the Child Behavior Checklist for Ages 1(1/2)-5 (CBCL/1(1/2)-5) and the Caregiver Report Form (C-TRF). Methods: The study was based on an age- and gender-stratified cohort sample of 1750 children aged 1(1/2)-5 years...... born at Aarhus University Hospital, Denmark. The CBCL/1(1/2)-5 and C-TRF were mailed to parents, who were asked to pass on the C-TRF to the preschool caregiver. The national standard register data gave access to information on socio-economic status, family type, ethnicity and parental educational level...

  19. Consistent realization of Celestial and Terrestrial Reference Frames

    Science.gov (United States)

    Kwak, Younghee; Bloßfeld, Mathis; Schmid, Ralf; Angermann, Detlef; Gerstl, Michael; Seitz, Manuela

    2018-03-01

    The Celestial Reference System (CRS) is currently realized only by Very Long Baseline Interferometry (VLBI) because it is the space geodetic technique that enables observations in that frame. In contrast, the Terrestrial Reference System (TRS) is realized by means of the combination of four space geodetic techniques: Global Navigation Satellite System (GNSS), VLBI, Satellite Laser Ranging (SLR), and Doppler Orbitography and Radiopositioning Integrated by Satellite. The Earth orientation parameters (EOP) are the link between the two types of systems, CRS and TRS. The EOP series of the International Earth Rotation and Reference Systems Service were combined of specifically selected series from various analysis centers. Other EOP series were generated by a simultaneous estimation together with the TRF while the CRF was fixed. Those computation approaches entail inherent inconsistencies between TRF, EOP, and CRF, also because the input data sets are different. A combined normal equation (NEQ) system, which consists of all the parameters, i.e., TRF, EOP, and CRF, would overcome such an inconsistency. In this paper, we simultaneously estimate TRF, EOP, and CRF from an inter-technique combined NEQ using the latest GNSS, VLBI, and SLR data (2005-2015). The results show that the selection of local ties is most critical to the TRF. The combination of pole coordinates is beneficial for the CRF, whereas the combination of Δ UT1 results in clear rotations of the estimated CRF. However, the standard deviations of the EOP and the CRF improve by the inter-technique combination which indicates the benefits of a common estimation of all parameters. It became evident that the common determination of TRF, EOP, and CRF systematically influences future ICRF computations at the level of several μas. Moreover, the CRF is influenced by up to 50 μas if the station coordinates and EOP are dominated by the satellite techniques.

  20. The INEL Tritium Research Facility

    International Nuclear Information System (INIS)

    Longhurst, G.R.

    1990-01-01

    The Tritium Research Facility (TRF) at the Idaho National Engineering Laboratory (INEL) is a small, multi-user facility dedicated to research into processes and phenomena associated with interaction of hydrogen isotopes with other materials. Focusing on bench-scale experiments, the main objectives include resolution of issues related to tritium safety in fusion reactors and the science and technology pertinent to some of those issues. In this report the TRF and many of its capabilities will be described. Work presently or recently underway there will be discussed, and the implications of that work to the development of fusion energy systems will be considered. (orig.)

  1. The INEL Tritium Research Facility

    Energy Technology Data Exchange (ETDEWEB)

    Longhurst, G.R. (Idaho National Engineering Lab., Idaho Falls (USA))

    1990-06-01

    The Tritium Research Facility (TRF) at the Idaho National Engineering Laboratory (INEL) is a small, multi-user facility dedicated to research into processes and phenomena associated with interaction of hydrogen isotopes with other materials. Focusing on bench-scale experiments, the main objectives include resolution of issues related to tritium safety in fusion reactors and the science and technology pertinent to some of those issues. In this report the TRF and many of its capabilities will be described. Work presently or recently underway there will be discussed, and the implications of that work to the development of fusion energy systems will be considered. (orig.).

  2. Chronic renal failure (CRF in children in Jugoslavia

    Directory of Open Access Journals (Sweden)

    Peco-Antić Amira

    2003-01-01

    Full Text Available The aim of this study was to analyse the demographic variables of chronic non-terminal (CRF and terminal (TRF renal failure patients (pts younger than 19 years treated in Serbia in June 2001. The prevalence of CRF pts was registered as 4,7 per million total population (pmtp or 14,1 per million child population (pmcp while corresponding values for TRF pts were 4,5 pmtp or 13,5 pmcp. The incidence of TRF pts during the period Jan.2000-Jan.2002 was 4,35 pmcp. Boys dominated only among CRF pts (34:14; 60,4% beeing between the ages of 6 and 19 yrs while at the time of diagnosis of HBI, 33,3 % of boys were yanger than 2 yrs.The causes of CRF were: reflux nephropathy 58,3%, congenital kidney disease 16,7%, familial/hereditary 14,6% glomerulonephritis 6,2% and Willms tu 4,1%. Reflux nephropathy was also the most common underlying disease of TRF accounted for 36,9% of total cases while glomerulonephritis was responsible for 23,9 %. Reflux nephropathy was associated with neural tube defect in 53,3% and with congenital lower urinary tract obstruction in 66,7%. The most of CRF (81,25% and TRF pts (95,6% were from Serbia, the others were from Monte Negro and Republic Srpska. The most of CRF (65% and TRF (80% pts were treated in University Children’s Hospital in Belgrade. Of CRF pts 46% had serum sreatinine 100-200 μmol/l, in 11% of pts it was 400-600 μmol/l and 2% of pts were in pre-terminal CRF. One third of CRF pts had proteinuria 150-500 mg/l, and second third had proteinuria greater of 1000 mg/l. Anemia was present in 54% of CRf pts, and arterial hypertension in 56%. Hemodialysis was dominant treatment modality for TRF pts and only 23,9% had functioning transplant. Conclusion: This is the first national study of demographic characteristics of pediatric CRF in Serbia. Since its prevalence is considerably lower than that in Western and North European countries the true prevalence is some what higher. The increasing incidence of pediatric TRF from 2

  3. Estudio de la interacción entre cuádruplex teloméricos y ligandos implicados en cáncer por SPR

    OpenAIRE

    Chaves Arquero, Belén; Santiveri, Clara M.; Campos Olivas, Ramón

    2015-01-01

    Los telómeros son considerados dianas terapéuticas ya que el mantenimiento de su secuencia por parte de la telomerasa es lo que caracteriza la inmortalidad de las células cancerígenas. Inicialmente, las estrategias terapéuticas se basaban en la telomerasa. En cambio, actualmente se están desarrollando estrategias centradas en los componentes que forman el telómero (shelterinas), como la proteína TRF1. Ensayos in vivo han demostrado que moléculas pequeñas consideradas como inhibidores de TRF1,...

  4. Effects of G-CSF on telomere lengths in PBMCs from human immunodeficiency virus-infected patients

    DEFF Research Database (Denmark)

    Aladdin, H; Ullum, H; Schjerling, P

    2000-01-01

    , and not in CD4+ T cells. In this double-blind placebo-controlled study, we investigated the effect of granulocyte colony stimulating factor (G-CSF) treatment combined with highly active antiretroviral therapy (HAART) on mean telomere length in peripheral blood mononuclear cells (PBMC). The terminal restriction...... fragment (TRF) length showed no changes during G-CSF treatment although the number of lymphocytes increased significantly. The mean TRF length correlated positively (R = 0.552, P = 0.009) and negatively (R = -0.503, P = 0.02) to the proportion of CD4+ memory and naïve cells, respectively. Our data suggest...

  5. Anticonvulsant effect of time-restricted feeding in a pilocarpine-induced seizure model: Metabolic and epigenetic implications.

    Directory of Open Access Journals (Sweden)

    Jorge eLandgrave-Gómez

    2016-01-01

    Full Text Available A new generation of antiepileptic drugs has emerged; however, one-third of epilepsy patients do not properly respond to pharmacological treatments. The purpose of the present study was to investigate whether time-restricted feeding has an anticonvulsant effect and whether this restrictive diet promotes changes in energy metabolism and epigenetic modifications in a pilocarpine-induced seizure model. To resolve our hypothesis, one group of rats had free access to food and water ad libitum (AL and a second group underwent a time-restricted feeding (TRF schedule. We used the lithium-pilocarpine model to induce status epilepticus (SE, and behavioral seizure monitoring was analyzed. Additionally, an electroencephalography (EEG recording was performed to verify the effect of TRF on cortical electrical activity after a pilocarpine injection. For biochemical analysis, animals were sacrificed 24 hours after SE and hippocampal homogenates were used to evaluate the proteins related to metabolism and chromatin structure. Our results showed that TRF had an anticonvulsant effect as measured by the prolonged latency of forelimb clonus seizure, a decrease in the seizure severity score and fewer animals reaching SE. Additionally, the power of the late phase EEG recordings in the AL group was significantly higher than the TRF group. Moreover, we found that TRF is capable of inducing alterations in signaling pathways that regulate energy metabolism, including an increase in the phosphorylation of AMP dependent kinase (AMPK and a decrease in the phosphorylation of Akt kinase. Furthermore, we found that TRF was able to significantly increase the beta hydroxybutyrate (β-HB concentration, an endogenous inhibitor of histone deacetylases (HDACs. Finally, we found a significant decrease in HDAC activity as well as an increase in acetylation on histone 3 (H3 in hippocampal homogenates from the TRF group. These findings suggest that alterations in energy metabolism and the

  6. Dynamics of the excited state intramolecular charge transfer

    International Nuclear Information System (INIS)

    Joo, T.; Kim, C.H.

    2006-01-01

    The 6-dodecanoyl-2-dimethylaminonaphtalene (laurdan), a derivative of 6-propanoyl- 2-dimethylaminonaphthalene (prodan), has been used as a fluorescent probe in cell imaging, especially in visualizing the lipid rafts by the generalized polarization (GP) images, where GP=(I 440 -I 490 )/(I 440 +I 490 ) with I being the fluorescence intensity. The fluorescence spectrum of laurdan is sensitive to its dipolar environment due to the intramolecular charge transfer (ICT) process in S 1 state, which results in a dual emission from the locally excited (LE) and the ICT states. The ICT process and the solvation of the ICT state are very sensitive to the dipolar nature of the environment. In this work, the ICT of laurdan in ethanol has been studied by femtosecond time resolved fluorescence (TRF), especially TRF spectra measurement without the conventional spectral reconstruction method. TRF probes the excited states exclusively, a unique advantage over the pump/probe transient absorption technique, although time resolution of the TRF is generally lower than transient absorption and the TRF spectra measurement was possible only though the spectral reconstruction. Over the years, critical advances in TRF technique have been made in our group to achieve <50 fs time resolution with direct full spectra measurement capability. Detailed ICT and the subsequent solvation processes can be visualized unambiguously from the TRF spectra. Fig. 1 shows the TRF spectra of laurdan in ethanol at several time delays. Surprisingly, two bands at 433 and 476 nm are clearly visible in the TRF spectra of laurdan even at T = 0 fs. As time increases, the band at 476 nm shifts to the red while its intensity increases. The band at 433 nm also shifts slightly to the red, but loses intensity as time increases. The intensity of the 476 nm band reaches maximum at around 5 ps, where it is roughly twice as intense as that at 0 fs, and stays constant until lifetime decay is noticeable. The spectra were fit by

  7. Experimental test of far-infrared polarimetry for Faraday rotation measurements on the TFR 600 Tokamak

    Energy Technology Data Exchange (ETDEWEB)

    Soltwisch, H [Kernforschungsanlage Juelich G.m.b.H. (Germany, F.R.). Inst. fuer Plasmaphysik; Association Euratom-Kernforschungsanlage Juelich G.m.b.H. (Germany, F.R.)); Equipe, T F.R. [Association Euratom-CEA sur la Fusion, Centre d' Etudes Nucleaires de Fontenay-aux-Roses, 92 (France). Dept. de Recherches sur la Fusion Controlee

    1981-09-01

    The results are reported on the feasibility of using far-infrared polarimetry for Faraday rotation diagnostic measurements on the TRF Tokamak. Precise quantitative results were not obtained but a satisfactory agreement with a simple theoretical model leads to a good understanding of the experimental limitations of the method.

  8. 75 FR 19444 - Self-Regulatory Organizations; The NASDAQ Stock Market LLC; Notice of Filing and Order Granting...

    Science.gov (United States)

    2010-04-14

    ... rapidly and profitably export the Project Boat technology to exploit the opportunities offered by... system as well as those reported to the FINRA/NASDAQ TRF. NASDAQ established two different pricing models... pricing discipline for the proprietary products themselves. Numerous exchanges compete with each other for...

  9. Evaluation of the protective effects of tocotrienol-rich fraction from palm oil on the dentate gyrus following chronic restraint stress in rats

    Directory of Open Access Journals (Sweden)

    Saiful Bhari Talip

    2013-06-01

    Full Text Available Exposure to chronic restraint stress has been shown to cause a number of morphological changes in the hippocampal formation of rats. Tocotrienol, an isoform of vitamin E, exhibits numerous health benefits, different from those of tocopherol. Recent studies have demonstrated that tocotrienol prevents stress-induced changes in the gastric mucosa, thus indicating that it may also protect other organs such as the brain from the damaging effects of stress. Therefore, the aim of the present study was to investigate the protective effect of tocotrienol-rich fraction (TRF extracted from palm oil on the dentate gyrus of rats following exposure to chronic restraint stress. Thirty-six male Sprague Dawley rats were divided into four groups: control, stress, tocotrienol and combination of stress and tocotrienol. Animals were stressed by restraining them for 5 hours every day for 21 consecutive days. TRF was administered via oral gavage at a dose of 200 mg/kg body weight. Our results showed that the plasma corticosterone level was significantly increased in response to stress, compared to the control. The results confirmed previous findings that chronic restraint stress suppresses cellular proliferation and reduces granule cell number in the dentate gyrus. However, TRF supplementation failed to prevent or minimize these stress-induced changes. Therefore, we conclude that TRF at the current dosage is not effective in preventing the morphological changes in the dentate gyrus induced by chronic restraint stress.

  10. Mapping surrogate gasoline compositions into RON/MON space

    NARCIS (Netherlands)

    Morgan, N.; Smallbone, A.; Bhave, A.; Kraft, M.; Cracknell, R.; Kalghatgi, G.T.

    2010-01-01

    In this paper, new experimentally determined octane numbers (RON and MON) of blends of a tri-component surrogate consisting of toluene, n-heptane, i-octane (called toluene reference fuel TRF) arranged in an augmented simplex design are used to derive a simple response surface model for the octane

  11. Telomere dysfunction and cell survival: roles for distinctTIN2-containing complexes

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sahn-Ho; Davalos, Albert R.; Heo, Seok-Jin; Rodier, Francis; Beausejour, Christian; Kaminker, Patrick; Campisi, Judith

    2006-11-07

    Telomeres are maintained by three DNA binding proteins, TRF1, TRF2 and POT1, and several associated factors. One factor, TIN2, binds TRF1 and TRF2 directly and POT1 indirectly. These and two other proteins form a soluble complex that may be the core telomere-maintenance complex. It is not clear whether subcomplexes exist or function in vivo. Here, we provide evidence for two TIN2 subcomplexes with distinct functions in human cells. TIN2 ablation by RNA interference caused telomere uncapping and p53-independent cell death in all cells tested. However, we isolated two TIN2 complexes from cell lysates, each selectively sensitive to a TIN2 mutant (TIN2-13, TIN2-15C). In cells with wild-type p53 function, TIN2-15C was more potent than TIN2-13 in causing telomere uncapping and eventual growth arrest. In cells lacking p53 function, TIN215C more than TIN2-13 caused genomic instability and cell death. Thus, TIN2 subcomplexes likely have distinct functions in telomere maintenance, and may provide selective targets for eliminating cells with mutant p53.

  12. Syndrome dimensions of the Child Behavior Checklist and the Teacher Report Form : a critical empirical evaluation

    NARCIS (Netherlands)

    Hartman, CA; Hox, J; Auerbach, J; Erol, N; Fonseca, AC; Mellenbergh, GJ; Novik, TS; Oosterlaan, J; Roussos, AC; Shalev, RS; Zilber, N; Sergeant, JA

    1999-01-01

    The construct representation of the cross-informant model of the Child Behavior Checklist (CBCL) and the Teacher Report Form (TRF) was evaluated using confirmatory factor analysis. Samples were collected in seven different countries. The results are based on 13,226 parent ratings and 8893 teacher

  13. Dissecting the telomere–inner nuclear membrane interface formed in meiosis

    Energy Technology Data Exchange (ETDEWEB)

    Pendlebury, Devon F.; Fujiwara, Yasuhiro; Tesmer, Valerie M.; Smith, Eric M.; Shibuya, Hiroki; Watanabe, Yoshinori; Nandakumar, Jayakrishnan

    2017-10-30

    Tethering telomeres to the inner nuclear membrane (INM) allows homologous chromosome pairing during meiosis. The meiosis-specific protein TERB1 binds the telomeric protein TRF1 to establish telomere–INM connectivity and is essential for mouse fertility. Here we solve the structure of the human TRF1–TERB1 interface to reveal the structural basis for telomere–INM linkage. Disruption of this interface abrogates binding and compromises telomere–INM attachment in mice. An embedded CDK-phosphorylation site within the TRF1-binding region of TERB1 provides a mechanism for cap exchange, a late-pachytene phenomenon involving the dissociation of the TRF1–TERB1 complex. Indeed, further strengthening this interaction interferes with cap exchange. Finally, our biochemical analysis implicates distinct complexes for telomere–INM tethering and chromosome-end protection during meiosis. Our studies unravel the structure, stoichiometry, and physiological implications underlying telomere–INM tethering, thereby providing unprecedented insights into the unique function of telomeres in meiosis.

  14. The temporal resolution factor with and without background noise

    NARCIS (Netherlands)

    Dreschler, W. A.; Leeuw, A. R.

    1990-01-01

    In this paper we discuss the measurement of the temporal resolution factor (TRF). The major merits of this measurement are that it is simple enough to be used clinically and that the results are, within certain limits, independent of presentation level. Addition of a continuous background noise has

  15. New Danish standardization of the Child Behaviour Checklist

    DEFF Research Database (Denmark)

    Henriksen, Jon Røikjær; Nielsen, Peter Fraas; Bilenberg, Niels

    2012-01-01

    In child mental health services, the Child Behaviour Checklist (CBCL) and related materials are internationally renowned psychometric questionnaires for assessment of children aged 6-16 years. The CBCL consists of three versions for different informants: the CBCL for parents, the Teacher's Report...... Form (TRF) and the Youth Self-Report (YSR) for 11-16-year-old children....

  16. The Effect of Error in Item Parameter Estimates on the Test Response Function Method of Linking.

    Science.gov (United States)

    Kaskowitz, Gary S.; De Ayala, R. J.

    2001-01-01

    Studied the effect of item parameter estimation for computation of linking coefficients for the test response function (TRF) linking/equating method. Simulation results showed that linking was more accurate when there was less error in the parameter estimates, and that 15 or 25 common items provided better results than 5 common items under both…

  17. Dissecting the telomere-inner nuclear membrane interface formed in meiosis.

    Science.gov (United States)

    Pendlebury, Devon F; Fujiwara, Yasuhiro; Tesmer, Valerie M; Smith, Eric M; Shibuya, Hiroki; Watanabe, Yoshinori; Nandakumar, Jayakrishnan

    2017-12-01

    Tethering telomeres to the inner nuclear membrane (INM) allows homologous chromosome pairing during meiosis. The meiosis-specific protein TERB1 binds the telomeric protein TRF1 to establish telomere-INM connectivity and is essential for mouse fertility. Here we solve the structure of the human TRF1-TERB1 interface to reveal the structural basis for telomere-INM linkage. Disruption of this interface abrogates binding and compromises telomere-INM attachment in mice. An embedded CDK-phosphorylation site within the TRF1-binding region of TERB1 provides a mechanism for cap exchange, a late-pachytene phenomenon involving the dissociation of the TRF1-TERB1 complex. Indeed, further strengthening this interaction interferes with cap exchange. Finally, our biochemical analysis implicates distinct complexes for telomere-INM tethering and chromosome-end protection during meiosis. Our studies unravel the structure, stoichiometry, and physiological implications underlying telomere-INM tethering, thereby providing unprecedented insights into the unique function of telomeres in meiosis.

  18. Tocotrienols Reverse Cardiovascular, Metabolic and Liver Changes in High Carbohydrate, High Fat Diet-Fed Rats

    Directory of Open Access Journals (Sweden)

    Weng-Yew Wong

    2012-10-01

    Full Text Available Tocotrienols have been reported to improve lipid profiles, reduce atherosclerotic lesions, decrease blood glucose and glycated haemoglobin concentrations, normalise blood pressure in vivo and inhibit adipogenesis in vitro, yet their role in the metabolic syndrome has not been investigated. In this study, we investigated the effects of palm tocotrienol-rich fraction (TRF on high carbohydrate, high fat diet-induced metabolic, cardiovascular and liver dysfunction in rats. Rats fed a high carbohydrate, high fat diet for 16 weeks developed abdominal obesity, hypertension, impaired glucose and insulin tolerance with increased ventricular stiffness, lower systolic function and reduced liver function. TRF treatment improved ventricular function, attenuated cardiac stiffness and hypertension, and improved glucose and insulin tolerance, with reduced left ventricular collagen deposition and inflammatory cell infiltration. TRF improved liver structure and function with reduced plasma liver enzymes, inflammatory cell infiltration, fat vacuoles and balloon hepatocytes. TRF reduced plasma free fatty acid and triglyceride concentrations but only omental fat deposition was decreased in the abdomen. These results suggest that tocotrienols protect the heart and liver, and improve plasma glucose and lipid profiles with minimal changes in abdominal obesity in this model of human metabolic syndrome.

  19. Tocotrienol-Rich Fraction Prevents Cell Cycle Arrest and Elongates Telomere Length in Senescent Human Diploid Fibroblasts

    Directory of Open Access Journals (Sweden)

    Suzana Makpol

    2011-01-01

    Full Text Available This study determined the molecular mechanisms of tocotrienol-rich fraction (TRF in preventing cellular senescence of human diploid fibroblasts (HDFs. Primary culture of HDFs at various passages were incubated with 0.5 mg/mL TRF for 24 h. Telomere shortening with decreased telomerase activity was observed in senescent HDFs while the levels of damaged DNA and number of cells in G0/G1 phase were increased and S phase cells were decreased. Incubation with TRF reversed the morphology of senescent HDFs to resemble that of young cells with decreased activity of SA-β-gal, damaged DNA, and cells in G0/G1 phase while cells in the S phase were increased. Elongated telomere length and restoration of telomerase activity were observed in TRF-treated senescent HDFs. These findings confirmed the ability of tocotrienol-rich fraction in preventing HDFs cellular ageing by restoring telomere length and telomerase activity, reducing damaged DNA, and reversing cell cycle arrest associated with senescence.

  20. Adolescent attachment, family functioning and depressive symptoms

    African Journals Online (AJOL)

    their sample, the depression rate was still higher in their adolescent sample compared ..... KwaZulu-Natal provincial Department of Education, as well as the. Ethics Review ... obtained from the relevant school authorities, teachers and parents. ..... Integrative guide for the 1991 CBCL 4-18, YSR, and TRF profiles. Burlington,.

  1. 75 FR 3768 - Self-Regulatory Organizations; Financial Industry Regulatory Authority, Inc.; Notice of Filing of...

    Science.gov (United States)

    2010-01-22

    ... ATS dark pool to more than one TRF, the data published on one Business Member's Web site may not... within a FINRA member's alternative trading system (``ATS'') dark pool and reported to a FINRA Trade... reporting the member's ATS dark pool transactions, and (2) adopt related Supplementary Material. \\3\\ The...

  2. 75 FR 11972 - Self-Regulatory Organizations; Financial Industry Regulatory Authority, Inc.; Order Approving...

    Science.gov (United States)

    2010-03-12

    ... Business Member's Web site may not reflect 100 percent of that member's volume for that ATS dark pool...' Web sites to see the total volume for any given ATS dark pool, and the TRF Business Members will make... they receive for each ATS dark pool on their Web site and must prominently disclose that the Web site...

  3. Th erapeutic potential of d-Th erapeutic potential of d-δ-tocotrienol rich fraction -tocotrienol rich fraction on excisional skin wounds in diabetic rats

    Directory of Open Access Journals (Sweden)

    Bijo Elsy

    2017-10-01

    Full Text Available Introduction: Long-standing hyperglycemia in addition to many of its associated complications also hampers normal wound healing which may be further aggravated in the presence of infection and oxidative stress. Therefore, antioxidant supplementation appears to be strategically relevant for wound healing. This study is designed to explore the therapeutic potential of d-δ-tocotrienol rich fraction (d-δ-TRF on skin wound healing in both healthy and diabetic rats. Materials and Methods: Diabetes was induced through single subcutaneous injection of alloxan at the dose of 100 mg/kg at hip region. 24 albino rats were divided into four groups; healthy control, diabetic control, healthy treated and diabetic treated. d-δ-TRF was administered to treated groups (200 mg/kg, orally, daily for 3 weeks. Full thickness excisional skin wounds were. Wound area was studied by assessing the morphological, histomorphological and histological features at weekly intervals and biochemical analyses were performed at the end of 3rd week. Results: The findings of present study revealed that d-δ-TRF accelerated the skin wound healing by means of early regeneration of both epidermal and dermal components; enhancement of serum protein synthesis, improvement of antioxidant status, maintenance of glycemic condition and controlling serum creatinine levels in diabetic rats. Conclusion: It is concluded that d-δ-TRF has significant therapeutic potency on the healing of skin wounds in both healthy and diabetics.

  4. Impact of probiotics on colonic microflora in patients with colitis: A prospective double blind randomised crossover study

    DEFF Research Database (Denmark)

    Ahmed, Jamil; Reddy, Bala S.; Mølbak, Lars

    2013-01-01

    restriction fragments (T-RF's) was identified in each patient. Dice cluster analysis showed that each patient had a unique microbial composition which did not change significantly at different time points in the study, irrespective of whether they had probiotics or the placebo. Probiotic organisms were...

  5. Conection between physical punishment of children and their agressive behavior

    Directory of Open Access Journals (Sweden)

    B.Torlak

    2008-08-01

    Full Text Available Aim Investigate influence of physical punishment of children to their aggressive, delinquent or asocialbehavior.Methods Data for this research were collected on a sample of 284 primary school students from SarajevoCanton being 11 to 14 years of age. Children filled in a Questionnaire for Students (designed by theauthor of this paper, and a Questionnaire for Youth YSR 6-18. Class teachers filled in a Questionnairefor Teachers, TRF 6-18.Results There was a statistically significant difference between children that were physically punishedon a scale of aggressiveness and delinquent behavior in favor of the children that were not physicallypunished (YSR-p=,016,TRF-p=,017. Physically punished children have shown tendency to aggressivebehavior and other behaviors ranging from running away from home and school and tendency of usinglies, up to delinquency (YSR-p=,040, TRF-p=,049. Correlation between negative attitude of parentstowards children including physical punishment and increased incidence of externalization of problemsin children is confirmed (YSR-p=046, TRF-p=007. Children that were physically punished are demonstratinghigher level of aggressiveness and delinquent behavior.Conclusion In addition to legal regulations, educational activities with parents and children are necessary,as well as teachers whose involvement is obligatory.

  6. The Dysregulation Profile in middle childhood and adolescence across reporters: factor structure, measurement invariance, and links with self-harm and suicidal ideation

    NARCIS (Netherlands)

    Deutz, Marike; Geeraerts, Sanne; van Baar, Anneloes; Dekovic, Maja; Prinzie, Peter

    2016-01-01

    Recently, a phenotype of severe dysregulation, the Dysregulation Profile (DP), has been identified. DP consists of elevated scores on the Anxious/Depressed (AD), Aggressive Behavior (AGG) and Attention Problems (AP) scales of the Child Behavior Checklist (CBCL), Teacher Report Form (TRF), or Youth

  7. Little ecological divergence associated with speciation in two African rain forest tree genera

    NARCIS (Netherlands)

    Couvreur, T.L.P.; Porter-Morgan, H.; Wieringa, J.J.; Chatrou, L.W.

    2011-01-01

    Background - The tropical rain forests (TRF) of Africa are the second largest block of this biome after the Amazon and exhibit high levels of plant endemism and diversity. Two main hypotheses have been advanced to explain speciation processes that have led to this high level of biodiversity:

  8. Normal telomere lengths in naive and memory CD4+ T cells in HIV type 1 infection: a mathematical interpretation

    NARCIS (Netherlands)

    Wolthers, K. C.; Noest, A. J.; Otto, S. A.; Miedema, F.; de Boer, R. J.

    1999-01-01

    To study CD4+ T cell productivity during HIV-1 infection, CD4+ T cell telomere lengths were measured. Cross-sectional and longitudinal analysis of HIV-1-infected individuals with CD4+ T cells counts >300 cells/mm3 showed normal average telomeric restriction fragment (TRF) length and normal

  9. Normal telomere lengths in naive and memory CD4 T cells in HIV type 1 infection : a mathematical interpretation

    NARCIS (Netherlands)

    Wolthers, K.C.; Noest, A.J.; Otto, S.A.; Miedema, F.; Boer, R.J. de

    1999-01-01

    To study CD4+ T cell productivity during HIV-1 infection, CD4+ T cell telomere lengths were measured. Cross-sectional and longitudinal analysis of HIV-1-infected individuals with CD4+ T cells counts >300 cells/mm3 showed normal average telomeric restriction fragment (TRF) length and normal

  10. Monte Carlo modeling of time-resolved fluorescence for depth-selective interrogation of layered tissue.

    Science.gov (United States)

    Pfefer, T Joshua; Wang, Quanzeng; Drezek, Rebekah A

    2011-11-01

    Computational approaches for simulation of light-tissue interactions have provided extensive insight into biophotonic procedures for diagnosis and therapy. However, few studies have addressed simulation of time-resolved fluorescence (TRF) in tissue and none have combined Monte Carlo simulations with standard TRF processing algorithms to elucidate approaches for cancer detection in layered biological tissue. In this study, we investigate how illumination-collection parameters (e.g., collection angle and source-detector separation) influence the ability to measure fluorophore lifetime and tissue layer thickness. Decay curves are simulated with a Monte Carlo TRF light propagation model. Multi-exponential iterative deconvolution is used to determine lifetimes and fractional signal contributions. The ability to detect changes in mucosal thickness is optimized by probes that selectively interrogate regions superficial to the mucosal-submucosal boundary. Optimal accuracy in simultaneous determination of lifetimes in both layers is achieved when each layer contributes 40-60% of the signal. These results indicate that depth-selective approaches to TRF have the potential to enhance disease detection in layered biological tissue and that modeling can play an important role in probe design optimization. Published by Elsevier Ireland Ltd.

  11. Time-Resolved Fluorescence of Water-Soluble Pyridinium Salt: Sensitive Detection of the Conformational Changes of Bovine Serum Albumin.

    Science.gov (United States)

    Li, Lei; Yi, Hua; Jia, Menghui; Chang, Mengfang; Zhou, Zhongneng; Zhang, Sanjun; Pan, Haifeng; Chen, Yan; Chen, Jinquan; Xu, Jianhua

    2016-06-20

    In this paper, we report a pyridinium salt "turn-on" fluorescent probe, 4-[2-(4-Dimethylamino-phenyl)-vinyl]-1-methylpyridinium iodide (p-DASPMI), and applied its time-resolved fluorescence (TRF) to monitor the protein conformational changes. Both the fluorescence lifetime and quantum yield (QY) of p-DASPMI were increased about two orders of magnitude after binding to the protein bovine serum albumin (BSA). The free p-DASPMI in solution presents an ultrashort fluorescence lifetime (12.4 ps), thus it does not interfere the detection of bound p-DASPMI which has nanosecond fluorescence lifetime. Decay-associated spectra (DAS) show that p-DASPMI molecules bind to subdomains IIA and IIIA of BSA. The TRF decay profiles of p-DASPMI can be described by multi-exponential decay function ([Formula: see text]), and the obtained parameters, such as lifetimes ([Formula: see text]), fractional amplitudes ([Formula: see text]), and fractional intensities ([Formula: see text]), may be used to deduce the conformational changes of BSA. The pH and Cu 2+ induced conformational changes of BSA were investigated through the TRF of p-DASPMI. The results show that the p-DASPMI is a candidate fluorescent probe in studying the conformational changes of proteins through TRF spectroscopy and microscopy in the visible range. © The Author(s) 2016.

  12. T cell telomere length in HIV-1 infection: no evidence for increased CD4+ T cell turnover

    NARCIS (Netherlands)

    Wolthers, K. C.; Bea, G.; Wisman, A.; Otto, S. A.; de Roda Husman, A. M.; Schaft, N.; de Wolf, F.; Goudsmit, J.; Coutinho, R. A.; van der Zee, A. G.; Meyaard, L.; Miedema, F.

    1996-01-01

    Progression to acquired immunodeficiency syndrome (AIDS) has been related to exhaustion of the regenerative capacity of the immune system resulting from high T cell turnover. Analysis of telomeric terminal restriction fragment (TRF) length, a marker for cellular replicative history, showed that

  13. Theory of Technical Systems--Educational Tool for Engineering

    Science.gov (United States)

    Eder, Wolfgang Ernst

    2016-01-01

    Hubka's theory of technical systems (TTS) is briefly outlined. It describes commonalities in all engineering devices, whatever their physical principles of action. This theory is based on a general transformation system (TrfS), which can be used to show engineering in the contexts of society, economics and historic developments. The life cycle of…

  14. Methanogenic community changes, and emissions of methane and other gases, during storage of acidified and untreated pig slurry

    DEFF Research Database (Denmark)

    Petersen, Søren O; Højberg, Ole; Poulsen, Morten

    2014-01-01

    H was nearly constant at values of 5.5 and 6.5. Ammonia losses were reduced by 84 and 49%, respectively, while CH4 emission with both acidification techniques was reduced by >90%. T-RFLP fingerprints showed little effect of acidification or storage time. A major T-RF of 105 bp could represent methanogens...

  15. The Effects of Digital Control on Longitudinal Autopilots for Bank-to-Turn and Skid-Turn Missiles.

    Science.gov (United States)

    1985-12-01

    Control of Dynamic Systems Addison-Wesley Publishing Company, 1961. 7. Karadimas , C., Design and Analysis of Discrete Lateral Autogilots for BTT...GREECE 8. LT Karadimas , Christos H.N 1 Kolokotroni 156 Piraeus GREECE ’-- 138 ............-.... *9. LT Karadimt trf s, Antont os HRN 201 Glenwood Circle

  16. Different bacterial communities associated with the roots and bulk sediment of the seagrass Zostera marina

    DEFF Research Database (Denmark)

    Jensen, Sheila Ingemann; Kühl, Michael; Priemé, Anders

    2007-01-01

    to Epsilonproteobacteria showed a relative mean distribution of between 5% and 11% in the root-associated communities of the youngest root bundle, in contrast to the bulk-sediment where this TRF only contributed Actinobacteria and Gammaproteobacteria also seemed important first root...

  17. Telomere binding protein TRB1 is associated with promoters of translation machinery genes in vivo

    Czech Academy of Sciences Publication Activity Database

    Schrumpfová, P.; Vychodilová, I.; Hapala, J.; Schorová, Š.; Dvořáček, Vojtěch; Fajkus, Jiří

    2016-01-01

    Roč. 90, 1-2 (2016), s. 189-206 ISSN 0167-4412 R&D Projects: GA ČR(CZ) GA13-06943S Institutional support: RVO:68081707 Keywords : Telomere repeat binding * ChIP-seq * Arabidopsis thaliana Subject RIV: BO - Biophysics Impact factor: 3.356, year: 2016

  18. Supra-optimal expression of the cold-regulated OsMyb4 transcription factor in transgenic rice changes the complexity of transcriptional network with major effects on stress tolerance and panicle development

    KAUST Repository

    Park, Myoungryoul; Yun, Kilyoung; Mohanty, Bijayalaxmi; Herath, Venura; Xu, Fuyu; Wijaya, Edward; Bajic, Vladimir B.; Yun, Songjoong; De Los Reyes, Benildo G.

    2010-01-01

    acid (JA), salicylic acid (SA), ethylene and reactive oxygen species (ROS) responsive genes. OsMyb4 network is independent of drought response element binding protein/C-repeat binding factor (DREB/CBF) and its sub-regulons operate with possible co

  19. Telomere length in normal and neoplastic canine tissues.

    Science.gov (United States)

    Cadile, Casey D; Kitchell, Barbara E; Newman, Rebecca G; Biller, Barbara J; Hetler, Elizabeth R

    2007-12-01

    To determine the mean telomere restriction fragment (TRF) length in normal and neoplastic canine tissues. 57 solid-tissue tumor specimens collected from client-owned dogs, 40 samples of normal tissue collected from 12 clinically normal dogs, and blood samples collected from 4 healthy blood donor dogs. Tumor specimens were collected from client-owned dogs during diagnostic or therapeutic procedures at the University of Illinois Veterinary Medical Teaching Hospital, whereas 40 normal tissue samples were collected from 12 control dogs. Telomere restriction fragment length was determined by use of an assay kit. A histologic diagnosis was provided for each tumor by personnel at the Veterinary Diagnostic Laboratory at the University of Illinois. Mean of the mean TRF length for 44 normal samples was 19.0 kilobases (kb; range, 15.4 to 21.4 kb), and the mean of the mean TRF length for 57 malignant tumors was 19.0 kb (range, 12.9 to 23.5 kb). Although the mean of the mean TRF length for tumors and normal tissues was identical, tumor samples had more variability in TRF length. Telomerase, which represents the main mechanism by which cancer cells achieve immortality, is an attractive therapeutic target. The ability to measure telomere length is crucial to monitoring the efficacy of telomerase inhibition. In contrast to many other mammalian species, the length of canine telomeres and the rate of telomeric DNA loss are similar to those reported in humans, making dogs a compelling choice for use in the study of human anti-telomerase strategies.

  20. School-associated problem behavior in childhood and adolescence and development of adult schizotypal symptoms: a follow-up of a clinical cohort.

    Science.gov (United States)

    Fagel, Selene; de Sonneville, Leo; van Engeland, Herman; Swaab, Hanna

    2014-01-01

    How school-associated behavioral problems in childhood and adolescence precede distinctive adult schizotypal symptoms was examined. Gender specific findings were explored. After 11.6 (SD = 3.1) years, 159 patients of the Department of Child and Adolescent Psychiatry of the University Medical Centre Utrecht, the Netherlands were reassessed for adult schizotypal symptoms. Severity of behavioral symptoms in childhood and adolescence using Teacher Report Form (TRF; Verhulst et al. 1997) and adult schizotypal symptoms using Schizotypal Personality Questionnaire-Revised (Raine in Schizophrenia Bulletin 17:555-564, 1991) were examined by Spearman's bivariate correlations. Multiple regression analyses were performed to determine the combined predictive value of significant TRF subscales for schizotypal symptomatology. Moderation was tested by adding the interactions of gender with TRF subscales to the models. Disregarding gender, correlational analyses revealed that TRF Total problems, in specific thought problems, social problems, and attentional problems were associated with disorganized schizotypal symptoms in adult life. TRF thought problems was also associated with future positive schizotypal symptoms. When gender was taken into account, for boys only thought problems was associated with adult positive schizotypal symptoms, whereas for girls externalizing problems, specifically attentional and aggressive problems, were associated with the higher levels of adult disorganized schizotypal symptoms. Moderated regression analyses provided trend significant evidence confirming that in girls externalizing problems were positively associated with general and disorganized schizotypal symptoms. When using teachers as informants, it was found that juvenile behavioral abnormalities were differentially associated with type of adult schizotypal symptoms, with these associations being further modified by gender.

  1. Eff ect of vitamin E isoforms on the primary intention Eff ect of vitamin E isoforms on the primary intention skin wound healing of diabetic rats

    Directory of Open Access Journals (Sweden)

    Bijo Elsy

    2017-10-01

    Full Text Available Introduction: Impaired wound healing events is a common complication in diabetes. One of the effective nutritional antioxidant on skin wound healing is vitamin E which contains saturated tocopherol and unsaturated tocotrienol forms. This present study is designed to explore the effect of different vitamin E isoforms on stitched skin wound in both healthy and diabetic rats. Materials and Methods: Forty eight albino rats were divided into eight groups; healthy control, diabetic control, healthy treated (d-α-tocopherol, d-δ-TRF and co-administrated and diabetic treated (d-αtocopherol, d-δ-TRF and co-administrated. Diabetes was induced through single subcutaneous injection of alloxan at the dose of 100 mg/kg. Treated groups were administered d-a-tocopherol (200 mg/kg, d-δ-TRF (200 mg/kg and co-administration (100 mg/kg of these two compounds each orally and daily for three weeks. A horizontal skin incision was made on right mid-thigh region at 2.95 ± 0.17cm in length and wound was closed with an absorbable suture. Results: Histopathological and histomorphological results at the end of 3rd week revealed that the d-δ-TRF treated groups promote the regeneration and reorganization of epidermal and dermal components in healing of primary intention more effectively than the d-α-tocopherol and co-administrated groups. Conclusion: It is concluded that among different vitamin E isoforms the d-δ-TRF appears to be a more effective nutritional antioxidant on skin wound healing in both healthy and diabetics.

  2. Human telomeric DNA: G-quadruplex, i-motif and Watson–Crick double helix

    Science.gov (United States)

    Phan, Anh Tuân; Mergny, Jean-Louis

    2002-01-01

    Human telomeric DNA composed of (TTAGGG/CCCTAA)n repeats may form a classical Watson–Crick double helix. Each individual strand is also prone to quadruplex formation: the G-rich strand may adopt a G-quadruplex conformation involving G-quartets whereas the C-rich strand may fold into an i-motif based on intercalated C·C+ base pairs. Using an equimolar mixture of the telomeric oligonucleotides d[AGGG(TTAGGG)3] and d[(CCCTAA)3CCCT], we defined which structures existed and which would be the predominant species under a variety of experimental conditions. Under near-physiological conditions of pH, temperature and salt concentration, telomeric DNA was predominantly in a double-helix form. However, at lower pH values or higher temperatures, the G-quadruplex and/or the i-motif efficiently competed with the duplex. We also present kinetic and thermodynamic data for duplex association and for G-quadruplex/i-motif unfolding. PMID:12409451

  3. Time-resolved fluorescence measurements using microlens array and area imaging devices.

    Science.gov (United States)

    Merk, Susanne; Lietz, Achim; Kroner, Margareta; Valler, Martin; Heilker, Ralf

    2004-02-01

    Time-resolved fluorescence (TRF) assay formats are frequently used technologies in high-throughput screening. In this article, we have characterised the novel Plate::Vision(2) 96-microlens array reader (Carl Zeiss Jena GmbH, Germany) and compared it to the novel LEADseeker Generation IV multimodality imaging system (LEADseeker Gen IV; Amersham Biosciences UK Ltd., UK) for applications in the TRF mode. In europium measurements using the TRF mode, the Plate::Vision displayed a limit of detection for europium of approximately 3 pM, which was comparable to two established TRF readers, the Discovery and the Victor V (both PerkinElmer Life Sciences Inc., USA). The LEADseeker's limit of detection only extended down to europium concentrations of approximately 10 pM in these experiments. For TRF resonance energy transfer (TR-FRET) experiments, a europium-biotin (Eu-biotin) conjugate was titrated with a streptavidin-allophycocyanin (SA-APC) conjugate. The Plate::Vision produced Z' values larger than 0.5 for the acceptor fluorophor emission with concentrations of Eu-biotin as low as 3 nM combined with 175 pM SA-APC. To achieve Z' values of at least 0.5 with the LEADseeker, concentrations of 10 nM Eu-biotin combined with SA-APC of at least 0.8 nM were required. In a drug screening application using TR-FRET, the energy transfer from a europium-labelled protein X (Eu-protein X) to a complex of biotinylated peptide Y with SA-APC was measured. Using the Plate::Vision, a Z' factor larger than 0.5 for the acceptor fluorophor emission was only obtained for a Eu-protein X concentration of at least 10 nM in combination with biotinylated peptide Y/SA-APC at saturating concentrations. Both the Plate::Vision and the LEADseeker show good quality results for applications in the TRF mode and enable an increased throughput based on their shortened measurement time in comparison to classic photomultiplier tube-based readers.

  4. Telomere dynamics in human mesenchymal stem cells after exposure to acute oxidative stress

    DEFF Research Database (Denmark)

    Harbo, M.; Koelvraa, S.; Serakinci, N.

    2012-01-01

    mesenchymal stem cells, either primary or hTERT immortalized, were exposed to sub-lethal doses of hydrogen peroxide, and the short term effect on telomere dynamics was monitored by Universal STELA and TRF measurements. Both telomere measures were then correlated with the percentage of senescent cells......A gradual shortening of telomeres due to replication can be measured using the standard telomere restriction fragments (TRF) assay and other methods by measuring the mean length of all the telomeres in a cell. In contrast, stress-induced telomere shortening, which is believed to be just...... estimated by senescence-associated beta-galactosidase staining. The exposure to acute oxidative stress resulted in an increased number of ultra-short telomeres, which correlated strongly with the percentage of senescent cells, whereas a correlation between mean telomere length and the percentage...

  5. Long-lived visible luminescence of UV LEDs and impact on LED excited time-resolved fluorescence applications

    International Nuclear Information System (INIS)

    Jin, D; Connally, R; Piper, J

    2006-01-01

    We report the results of a detailed study of the spectral and temporal properties of visible emission from three different GaN-based ultraviolet (UV) light emitting diodes (UV LEDs). The primary UV emission in the 360-380 nm band decays rapidly (less than 1 μs) following switch-off; however, visible luminescence (470-750 nm) with a decay lifetime of tens of microseconds was observed at approximately 10 -4 of the UV intensity. For applications of UV LEDs in time-resolved fluorescence (TRF) employing lanthanide chelates, the visible luminescence from the LEDs competes with the target Eu 3+ or Tb 3+ fluorescence in both spectral and temporal domains. A UV band-pass filter (Schott UG11 glass) was therefore used to reduce the visible luminescence of the UV LEDs by three orders of magnitude relative to UV output to yield a practical excitation source for TRF

  6. Generalized helicity and Beltrami fields

    International Nuclear Information System (INIS)

    Buniy, Roman V.; Kephart, Thomas W.

    2014-01-01

    We propose covariant and non-abelian generalizations of the magnetic helicity and Beltrami equation. The gauge invariance, variational principle, conserved current, energy–momentum tensor and choice of boundary conditions elucidate the subject. In particular, we prove that any extremal of the Yang–Mills action functional 1/4 ∫ Ω trF μν F μν d 4 x subject to the local constraint ε μναβ trF μν F αβ =0 satisfies the covariant non-abelian Beltrami equation. -- Highlights: •We introduce the covariant non-abelian helicity and Beltrami equation. •The Yang–Mills action and instanton term constraint lead to the Beltrami equation. •Solutions of the Beltrami equation conserve helicity

  7. Daily Eating Patterns and Their Impact on Health and Disease.

    Science.gov (United States)

    Zarrinpar, Amir; Chaix, Amandine; Panda, Satchidananda

    2016-02-01

    Cyclical expression of cell-autonomous circadian clock components and key metabolic regulators coordinate often discordant and distant cellular processes for efficient metabolism. Perturbation of these cycles, either by genetic manipulation, disruption of light/dark cycles, or, most relevant to the human population, via eating patterns, contributes to obesity and dysmetabolism. Time-restricted feeding (TRF), during which time of access to food is restricted to a few hours, without caloric restriction, supports robust metabolic cycles and protects against nutritional challenges that predispose to obesity and dysmetabolism. The mechanism by which TRF imparts its benefits is not fully understood but likely involves entrainment of metabolically active organs through gut signaling. Understanding the relationship of feeding pattern and metabolism could yield novel therapies for the obesity pandemic. Copyright © 2015. Published by Elsevier Ltd.

  8. Different requirements of functional telomeres in neural stem cells and terminally differentiated neurons.

    Science.gov (United States)

    Lobanova, Anastasia; She, Robert; Pieraut, Simon; Clapp, Charlie; Maximov, Anton; Denchi, Eros Lazzerini

    2017-04-01

    Telomeres have been studied extensively in peripheral tissues, but their relevance in the nervous system remains poorly understood. Here, we examine the roles of telomeres at distinct stages of murine brain development by using lineage-specific genetic ablation of TRF2, an essential component of the shelterin complex that protects chromosome ends from the DNA damage response machinery. We found that functional telomeres are required for embryonic and adult neurogenesis, but their uncapping has surprisingly no detectable consequences on terminally differentiated neurons. Conditional knockout of TRF2 in post-mitotic immature neurons had virtually no detectable effect on circuit assembly, neuronal gene expression, and the behavior of adult animals despite triggering massive end-to-end chromosome fusions across the brain. These results suggest that telomeres are dispensable in terminally differentiated neurons and provide mechanistic insight into cognitive abnormalities associated with aberrant telomere length in humans. © 2017 Lobanova et al.; Published by Cold Spring Harbor Laboratory Press.

  9. Spectral characterization of crude oil using fluorescence (synchronous and time-resolved) and NIR (Near Infrared Spectroscopy); Caracterizacao espectral do petroleo utilizando fluorescencia (sincronizada e resolvida no tempo) e NIR (Near Infrared Spectroscopy)

    Energy Technology Data Exchange (ETDEWEB)

    Falla Sotelo, F.; Araujo Pantoja, P.; Lopez-Gejo, J.; Le Roux, G.A.C.; Nascimento, C.A.O. [Universidade de Sao Paulo (USP), SP (Brazil). Dept. de Engenharia Quimica. Lab. de Simulacao e Controle de Processos; Quina, F.H. [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Quimica. Centro de Capacitacao e Pesquisa em Meio Ambiente (CEPEMA)

    2008-07-01

    The objective of the present work is to evaluate the performance of two spectroscopic techniques employed in the crude oil characterization: NIR spectroscopy and fluorescence spectroscopy (Synchronous fluorescence - SF and Time Resolved Fluorescence - TRF) for the development of correlation models between spectral profiles of crude oil samples and both physical properties (viscosity and API density) and physico-chemical properties (SARA analysis: Saturated, Aromatic, Resins and Asphaltenes). The better results for viscosity and density were obtained using NIR whose prediction capacity was good (1.5 cP and 0.5 deg API, respectively). For SARA analysis, fluorescence spectroscopy revealed its potential in the model calibration showing good results (R2 coefficients greater than 0.85). TRF spectroscopy had better performance than SF spectroscopy. (author)

  10. THE KNOWLEDGE MANAGEMENT FOR BEST PRACTICES SHARING IN A DATABASE AT THE TRIBUNAL REGIONAL FEDERAL DA PRIMEIRA REGIÃO

    Directory of Open Access Journals (Sweden)

    Márcia Mazo Santos de Miranda

    2010-08-01

    Full Text Available A quick, effective and powerful alternative for knowledge management is the systematic sharing of best practices. This study identified in the literature recommendations for structuring a best practices database and summarized the benefits of its deployment to the Tribunal Regional Federal da Primeira Região TRF - 1ª Região. A It was conducted a quantitative research was then carried out, with the distribuition of where questionnaires were distributed to federal judges of the TRF- 1ª Região, which was divided into 4 parts: magistrate profile, flow of knowledge / information, internal environment, organizational facilitator. As a result, we identified the need to have a best practices database in the Institution for the organizational knowledge identification, transfer and sharing. The conclusion presents recommendations for the development of the database and highlights its importance for knowledge management in an organization.

  11. Fasting, circadian rhythms, and time restricted feeding in healthy lifespan

    Science.gov (United States)

    Longo, Valter D.; Panda, Satchidananda

    2016-01-01

    Summary Feeding in most animals is confined to a defined period, leaving short periods of fasting that coincide with sleep. Fasting enables organisms to enter alternative metabolic phases, which rely less on glucose and more on ketone body-like carbon sources. Both intermittent and periodic fasting result in benefits ranging from prevention to the enhanced treatment of diseases. Similarly, time-restricted feeding (TRF), in which feeding time is restricted to certain hours of the day, allows the daily fasting period to last >12 h, thus imparting pleiotropic benefits in multiple organisms. Understanding the mechanistic link between nutrients and the fasting benefits is leading to the identification of fasting mimicking diets (FMDs) that achieve changes similar to those caused by fasting. Given the pleiotropic and sustained benefits of TRF and FMD, both basic science and translational research are warranted to develop fasting-associated interventions into effective and inexpensive treatments with the potential to improve healthspan. PMID:27304506

  12. Radiological safety aspects during handling of adjuster rod (cobalt-60) samples at Cirus

    International Nuclear Information System (INIS)

    Yadav, R.K.B.; Prasad, S.K.; Khuspe, R.R.; Babu, K.S.; Kamble, M.K.; Deshpande, S.B.; Srivastava, Alok; Ramesh, N.; Sharma, R.C.

    2008-01-01

    Cobalt -60 produced from the irradiation of the 59 Co capsules in adjuster rod of Cirus is used in many medical and industrial applications such as radiation therapy, sterilization of agricultural products, sterilization of medical equipment, industrial gamma exposure devices, nucleonic gauging sources etc. Two adjuster rods used for adjusting small reactivity loads (3.5 mk at 40 MW) in reactor contain thirty 59 Co slugs each. The 59 Co slugs are in the form of small capsules. These slugs were irradiated for nearly 3 years of full power operation. Since the amount of activity handled in the adjuster rod is very high (35 - 40 kCi), adequate radiological safety coverage is to be provided during the transfer of adjuster rod from the reactor pile to Tray Rod Facility (TRF) and during remote handling and loading of adjuster rod samples in shipping flask at TRF. Radiation fields observed are 0.4 - 3.0 Gy/h. In Cirus, falling of Adjuster rod from vertical flask at top of pile could result in declaration of evacuation emergency due to potential hazards associated with the high radiation field (∼26 Gy/h at 1m) of 60 Co slugs. Entire process of remote handling is viewed through lead glass and movement of samples is carried out with the help of a pair of master slave manipulator (MSM) arms. TRF with full samples give a radiation background of 0.2 - 20 mGy/h, requiring control of movement of personnel in the area. This paper explains in detail about the health physics aspects during handling of adjuster rod from reactor pile to TRF and back to pile, unloading and loading of radioisotope samples, operational radiation protection experience gained and collective dose consumed for the above transfer and loading job. The collective dose consumed in the handling of two adjuster rods is 8.7 per-mSv. (author)

  13. The Tocotrienol-Rich Fraction Is Superior to Tocopherol in Promoting Myogenic Differentiation in the Prevention of Replicative Senescence of Myoblasts.

    Directory of Open Access Journals (Sweden)

    Shy Cian Khor

    Full Text Available Aging results in a loss of muscle mass and strength. Myoblasts play an important role in maintaining muscle mass through regenerative processes, which are impaired during aging. Vitamin E potentially ameliorates age-related phenotypes. Hence, this study aimed to determine the effects of the tocotrienol-rich fraction (TRF and α-tocopherol (ATF in protecting myoblasts from replicative senescence and promoting myogenic differentiation. Primary human myoblasts were cultured into young and senescent stages and were then treated with TRF or ATF for 24 h, followed by an analysis of cell proliferation, senescence biomarkers, cellular morphology and differentiation. Our data showed that replicative senescence impaired the normal regenerative processes of myoblasts, resulting in changes in cellular morphology, cell proliferation, senescence-associated β-galactosidase (SA-β-gal expression, myogenic differentiation and myogenic regulatory factors (MRFs expression. Treatment with both TRF and ATF was beneficial to senescent myoblasts in reclaiming the morphology of young cells, improved cell viability and decreased SA-β-gal expression. However, only TRF treatment increased BrdU incorporation in senescent myoblasts, as well as promoted myogenic differentiation through the modulation of MRFs at the mRNA and protein levels. MYOD1 and MYOG gene expression and myogenin protein expression were modulated in the early phases of myogenic differentiation. In conclusion, the tocotrienol-rich fraction is superior to α-tocopherol in ameliorating replicative senescence-related aberration and promoting differentiation via modulation of MRFs expression, indicating vitamin E potential in modulating replicative senescence of myoblasts.

  14. Derivation of the canopy conductance from surface temperature and spectral indices for estimating evapotranspiration in semiarid vegetation; Monitorizacion de conductancia en vegetacion semiarida a partir de indices espectrales y temperatura de supeficie

    Energy Technology Data Exchange (ETDEWEB)

    Morillas, L.; Garcia, M.; Zarco-Tejada, P.; Ladron de Guevara, M.; Villagarcia, L.; Were, A.; Domingo, F.

    2009-07-01

    This work evaluates the possibilities for estimating stomata conductance (C) and leaf transpiration (Trf) at the ecosystem scale from radiometric indices and surface temperature. The relationships found between indices and the transpiration component of the water balance in a semiarid tussock ecosystem in SE Spain are discussed. Field data were collected from spring 2008 until winter 2009 in order to observe the annual variability of the relationships and the behaviour of spectral indices and surface temperature. (Author) 11 refs.

  15. Derivation of the canopy conductance from surface temperature and spectral indices for estimating evapotranspiration in semiarid vegetation

    International Nuclear Information System (INIS)

    Morillas, L.; Garcia, M.; Zarco-Tejada, P.; Ladron de Guevara, M.; Villagarcia, L.; Were, A.; Domingo, F.

    2009-01-01

    This work evaluates the possibilities for estimating stomata conductance (C) and leaf transpiration (Trf) at the ecosystem scale from radiometric indices and surface temperature. The relationships found between indices and the transpiration component of the water balance in a semiarid tussock ecosystem in SE Spain are discussed. Field data were collected from spring 2008 until winter 2009 in order to observe the annual variability of the relationships and the behaviour of spectral indices and surface temperature. (Author) 11 refs.

  16. First direct landscape-scale measurement of tropical rain forest Leaf Area Index, a key driver of global primary productivity

    Science.gov (United States)

    David B. Clark; Paulo C. Olivas; Steven F. Oberbauer; Deborah A. Clark; Michael G. Ryan

    2008-01-01

    Leaf Area Index (leaf area per unit ground area, LAI) is a key driver of forest productivity but has never previously been measured directly at the landscape scale in tropical rain forest (TRF). We used a modular tower and stratified random sampling to harvest all foliage from forest floor to canopy top in 55 vertical transects (4.6 m2) across 500 ha of old growth in...

  17. Evaluation of genome damage and transcription profile of DNA damage/repair response genes in peripheral blood mononuclear cells exposed to low dose radiation

    International Nuclear Information System (INIS)

    Soren, D.C.; Saini, Divyalakshmi; Das, Birajalaxmi

    2016-01-01

    Humans are exposed to various physical and chemical mutagens in their life time. Physical mutagens, like ionizing radiation (IR), may induce adverse effect at high acute dose exposures in human cells. However, there are inconsistent results on the effect of low dose radiation exposure in human cells. There are a variety of DNA damage endpoints to evaluate the effect of low dose radiation in human cells. DNA damage response (DDR) may lead to changes in expression profile of many genes. In the present study, an attempt has been made to evaluate genome damage at low dose IR exposure in human blood lymphocytes. Cytochalasin blocked micronuclei (CBMN) assay has been used to determine the frequency of micronuclei in binucleated cells in PBMCs exposed to IR. Transcription profile of ATM, P53, GADD45A, CDKN1A, TRF1 and TRF2 genes was studied using real time quantitative PCR. Venous blood samples collected from 10 random healthy donors were irradiated with different doses of γ-radiation ( 137 Cs) along with sham irradiated control. Whole blood culture was set up using microculture technique. Blood samples were stimulated with phytohemagglutinin, and CBMN assay was performed. An average of 2,500 binucleated cells was scored for each dose point. For gene expression analysis, total RNA was isolated, cDNA was prepared, and gene expression analysis for ATM, P53, CDKN1A, GADD45A, TRF1 and TRF2 was done using real time PCR. Our results revealed no significant increase in the frequency of MN up to 100 mGy as compared to control. However, no significant alteration in gene expression profile was observed. In conclusion, no significant dose response was observed at the frequency of MN as well as the expression profile of DDR/repair genes, suggesting low dose radiation did not induce significant DNA damage at these acute dose exposures. (author)

  18. Photobiomodulation effects on mRNA levels from genomic and chromosome stabilization genes in injured muscle.

    Science.gov (United States)

    da Silva Neto Trajano, Larissa Alexsandra; Trajano, Eduardo Tavares Lima; da Silva Sergio, Luiz Philippe; Teixeira, Adilson Fonseca; Mencalha, Andre Luiz; Stumbo, Ana Carolina; de Souza da Fonseca, Adenilson

    2018-04-26

    Muscle injuries are the most prevalent type of injury in sports. A great number of athletes have relapsed in muscle injuries not being treated properly. Photobiomodulation therapy is an inexpensive and safe technique with many benefits in muscle injury treatment. However, little has been explored about the infrared laser effects on DNA and telomeres in muscle injuries. Thus, the aim of this study was to evaluate photobiomodulation effects on mRNA relative levels from genes related to telomere and genomic stabilization in injured muscle. Wistar male rats were randomly divided into six groups: control, laser 25 mW, laser 75 mW, injury, injury laser 25 mW, and injury laser 75 mW. Photobiomodulation was performed with 904 nm, 3 J/cm 2 at 25 or 75 mW. Cryoinjury was induced by two applications of a metal probe cooled in liquid nitrogen directly on the tibialis anterior muscle. After euthanasia, skeletal muscle samples were withdrawn and total RNA extracted for evaluation of mRNA levels from genomic (ATM and p53) and chromosome stabilization (TRF1 and TRF2) genes by real-time quantitative polymerization chain reaction. Data show that photobiomodulation reduces the mRNA levels from ATM and p53, as well reduces mRNA levels from TRF1 and TRF2 at 25 and 75 mW in injured skeletal muscle. In conclusion, photobiomodulation alters mRNA relative levels from genes related to genomic and telomere stabilization in injured skeletal muscle.

  19. Report of Findings: Lake Superior Classified Barrel Disposal Site. Defense Environmental Restoration Program for Formerly Used Defense Sites. Project No. E05MN025501

    Science.gov (United States)

    1991-08-01

    Anny Coa" of EFnser. Teciiniciam are stil wairhing on water, -ml taken trom near the arma where several barrel werefm e Saitw- day. tbiY1 Y0wL do" wy...Army Corps Tha trfPDiCs:: *thes barres bocame~ enang..2es deunane. r hcea-;:. scra ( letal of comerc fiom - 0 cone of tssreC rlns ZeIi. -ae spwee scapes

  20. Chalcone-imidazolone conjugates induce apoptosis through DNA damage pathway by affecting telomeres

    Directory of Open Access Journals (Sweden)

    Kamal Ahmed

    2011-04-01

    Full Text Available Abstract Background Breast cancer is one of the most prevalent cancers in the world and more than one million women are diagnosed leading to 410,000 deaths every year. In our previous studies new chalcone-imidazolone conjugates were prepared and evaluated for their anticancer activity in a panel of 53 human tumor cell lines and the lead compounds identified were 6 and 8. This prompted us to investigate the mechanism of apoptotic event. Results Involvement of pro-apoptotic protein (Bax, active caspase-9 and cleavage of retinoblastoma protein was studied. Interestingly, the compounds caused upregulation of p21, check point proteins (Chk1, Chk2 and as well as their phosphorylated forms which are known to regulate the DNA damage pathway. Increased p53BP1 foci by immunolocalisation studies and TRF1 suggested the possible involvement of telomere and associated proteins in the apoptotic event. The telomeric protein such as TRF2 which is an important target for anticancer therapy against human breast cancer was extensively studied along with proteins involved in proper functioning of telomeres. Conclusions The apoptotic proteins such as Bax, active caspase-9 and cleaved RB are up-regulated in the compound treated cells revealing the apoptotic nature of the compounds. Down regulation of TRF2 and upregulation of the TRF1 as well as telomerase assay indicated the decrease in telomeric length revealing telomeric dysfunction and thereby controlling the rapid rate of cell proliferation. In summary, chalcone-imidazolone conjugates displayed significant DNA damage activity particularly at telomeres and caused both apoptosis and senescence-like growth arrest which suggested that these compounds have potential activity against breast carcinoma.

  1. Determining Tocotrienol Rich Fraction Effects on Laying, Mortality and Egg Quality Parameters

    OpenAIRE

    Mardhati, M.; Farah Nurshahida, M.S.; Osman, A.; Wan Nooraida, W.M.

    2016-01-01

    The public interest and awareness in healthier lifestyle has increased the demand of functional food, nutraceuticals and designer foods, which has resulted in the creation of new foods in marketplace, for instance designer and specialty eggs. A feeding trial was carried out to evaluate the laying performance of H&N layer chickens fed diets supplemented with tocotrienol rich fraction (TRF), as well as to quantify tocopherol and tocotrienol accumulation in eggs and their effect on egg quality. ...

  2. How reliable and valid is the teacher version of the Strengths and Difficulties Questionnaire in primary school children?

    Science.gov (United States)

    van den Heuvel, Meta; Jansen, Danielle E M C; Stewart, Roy E; Smits-Engelsman, Bouwien C M; Reijneveld, Sijmen A; Flapper, Boudien C T

    2017-01-01

    The Strengths and Difficulties Questionnaire (SDQ) is validated for parents, but not yet for teachers in a broad age range of children. We conducted a cross-sectional study with 4-10 years old school children to investigate if the SDQ-T can be used instead of the validated but lengthy Teacher's Report Form (TRF) to acquire information about emotional and behavioral problems in the school community. Teachers of 453 children from primary schools were approached. Teachers of 394 children (response rate 86.9%) with a mean age of 7.1 years filled in the SDQ-T (n = 387), the TRF (n = 349) or both (n = 342). We assessed reliability by calculating internal consistency and concurrent validity (using correlation coefficients, sensitivity, specificity) of the SDQ-T compared with the TRF. Internal consistency of the SDQ-T Total Difficulties Score (SDQ-T TDS; Cronbach α = 0.80), hyperactivity/ inattention- (α = 0.86) and prosocial behavior (α = 0.81) was very good. Concurrent validity demonstrated a strong correlation of all subscales of the SDQ-T with the corresponding scale on the TRF (range 0.54-0.73), except for peer problems (0.46). Using a SDQ-T TDS cut-off score > 14, the SDQ-T had a good sensitivity (90%) and specificity (94%). The good reliability, validity and brevity of the SDQ-T make it an easily applicable questionnaire for obtaining information about emotional and behavioral problems from teachers in primary school children.

  3. How reliable and valid is the teacher version of the Strengths and Difficulties Questionnaire in primary school children?

    Directory of Open Access Journals (Sweden)

    Meta van den Heuvel

    Full Text Available The Strengths and Difficulties Questionnaire (SDQ is validated for parents, but not yet for teachers in a broad age range of children. We conducted a cross-sectional study with 4-10 years old school children to investigate if the SDQ-T can be used instead of the validated but lengthy Teacher's Report Form (TRF to acquire information about emotional and behavioral problems in the school community.Teachers of 453 children from primary schools were approached. Teachers of 394 children (response rate 86.9% with a mean age of 7.1 years filled in the SDQ-T (n = 387, the TRF (n = 349 or both (n = 342. We assessed reliability by calculating internal consistency and concurrent validity (using correlation coefficients, sensitivity, specificity of the SDQ-T compared with the TRF.Internal consistency of the SDQ-T Total Difficulties Score (SDQ-T TDS; Cronbach α = 0.80, hyperactivity/ inattention- (α = 0.86 and prosocial behavior (α = 0.81 was very good. Concurrent validity demonstrated a strong correlation of all subscales of the SDQ-T with the corresponding scale on the TRF (range 0.54-0.73, except for peer problems (0.46. Using a SDQ-T TDS cut-off score > 14, the SDQ-T had a good sensitivity (90% and specificity (94%.The good reliability, validity and brevity of the SDQ-T make it an easily applicable questionnaire for obtaining information about emotional and behavioral problems from teachers in primary school children.

  4. On the Assessment of Global Terrestrial Reference Frame Temporal Variations

    Science.gov (United States)

    Ampatzidis, Dimitrios; Koenig, Rolf; Zhu, Shengyuan

    2015-04-01

    Global Terrestrial Reference Frames (GTRFs) as the International Terrestrial Reference Frame (ITRF) provide reliable 4-D position information (3-D coordinates and their evolution through time). The given 3-D velocities play a significant role in precise position acquisition and are estimated from long term coordinate time series from the space-geodetic techniques DORIS, GNSS, SLR, and VLBI. GTRFs temporal evolution is directly connected with their internal stability: The more intense and inhomogeneous velocity field, the less stable TRF is derived. The assessment of the quality of the GTRF is mainly realized by comparing it to each individual technique's reference frame. E.g the comparison of GTRFs to SLR-only based TRF gives the sense of the ITRF stability with respect to the Geocenter and scale and their associated rates respectively. In addition, the comparison of ITRF to the VLBI-only based TRF can be used for the scale validation. However, till now there is not any specified methodology for the total assessment (in terms of origin, orientation and scale respectively) of the temporal evolution and GTRFs associated accuracy. We present a new alternative diagnostic tool for the assessment of GTRFs temporal evolution based on the well-known time-dependent Helmert type transformation formula (three shifts, three rotations and scale rates respectively). The advantage of the new methodology relies on the fact that it uses the full velocity field of the TRF and therefore all points not just the ones common to different techniques. It also examines simultaneously rates of origin, orientation and scale. The methodology is presented and implemented to the two existing GTRFs on the market (ITRF and DTRF which is computed from DGFI) , the results are discussed. The results also allow to compare directly each GTRF dynamic behavior. Furthermore, the correlations of the estimated parameters can also provide useful information to the proposed GTRFs assessment scheme.

  5. Electrosynthesized MIPs for transferrin: Plastibodies or nano-filters?

    Science.gov (United States)

    Zhang, Xiaorong; Yarman, Aysu; Erdossy, Júlia; Katz, Sagie; Zebger, Ingo; Jetzschmann, Katharina J; Altintas, Zeynep; Wollenberger, Ulla; Gyurcsányi, Róbert E; Scheller, Frieder W

    2018-05-15

    Molecularly imprinted polymer (MIP) nanofilms for transferrin (Trf) have been synthesized on gold surfaces by electro-polymerizing the functional monomer scopoletin in the presence of the protein target or around pre-adsorbed Trf. As determined by atomic force microscopy (AFM) the film thickness was comparable with the molecular dimension of the target. The target (re)binding properties of the electro-synthesized MIP films was evaluated by cyclic voltammetry (CV) and square wave voltammetry (SWV) through the target-binding induced permeability changes of the MIP nanofilms to the ferricyanide redox marker, as well as by surface plasmon resonance (SPR) and surface enhanced infrared absorption spectroscopy (SEIRAS) of the immobilized protein molecules. For Trf a linear concentration dependence in the lower micromolar range and an imprinting factor of ~5 was obtained by SWV and SPR. Furthermore, non-target proteins including the iron-free apo-Trf were discriminated by pronounced size and shape specificity. Whilst it is generally assumed that the rebinding of the target or of cross-reacting proteins exclusively takes place at the polymer here we considered also the interaction of the protein molecules with the underlying gold transducers. We demonstrate by SWV that adsorption of proteins suppresses the signal of the redox marker even at the bare gold surface and by SEIRAS that the treatment of the MIP with proteinase K or NaOH only partially removes the target protein. Therefore, we conclude that when interpreting binding of proteins to directly MIP-covered gold electrodes the interactions between the protein and the gold surface should also be considered. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. Heterogeneity in the kinetics of nuclear proteins and trajectories of substructures associated with heterochromatin

    Czech Academy of Sciences Publication Activity Database

    Stixová, Lenka; Bártová, Eva; Matula, Pa.; Daněk, O.; Legartová, Soňa; Kozubek, Stanislav

    2011-01-01

    Roč. 4, č. 5 (2011), s. 1-17 ISSN 1756-8935 R&D Projects: GA MŠk(CZ) LC06027; GA MŠk(CZ) LC535; GA MŠk(CZ) ME 919; GA ČR(CZ) GAP302/10/1022 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : HP1 * BMI1 * TRF1 Subject RIV: BO - Biophysics Impact factor: 4.462, year: 2011

  7. Precise Orbital and Geodetic Parameter Estimation using SLR Observations for ILRS AAC

    Directory of Open Access Journals (Sweden)

    Young-Rok Kim

    2013-12-01

    Full Text Available In this study, we present results of precise orbital geodetic parameter estimation using satellite laser ranging (SLR observations for the International Laser Ranging Service (ILRS associate analysis center (AAC. Using normal point observations of LAGEOS-1, LAGEOS-2, ETALON-1, and ETALON-2 in SLR consolidated laser ranging data format, the NASA/ GSFC GEODYN II and SOLVE software programs were utilized for precise orbit determination (POD and finding solutions of a terrestrial reference frame (TRF and Earth orientation parameters (EOPs. For POD, a weekly-based orbit determination strategy was employed to process SLR observations taken from 20 weeks in 2013. For solutions of TRF and EOPs, loosely constrained scheme was used to integrate POD results of four geodetic SLR satellites. The coordinates of 11 ILRS core sites were determined and daily polar motion and polar motion rates were estimated. The root mean square (RMS value of post-fit residuals was used for orbit quality assessment, and both the stability of TRF and the precision of EOPs by external comparison were analyzed for verification of our solutions. Results of post-fit residuals show that the RMS of the orbits of LAGEOS-1 and LAGEOS-2 are 1.20 and 1.12 cm, and those of ETALON-1 and ETALON-2 are 1.02 and 1.11 cm, respectively. The stability analysis of TRF shows that the mean value of 3D stability of the coordinates of 11 ILRS core sites is 7.0 mm. An external comparison, with respect to International Earth rotation and Reference systems Service (IERS 08 C04 results, shows that standard deviations of polar motion Xp and Yp are 0.754 milliarcseconds (mas and 0.576 mas, respectively. Our results of precise orbital and geodetic parameter estimation are reasonable and help advance research at ILRS AAC.

  8. Thujone-Rich Fraction of Thuja occidentalis Demonstrates Major Anti-Cancer Potentials: Evidences from In Vitro Studies on A375 Cells

    OpenAIRE

    Biswas, Raktim; Mandal, Sushil Kumar; Dutta, Suman; Bhattacharyya, Soumya Sundar; Boujedaini, Naoual; Khuda-Bukhsh, Anisur Rahman

    2011-01-01

    Crude ethanolic extract of Thuja occidentalis (Fam: Cupressaceae) is used as homeopathic mother tincture (TOΦ) to treat various ailments, particularly moles and tumors, and also used in various other systems of traditional medicine. Anti-proliferative and apoptosis-inducing properties of TOΦ and the thujone-rich fraction (TRF) separated from it have been evaluated for their possible anti-cancer potentials in the malignant melanoma cell line A375. On initial trial by S-diphenyltetrazolium brom...

  9. Fasting, circadian rhythms, and time restricted feeding in healthy lifespan

    OpenAIRE

    Longo, Valter D.; Panda, Satchidananda

    2016-01-01

    Feeding in most animals is confined to a defined period, leaving short periods of fasting that coincide with sleep. Fasting enables organisms to enter alternative metabolic phases, which rely less on glucose and more on ketone body-like carbon sources. Both intermittent and periodic fasting result in benefits ranging from prevention to the enhanced treatment of diseases. Similarly, time-restricted feeding (TRF), in which feeding time is restricted to certain hours of the day, allows the daily...

  10. Solanum tuberosum y Solanum commersonii, sobre-expresando el gen ScCBFI, mejoraron su tolerancia a sequía en condiciones de cultivo in vitro

    OpenAIRE

    Pino, María Teresa; Ávila, Andrea; Molina, Andrea; Jeknic, Zoran; Chen, Tony H.H

    2013-01-01

    Cultivated potato crops are sensitive to drought stress, reducing yield and tuber quality when the soil water potential drops to -0.3 MPa. However, drought not only affects plant growth and physiological activity, but this stress also induces biochemical and molecular changes in the cellular gene expression profile. triggering genes that play a direct role in plant protection and gene regulation. The genes involved in regulation include C-repeat Binding Factors/Dehydration responsive element ...

  11. RNA-processing proteins regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA.

    Science.gov (United States)

    Manfrini, Nicola; Trovesi, Camilla; Wery, Maxime; Martina, Marina; Cesena, Daniele; Descrimes, Marc; Morillon, Antonin; d'Adda di Fagagna, Fabrizio; Longhese, Maria Pia

    2015-02-01

    Eukaryotic cells respond to DNA double-strand breaks (DSBs) by activating a checkpoint that depends on the protein kinases Tel1/ATM and Mec1/ATR. Mec1/ATR is activated by RPA-coated single-stranded DNA (ssDNA), which arises upon nucleolytic degradation (resection) of the DSB. Emerging evidences indicate that RNA-processing factors play critical, yet poorly understood, roles in genomic stability. Here, we provide evidence that the Saccharomyces cerevisiae RNA decay factors Xrn1, Rrp6 and Trf4 regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA. The lack of Xrn1 inhibits ssDNA generation at the DSB by preventing the loading of the MRX complex. By contrast, DSB resection is not affected in the absence of Rrp6 or Trf4, but their lack impairs the recruitment of RPA, and therefore of Mec1, to the DSB. Rrp6 and Trf4 inactivation affects neither Rad51/Rad52 association nor DSB repair by homologous recombination (HR), suggesting that full Mec1 activation requires higher amount of RPA-coated ssDNA than HR-mediated repair. Noteworthy, deep transcriptome analyses do not identify common misregulated gene expression that could explain the observed phenotypes. Our results provide a novel link between RNA processing and genome stability. © 2014 The Authors.

  12. Vitamin E-Mediated Modulation of Glutamate Receptor Expression in an Oxidative Stress Model of Neural Cells Derived from Embryonic Stem Cell Cultures

    Directory of Open Access Journals (Sweden)

    Afifah Abd Jalil

    2017-01-01

    Full Text Available Glutamate is the primary excitatory neurotransmitter in the central nervous system. Excessive concentrations of glutamate in the brain can be excitotoxic and cause oxidative stress, which is associated with Alzheimer’s disease. In the present study, the effects of vitamin E in the form of tocotrienol-rich fraction (TRF and alpha-tocopherol (α-TCP in modulating the glutamate receptor and neuron injury markers in an in vitro model of oxidative stress in neural-derived embryonic stem (ES cell cultures were elucidated. A transgenic mouse ES cell line (46C was differentiated into a neural lineage in vitro via induction with retinoic acid. These cells were then subjected to oxidative stress with a significantly high concentration of glutamate. Measurement of reactive oxygen species (ROS was performed after inducing glutamate excitotoxicity, and recovery from this toxicity in response to vitamin E was determined. The gene expression levels of glutamate receptors and neuron-specific enolase were elucidated using real-time PCR. The results reveal that neural cells derived from 46C cells and subjected to oxidative stress exhibit downregulation of NMDA, kainate receptor, and NSE after posttreatment with different concentrations of TRF and α-TCP, a sign of neurorecovery. Treatment of either TRF or α-TCP reduced the levels of ROS in neural cells subjected to glutamate-induced oxidative stress; these results indicated that vitamin E is a potent antioxidant.

  13. Influence of timing and dose of thyroid hormone replacement on mental, psychomotor, and behavioral development in children with congenital hypothyroidism.

    Science.gov (United States)

    Bongers-Schokking, Jacoba J; de Muinck Keizer-Schrama, Sabine M P F

    2005-12-01

    To evaluate the influence of initial and postinitial treatment factors on cognitive, psychomotor, and psychological outcome in schoolchildren with congenital hypothyroidism (CH). We studied 45 patients (19 with severe CH and 26 with mild CH) and 37 control children by correlating initial and postinitial treatment factors (free thyroxine and thyroid-stimulating hormone [TSH] concentrations, and the percentage of overtreatment and undertreatment periods) with the results of neuropsychological tests and behavior (as reported on the Teacher Report Form [TRF]). The global IQ of the children with CH was comparable to that of the controls; visuomotor and verbal scores were lower, and total TRF scores were higher. Ethnic group, previous development, and overtreatment predicted IQ and verbal scores, with higher scores seen for the overtreated patients than for the control children and those patients who had not been overtreated. As initial treatment was less satisfactory, total TRF scores were higher. Our study suggests that initial and postinitial suboptimal treatment of CH leads to abnormalities in IQ and specific fields. Overtreatment may advance cognitive development in 5-1/2- to 7-year-olds. Suboptimal initial treatment may lead to behavioral problems. We recommend that TSH concentrations be maintained within the normal range in patients with CH.

  14. Determining Tocotrienol Rich Fraction Effects on Laying, Mortality and Egg Quality Parameters

    Directory of Open Access Journals (Sweden)

    Mardhati, M.

    2016-12-01

    Full Text Available The public interest and awareness in healthier lifestyle has increased the demand of functional food, nutraceuticals and designer foods, which has resulted in the creation of new foods in marketplace, for instance designer and specialty eggs. A feeding trial was carried out to evaluate the laying performance of H&N layer chickens fed diets supplemented with tocotrienol rich fraction (TRF, as well as to quantify tocopherol and tocotrienol accumulation in eggs and their effect on egg quality. A total of 300 H&N laying hens (17-wk old were randomly allocated to 50 battery cages with 6 birds per cage. The birds were assigned equally to 6 different treatment diets; commercial diet (F1, corn-soy based diet (F2, corn-soy based diet added with 25ppm (F3, 50ppm (F4, 75ppm (F5 and 100ppm (F6 of tocotrienol rich fraction (TRF. Egg production and egg quality measurements were not significantly different (P>0.05 between treatments. Egg yolk from hens fed TRF supplemented diets contained more tocotrienol compared to those fed corn-soy based diet (P0.05. As feeding time progressed, the tocotrienol content tended to accumulate in the egg yolks (P<0.05.

  15. Effectiveness of Behavioral Parent Therapy in Preschool Children With Attention-Deficit Hyperactivity Disorder

    Directory of Open Access Journals (Sweden)

    Huei-Lin Huang

    2009-07-01

    Full Text Available The purpose of this study was to assess the effectiveness of a behavioral parent therapy (BPT program in children with attention-deficit hyperactivity disorder (ADHD using multidimensional evaluations, the Child Behavior Checklist (CBCL and the Teacher Report Form (TRF. Between 2001 and 2005, the parents of 21 preschool children with ADHD were divided into six groups and participated in a series of 11 BPT sessions. Before and after BPT, the parents completed the CBCL, and the teachers completed the TRF. The behavioral and emotional problems of the children showed improvement after the BPT sessions, specifically for the following categories: internalizing problems, anxious/depressed syndromes, somatic complaints, externalizing problems, rule-breaking behaviors, aggressive behaviors, social problems, thought problems, and attention problems. In the DSM-oriented scale of the CBCL, affective problems, anxiety problems, somatic problems, ADHD problems, oppositional defiant disorder problems, and conduct disorder problems showed significant improvements. On the DSM scale of the TRF, Inattention syndrome improved significantly after the BPT sessions, while other syndromes showed non-significant changes. We conclude that the BPT program significantly improved the children's behavioral problems at home and inattention problems at school.

  16. Higgs Amplitudes from N=4 Supersymmetric Yang-Mills Theory.

    Science.gov (United States)

    Brandhuber, Andreas; Kostacińska, Martyna; Penante, Brenda; Travaglini, Gabriele

    2017-10-20

    Higgs plus multigluon amplitudes in QCD can be computed in an effective Lagrangian description. In the infinite top-mass limit, an amplitude with a Higgs boson and n gluons is computed by the form factor of the operator TrF^{2}. Up to two loops and for three gluons, its maximally transcendental part is captured entirely by the form factor of the protected stress tensor multiplet operator T_{2} in N=4 supersymmetric Yang-Mills theory. The next order correction involves the calculation of the form factor of the higher-dimensional, trilinear operator TrF^{3}. We present explicit results at two loops for three gluons, including the subleading transcendental terms derived from a particular descendant of the Konishi operator that contains TrF^{3}. These are expressed in terms of a few universal building blocks already identified in earlier calculations. We show that the maximally transcendental part of this quantity, computed in nonsupersymmetric Yang-Mills theory, is identical to the form factor of another protected operator, T_{3}, in the maximally supersymmetric theory. Our results suggest that the maximally transcendental part of Higgs amplitudes in QCD can be entirely computed through N=4 super Yang-Mills theory.

  17. Reliability and validity of teacher-rated symptoms of oppositional defiant disorder and conduct disorder in a clinical sample.

    Science.gov (United States)

    Ise, Elena; Görtz-Dorten, Anja; Döpfner, Manfred

    2014-01-01

    It is recommended to use information from multiple informants when making diagnostic decisions concerning oppositional defiant disorder (ODD) and conduct disorder (CD). The purpose of this study was to investigate the reliability and validity of teacher-rated symptoms of ODD and CD in a clinical sample. The sample comprised 421 children (84% boys; 6-17 years) diagnosed with ODD, CD, and/or attention deficit hyperactivity disorder (ADHD). Teachers completed a standardized ODD/CD symptom rating scale and the Teacher Report Form (TRF). The reliability (internal consistency) of the symptom rating scale was high (α = 0.90). Convergent and divergent validity were demonstrated by substantial correlations with similar TRF syndrome scales and low-to-moderate correlations with dissimilar TRF scales. Discriminant validity was shown by the ability of the symptom rating scale to differentiate between children with ODD/CD and those with ADHD. Factorial validity was demonstrated by principal component analysis, which produced a two-factor solution that is largely consistent with the two-dimensional model of ODD and CD proposed by the Diagnostic and Statistical Manual of Mental Disorders (DSM)-IV-TR, although some CD symptoms representing aggressive behavior loaded on the ODD dimension. These findings suggest that DSM-IV-TR-based teacher rating scales are useful instruments for assessing disruptive behavior problems in children and adolescents.

  18. Higgs amplitudes from supersymmetric form factors Part I: $\\mathcal{N}=4$ super Yang-Mills arXiv

    CERN Document Server

    Brandhuber, Andreas; Penante, Brenda; Travaglini, Gabriele

    In the large top-mass limit, Higgs plus multi-gluon amplitudes in QCD can be computed using an effective field theory. This approach turns the computation of such amplitudes into that of form factors of operators of increasing classical dimension. In this paper we focus on the first finite top-mass correction, arising from the operator ${\\rm Tr}(F^3)$, up to two loops and three gluons. Setting up the calculation in the maximally supersymmetric theory requires identification of an appropriate supersymmetric completion of ${\\rm Tr}(F^3)$, which we recognise as a descendant of the Konishi operator. We provide detailed computations for both this operator and the component operator ${\\rm Tr}(F^3)$, preparing the ground for the calculation in $\\mathcal{N}<4$, to be detailed in a companion paper. Our results for both operators are expressed in terms of a few universal functions of transcendental degree four and below, some of which have appeared in other contexts, hinting at universality of such quantities. An im...

  19. Behaviour problems in children with dyslexia.

    Science.gov (United States)

    Heiervang, E; Stevenson, J; Lund, A; Hugdahl, K

    2001-01-01

    The association between behaviour problems and dyslexia was assessed in a population sample of 10- to 12-year-old children. Twenty-five dyslexic children and a matched control group were recruited through a screening in primary schools in the city of Bergen, Norway. For the assessment of behaviour problems the Child Behavior Checklist (CBCL), Teacher Self Report (TRF), and Youth Self Report (YSR) were filled out by parents, teachers, and children, respectively. Information on health and developmental factors were obtained from parents on a separate questionnaire designed for the study. The dyslexic group had significantly more behaviour problems than the control group according to both the CBCL and the TRF. On the YSR there was no significant difference between the groups. Dyslexic children had higher CBCL and TRF scores on the Total Behaviour Problem scale, the Internalizing and Externalizing subdomains, and the Attention problem subscale. The groups differed in social background, prenatal risk factors, birth weight, preschool language problems, and IQ, but these variables showed no relationship to the level of behaviour problems in the present sample. We conclude that pre-adolescent dyslexic children show a wide range of behaviour problems that cannot be attributed to social or developmental background variables.

  20. Quantitative Time-Resolved Fluorescence Imaging of Androgen Receptor and Prostate-Specific Antigen in Prostate Tissue Sections.

    Science.gov (United States)

    Krzyzanowska, Agnieszka; Lippolis, Giuseppe; Helczynski, Leszek; Anand, Aseem; Peltola, Mari; Pettersson, Kim; Lilja, Hans; Bjartell, Anders

    2016-05-01

    Androgen receptor (AR) and prostate-specific antigen (PSA) are expressed in the prostate and are involved in prostate cancer (PCa). The aim of this study was to develop reliable protocols for reproducible quantification of AR and PSA in benign and malignant prostate tissue using time-resolved fluorescence (TRF) imaging techniques. AR and PSA were detected with TRF in tissue microarrays from 91 PCa patients. p63/ alpha-methylacyl-CoA racemase (AMACR) staining on consecutive sections was used to categorize tissue areas as benign or cancerous. Automated image analysis was used to quantify staining intensity. AR intensity was significantly higher in AMACR+ and lower in AMACR- cancer areas as compared with benign epithelium. The PSA intensity was significantly lower in cancer areas, particularly in AMACR- glands. The AR/PSA ratio varied significantly in the AMACR+ tumor cells as compared with benign glands. There was a trend of more rapid disease progression in patients with higher AR/PSA ratios in the AMACR- areas. This study demonstrates the feasibility of developing reproducible protocols for TRF imaging and automated image analysis to study the expression of AR and PSA in benign and malignant prostate. It also highlighted the differences in AR and PSA protein expression within AMACR- and AMACR+ cancer regions. © 2016 The Histochemical Society.

  1. Post-PCR detection of nucleic acids using metalloporphyrin labels and time-resolved fluorescence

    International Nuclear Information System (INIS)

    O'Shea, Desmond J.; O'Sullivan, Paul J.; Ponomarev, Gelii V.; Papkovsky, Dmitri B.

    2005-01-01

    Phosphorescent platinum(II)-coproporphyrin label (PtCP) was evaluated in post-PCR detection of nucleic acids by time-resolved fluorescence (TR-F) using three common formats. PtCP-labelled oligonucleotide primers and PtCP-dUTP were incorporated in a PCR to produce labelled amplified target -173 or 305 bp DNA. Alternatively, aminoallyl-dUTP was incorporated in a PCR and the product was subsequently labelled with PtCP. The resulting PCR mixtures containing labelled dsDNA were separated on 1.5% agarose gels and then analysed by ethidium bromide staining and by direct detection of PtCP label on a commercial TR-F plate reader Victor 2 (Perkin Elmer Life Sciences) used in scanning mode. In all cases label incorporation and high yields of amplified DNA were observed. Direct TR-F detection of PtCP-labelled DNA from a gel provided high sensitivity and signal to noise ratio, with limits of detection in the range of 9-22 pg for all three formats. The sensitivity achieved with PtCP label was considerably better than that achieved with ethidium bromide staining (∼1 ng of dsDNA) or with conventional fluorescent label FITC. Neither the FITC label nor ethidium bromide staining interfered with PtCP detection, thus allowing multiplexed detection

  2. Karyotypic characterization of Trachemys dorbigni (Testudines: Emydidae) and Chelonoidis (Geochelone) donosobarrosi (Testudines: Testudinidae), two species of Cryptodiran turtles from Argentina.

    Science.gov (United States)

    Martinez, Pablo A; Boeris, Juan M; Sánchez, Julieta; Pastori, María C; Bolzán, Alejandro D; Ledesma, Mario A

    2009-12-01

    We describe for the first time the karyotypes of two species of Cryptodiran turtles from Argentina, namely, Trachemys dorbigni (Emydidae) and Chelonoidis (Geochelone) donosobarrosi (Testudinidae). The karyotype of T. dorbigni (2n = 50) consists of 13 pairs of macrochromosomes and 12 pairs of microchromosomes, whereas the karyotype of C. donosobarrosi (2n = 52) consists of 11 pairs of macrochromosomes and 15 pairs of microchromosomes. Fluorescence in situ hybridization (FISH) with a (TTAGGG)n telomeric probe showed that the chromosomes of these species have four telomeric signals, two at each end, indicating that none of the chromosomes of T. dorbigni and C. donosobarrosi are telocentric. The fact that no interstitial telomeric signals were observed after FISH, suggests that interstitial telomeric sequences did not have a major role in the chromosomal evolution of these species. Additional data will be needed to elucidate if interstitial telomeric sequences have a major role in the karyotypic evolution of Testudines.

  3. Karyotypic variation of Glanidium ribeiroi Haseman, 1911 (Siluriformes, Auchenipteridae along the Iguazu river basin

    Directory of Open Access Journals (Sweden)

    R. L. Lui

    Full Text Available Abstract The Iguazu river is a tributary of the left margin of the Paraná river, isolated from this basin about 22 million years ago with the appearance of the Iguazu Falls. The Iguazu river is characterized by high endemism due to two factors: its rugged topography and the old isolation caused by formation of the Iguazu Falls. This study analyzed cytogenetically a population of Glanidium ribeiroi collected in a region at the final stretch of this basin, by Giemsa staining, C-banding, impregnation by silver nitrate, and FISH with probes of 5S rDNA, 18S rDNA, telomeric sequence [TTAGGG]n, and [GATA]n repeats. The diploid number was equal to 58 chromosomes. The heterochromatin was present in the terminal region of almost all chromosomes. The Ag-NORs were simple and presented interstitially on the short arm of the submetacentric pair 14, which was confirmed by FISH with 18S rDNA probe. The 5S rDNA-FISH marked only the submetacentric pair 16 on the long arm in interstitial position. The FISH with [TTAGGG]n probe presented all telomeres labeled as expected, with an absence of Interstitial Telomeric Sequence (ITS. The repetitive [GATA]n sequence was dispersed throughout the genome, with preferential location in the terminal region of all chromosomes. The data obtained are discussed herein with other species of Auchenipteridae, and other previously analyzed populations of G. ribeiroi from the Iguazu river, verifying differences among these populations, which should be mainly related to the rugged topography of this basin.

  4. A Terrestrial Reference Frame realised on the observation level using a GPS-LEO satellite constellation

    Science.gov (United States)

    Koenig, Daniel

    2018-02-01

    Applying a one-step integrated process, i.e. by simultaneously processing all data and determining all satellite orbits involved, a Terrestrial Reference Frame (TRF) consisting of a geometric as well as a dynamic part has been determined at the observation level using the EPOS-OC software of Deutsches GeoForschungsZentrum. The satellite systems involved comprise the Global Positioning System (GPS) as well as the twin GRACE spacecrafts. Applying a novel approach, the inherent datum defect has been overcome empirically. In order not to rely on theoretical assumptions this is done by carrying out the TRF estimation based on simulated observations and using the associated satellite orbits as background truth. The datum defect is identified here as the total of all three translations as well as the rotation about the z-axis of the ground station network leading to a rank-deficient estimation problem. To rectify this singularity, datum constraints comprising no-net translation (NNT) conditions in x, y, and z as well as a no-net rotation (NNR) condition about the z-axis are imposed. Thus minimally constrained, the TRF solution covers a time span of roughly a year with daily resolution. For the geometric part the focus is put on Helmert transformations between the a priori and the estimated sets of ground station positions, and the dynamic part is represented by gravity field coefficients of degree one and two. The results of a reference solution reveal the TRF parameters to be estimated reliably with high precision. Moreover, carrying out a comparable two-step approach using the same data and models leads to parameters and observational residuals of worse quality. A validation w.r.t. external sources shows the dynamic origin to coincide at a level of 5 mm or better in x and y, and mostly better than 15 mm in z. Comparing the derived GPS orbits to IGS final orbits as well as analysing the SLR residuals for the GRACE satellites reveals an orbit quality on the few cm level

  5. Study on computer-aided diagnosis of hepatic MR imaging and mammography

    International Nuclear Information System (INIS)

    Zhang Xuejun

    2005-01-01

    It is well known that the liver is an organ easily attacked by diseases. The purpose of this study is to develop a computer-aided diagnosis (CAD) scheme for helping radiologists to differentiate hepatic diseases more efficiently. Our software named LIVERANN integrated the magnetic resonance (MR) imaging findings with different pulse sequences to classify the five categories of hepatic diseases by using the artificial neural network (ANN) method. The intensity and homogeneity within the region of interest (ROI) delineated by a radiologist were automatically calculated to obtain numerical data by the program for input signals to the ANN. Outputs were the five pathological categories of hepatic diseases (hepatic cyst, hepatocellular carcinoma, dysplasia in cirrhosis, cavernous hemangioma, and metastasis). The experiment demonstrated a testing accuracy of 93% from 80 patients. In order to differentiate the cirrhosis from normal liver, the volume ratio of left to whole (LTW) was proposed to quantify the degree of cirrhosis by three-dimensional (3D) volume analysis. The liver region was firstly extracted from computed tomography (CT) or MR slices based on edge detection algorithms, and then separated into left lobe and right lobe by the hepatic umbilical fissure. The volume ratio of these two parts showed that the LTW ratio in the liver was significantly improved in the differentiation performance, with (25.6%±4.3%) in cirrhosis versus the normal liver (16.4%±5.4%). In addition, the application of the ANN method for detecting clustered microcalcifications in masses on mammograms was described here as well. A new structural ANN, so-called a shift-invariant artificial neural network (SIANN), was integrated with our triple-ring filter (TRF) method in our CAD system. As the result, the sensitivity of detecting clusters was improved from 90% by our previous TRF method to 95% by using both SIANN and TRF

  6. Effects of intermittent fasting on glucose and lipid metabolism.

    Science.gov (United States)

    Antoni, Rona; Johnston, Kelly L; Collins, Adam L; Robertson, M Denise

    2017-08-01

    Two intermittent fasting variants, intermittent energy restriction (IER) and time-restricted feeding (TRF), have received considerable interest as strategies for weight-management and/or improving metabolic health. With these strategies, the pattern of energy restriction and/or timing of food intake are altered so that individuals undergo frequently repeated periods of fasting. This review provides a commentary on the rodent and human literature, specifically focusing on the effects of IER and TRF on glucose and lipid metabolism. For IER, there is a growing evidence demonstrating its benefits on glucose and lipid homeostasis in the short-to-medium term; however, more long-term safety studies are required. Whilst the metabolic benefits of TRF appear quite profound in rodents, findings from the few human studies have been mixed. There is some suggestion that the metabolic changes elicited by these approaches can occur in the absence of energy restriction, and in the context of IER, may be distinct from those observed following similar weight-loss achieved via modest continuous energy restriction. Mechanistically, the frequently repeated prolonged fasting intervals may favour preferential reduction of ectopic fat, beneficially modulate aspects of adipose tissue physiology/morphology, and may also impinge on circadian clock regulation. However, mechanistic evidence is largely limited to findings from rodent studies, thus necessitating focused human studies, which also incorporate more dynamic assessments of glucose and lipid metabolism. Ultimately, much remains to be learned about intermittent fasting (in its various forms); however, the findings to date serve to highlight promising avenues for future research.

  7. Laminar burning velocities at elevated pressures for gasoline and gasoline surrogates associated with RON

    KAUST Repository

    Mannaa, Ossama

    2015-06-01

    The development and validation of a new gasoline surrogate using laminar flame speed as a target parameter is presented. Laminar burning velocities were measured using a constant-volume spherical vessel with ignition at the center of the vessel. Tested fuels included iso-octane, n-heptane, toluene, various mixtures of primary reference fuels (PRFs) and toluene reference fuels (TRFs) and three gasoline fuels of 70, 85 and 95 RON (FACE J, C and F) at the initial temperature of 358K and pressures up to 0.6MPa in the equivalence ratio ranging from 0.8 to 1.6. Normalized laminar burning velocity data were mapped into a tri-component mixture space at different experimental conditions to allocate different gasoline surrogates for different gasoline fuels, having RON of 70, 85 and 95. The surrogates of TRF-70-4 (17.94% iso-C8H18 +42.06% n-C7H16 +40% C7H8), TRF-85-1 (77.4% iso-C8H18 +17.6% n-C7H16 +5% C7H8), and TRF-95-1 (88.47% iso-C8H18 +6.53% n-C7H16 +5% C7H8) of RON 70, 85 and 95, respectively, are shown to successfully emulate the burning rate characteristics of the gasoline fuels associated with these RONs under the various experimental conditions investigated. An empirical correlation was derived to obtain laminar burning velocities at pressures that are experimentally unattainable as high as 3.0MPa. Laminar burning velocities were comparable to the simulated values for lean and stoichiometric flames but they were relatively higher than the simulated values for rich flames. A flame instability assessment was conducted by determining Markstein length, critical Pecklet number, and critical Karlovitz number at the onset of flame instability.

  8. Standard deviation of carotid young's modulus and presence or absence of plaque improves prediction of coronary heart disease risk.

    Science.gov (United States)

    Niu, Lili; Zhang, Yanling; Qian, Ming; Xiao, Yang; Meng, Long; Zheng, Rongqin; Zheng, Hairong

    2017-11-01

    The stiffness of large arteries and the presence or absence of plaque are associated with coronary heart disease (CHD). Because arterial walls are biologically heterogeneous, the standard deviation of Young's modulus (YM-std) of the large arteries may better predict coronary atherosclerosis. However, the role of YM-std in the occurrence of coronary events has not been addressed so far. Therefore, this study investigated whether the carotid YM-std and the presence or absence of plaque improved CHD risk prediction. One hundred and three patients with CHD (age 66 ± 11 years) and 107 patients at high risk of atherosclerosis (age 61 ± 7 years) were recruited. Carotid YM was measured by the vessel texture matching method, and YM-std was calculated. Carotid intima-media thickness was measured by the MyLab 90 ultrasound Platform employed dedicated software RF-tracking technology. In logistic regression analysis, YM-std (OR = 1·010; 95% CI = 1·003-1·016), carotid plaque (OR = 16·759; 95% CI = 3·719-75·533) and YM-std plus plaque (OR = 0·989; 95% CI = 0·981-0·997) were independent predictors of CHD. The traditional risk factors (TRF) plus YM-std plus plaque model showed a significant improvement in area under the receiver-operating characteristic curve (AUC), which increased from 0·717 (TRF only) to 0·777 (95% CI for the difference in adjusted AUC: 0·010-0·110). Carotid YM-std is a powerful independent predictor of CHD. Adding plaque and YM-std to TRF improves CHD risk prediction. © 2016 Scandinavian Society of Clinical Physiology and Nuclear Medicine. Published by John Wiley & Sons Ltd.

  9. Age, sex, and telomere dynamics in a long-lived seabird with male-biased parental care.

    Directory of Open Access Journals (Sweden)

    Rebecca C Young

    Full Text Available The examination of telomere dynamics is a recent technique in ecology for assessing physiological state and age-related traits from individuals of unknown age. Telomeres shorten with age in most species and are expected to reflect physiological state, reproductive investment, and chronological age. Loss of telomere length is used as an indicator of biological aging, as this detrimental deterioration is associated with lowered survival. Lifespan dimorphism and more rapid senescence in the larger, shorter-lived sex are predicted in species with sexual size dimorphism, however, little is known about the effects of behavioral dimorphism on senescence and life history traits in species with sexual monomorphism. Here we compare telomere dynamics of thick-billed murres (Urialomvia, a species with male-biased parental care, in two ways: 1 cross-sectionally in birds of known-age (0-28 years from one colony and 2 longitudinally in birds from four colonies. Telomere dynamics are compared using three measures: the telomere restriction fragment (TRF, a lower window of TRF (TOE, and qPCR. All showed age-related shortening of telomeres, but the TRF measure also indicated that adult female murres have shorter telomere length than adult males, consistent with sex-specific patterns of ageing. Adult males had longer telomeres than adult females on all colonies examined, but chick telomere length did not differ by sex. Additionally, inter-annual telomere changes may be related to environmental conditions; birds from a potentially low quality colony lost telomeres, while those at more hospitable colonies maintained telomere length. We conclude that sex-specific patterns of telomere loss exist in the sexually monomorphic thick-billed murre but are likely to occur between fledging and recruitment. Longer telomeres in males may be related to their homogamous sex chromosomes (ZZ or to selection for longer life in the care-giving sex. Environmental conditions appeared to

  10. Differences in gene expression of cells growing in conventional 2D versus 3D cell culture

    International Nuclear Information System (INIS)

    Zschenker, Oliver; Cordes, Nils; Streichert, Thomas

    2009-01-01

    Full text: Telomeres are DNA protein complexes on the ends of chromosomes that distinguish the ends of chromosomes from double strand breaks and prevent degradation or fusion by nonhomologous end-joining. The loss of telomeres is associated with a loss of heterochromatic features leading to a less compact chromatin structure which allows e.g. DNA repair proteins to get better access to the site of the DNA damage and facilitate chromosome fusions. Telomerase is an enzyme that can counteract the loss of telomeres by adding telomeric repeats on the ends of chromosomes. Since telomerase is active in most tumor cells, telomerase is suggested to be the reason for the unlimited number of cell divisions of cancer cells. TRF2 is one of the most important proteins of the Shelterin complex protecting the telomeres from shortening by inhibiting ATM which is up-stream of the DNA repair mechanisms. Thus, we are concentrating on TRF2 and telomerase to investigate the differences in DNA repair in telomeric (heterochromatic) versus euchromatic regions. Human cancer cells with differences in status of p53 and telomerase like A549, UT-SCC15 and FaDu cells are used. Without any treatment, FaDu cells express high levels of telomerase and TRF2 in conventional 2D cell culture which is in contrast to e.g. A549. We found that telomerase is even higher expressed in 3D than in 2D cell culture. To connect telomere associated processes to both repair of radiogenic DNA damage/lesions and to cell-extracellular matrix interactions, we performed whole genome microarray analysis. By comparing the differential expression of genes associated with these three cell functions, we intend to yield new molecular insight into radiotherapy relevant tumor characteristics, particularly radioresistance and DNA damage response network processing. (author)

  11. Genetic moderation of child maltreatment effects on depression and internalizing symptoms by 5-HTTLPR, BDNF, NET, and CRHR1 genes in African-American children

    Science.gov (United States)

    Cicchetti, Dante; Rogosch, Fred A.

    2014-01-01

    Genetic moderation of the effects of child maltreatment on depression and internalizing symptoms was investigated in a sample of low-income maltreated and nonmaltreated African-American children (N = 1,096). Lifetime child maltreatment experiences were independently coded from Child Protective Services records and maternal report. Child depression and internalizing problems were assessed in the context of a summer research camp by self-report (Children’s Depression Inventory, CDI) and adult counselor-report (Teacher Report Form, TRF). DNA was obtained from buccal cell or saliva samples and genotyped for polymorphisms of the following genes: 5-HTTLPR, BDNF, NET, and CRHR1. ANCOVAs with age and gender as covariates were conducted, with maltreatment status and respective polymorphism as main effects and their GxE interactions. Maltreatment consistently was associated with higher CDI and TRF symptoms. Results for child self-report symptoms indicated a GxE interaction for BDNF and maltreatment. Additionally, BDNF and tri-allelic 5-HTTLPR interacted with child maltreatment in a GxGxE interaction. Analyses for counselor-report of child anxiety/depression symptoms on the TRF indicated moderation of child maltreatment effects by tri-allelic 5-HTTLPR. These effects were elaborated based on variation in developmental timing of maltreatment experiences. NET was found to further moderate the GxE interaction of 5-HTTLPR and maltreatment status revealing a GxGxE interaction. This GxGxE was extended by consideration of variation in maltreatment subtype experiences. Finally, GxGxE effects were observed for the co-action of BDNF and the CRHR1 haplotype. The findings illustrate the variable influence of specific genotypes in GxE interactions based on variation in maltreatment experiences and the importance of a multi-genic approach for understanding influences on depression and internalizing symptoms among African-American children. PMID:25422957

  12. Bacterial communities associated with the digestive tract of the predatory ground beetle, Poecilus chalcites, and their response to laboratory rearing and antibiotic treatment

    Energy Technology Data Exchange (ETDEWEB)

    Michael Lehman

    2008-06-01

    Ground beetles such as Poecilus chalcites (Coleoptera: Carabidae) are beneficial insects in agricultural systems where they contribute to the control of insect and weed pests. We assessed the complexity of bacterial communities occurring in the digestive tracts of field-collected P. chalcites using terminal restriction fragment length polymorphism analyses of polymerase chain reaction-amplified 16S rRNA genes. Bacterial identification was performed by the construction of 16S rRNA gene clone libraries and sequence analysis. Intestinal bacteria in field-collected beetles were then compared to those from groups of beetles that were reared in the lab on an artificial diet with and without antibiotics. Direct cell counts estimated 1.5 × 10S bacteria per milliliter of gut. The digestive tract of field-collected P. chalcites produced an average of 4.8 terminal restriction fragments (tRF) for each beetle. The most abundant clones were affiliated with the genus Lactobacillus, followed by the taxa Enterobacteriaceae, Clostridia, and Bacteriodetes. The majority of the sequences recovered were closely related to those reported from other insect gastrointestinal tracts. Lab-reared beetles produced fewer tRF, an average of 3.1 per beetle, and a reduced number of taxa with a higher number of clones from the family Enterobacteriaceae compared to the field-collected beetles. Antibiotic treatment significantly (p < 0.05) reduced the number of tRF per beetle and selected for a less diverse set of bacterial taxa. We conclude that the digestive tract of P. chalcites is colonized by a simple community of bacteria that possess autochthonous characteristics. Laboratory-reared beetles harbored the most common bacteria found in field-collected beetles, and these bacterial communities may be manipulated in the laboratory with the addition of antibiotics to the diet to allow study of functional roles.

  13. Troca rápida de ferramentas: proposta metodológica e estudo de caso Rapid exchange of tools: method steps and case study

    Directory of Open Access Journals (Sweden)

    Flávio Sanson Fogliatto

    2003-08-01

    Full Text Available A troca rápida de ferramentas (TRF tem por objetivo reduzir o tempo de preparação (ou setup de equipamentos, minimizando períodos não-produtivos no chão-de-fábrica. Como conseqüência, é possível a redução do tamanho dos lotes de produção na manufatura. A TRF fundamenta-se em técnicas que enfatizam o trabalho cooperativo em equipe e a proposição de formas criativas de melhoria de processos. Este artigo apresenta uma proposta metodológica para a TRF, constituída dos seguintes passos: definição do projeto, planejamento das atividades, treinamento da equipe de implantação, implantação propriamente dita, acompanhamento e consolidação. Um estudo de caso desenvolvido na indústria moveleira ilustra a metodologia proposta.Single-minute exchange of die (SMED and alternative methodologies aim at reducing the time needed for equipment setup, minimizing non-productive periods in the shop floor. That in turns enables production of increasingly smaller lot sizes. SMED is based on a number of improvement techniques that emphasize cooperative teamwork and the proposal of creative ways to improve existing processes. This paper presents a methodology for rapid exchange of die, comprised of the following steps: project definition, planning of activities, personnel training in SMED, implementation per se and follow-up and consolidation. A case study from the furniture industry illustrates the methodology steps.

  14. Regenerative Potential of D-δ-Tocotrienol Rich Fraction on Crushed Skeletal Muscle of Diabetic Rats

    Directory of Open Access Journals (Sweden)

    Bijo Elsy

    2017-06-01

    Full Text Available Background: Delayed muscle growth and regeneration of skeletal muscle in diabetics is believed to be due to diabetic myopathy because of alteration in the skeletal muscle homeostatis. Since vitamin E is a natural antioxidant and is also important for the integrity of sarcolemma, the present study was designed to explore the muscle regenerative potency of d-δ-tocotrienol-rich fraction (d-δ-TRF on crushed skeletal muscle in healthy and diabetic rats. Materials and Methods: Diabetes was induced through single subcutaneous injection of alloxan (100 mg/kg. Twenty-four albino rats were divided into four groups; healthy control, diabetic control, healthy treated, and diabetic treated. Treated groups received injections orally, daily (200 mg/kg for 3 weeks. A horizontal skin incision was made on the shaved right mid-thigh region, by splitting the fascia between gluteus maximus and tensor fascia lata, and gluteus maximus was crushed with Kocher’s forceps. Skin wound was closed with an absorbable suture. The crushed muscle changes were studied by assessing the histopathological features, histomorphological measurements, and biochemical analyses on 3rd week following induction of injury. One-way “ANOVA” followed by Tukey’s test and Student t-test were used for statistical analysis of data. Results: Results obtained through various methods indicate that the d-δ-TRF treated groups have controlled glycemic status, improved antioxidant capacity, faster revascularization, re-innervation, regeneration of myofibers, and connective tissue remodeling. Conclusion: It is, therefore, concluded that the d-δ-TRF is a beneficial nutritional adjuvant for skeletal muscles’ structural and functional recovery after crushed injury in both healthy and diabetics. [J Interdiscip Histopathol 2017; 5(2.000: 36-42

  15. Design of peptide substrates for nanosecond time-resolved fluorescence assays of proteases: 2,3-diazabicyclo[2.2.2]oct-2-ene as a noninvasive fluorophore.

    Science.gov (United States)

    Hennig, Andreas; Florea, Mara; Roth, Doris; Enderle, Thilo; Nau, Werner M

    2007-01-15

    Fluorescence protease assays were investigated with peptide substrates containing a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) as a fluorescent amino acid. The special characteristic of the fluorophore Dbo is its exceedingly long fluorescence lifetime (ca. 300 ns in water under air), which allows the use of nanosecond time-resolved fluorescence (Nano-TRF) detection to efficiently suppress shorter-lived background emission. In addition, the natural amino acids tryptophan and tyrosine can be employed as intramolecular fluorescence quenchers, which facilitates substrate design. Fourteen synthetic peptide substrates (composed of 2-19 amino acids) and five enzymes (trypsin, pepsin, carboxypeptidase A, leucine aminopeptidase, and chymotrypsin) were investigated and, in all 28 examined combinations, enzymatic activity was detected by monitoring the increase in steady state fluorescence with time and determining the reaction rates as kcat/Km values, which ranged from 0.2 to 80x10(6) M-1 min-1. The results suggest an excellent compatibility of the very small and hydrophilic fluorescent probe Dbo with solid-phase peptide synthesis and the investigated proteases. For all 14 peptides the fluorescence lifetimes before and after enzymatic cleavage were measured and Nano-TRF measurements were performed in 384-well microplates. The fluorescence lifetimes of the different peptides provide the basis for the rational design of Dbo-based fluorescent substrates for protease assays. Measurements in Nano-TRF mode revealed, in addition to efficient suppression of background fluorescence, an increased differentiation between cleaved and uncleaved substrate. The Dbo-based assays can be adapted for high-throughput screening.

  16. The influence of n-butanol blending on the ignition delay times of gasoline and its surrogate at high pressures

    KAUST Repository

    Agbro, Edirin

    2016-09-24

    The influence of blending n-butanol at 20% by volume on the ignition delay times for a reference gasoline was studied in a rapid compression machine (RCM) for stoichiometric fuel/air mixtures at 20 bar and 678-858 K. Delay times for the blend lay between those of stoichiometric gasoline and stoichiometric n-butanol across the temperature range studied. At lower temperatures, delays for the blend were however, much closer to those of n-butanol than gasoline despite n-butanol being only 20% of the mixture. Under these conditions n-butanol acted as an octane enhancer over and above what might be expected from a simple linear blending law. The ability of a gasoline surrogate, based on a toluene reference fuel (TRF), to capture the main trends of the gasoline/n-butanol blending behaviour was also tested within the RCM. The 3-component TRF based on a mixture of toluene, n-heptane and iso-octane was able to capture the trends well across the temperature range studied. Simulations of ignition delay times were also performed using a detailed blended n-butanol/TRF mechanism based on the adiabatic core assumption and volume histories from the experimental data. Overall, the model captured the main features of the blending behaviour, although at the lowest temperatures, predicted ignition delays for stoichiometric n-butanol were longer than those observed. A brute-force local sensitivity analysis was performed to evaluate the main chemical processes driving the ignition behaviour of the TRF, n-butanol and blended fuels. The reactions of fuel + OH dominated the sensitivities at lower temperatures, with H abstraction from n-butanol from a and 7 sites being key for both the n-butanol and the blend. At higher temperatures the decomposition of H2O2 and reactions of HO2 and that of formaldehyde with OH became critical, in common with the ignition behaviour of other fiiels. Remaining uncertainties in the rates of these key reactions are discussed. Crown Copyright (C) 2016 Published

  17. Gravity changes in mid-west Greenland from GOCE gravity model and gradient data using ground and airborne gravity

    DEFF Research Database (Denmark)

    Tscherning, Carl Christian; Herceg, Matija; Fredenslund Levinsen, Joanna

    GOCE TRF (terrestrial reference frame) vertical anomalous gradients (Tzz) from two periods have been used to determine gravity anomalies changes in mid-west Greenland, where a large mass-loss has been detected using GRACE (Fig. 1). As additional data were used the GOCE DIR-3 model and ground...... gravity at the coast on solid rock, where no mass loss is expected. The methods of Least-Squares Collocation (LSC) and the Reduced Point Mass (RPM) methods have been used, however only LSC included the ground data....

  18. A teoria das restrições como balizadora das ações visando a troca rápida de ferramentas

    Directory of Open Access Journals (Sweden)

    José Antônio Valle Antunes Jr

    1993-12-01

    Full Text Available O presente artigo propõe a sinergia entre duas técnicas famosas no contexto da gestão industrial: a Troca Rápida de Ferramenta (TRF e a Teoria das Restrições (Theory of Constraints TOC. Para tanto, inicialmente, discute-se a problemática macroeconômica e sua vinculação com o aspecto microeconômico da TRF. Posteriormente, discute-se as principais vantagens potenciais decorrentes daredução dos custos e dos tempos de preparação. Na seqüência, revisa-se o método da TRF, pontuando-se que a abordagem apresenta uma deficiência no que concerne à apresentação de um procedimento de priorização que permita um plano lógico de ataque à questão. Neste ponto do texto, propõe-se a TOC como balizadora das ações visando a Troca Rápida de Ferramentas. Após uma breve revisão teórica da TOC, apresenta-se uma discussão sobre a possível sinergia da mesma com a TRF. Finalmente, apresenta-se um exemplo didático e as conclusões finais do artigo.This paper presents the relationship between two well-known techniques with in lndustrial Management: SMED and the Theory of Constraints (TOC. As a first step, it is discussed the macroeconomics environment and its relationship with microeconomics aspects of the SMED. After that, the main advantages ofcost and set-up time reductions are discussed and the TOC is proposed as away to define priorities to SMED activities. Both, SMED concepts and TOC concepts are presented in this paper. Finally, an example is built up to show the synergy between the two techniques and a conclusion is presented.

  19. Effect of Yunpi Huoxue soup combined chemotherapy on T lymphocyte subsets and nutritional status in patients with advanced gastric cancer

    Directory of Open Access Journals (Sweden)

    Pei Xiang

    2016-08-01

    Full Text Available Objective: To observe the effect of Yunpi Huoxue soup combined with chemotherapy on T lymphocyte subsets and nutritional status in patients with advanced gastric cancer. Methods: A total of 94 cases patients with advanced gastric cancer were randomly divided into the treatment group (49 cases and the control group (45 cases according to the results of the draw. The control group was given chemotherapy, the treatment group was given Yunpi Huoxue soup on the basis of the control group. Treated for 6 weeks, observed the changes of T cell subsets (CD3, CD4, CD8 and CD4/CD8 and nutrition indexes: total protein (TP, albumin (ALB, prealbumin (PA and transferrin (TRF in the two groups. Results: After treatment, CD3, CD4, CD8 and CD4/CD8 in the treatment group were (57.38±4.03, (31.63±4.26, (30.82±3.52 and (1.16±0.20 respectively, there were no significant differences compared with before treatment; After treatment, the levels of CD3, CD4, CD8 and CD4/CD8 in the control group were significantly lower than those before treatment, and the differences were statistically significant; After treatment, the levels of CD3, CD4, CD8 and CD4/CD8 in the treatment group were significant higher than those in the control group after treatment, and the differences were statistically significant. After treatment, TP, ALB, PA and TRF in the treatment group were(54.22±5.93 g/L, (32.47±4.97 g/L, (2.52±0.43 g/L and (1.66±0.40 g/L respectively, there were no significant differences compared with before treatment; After treatment, the levels of TP, ALB, PA and TRF in the control group were significantly lower than those before treatment; After treatment, the levels of TP, ALB, PA and TRF in the treatment group were significant higher than those in the control group after treatment, and the differences were statistically significant. Conclusion: When chemotherapy for patients with advanced gastric cancer, Yunpi Huoxue soup is helpful to maintain the immune function and

  20. Soluble serum Klotho levels in healthy subjects

    DEFF Research Database (Denmark)

    Pedersen, Lise Mariager; Pedersen, Susanne Møller; Brasen, Claus Lohman

    2013-01-01

    Klotho concentrations were determined in 120 healthy adults aged 19-66years. Blood samples were collected, and stored sera were assayed for Klotho according to age and gender. In addition several other clinical and laboratory characteristics were determined in the cohort and compared to the levels...... genders. Concentrations of serum Klotho were independently associated with estimated glomerular filtration rate (eGFR) and body weight using TRF whereas serum Klotho concentrations were associated with age using the ELISA. CONCLUSION: Comparison of two different immunoassays for serum Klotho indicate...

  1. Machine learning models in breast cancer survival prediction.

    Science.gov (United States)

    Montazeri, Mitra; Montazeri, Mohadeseh; Montazeri, Mahdieh; Beigzadeh, Amin

    2016-01-01

    Breast cancer is one of the most common cancers with a high mortality rate among women. With the early diagnosis of breast cancer survival will increase from 56% to more than 86%. Therefore, an accurate and reliable system is necessary for the early diagnosis of this cancer. The proposed model is the combination of rules and different machine learning techniques. Machine learning models can help physicians to reduce the number of false decisions. They try to exploit patterns and relationships among a large number of cases and predict the outcome of a disease using historical cases stored in datasets. The objective of this study is to propose a rule-based classification method with machine learning techniques for the prediction of different types of Breast cancer survival. We use a dataset with eight attributes that include the records of 900 patients in which 876 patients (97.3%) and 24 (2.7%) patients were females and males respectively. Naive Bayes (NB), Trees Random Forest (TRF), 1-Nearest Neighbor (1NN), AdaBoost (AD), Support Vector Machine (SVM), RBF Network (RBFN), and Multilayer Perceptron (MLP) machine learning techniques with 10-cross fold technique were used with the proposed model for the prediction of breast cancer survival. The performance of machine learning techniques were evaluated with accuracy, precision, sensitivity, specificity, and area under ROC curve. Out of 900 patients, 803 patients and 97 patients were alive and dead, respectively. In this study, Trees Random Forest (TRF) technique showed better results in comparison to other techniques (NB, 1NN, AD, SVM and RBFN, MLP). The accuracy, sensitivity and the area under ROC curve of TRF are 96%, 96%, 93%, respectively. However, 1NN machine learning technique provided poor performance (accuracy 91%, sensitivity 91% and area under ROC curve 78%). This study demonstrates that Trees Random Forest model (TRF) which is a rule-based classification model was the best model with the highest level of

  2. Direct comparison of flow-FISH and qPCR as diagnostic tests for telomere length measurement in humans.

    Directory of Open Access Journals (Sweden)

    Fernanda Gutierrez-Rodrigues

    Full Text Available Telomere length measurement is an essential test for the diagnosis of telomeropathies, which are caused by excessive telomere erosion. Commonly used methods are terminal restriction fragment (TRF analysis by Southern blot, fluorescence in situ hybridization coupled with flow cytometry (flow-FISH, and quantitative PCR (qPCR. Although these methods have been used in the clinic, they have not been comprehensively compared. Here, we directly compared the performance of flow-FISH and qPCR to measure leukocytes' telomere length of healthy individuals and patients evaluated for telomeropathies, using TRF as standard. TRF and flow-FISH showed good agreement and correlation in the analysis of healthy subjects (R(2 = 0.60; p<0.0001 and patients (R(2 = 0.51; p<0.0001. In contrast, the comparison between TRF and qPCR yielded modest correlation for the analysis of samples of healthy individuals (R(2 = 0.35; p<0.0001 and low correlation for patients (R(2 = 0.20; p = 0.001; Bland-Altman analysis showed poor agreement between the two methods for both patients and controls. Quantitative PCR and flow-FISH modestly correlated in the analysis of healthy individuals (R(2 = 0.33; p<0.0001 and did not correlate in the comparison of patients' samples (R(2 = 0.1, p = 0.08. Intra-assay coefficient of variation (CV was similar for flow-FISH (10.8 ± 7.1% and qPCR (9.5 ± 7.4%; p = 0.35, but the inter-assay CV was lower for flow-FISH (9.6 ± 7.6% vs. 16 ± 19.5%; p = 0.02. Bland-Altman analysis indicated that flow-FISH was more precise and reproducible than qPCR. Flow-FISH and qPCR were sensitive (both 100% and specific (93% and 89%, respectively to distinguish very short telomeres. However, qPCR sensitivity (40% and specificity (63% to detect telomeres below the tenth percentile were lower compared to flow-FISH (80% sensitivity and 85% specificity. In the clinical setting, flow-FISH was more accurate, reproducible, sensitive, and specific in the measurement of human

  3. An Overview of the Common Fluid Models Used in Fluid-Structure Interactions

    Science.gov (United States)

    1991-08-05

    inverse transform of (63) and us- ing the relation f (t - a) = exp (iwa)f(0) (64) for arbitrary (reasonably behaved) f, with f = C and a = r/c since...lri( r - ) - F -la* exp(ikr)dS’ using (53). The inverse transform of this equation gives 3 rV (Xi, t) = 2Vi c + T r3’fV+(- C (an ))dS’ (72) upon...T, when it exists, has the property rl-Tf= T=rf = f for a generic function f, the Laplace transform and inverse transform being examples of such a T

  4. Telomere shortening in the colonic mucosa of patients with ulcerative colitis.

    Science.gov (United States)

    Kinouchi, Y; Hiwatashi, N; Chida, M; Nagashima, F; Takagi, S; Maekawa, H; Toyota, T

    1998-06-01

    Telomere length in human somatic cells gradually decreases with the number of cell divisions and is regarded as a marker of somatic cell turnover. Mucosal cells of the affected colon show rapid turnover in individuals with active ulcerative colitis (UC). Telomere length was determined by Southern blot analysis of terminal restriction fragments (TRFs) from the colonic mucosa of 17 patients with UC in remission, two of whom showed dysplasia, and 17 control subjects without colitis. For each individual, mean TRF length was compared between rectal mucosa and unaffected cecal mucosa. The mean TRF length of the rectal mucosa was significantly less than that of cecal mucosa in UC patients (7.87 +/- 0.36kb versus 8.77 +/- 0.21 kb; P = 0.0015, Wilcoxon signed rank test), whereas no significant difference was detected in the control subjects. The extent of telomere shortening was 10.6 +/- 3.35% in UC patients, compared with 0.8 +/- 0.64% in noncolitis controls (P = 0.0024, Mann-Whitney U-test). Four UC patients, two of whom had dysplasia, showed telomere shortening of more than 20% in the rectal mucosa. These observations suggest that telomere shortening in the colonic mucosa of individuals with UC may represent the history of mucosal inflammation during disease of long duration, and that it may contribute to aneuploidy in UC.

  5. Chitinase genes revealed and compared in bacterial isolates, DNA extracts and a metagenomic library from a phytopathogen suppressive soil

    Energy Technology Data Exchange (ETDEWEB)

    Hjort, K.; Bergstrom, M.; Adesina, M.F.; Jansson, J.K.; Smalla, K.; Sjoling, S.

    2009-09-01

    Soil that is suppressive to disease caused by fungal pathogens is an interesting source to target for novel chitinases that might be contributing towards disease suppression. In this study we screened for chitinase genes, in a phytopathogen-suppressive soil in three ways: (1) from a metagenomic library constructed from microbial cells extracted from soil, (2) from directly extracted DNA and (3) from bacterial isolates with antifungal and chitinase activities. Terminal-restriction fragment length polymorphism (T-RFLP) of chitinase genes revealed differences in amplified chitinase genes from the metagenomic library and the directly extracted DNA, but approximately 40% of the identified chitinase terminal-restriction fragments (TRFs) were found in both sources. All of the chitinase TRFs from the isolates were matched to TRFs in the directly extracted DNA and the metagenomic library. The most abundant chitinase TRF in the soil DNA and the metagenomic library corresponded to the TRF{sup 103} of the isolate, Streptomyces mutomycini and/or Streptomyces clavifer. There were good matches between T-RFLP profiles of chitinase gene fragments obtained from different sources of DNA. However, there were also differences in both the chitinase and the 16S rRNA gene T-RFLP patterns depending on the source of DNA, emphasizing the lack of complete coverage of the gene diversity by any of the approaches used.

  6. A study on the primary requirement for the safety of the Wolsong tritium removal facility

    International Nuclear Information System (INIS)

    Hwang, K. H.; Lee, K. J.; Jeong, C. W.

    2001-01-01

    Owing to the using a heavy water as a moderator and a coolant in Heavy water reactor, A large mount of tritium is produced due to a reaction of deuterium with neutron in the reactor and some of tritium is released to the environment. In Wolsong, 4 units (CANDU-600 type) Heavy water reactor is in operation. And the generated amount of tritium is increased with the increase of operational year of the Wolsong nuclear reactor. Decommissioning of the Wolsong unit 1 is expected to start at 2013. Before 2013, to reduce the workers internal radiation doses and environmental release of tritium, Tritium Removal Facility (TRF) is required and should be operated. Wolsong TRF (WTRF) is under developing stage by Korea Electric Power Corporation(KEPCO)and scheduled to start operation about 2006. Once the facility begins operation it can be contributed to the greatly reduction of tritium release to the environment and worker's expose. In this situation, study about the safety assessment method and regulatory requirement is essential for safety insurance of WTRF. And this helps the safety acquirement, successful operation and reliance of WTRF

  7. Laminar Flame Speeds of Gasoline Surrogates Measured with the Flat Flame Method

    KAUST Repository

    Liao, Y.-H.

    2016-01-27

    © 2016 American Chemical Society. The adiabatic, laminar flame speeds of gasoline surrogates at atmospheric pressure over a range of equivalence ratios of = 0.8-1.3 and unburned gas temperatures of 298-400 K are measured with the flat flame method, which produces a one-dimensional flat flame free of stretch. Surrogates used in the current work are the primary reference fuels (PRFs, mixtures of n-heptane and isooctane), the toluene reference fuels (TRFs, mixtures of toluene and PRFs), and the ethanol reference fuels (ERFs, mixtures of ethanol and PRFs). In general, there is good agreement between the present work and the literature data for single-component fuel and PRF mixtures. Surrogates of TRF mixtures are found to exhibit comparable flame speeds to a real gasoline, while there is discrepancy observed between isooctane and gasoline. Moreover, the laminar flame speeds of TRF mixtures with similar fractions of n-heptane are found to be insensitive to the quantity of toluene in the mixture. Mixtures of ERFs exhibit comparable flame speeds to those of TRFs with similar mole fractions of n-heptane and isooctane.

  8. Thresholding of auditory cortical representation by background noise

    Science.gov (United States)

    Liang, Feixue; Bai, Lin; Tao, Huizhong W.; Zhang, Li I.; Xiao, Zhongju

    2014-01-01

    It is generally thought that background noise can mask auditory information. However, how the noise specifically transforms neuronal auditory processing in a level-dependent manner remains to be carefully determined. Here, with in vivo loose-patch cell-attached recordings in layer 4 of the rat primary auditory cortex (A1), we systematically examined how continuous wideband noise of different levels affected receptive field properties of individual neurons. We found that the background noise, when above a certain critical/effective level, resulted in an elevation of intensity threshold for tone-evoked responses. This increase of threshold was linearly dependent on the noise intensity above the critical level. As such, the tonal receptive field (TRF) of individual neurons was translated upward as an entirety toward high intensities along the intensity domain. This resulted in preserved preferred characteristic frequency (CF) and the overall shape of TRF, but reduced frequency responding range and an enhanced frequency selectivity for the same stimulus intensity. Such translational effects on intensity threshold were observed in both excitatory and fast-spiking inhibitory neurons, as well as in both monotonic and nonmonotonic (intensity-tuned) A1 neurons. Our results suggest that in a noise background, fundamental auditory representations are modulated through a background level-dependent linear shifting along intensity domain, which is equivalent to reducing stimulus intensity. PMID:25426029

  9. Effects of naloxone opiate blockade on the immunomodulation induced by exercise in rats.

    Science.gov (United States)

    Bouix, O; elMezouini, M; Orsetti, A

    1995-01-01

    The purpose of this study was to examine the possible involvement of the endogenous opiate system in the changes in immune competence induced by isolated exercise. Male untrained rats were subjected to a 2.5 hours swimming exercise bout. Animals were killed 15 min after the end of the exercise. The concentration of leukocytes, lymphocytes, monocytes and granulocytes and T4 (T-helper), T8 (T-suppressor/cytotoxic), interleukin-2 receptor (IL-2R) and transferrin receptor (TrfR) positive lymphocytes were determined both in peripheral blood and spleen by flow cytometric analysis. Exercise resulted in a significant decrease in 1) blood lymphocyte and splenic granulocyte number (p exercising rats induced a decrease in the concentration and proportion of T8 positive lymphocytes, thereby restoring a normal T4/T8 ratio both in peripheral blood and spleen. Naloxone had no effect in control animals. The concentration and proportion of IL-2R and TrfR positive lymphocytes were not affected by naloxone. The mechanisms of the immunomodulation induced by isolated intense exercise are unclear. These data suggest that endogenous opiates participate in the alteration of cell-mediated immunity associated with exercise by modulating the T8 (suppressor/cytotoxic)-cell activity.

  10. Expression of Telomere-Associated Proteins is Interdependent to Stabilize Native Telomere Structure and Telomere Dysfunction by G-Quadruplex Ligand Causes TERRA Upregulation.

    Science.gov (United States)

    Sadhukhan, Ratan; Chowdhury, Priyanka; Ghosh, Sourav; Ghosh, Utpal

    2018-06-01

    Telomere DNA can form specialized nucleoprotein structure with telomere-associated proteins to hide free DNA ends or G-quadruplex structures under certain conditions especially in presence of G-quadruplex ligand. Telomere DNA is transcribed to form non-coding telomere repeat-containing RNA (TERRA) whose biogenesis and function is poorly understood. Our aim was to find the role of telomere-associated proteins and telomere structures in TERRA transcription. We silenced four [two shelterin (TRF1, TRF2) and two non-shelterin (PARP-1, SLX4)] telomere-associated genes using siRNA and verified depletion in protein level. Knocking down of one gene modulated expression of other telomere-associated genes and increased TERRA from 10q, 15q, XpYp and XqYq chromosomes in A549 cells. Telomere was destabilized or damaged by G-quadruplex ligand pyridostatin (PDS) and bleomycin. Telomere dysfunction-induced foci (TIFs) were observed for each case of depletion of proteins, treatment with PDS or bleomycin. TERRA level was elevated by PDS and bleomycin treatment alone or in combination with depletion of telomere-associated proteins.

  11. Thresholding of auditory cortical representation by background noise.

    Science.gov (United States)

    Liang, Feixue; Bai, Lin; Tao, Huizhong W; Zhang, Li I; Xiao, Zhongju

    2014-01-01

    It is generally thought that background noise can mask auditory information. However, how the noise specifically transforms neuronal auditory processing in a level-dependent manner remains to be carefully determined. Here, with in vivo loose-patch cell-attached recordings in layer 4 of the rat primary auditory cortex (A1), we systematically examined how continuous wideband noise of different levels affected receptive field properties of individual neurons. We found that the background noise, when above a certain critical/effective level, resulted in an elevation of intensity threshold for tone-evoked responses. This increase of threshold was linearly dependent on the noise intensity above the critical level. As such, the tonal receptive field (TRF) of individual neurons was translated upward as an entirety toward high intensities along the intensity domain. This resulted in preserved preferred characteristic frequency (CF) and the overall shape of TRF, but reduced frequency responding range and an enhanced frequency selectivity for the same stimulus intensity. Such translational effects on intensity threshold were observed in both excitatory and fast-spiking inhibitory neurons, as well as in both monotonic and nonmonotonic (intensity-tuned) A1 neurons. Our results suggest that in a noise background, fundamental auditory representations are modulated through a background level-dependent linear shifting along intensity domain, which is equivalent to reducing stimulus intensity.

  12. Effects of internal displacement and resettlement on the mental health of Turkish children and adolescents.

    Science.gov (United States)

    Erol, Neşe; Simşek, Zeynep; Oner, Ozgür; Munir, Kerim

    2005-03-01

    To evaluate the effects of internal displacement and resettlement within Turkey on the emotional and behavioral profile of children, age 5-18 after controlling for possible confounding and demographic variables. We conducted a national population survey using a self-weighted, equal probability sample. We compared the CBCL, TRF and YSR responses regarding children with (n = 1644) and without (n = 1855) experience of internal displacement. We examined the effects of gender, age, paternal employment, resettlement, urban residence and physical illness. The children and adolescents with internal displacement had significantly higher internalizing, externalizing and total problem scores on the CBCL and YSR, and higher internalizing scores on the TRF. The effect of displacement was related to higher internalizing problems when factors like physical illness, child age, child gender and urban residence were accounted. The overall effect was small explaining only 0.1-1.5% of the total variance by parent reports, and not evident by teacher reports. To our knowledge the present study is the first to examine Turkish children and adolescents with and without experience of internal displacement. The results are consistent with previous immigration studies: child age, gender, presence of physical illness and urban residence were more important predictors of internalization and externalization problem scores irrespective of informant source.

  13. A stochastic space-time model for intermittent precipitation occurrences

    KAUST Repository

    Sun, Ying; Stein, Michael L.

    2016-01-01

    Modeling a precipitation field is challenging due to its intermittent and highly scale-dependent nature. Motivated by the features of high-frequency precipitation data from a network of rain gauges, we propose a threshold space-time t random field (tRF) model for 15-minute precipitation occurrences. This model is constructed through a space-time Gaussian random field (GRF) with random scaling varying along time or space and time. It can be viewed as a generalization of the purely spatial tRF, and has a hierarchical representation that allows for Bayesian interpretation. Developing appropriate tools for evaluating precipitation models is a crucial part of the model-building process, and we focus on evaluating whether models can produce the observed conditional dry and rain probabilities given that some set of neighboring sites all have rain or all have no rain. These conditional probabilities show that the proposed space-time model has noticeable improvements in some characteristics of joint rainfall occurrences for the data we have considered.

  14. A stochastic space-time model for intermittent precipitation occurrences

    KAUST Repository

    Sun, Ying

    2016-01-28

    Modeling a precipitation field is challenging due to its intermittent and highly scale-dependent nature. Motivated by the features of high-frequency precipitation data from a network of rain gauges, we propose a threshold space-time t random field (tRF) model for 15-minute precipitation occurrences. This model is constructed through a space-time Gaussian random field (GRF) with random scaling varying along time or space and time. It can be viewed as a generalization of the purely spatial tRF, and has a hierarchical representation that allows for Bayesian interpretation. Developing appropriate tools for evaluating precipitation models is a crucial part of the model-building process, and we focus on evaluating whether models can produce the observed conditional dry and rain probabilities given that some set of neighboring sites all have rain or all have no rain. These conditional probabilities show that the proposed space-time model has noticeable improvements in some characteristics of joint rainfall occurrences for the data we have considered.

  15. E-type cyclins modulate telomere integrity in mammalian male meiosis.

    Science.gov (United States)

    Manterola, Marcia; Sicinski, Piotr; Wolgemuth, Debra J

    2016-06-01

    We have shown that E-type cyclins are key regulators of mammalian male meiosis. Depletion of cyclin E2 reduced fertility in male mice due to meiotic defects, involving abnormal pairing and synapsis, unrepaired DNA, and loss of telomere structure. These defects were exacerbated by additional loss of cyclin E1, and complete absence of both E-type cyclins produces a meiotic catastrophe. Here, we investigated the involvement of E-type cyclins in maintaining telomere integrity in male meiosis. Spermatocytes lacking cyclin E2 and one E1 allele (E1+/-E2-/-) displayed a high rate of telomere abnormalities but can progress to pachytene and diplotene stages. We show that their telomeres exhibited an aberrant DNA damage repair response during pachynema and that the shelterin complex proteins TRF2 and RAP2 were significantly decreased in the proximal telomeres. Moreover, the insufficient level of these proteins correlated with an increase of γ-H2AX foci in the affected telomeres and resulted in telomere associations involving TRF1 and telomere detachment in later prophase-I stages. These results suggest that E-type cyclins are key modulators of telomere integrity during meiosis by, at least in part, maintaining the balance of shelterin complex proteins, and uncover a novel role of E-type cyclins in regulating chromosome structure during male meiosis.

  16. Tritium inventory prediction in a CANDU plant

    International Nuclear Information System (INIS)

    Song, M.J.; Son, S.H.; Jang, C.H.

    1995-01-01

    The flow of tritium in a CANDU nuclear power plant was modeled to predict tritium activity build-up. Predictions were generally in good agreement with field measurements for the period 1983--1994. Fractional contributions of coolant and moderator systems to the environmental tritium release were calculated by least square analysis using field data from the Wolsong plant. From the analysis, it was found that: (1) about 94% of tritiated heavy water loss came from the coolant system; (2) however, about 64% of environmental tritium release came from the moderator system. Predictions of environmental tritium release were also in good agreement with field data from a few other CANDU plants. The model was used to calculate future tritium build-up and environmental tritium release at Wolsong site, Korea, where one unit is operating and three more units are under construction. The model predicts the tritium inventory at Wolsong site to increase steadily until it reaches the maximum of 66.3 MCi in the year 2026. The model also predicts the tritium release rate to reach a maximum of 79 KCi/yr in the year 2012. To reduce the tritium inventory at Wolsong site, construction of a tritium removal facility (TRF) is under consideration. The maximum needed TRF capacity of 8.7 MCi/yr was calculated to maintain tritium concentration effectively in CANDU reactors

  17. Transfer RNA Derived Small RNAs Targeting Defense Responsive Genes Are Induced during Phytophthora capsici Infection in Black Pepper (Piper nigrum L.).

    Science.gov (United States)

    Asha, Srinivasan; Soniya, Eppurath V

    2016-01-01

    Small RNAs derived from transfer RNAs were recently assigned as potential gene regulatory candidates for various stress responses in eukaryotes. In this study, we report on the cloning and identification of tRNA derived small RNAs from black pepper plants in response to the infection of the quick wilt pathogen, Phytophthora capsici. 5'tRFs cloned from black pepper were validated as highly expressed during P. capsici infection. A high-throughput systematic analysis of the small RNAome (sRNAome) revealed the predominance of 5'tRFs in the infected leaf and root. The abundance of 5'tRFs in the sRNAome and the defense responsive genes as their potential targets indicated their regulatory role during stress response in black pepper. The 5'Ala(CGC) tRF mediated cleavage was experimentally mapped at the tRF binding sites on the mRNA targets of Non-expresser of pathogenesis related protein (NPR1), which was down-regulated during pathogen infection. Comparative sRNAome further demonstrated sequence conservation of 5'Ala tRFs across the angiosperm plant groups, and many important genes in the defense response were identified in silico as their potential targets. Our findings uncovered the diversity, differential expression and stress responsive functional role of tRNA-derived small RNAs during Phytophthora infection in black pepper.

  18. Protective role of klotho protein on epithelial cells upon co-culture with activated or senescent monocytes

    Energy Technology Data Exchange (ETDEWEB)

    Mytych, Jennifer, E-mail: jennifermytych@gmail.com [Institute of Applied Biotechnology and Basic Sciences, University of Rzeszow, Werynia 502, 36-100 Kolbuszowa (Poland); Centre of Applied Biotechnology and Basic Sciences, University of Rzeszow, Werynia 502, 36-100 Kolbuszowa (Poland); Wos, Izabela; Solek, Przemyslaw; Koziorowski, Marek [Institute of Applied Biotechnology and Basic Sciences, University of Rzeszow, Werynia 502, 36-100 Kolbuszowa (Poland); Centre of Applied Biotechnology and Basic Sciences, University of Rzeszow, Werynia 502, 36-100 Kolbuszowa (Poland)

    2017-01-15

    Monocytes ensure proper functioning and maintenance of epithelial cells, while good condition of monocytes is a key factor of these interactions. Although, it was shown that in some circumstances, a population of altered monocytes may appear, there is no data regarding their effect on epithelial cells. In this study, using direct co-culture model with LPS-activated and Dox-induced senescent THP-1 monocytes, we reported for the first time ROS-induced DNA damage, reduced metabolic activity, proliferation inhibition and cell cycle arrest followed by p16-, p21- and p27-mediated DNA damage response pathways activation, premature senescence and apoptosis induction in HeLa cells. Also, we show that klotho protein possessing anti-aging and anti-inflammatory characteristics reduced cytotoxic and genotoxic events by inhibition of insulin/IGF-IR and downregulation of TRF1 and TRF2 proteins. Therefore, klotho protein could be considered as a protective factor against changes caused by altered monocytes in epithelial cells. - Highlights: • Activated and senescent THP-1 monocytes induced cyto- and genotoxicity in HeLa cells. • Altered monocytes provoked oxidative and nitrosative stress-induced DNA damage. • DNA damage activated DDR pathways and lead to premature senescence and apoptosis. • Klotho reduced ROS/RNS-mediated toxicity through insulin/IGF-IR pathway inhibition. • Klotho protects HeLa cells from cyto- and genotoxicity induced by altered monocytes.

  19. The Total Risk Analysis of Large Dams under Flood Hazards

    Directory of Open Access Journals (Sweden)

    Yu Chen

    2018-02-01

    Full Text Available Dams and reservoirs are useful systems in water conservancy projects; however, they also pose a high-risk potential for large downstream areas. Flood, as the driving force of dam overtopping, is the main cause of dam failure. Dam floods and their risks are of interest to researchers and managers. In hydraulic engineering, there is a growing tendency to evaluate dam flood risk based on statistical and probabilistic methods that are unsuitable for the situations with rare historical data or low flood probability, so a more reasonable dam flood risk analysis method with fewer application restrictions is needed. Therefore, different from previous studies, this study develops a flood risk analysis method for large dams based on the concept of total risk factor (TRF used initially in dam seismic risk analysis. The proposed method is not affected by the adequacy of historical data or the low probability of flood and is capable of analyzing the dam structure influence, the flood vulnerability of the dam site, and downstream risk as well as estimating the TRF of each dam and assigning corresponding risk classes to each dam. Application to large dams in the Dadu River Basin, Southwestern China, demonstrates that the proposed method provides quick risk estimation and comparison, which can help local management officials perform more detailed dam safety evaluations for useful risk management information.

  20. Estimating the Celestial Reference Frame via Intra-Technique Combination

    Science.gov (United States)

    Iddink, Andreas; Artz, Thomas; Halsig, Sebastian; Nothnagel, Axel

    2016-12-01

    One of the primary goals of Very Long Baseline Interferometry (VLBI) is the determination of the International Celestial Reference Frame (ICRF). Currently the third realization of the internationally adopted CRF, the ICRF3, is under preparation. In this process, various optimizations are planned to realize a CRF that does not benefit only from the increased number of observations since the ICRF2 was published. The new ICRF can also benefit from an intra-technique combination as is done for the Terrestrial Reference Frame (TRF). Here, we aim at estimating an optimized CRF by means of an intra-technique combination. The solutions are based on the input to the official combined product of the International VLBI Service for Geodesy and Astrometry (IVS), also providing the radio source parameters. We discuss the differences in the setup using a different number of contributions and investigate the impact on TRF and CRF as well as on the Earth Orientation Parameters (EOPs). Here, we investigate the differences between the combined CRF and the individual CRFs from the different analysis centers.

  1. Analysis of Global Solar Irradiance over Climatic Zones in Nigeria for Solar Energy Applications

    Directory of Open Access Journals (Sweden)

    Adekunle Ayodotun Osinowo

    2015-01-01

    Full Text Available Satellite derived solar irradiance over 25 locations in the 5 climatic zones of Nigeria (tropical rainforest TRF, Guinea savannah GS, Sahel savannah SHS, Sudan savannah SUS, and Mangrove swamp forest MSF was analyzed. To justify its use, the satellite data was tested for goodness of agreement with ground measured solar radiation data using 26-year mean monthly and daily data over 16 locations in the 5 climatic zones. The well-known R2, RMSE, MBE, and MPE statistical tests were used and good agreement was found. The 25 locations were grouped into the 5 climatic zones. Frequency distribution of global solar irradiance was done for each of the climatic zones. This showed that 46.88%, and 40.6% of the number of days (9794 over TRF and MSF, respectively, had irradiation within the range of 15.01–20.01 MJ/m2/day. For the GS, SHS, and SUS, 46.19%, 55.84% and 58.53% of the days had total irradiation within the range of 20.01–25.01 MJ/m2/day, respectively. Generally, in all the climatic zones, coefficients of variation of solar radiation were high and mean values were low in July and August. Contour maps showed that high and low values of global solar irradiance and clearness index were observed in the Northern and Southern locations of Nigeria, respectively.

  2. NEIL3 Repairs Telomere Damage during S Phase to Secure Chromosome Segregation at Mitosis

    Directory of Open Access Journals (Sweden)

    Jia Zhou

    2017-08-01

    Full Text Available Oxidative damage to telomere DNA compromises telomere integrity. We recently reported that the DNA glycosylase NEIL3 preferentially repairs oxidative lesions in telomere sequences in vitro. Here, we show that loss of NEIL3 causes anaphase DNA bridging because of telomere dysfunction. NEIL3 expression increases during S phase and reaches maximal levels in late S/G2. NEIL3 co-localizes with TRF2 and associates with telomeres during S phase, and this association increases upon oxidative stress. Mechanistic studies reveal that NEIL3 binds to single-stranded DNA via its intrinsically disordered C terminus in a telomere-sequence-independent manner. Moreover, NEIL3 is recruited to telomeres through its interaction with TRF1, and this interaction enhances the enzymatic activity of purified NEIL3. Finally, we show that NEIL3 interacts with AP Endonuclease 1 (APE1 and the long-patch base excision repair proteins PCNA and FEN1. Taken together, we propose that NEIL3 protects genome stability through targeted repair of oxidative damage in telomeres during S/G2 phase.

  3. Darlington tritium removal facility and station upgrading plant dynamic process simulation

    International Nuclear Information System (INIS)

    Busigin, A.; Williams, G. I. D.; Wong, T. C. W.; Kulczynski, D.; Reid, A.

    2008-01-01

    Ontario Power Generation Nuclear (OPGN) has a 4 x 880 MWe CANDU nuclear station at its Darlington Nuclear Div. located in Bowmanville. The station has been operating a Tritium Removal Facility (TRF) and a D 2 O station Upgrading Plant (SUP) since 1989. Both facilities were designed with a Distributed Control System (DCS) and programmable logic controllers (PLC) for process control. This control system was replaced with a DCS only, in 1998. A dynamic plant simulator was developed for the Darlington TRF (DTRF) and the SUP, as part of the computer control system replacement. The simulator was used to test the new software, required to eliminate the PLCs. The simulator is now used for operator training and testing of process control software changes prior to field installation. Dynamic simulation will be essential for the ITER isotope separation system, where the process is more dynamic than the relatively steady-state DTRF process. This paper describes the development and application of the DTRF and SUP dynamic simulator, its benefits, architecture, and the operational experience with the simulator. (authors)

  4. HPLC Separation of Vitamin E and Its Oxidation Products and Effects of Oxidized Tocotrienols on the Viability of MCF-7 Breast Cancer Cells in Vitro.

    Science.gov (United States)

    Drotleff, Astrid M; Büsing, Anne; Willenberg, Ina; Empl, Michael T; Steinberg, Pablo; Ternes, Waldemar

    2015-10-14

    Tocotrienols, a vitamin E subgroup, exert potent anticancer effects, but easily degrade due to oxidation. Eight vitamin E reference compounds, α-, β-, γ-, or δ-tocopherols or -tocotrienols, were thermally oxidized in n-hexane. The corresponding predominantly dimeric oxidation products were separated from the parent compounds by diol-modified normal-phase HPLC-UV and characterized by mass spectroscopy. The composition of test compounds, that is, α-tocotrienol, γ-tocotrienol, or palm tocotrienol-rich fraction (TRF), before and after thermal oxidation was determined by HPLC-DAD, and MCF-7 cells were treated with both nonoxidized and oxidized test compounds for 72 h. Whereas all nonoxidized test compounds (0-100 μM) led to dose-dependent decreases in cell viability, equimolar oxidized α-tocotrienol had a weaker effect, and oxidized TRF had no such effect. However, the IC50 value of oxidized γ-tocotrienol was lower (85 μM) than that of nonoxidized γ-tocotrienol (134 μM), thereby suggesting that γ-tocotrienol oxidation products are able to reduce tumor cell viability in vitro.

  5. Genomic Instability: The Driving Force behind Refractory/Relapsing Hodgkin’s Lymphoma

    International Nuclear Information System (INIS)

    Knecht, Hans; Righolt, Christiaan; Mai, Sabine

    2013-01-01

    In classical Hodgkin’s lymphoma (HL) the malignant mononuclear Hodgkin (H) and multinuclear, diagnostic Reed-Sternberg (RS) cells are rare and generally make up <3% of the total cellular mass of the affected lymph nodes. During recent years, the introduction of laser micro-dissection techniques at the single cell level has substantially improved our understanding of the molecular pathogenesis of HL. Gene expression profiling, comparative genomic hybridization analysis, micro-RNA expression profiling and viral oncogene sequencing have deepened our knowledge of numerous facets of H- and RS-cell gene expression deregulation. The question remains whether disturbed signaling pathways and deregulated transcription factors are at the origin of refractory/relapsing Hodgkin’s lymphoma or whether these hallmarks are at least partially related to another major factor. We recently showed that the 3D nuclear organization of telomeres and chromosomes marked the transition from H- to RS-cells in HL cell lines. This transition is associated with progression of telomere dysfunction, shelterin disruption and progression of complex chromosomal rearrangements. We reported analogous findings in refractory/relapsing HL and identified the shelterin proteins TRF1, TRF2 and POT1 as targets of the LMP1 oncogene in post-germinal center B-cells. Here we summarize our findings, including data not previously published, and propose a model in which progressive disruption of nuclear integrity, a form of genomic instability, is the key-player in refractory/relapsing HL. Therapeutic approaches should take these findings into account

  6. Identification and Mapping of Simple Sequence Repeat Markers from Common Bean (Phaseolus vulgaris L. Bacterial Artificial Chromosome End Sequences for Genome Characterization and Genetic–Physical Map Integration

    Directory of Open Access Journals (Sweden)

    Juana M. Córdoba

    2010-11-01

    Full Text Available Microsatellite markers or simple sequence repeat (SSR loci are useful for diversity characterization and genetic–physical mapping. Different in silico microsatellite search methods have been developed for mining bacterial artificial chromosome (BAC end sequences for SSRs. The overall goal of this study was genome characterization based on SSRs in 89,017 BAC end sequences (BESs from the G19833 common bean ( L. library. Another objective was to identify new SSR taking into account three tandem motif identification programs (Automated Microsatellite Marker Development [AMMD], Tandem Repeats Finder [TRF], and SSRLocator [SSRL]. Among the microsatellite search engines, SSRL identified the highest number of SSRs; however, when primer design was attempted, the number dropped due to poor primer design regions. Automated Microsatellite Marker Development software identified many SSRs with valuable AT/TA or AG/TC motifs, while TRF found fewer SSRs and produced no primers. A subgroup of 323 AT-rich, di-, and trinucleotide SSRs were selected from the AMMD results and used in a parental survey with DOR364 and G19833, of which 75 could be mapped in the corresponding population; these represented 4052 BAC clones. Together with 92 previously mapped BES- and 114 non-BES-derived markers, a total of 280 SSRs were included in the polymerase chain reaction (PCR-based map, integrating a total of 8232 BAC clones in 162 contigs from the physical map.

  7. Nuclear fusion: technology development and achievements

    International Nuclear Information System (INIS)

    Ana, G.; Brad, S.; Lazar, A.; Spiridon, I.; Vijulie, M.

    2009-01-01

    The strategy for reducing the enhanced greenhouse effect, affecting our planet and the increasing energy demand caused by global growth of population, as well, is, certainly, that of adopting, all the three long-term carbon-free options for energy: renewable sources, fission and fusion reactions. All of them should be further explored and developed for the welfare of future generations, thus leaving them the option of a clean and green energy. From all those available options, the fusion is viewed as an energy source that would be effective in solving future demands, referring to the amount of the energy that can be produced taking into account spent fuel to obtain it. The slow (but steady) progress of fusion development linked with the need for large and expensive experimental devices is remarkable; all available technical and scientific information from experiment shows that progress is being made towards a successful reactor (ITER). The desired objective of this intensive research is obtaining of an industrial reactor able to cover energy future requirements. For the construction of this desired reactor, special designs systems are required. The TRF (TRITIUM RECOVERY FACILITY) is one of the installations that have a major importance maintaining an efficient fuel cycle of the reactor in proper function stage. A test model of TRF for implementation in reactor systems, proposed and developed at ICIT Ramnicu Valcea is presented. (authors)

  8. Protective role of klotho protein on epithelial cells upon co-culture with activated or senescent monocytes

    International Nuclear Information System (INIS)

    Mytych, Jennifer; Wos, Izabela; Solek, Przemyslaw; Koziorowski, Marek

    2017-01-01

    Monocytes ensure proper functioning and maintenance of epithelial cells, while good condition of monocytes is a key factor of these interactions. Although, it was shown that in some circumstances, a population of altered monocytes may appear, there is no data regarding their effect on epithelial cells. In this study, using direct co-culture model with LPS-activated and Dox-induced senescent THP-1 monocytes, we reported for the first time ROS-induced DNA damage, reduced metabolic activity, proliferation inhibition and cell cycle arrest followed by p16-, p21- and p27-mediated DNA damage response pathways activation, premature senescence and apoptosis induction in HeLa cells. Also, we show that klotho protein possessing anti-aging and anti-inflammatory characteristics reduced cytotoxic and genotoxic events by inhibition of insulin/IGF-IR and downregulation of TRF1 and TRF2 proteins. Therefore, klotho protein could be considered as a protective factor against changes caused by altered monocytes in epithelial cells. - Highlights: • Activated and senescent THP-1 monocytes induced cyto- and genotoxicity in HeLa cells. • Altered monocytes provoked oxidative and nitrosative stress-induced DNA damage. • DNA damage activated DDR pathways and lead to premature senescence and apoptosis. • Klotho reduced ROS/RNS-mediated toxicity through insulin/IGF-IR pathway inhibition. • Klotho protects HeLa cells from cyto- and genotoxicity induced by altered monocytes.

  9. Evanescent Wave Fiber Optic Biosensor for Salmonella Detection in Food

    Directory of Open Access Journals (Sweden)

    Arun K. Bhunia

    2009-07-01

    Full Text Available Salmonella enterica is a major food-borne pathogen of world-wide concern. Sensitive and rapid detection methods to assess product safety before retail distribution are highly desirable. Since Salmonella is most commonly associated with poultry products, an evanescent wave fiber-optic assay was developed to detect Salmonella in shell egg and chicken breast and data were compared with a time-resolved fluorescence (TRF assay. Anti-Salmonella polyclonal antibody was immobilized onto the surface of an optical fiber using biotin-avidin interactions to capture Salmonella. Alexa Fluor 647-conjugated antibody (MAb 2F-11 was used as the reporter. Detection occurred when an evanescent wave from a laser (635 nm excited the Alexa Fluor and the fluorescence was measured by a laser-spectrofluorometer at 710 nm. The biosensor was specific for Salmonella and the limit of detection was established to be 103 cfu/mL in pure culture and 104 cfu/mL with egg and chicken breast samples when spiked with 102 cfu/mL after 2–6 h of enrichment. The results indicate that the performance of the fiber-optic sensor is comparable to TRF, and can be completed in less than 8 h, providing an alternative to the current detection methods.

  10. Characterization of bacterial consortia capable of degrading 4-chlorobenzoate and 4-bromobenzoate under denitrifying conditions.

    Science.gov (United States)

    Song, Bongkeun; Kerkhof, Lee J; Häggblom, Max M

    2002-08-06

    4-Chlorobenzoate and 4-bromobenzoate were readily degraded in denitrifying enrichment cultures established with river sediment, estuarine sediment or agricultural soil as inoculum. Stable denitrifying consortia were obtained and maintained by serial dilution and repeated feeding of substrates. Microbial community analyses were performed to characterize the 4-chlorobenzoate and 4-bromobenzoate degrading consortia with terminal restriction fragment length polymorphism (T-RFLP) and cloning of 16S rRNA genes from the cultures. Interestingly, two major terminal restriction fragments (T-RFs) in the 4-chlorobenzoate degrading consortia and one T-RF in the 4-bromobenzoate utilizing consortium were observed from T-RFLP analysis regardless of their geographical and ecological origins. The two T-RFs (clones 4CB1 and 4CB2) in 4-chlorobenzoate degrading consortia were identified as members of the beta-subunit of the Proteobacteria on the basis of 16S rRNA sequencing analysis. Phylogenetic analysis of 16S rRNA genes showed that clone 4CB1 was closely related to Thauera aromatica while clone 4CB2 was distantly related to the genera Limnobacter and Ralstonia. The 4-bromobenzoate utilizing consortium mainly consisted of one T-RF, which was identical to clone 4CB2 in spite of different enrichment substrate. This suggests that degradation of 4-chlorobenzoate and 4-bromobenzoate under denitrifying conditions was mediated by bacteria belonging to the beta-subunit of the Proteobacteria.

  11. Intracellular O2 sensing probe based on cell-penetrating phosphorescent nanoparticles.

    Science.gov (United States)

    Fercher, Andreas; Borisov, Sergey M; Zhdanov, Alexander V; Klimant, Ingo; Papkovsky, Dmitri B

    2011-07-26

    A new intracellular O(2) (icO(2)) sensing probe is presented, which comprises a nanoparticle (NP) formulation of a cationic polymer Eudragit RL-100 and a hydrophobic phosphorescent dye Pt(II)-tetrakis(pentafluorophenyl)porphyrin (PtPFPP). Using the time-resolved fluorescence (TR-F) plate reader set-up, cell loading was investigated in detail, particularly the effects of probe concentration, loading time, serum content in the medium, cell type, density, etc. The use of a fluorescent analogue of the probe in conjunction with confocal microscopy and flow cytometry analysis, revealed that cellular uptake of the NPs is driven by nonspecific energy-dependent endocytosis and that the probe localizes inside the cell close to the nucleus. Probe calibration in biological environment was performed, which allowed conversion of measured phosphorescence lifetime signals into icO(2) concentration (μM). Its analytical performance in icO(2) sensing experiments was demonstrated by monitoring metabolic responses of mouse embryonic fibroblast cells under ambient and hypoxic macroenvironment. The NP probe was seen to generate stable and reproducible signals in different types of mammalian cells and robust responses to their metabolic stimulation, thus allowing accurate quantitative analysis. High brightness and photostability allow its use in screening experiments with cell populations on a commercial TR-F reader, and for single cell analysis on a fluorescent microscope.

  12. Oxidative stress induces senescence in human mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Brandl, Anita [Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042 Regensburg (Germany); Meyer, Matthias; Bechmann, Volker [Department of Trauma Surgery, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042 Regensburg (Germany); Nerlich, Michael [Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042 Regensburg (Germany); Angele, Peter, E-mail: Peter.Angele@klinik.uni-regensburg.de [Department of Trauma Surgery, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042 Regensburg (Germany)

    2011-07-01

    Mesenchymal stem cells (MSCs) contribute to tissue repair in vivo and form an attractive cell source for tissue engineering. Their regenerative potential is impaired by cellular senescence. The effects of oxidative stress on MSCs are still unknown. Our studies were to investigate into the proliferation potential, cytological features and the telomere linked stress response system of MSCs, subject to acute or prolonged oxidant challenge with hydrogen peroxide. Telomere length was measured using the telomere restriction fragment assay, gene expression was determined by rtPCR. Sub-lethal doses of oxidative stress reduced proliferation rates and induced senescent-morphological features and senescence-associated {beta}-galactosidase positivity. Prolonged low dose treatment with hydrogen peroxide had no effects on cell proliferation or morphology. Sub-lethal and prolonged low doses of oxidative stress considerably accelerated telomere attrition. Following acute oxidant insult p21 was up-regulated prior to returning to initial levels. TRF1 was significantly reduced, TRF2 showed a slight up-regulation. SIRT1 and XRCC5 were up-regulated after oxidant insult and expression levels increased in aging cells. Compared to fibroblasts and chondrocytes, MSCs showed an increased tolerance to oxidative stress regarding proliferation, telomere biology and gene expression with an impaired stress tolerance in aged cells.

  13. FR-like EBNA1 binding repeats in the human genome

    International Nuclear Information System (INIS)

    D'Herouel, Aymeric Fouquier; Birgersdotter, Anna; Werner, Maria

    2010-01-01

    Epstein-Barr virus (EBV) is widely spread in the human population. EBV nuclear antigen 1 (EBNA1) is a transcription factor that activates viral genes and is necessary for viral replication and partitioning, which binds the EBV genome cooperatively. We identify similar EBNA1 repeat binding sites in the human genome using a nearest-neighbor positional weight matrix. Previously experimentally verified EBNA1 sites in the human genome are successfully recovered by our approach. Most importantly, 40 novel regions are identified in the human genome, constituted of tandemly repeated binding sites for EBNA1. Genes located in the vicinity of these regions are presented as possible targets for EBNA1-mediated regulation. Among these, four are discussed in more detail: IQCB1, IMPG1, IRF2BP2 and TPO. Incorporating the cooperative actions of EBNA1 is essential when identifying regulatory regions in the human genome and we believe the findings presented here are highly valuable for the understanding of EBV-induced phenotypic changes.

  14. Impact of ITRS 2014 realizations on altimeter satellite precise orbit determination

    Science.gov (United States)

    Zelensky, Nikita P.; Lemoine, Frank G.; Beckley, Brian D.; Chinn, Douglas S.; Pavlis, Despina E.

    2018-01-01

    This paper evaluates orbit accuracy and systematic error for altimeter satellite precise orbit determination on TOPEX, Jason-1, Jason-2 and Jason-3 by comparing the use of four SLR/DORIS station complements from the International Terrestrial Reference System (ITRS) 2014 realizations with those based on ITRF2008. The new Terrestrial Reference Frame 2014 (TRF2014) station complements include ITRS realizations from the Institut National de l'Information Géographique et Forestière (IGN) ITRF2014, the Jet Propulsion Laboratory (JPL) JTRF2014, the Deutsche Geodätisches Forschungsinstitut (DGFI) DTRF2014, and the DORIS extension to ITRF2014 for Precise Orbit Determination, DPOD2014. The largest source of error stems from ITRF2008 station position extrapolation past the 2009 solution end time. The TRF2014 SLR/DORIS complement impact on the ITRF2008 orbit is only 1-2 mm RMS radial difference between 1992-2009, and increases after 2009, up to 5 mm RMS radial difference in 2016. Residual analysis shows that station position extrapolation error past the solution span becomes evident even after two years, and will contribute to about 3-4 mm radial orbit error after seven years. Crossover data show the DTRF2014 orbits are the most accurate for the TOPEX and Jason-2 test periods, and the JTRF2014 orbits for the Jason-1 period. However for the 2016 Jason-3 test period only the DPOD2014-based orbits show a strong and statistically significant margin of improvement. The positive results with DTRF2014 suggest the new approach to correct station positions or normal equations for non-tidal loading before combination is beneficial. We did not find any compelling POD advantage in using non-linear over linear station velocity models in our SLR & DORIS orbit tests on the Jason satellites. The JTRF2014 proof-of-concept ITRS realization demonstrates the need for improved SLR+DORIS orbit centering when compared to the Ries (2013) CM annual model. Orbit centering error is seen as an annual

  15. Biolistic co-transformation of Metarhizium anisopliae var. acridum strain CG423 with green fluorescent protein and resistance to glufosinate ammonium.

    Science.gov (United States)

    Inglis, P W; Aragão, F J; Frazão, H; Magalhães, B P; Valadares-Inglis, M C

    2000-10-15

    Metarhizium anisopliae var. acridum (syn. M. flavoviride) is recognized as a highly specific and virulent mycopathogen of locusts and grasshoppers and is currently being developed as a biological control agent for this group of insects in Brazil. Intact conidia of M. anisopliae var. acridum strain CG423 were transformed using microparticle bombardment. Plasmids used were: (1) pBARKS1 carrying the bar gene of Streptomyces hygroscopicus fused to the Aspergillus nidulans trpC promoter, encoding resistance to glufosinate ammonium (or phosphinothricin) and modified by addition of the telomeric repeat (TTAGGG)(18) of Fusarium oxysporum and 2.pEGFP/gpd/tel carrying a red-shifted variant gene for Aequorea victoria green fluorescent protein (EGFP) which we have fused to the A. nidulans gpd promoter and trpC terminator. Highly fluorescent co-transformants were selected on solid minimal medium containing 100 microg ml(-1) glufosinate ammonium using an inverted microscope with 450-490 nm excitation/510 nm emission filter set. Southern blot analysis of co-transformants revealed varying multiple chromosomal integrations of both bar and egfp genes at both telomeric and non-telomeric loci. Transformants retained pathogenicity in bioassays against Rhammatocerus schistocercoides and showed unaltered lack of pathogenicity against larvae of the non-target insect Anticarsia gemmatalis. One co-transformant from four tested, however, showed a significant, but non-dose-dependent, elevation in virulence against Tenebrio molitor.

  16. A Shld1-controlled POT1a provides support for repression of ATR signaling at telomeres through RPA exclusion.

    Science.gov (United States)

    Gong, Yi; de Lange, Titia

    2010-11-12

    We previously proposed that POT1 prevents ATR signaling at telomeres by excluding RPA from the single-stranded TTAGGG repeats. Here, we use a Shld1-stabilized degron-POT1a fusion (DD-POT1a) to study the telomeric ATR kinase response. In the absence of Shld1, DD-POT1a degradation resulted in rapid and reversible activation of the ATR pathway in G1 and S/G2. ATR signaling was abrogated by shRNAs to ATR and TopBP1, but shRNAs to the ATM kinase or DNA-PKcs did not affect the telomere damage response. Importantly, ATR signaling in G1 and S/G2 was reduced by shRNAs to RPA. In S/G2, RPA was readily detectable at dysfunctional telomeres, and both POT1a and POT1b were required to exclude RPA and prevent ATR activation. In G1, the accumulation of RPA at dysfunctional telomeres was strikingly less, and POT1a was sufficient to repress ATR signaling. These results support an RPA exclusion model for the repression of ATR signaling at telomeres. Copyright © 2010 Elsevier Inc. All rights reserved.

  17. RPA and POT1: friends or foes at telomeres?

    Science.gov (United States)

    Flynn, Rachel Litman; Chang, Sandy; Zou, Lee

    2012-02-15

    Telomere maintenance in cycling cells relies on both DNA replication and capping by the protein complex shelterin. Two single-stranded DNA (ssDNA)-binding proteins, replication protein A (RPA) and protection of telomere 1 (POT1) play critical roles in DNA replication and telomere capping, respectively. While RPA binds to ssDNA in a non-sequence-specific manner, POT1 specifically recognizes singlestranded TTAGGG telomeric repeats. Loss of POT1 leads to aberrant accumulation of RPA at telomeres and activation of the ataxia telangiectasia and Rad3-related kinase (ATR)-mediated checkpoint response, suggesting that POT1 antagonizes RPA binding to telomeric ssDNA. The requirement for both POT1 and RPA in telomere maintenance and the antagonism between the two proteins raises the important question of how they function in concert on telomeric ssDNA. Two interesting models were proposed by recent studies to explain the regulation of POT1 and RPA at telomeres. Here, we discuss how these models help unravel the coordination, and also the antagonism, between POT1 and RPA during the cell cycle.

  18. Tumour-cell apoptosis after cisplatin treatment is not telomere dependent.

    Science.gov (United States)

    Jeyapalan, Jessie C; Saretzki, Gabriele; Leake, Alan; Tilby, Michael J; von Zglinicki, Thomas

    2006-06-01

    Cisplatin is a major chemotherapeutic agent, especially for the treatment of neuroblastoma. Telomeres with their sequence (TTAGGG)n are probable targets for cisplatin intrastrand cross-linking, but the role of telomeres in mediating cisplatin cytotoxicity is not clear. After exposure to cisplatin as single dose or continuous treatment, we found no loss of telomeres in either SHSY5Y neuroblastoma cells (telomere length, approximately 4 kbp), HeLa 229 cells (telomere length, 20 kbp) or in the acute lymphoblastic T cell line 1301 (telomere length, approximately 80 kbp). There was no induction of telomeric single strand breaks, telomeric overhangs were not degraded and telomerase activity was down-regulated only after massive onset of apoptosis. In contrast, cisplatin induced a delayed formation of DNA strand breaks and induced DNA damage foci containing gamma-H2A.X at nontelomeric sites. Interstitial DNA damage appears to be more important than telomere loss or telomeric damage as inducer of the signal pathway towards apoptosis and/or growth arrest in cisplatin-treated tumour cells.

  19. Effects of Unpredictable Variable Prenatal Stress (UVPS) on Bdnf DNA Methylation and Telomere Length in the Adult Rat Brain

    Science.gov (United States)

    Blaze, Jennifer; Asok, A.; Moyer, E. L.; Roth, T. L.; Ronca, A. E.

    2015-01-01

    In utero exposure to stress can shape neurobiological and behavioral outcomes in offspring, producing vulnerability to psychopathology later in life. Animal models of prenatal stress likewise have demonstrated long-­-term alterations in brain function and behavioral deficits in offspring. For example, using a rodent model of unpredictable variable prenatal stress (UVPS), in which dams are exposed to unpredictable, variable stress across pregnancy, we have found increased body weight and anxiety-­-like behavior in adult male, but not female, offspring. DNA methylation (addition of methyl groups to cytosines which normally represses gene transcription) and changes in telomere length (TTAGGG repeats on the ends of chromosomes) are two molecular modifications that result from stress and could be responsible for the long-­-term effects of UVPS. Here, we measured methylation of brain-­-derived neurotrophic factor (bdnf), a gene important in development and plasticity, and telomere length in the brains of adult offspring from the UVPS model. Results indicate that prenatally stressed adult males have greater methylation in the medial prefrontal cortex (mPFC) compared to non-­-stressed controls, while females have greater methylation in the ventral hippocampus compared to controls. Further, prenatally stressed males had shorter telomeres than controls in the mPFC. These findings demonstrate the ability of UVPS to produce epigenetic alterations and changes in telomere length across behaviorally-­-relevant brain regions, which may have linkages to the phenotypic outcomes.

  20. Intrauterine Exposure to Maternal Stress Alters Bdnf IV DNA Methylation and Telomere Length in the Brain of Adult Rat Offspring

    Science.gov (United States)

    Blaze, Jennifer; Asok, Arun; Borrelli, Kristyn; Tulbert, Christine; Bollinger, Justin; Ronca Finco, April E.; Roth, Tania L.

    2017-01-01

    DNA methylation (addition of methyl groups to cytosines which normally represses gene transcription) and changes in telomere length (TTAGGG repeats on the ends of chromosomes) are two molecular modifications that result from stress and could contribute to the long-term effects of intrauterine exposure to maternal stress on offspring behavioral outcomes. Here, we measured methylation of Brain-derived neurotrophic factor (Bdnf), a gene important in development and plasticity, and telomere length in the brains of adult rat male and female offspring whose mothers were exposed to unpredictable and variable stressors throughout gestation. Males exposed to prenatal stress had greater methylation (Bdnf IV) in the medial prefrontal cortex (mPFC) compared to non-stressed controls. Further, prenatally-stressed males had shorter telomeres than controls in the mPFC. This study provides the first evidence in a rodent model of an association between prenatal stress exposure and subsequent shorter brain telomere length. Together findings indicate a long-term impact of prenatal stress on DNA methylation and telomere biology with relevance for behavioral and health outcomes, and contribute to a growing literature linking stress to intergenerational epigenetic alterations and changes in telomere length.

  1. Comparative cytogenetics of six Indo-Pacific moray eels (Anguilliformes: Muraenidae) by chromosomal banding and fluorescence in situ hybridization.

    Science.gov (United States)

    Coluccia, E; Deidda, F; Cannas, R; Lobina, C; Cuccu, D; Deiana, A M; Salvadori, S

    2015-09-01

    A comparative cytogenetic analysis, using both conventional staining techniques and fluorescence in situ hybridization, of six Indo-Pacific moray eels from three different genera (Gymnothorax fimbriatus, Gymnothorax flavimarginatus, Gymnothorax javanicus, Gymnothorax undulatus, Echidna nebulosa and Gymnomuraena zebra), was carried out to investigate the chromosomal differentiation in the family Muraenidae. Four species displayed a diploid chromosome number 2n = 42, which is common among the Muraenidae. Two other species, G. javanicus and G. flavimarginatus, were characterized by different chromosome numbers (2n = 40 and 2n = 36). For most species, a large amount of constitutive heterochromatin was detected in the chromosomes, with species-specific C-banding patterns that enabled pairing of the homologous chromosomes. In all species, the major ribosomal genes were localized in the guanine-cytosine-rich region of one chromosome pair, but in different chromosomal locations. The (TTAGGG)n telomeric sequences were mapped onto chromosomal ends in all muraenid species studied. The comparison of the results derived from this study with those available in the literature confirms a substantial conservation of the diploid chromosome number in the Muraenidae and supports the hypothesis that rearrangements have occurred that have diversified their karyotypes. Furthermore, the finding of two species with different diploid chromosome numbers suggests that additional chromosomal rearrangements, such as Robertsonian fusions, have occurred in the karyotype evolution of the Muraenidae. © 2015 The Fisheries Society of the British Isles.

  2. Karyological characterization and identification of four repetitive element groups (the 18S – 28S rRNA gene, telomeric sequences, microsatellite repeat motifs, Rex retroelements) of the Asian swamp eel (Monopterus albus)

    Science.gov (United States)

    Suntronpong, Aorarat; Thapana, Watcharaporn; Twilprawat, Panupon; Prakhongcheep, Ornjira; Somyong, Suthasinee; Muangmai, Narongrit; Surin Peyachoknagul; Srikulnath, Kornsorn

    2017-01-01

    Abstract Among teleost fishes, Asian swamp eel (Monopterus albus Zuiew, 1793) possesses the lowest chromosome number, 2n = 24. To characterize the chromosome constitution and investigate the genome organization of repetitive sequences in M. albus, karyotyping and chromosome mapping were performed with the 18S – 28S rRNA gene, telomeric repeats, microsatellite repeat motifs, and Rex retroelements. The 18S – 28S rRNA genes were observed to the pericentromeric region of chromosome 4 at the same position with large propidium iodide and C-positive bands, suggesting that the molecular structure of the pericentromeric regions of chromosome 4 has evolved in a concerted manner with amplification of the 18S – 28S rRNA genes. (TTAGGG)n sequences were found at the telomeric ends of all chromosomes. Eight of 19 microsatellite repeat motifs were dispersedly mapped on different chromosomes suggesting the independent amplification of microsatellite repeat motifs in M. albus. Monopterus albus Rex1 (MALRex1) was observed at interstitial sites of all chromosomes and in the pericentromeric regions of most chromosomes whereas MALRex3 was scattered and localized to all chromosomes and MALRex6 to several chromosomes. This suggests that these retroelements were independently amplified or lost in M. albus. Among MALRexs (MALRex1, MALRex3, and MALRex6), MALRex6 showed higher interspecific sequence divergences from other teleost species in comparison. This suggests that the divergence of Rex6 sequences of M. albus might have occurred a relatively long time ago. PMID:29093797

  3. Telomeres, telomerase and premature ovarian failure

    Directory of Open Access Journals (Sweden)

    Renata Košir Pogačnik

    2011-11-01

    Full Text Available Telomeres are specialized structures at the ends of chromosomes, consisting of six repeated nucleotides in TTAGGG sequence. Genome stability is partly maintained by the architecture of telomeres and is gradually lost as telomeres progressively shorten with each cell replication. Critically shortened telomeres are recognized by DNA repair mechanisms as DNA damage and the cell replication cycle stops. The cell eventually dies or undergoes cell apoptosis. Telomere represents a cellular marker of biological age and are therefore also called cell mitotic clock. The enzyme that counteracts telomere shortening by adding nucleotides to the 3’ end of DNA strand is called telomerase. It is composed of the RNA subunit (TR, which is special type of messenger RNA (mRNA, the catalytic protein subunit (TERT, which works as a reverse transcriptase and numerous additional proteins. Telomerase is active in some germline, epithelial and haemopoietic cells, but in most somatic cells the activity is undetectable. In literature, the length of telomeres is closely connected with premature ovarian failure (POF. POF is generally defined as the onset of menopause before the age of 40. The causes of disease are genetical, autoimmune, iatrogenic or if we cannot establish the cause – idiopathic. A lot of studies examined correlation between idiopathic POF, length of telomeres and telomerase activity. The studies mostly show that women with POF have shortened telomeres and decreased activity of telomerase as compared to healthy women.

  4. Thermodynamic fingerprints of ligand binding to human telomeric G-quadruplexes.

    Science.gov (United States)

    Bončina, Matjaž; Podlipnik, Črtomir; Piantanida, Ivo; Eilmes, Julita; Teulade-Fichou, Marie-Paule; Vesnaver, Gorazd; Lah, Jurij

    2015-12-02

    Thermodynamic studies of ligand binding to human telomere (ht) DNA quadruplexes, as a rule, neglect the involvement of various ht-DNA conformations in the binding process. Therefore, the thermodynamic driving forces and the mechanisms of ht-DNA G-quadruplex-ligand recognition remain poorly understood. In this work we characterize thermodynamically and structurally binding of netropsin (Net), dibenzotetraaza[14]annulene derivatives (DP77, DP78), cationic porphyrin (TMPyP4) and two bisquinolinium ligands (Phen-DC3, 360A-Br) to the ht-DNA fragment (Tel22) AGGG(TTAGGG)3 using isothermal titration calorimetry, CD and fluorescence spectroscopy, gel electrophoresis and molecular modeling. By global thermodynamic analysis of experimental data we show that the driving forces characterized by contributions of specific interactions, changes in solvation and conformation differ significantly for binding of ligands with low quadruplex selectivity over duplexes (Net, DP77, DP78, TMPyP4; KTel22 ≈ KdsDNA). These contributions are in accordance with the observed structural features (changes) and suggest that upon binding Net, DP77, DP78 and TMPyP4 select hybrid-1 and/or hybrid-2 conformation while Phen-DC3 and 360A-Br induce the transition of hybrid-1 and hybrid-2 to the structure with characteristics of antiparallel or hybrid-3 type conformation. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. MicroRNA Regulation of Telomerase Reverse Transcriptase (TERT: Micro Machines Pull Strings of Papier-Mâché Puppets

    Directory of Open Access Journals (Sweden)

    Ammad Ahmad Farooqi

    2018-04-01

    Full Text Available Substantial fraction of high-quality information is continuously being added into the existing pool of knowledge related to the biology of telomeres. Based on the insights gleaned from decades of research, it is clear that chromosomal stability needs a highly controlled and dynamic balance of DNA gain and loss in each terminal tract of telomeric repeats. Telomeres are formed by tandem repeats of TTAGGG sequences, which are gradually lost with each round of division of the cells. Targeted inhibition of telomerase to effectively induce apoptosis in cancer cells has attracted tremendous attention and overwhelmingly increasingly list of telomerase inhibitors truthfully advocates pharmacological significance of telomerase. Telomerase reverse transcriptase (TERT is a multi-talented and catalytically active component of the telomerase-associated protein machinery. Different proteins of telomerase-associated machinery work in a synchronized and orchestrated manner to ensure proper maintenance of telomeric length of chromosomes. Rapidly emerging scientific findings about regulation of TERT by microRNAs has revolutionized our understanding related to the biology of telomeres and telomerase. In this review, we have comprehensively discussed how different miRNAs regulate TERT in different cancers. Use of miRNA-based therapeutics against TERT in different cancers needs detailed research in preclinical models for effective translation of laboratory findings to clinically effective therapeutics.

  6. Cytogenetic characterisation of the razor shells Ensis directus (Conrad, 1843) and E. minor (Chenu, 1843) (Mollusca: Bivalvia)

    Science.gov (United States)

    González-Tizón, Ana M.; Rojo, Verónica; Vierna, Joaquín; Jensen, K. Thomas; Egea, Emilie; Martínez-Lage, Andrés

    2013-03-01

    The European razor shell Ensis minor (Chenu 1843) and the American E. directus (Conrad 1843) have a diploid chromosome number of 38 and remarkable differences in their karyotypes: E. minor has four metacentric, one metacentric-submetacentric, five submetacentric, one subtelocentric and eight telocentric chromosome pairs, whereas E. directus has three metacentric, two metacentric-submetacentric, six submetacentric, six subtelocentric and two telocentric pairs. Fluorescent in situ hybridisation (FISH) using a major ribosomal DNA probe located the major ribosomal genes on one submetacentric chromosome pair in both species; FISH with a 5S ribosomal DNA (5S rDNA) probe rendered one chromosomal (weak) signal for E. minor and no signal for E. directus, supporting a more dispersed organisation of 5S rDNA compared to the major ribosomal genes. The vertebrate telomeric sequence (TTAGGG) n was located on both ends of each chromosome, and no interstitial signals were detected. In this work, a comparative karyological analysis was also performed between the four Ensis species analysed revealing that the three European species studied so far, namely E. minor, E. siliqua (Linné 1758) and E. magnus Schumacher 1817 show more similarities among them than compared to the American species E. directus. In addition, clear karyotype differences were found between the morphologically similar species E. minor and E. siliqua.

  7. Molecular structures of centromeric heterochromatin and karyotypic evolution in the Siamese crocodile (Crocodylus siamensis) (Crocodylidae, Crocodylia).

    Science.gov (United States)

    Kawagoshi, Taiki; Nishida, Chizuko; Ota, Hidetoshi; Kumazawa, Yoshinori; Endo, Hideki; Matsuda, Yoichi

    2008-01-01

    Crocodilians have several unique karyotypic features, such as small diploid chromosome numbers (30-42) and the absence of dot-shaped microchromosomes. Of the extant crocodilian species, the Siamese crocodile (Crocodylus siamensis) has no more than 2n = 30, comprising mostly bi-armed chromosomes with large centromeric heterochromatin blocks. To investigate the molecular structures of C-heterochromatin and genomic compartmentalization in the karyotype, characterized by the disappearance of tiny microchromosomes and reduced chromosome number, we performed molecular cloning of centromeric repetitive sequences and chromosome mapping of the 18S-28S rDNA and telomeric (TTAGGG)( n ) sequences. The centromeric heterochromatin was composed mainly of two repetitive sequence families whose characteristics were quite different. Two types of GC-rich CSI-HindIII family sequences, the 305 bp CSI-HindIII-S (G+C content, 61.3%) and 424 bp CSI-HindIII-M (63.1%), were localized to the intensely PI-stained centric regions of all chromosomes, except for chromosome 2 with PI-negative heterochromatin. The 94 bp CSI-DraI (G+C content, 48.9%) was tandem-arrayed satellite DNA and localized to chromosome 2 and four pairs of small-sized chromosomes. The chromosomal size-dependent genomic compartmentalization that is supposedly unique to the Archosauromorpha was probably lost in the crocodilian lineage with the disappearance of microchromosomes followed by the homogenization of centromeric repetitive sequences between chromosomes, except for chromosome 2.

  8. Telomere elongation in immortal human cells without detectable telomerase activity.

    Science.gov (United States)

    Bryan, T M; Englezou, A; Gupta, J; Bacchetti, S; Reddel, R R

    1995-09-01

    Immortalization of human cells is often associated with reactivation of telomerase, a ribonucleoprotein enzyme that adds TTAGGG repeats onto telomeres and compensates for their shortening. We examined whether telomerase activation is necessary for immortalization. All normal human fibroblasts tested were negative for telomerase activity. Thirteen out of 13 DNA tumor virus-transformed cell cultures were also negative in the pre-crisis (i.e. non-immortalized) stage. Of 35 immortalized cell lines, 20 had telomerase activity as expected, but 15 had no detectable telomerase. The 15 telomerase-negative immortalized cell lines all had very long and heterogeneous telomeres of up to 50 kb. Hybrids between telomerase-negative and telomerase-positive cells senesced. Two senescent hybrids demonstrated telomerase activity, indicating that activation of telomerase is not sufficient for immortalization. Some hybrid clones subsequently recommenced proliferation and became immortalized either with or without telomerase activity. Those without telomerase activity also had very long and heterogeneous telomeres. Taken together, these data suggest that the presence of lengthened or stabilized telomeres is necessary for immortalization, and that this may be achieved either by the reactivation of telomerase or by a novel and as yet unidentified mechanism.

  9. The telomere length dynamic and methods of its assessment.

    Science.gov (United States)

    Lin, Kah-Wai; Yan, Ju

    2005-01-01

    Human telomeres are composed of long repeating sequences of TTAGGG, associated with a variety of telomere-binding proteins. Its function as an end-protector of chromosomes prevents the chromosome from end-to-end fusion, recombination and degradation. Telomerase acts as reverse transcriptase in the elongation of telomeres, which prevent the loss of telomeres due to the end replication problems. However, telomerase activity is detected at low level in somatic cells and high level in embryonic stem cells and tumor cells. It confers immortality to embryonic stem cells and tumor cells. In most tumor cells, telomeres are extremely short and stable. Telomere length is an important indicator of the telomerase activity in tumor cells and it may be used in the prognosis of malignancy. Thus, the assessment of telomeres length is of great experimental and clinical significance. This review describes the role of telomere and telomerase in cancer pathogenesis and the dynamics of the telomeres length in different cell types. The various methods of measurement of telomeres length, i.e. southern blot, hybridization protection assay, fluorescence in situ hybridization, primed in situ, quantitative PCR and single telomere length analysis are discussed. The principle and comparative evaluation of these methods are reviewed. The detection of G-strand overhang by telomeric-oligonucleotide ligation assay, primer extension/nick translation assay and electron microscopy are briefly discussed.

  10. Mental health needs and availability of mental health care for children and adolescents with intellectual disability in Berlin.

    Science.gov (United States)

    Soltau, B; Biedermann, J; Hennicke, K; Fydrich, T

    2015-11-01

    The increased risk of mental health problems in children and adolescents with intellectual disability (ID) has been reported in several studies. However, almost no research has been conducted on parents' experiences with the general mental health system. We have investigated the prevalence of emotional and behavioural problems in children with ID as well as the availability and quality of mental health care from the parents' point of view. Teachers of specialised schools for ID in Berlin were asked to complete the Teacher's Report Form (TRF) of the Child Behavior Checklist. Information was collected for 1226 children and adolescents aged 6-18 years with mild to profound ID (response 70.5%). The availability and quality of mental health care was assessed by a questionnaire given to parents who had already been seeking help for their children. A total of 330 parents completed the questionnaires (response 62.0%). In addition to univariate analysis, we conducted multiple logistic regressions regarding the psychopathology reported by teachers (TRF-syndrome scales) and difficulties concerning mental health care reported by parents for a paired sample of 308 children. Overall, 52.4% of the children and adolescents with ID had a total problem score on the TRF in the deviant range (47.1% when eliminating four items reflecting cognitive deficits). Compared with the general population normative sample of children, this is a three-time higher prevalence. The most striking problems were thought problems (schizoid and obsessive-compulsive), aggressive behaviour, attention problems and social problems. Parents whose children had more severe behavioural or emotional dysfunction reported more difficulties with the mental health system. From the parents' point of view, mental health professionals frequently did not feel responsible or were not sufficiently skilled for the treatment of children with ID. As a consequence, 96% of all parents were longing for specialised in- and

  11. Safety Assessment of Tocotrienol Supplementation in Subjects with Metabolic Syndrome: A Randomised Control Trial

    International Nuclear Information System (INIS)

    Lin, G.Y.; Ming, L.O.; Nesaratnam, K.; Kim-Tiu, T.; Selvaduray, K.R.; Meganathan, P.; Yen, F.J.

    2016-01-01

    Previous studies have reported that tocotrienols (T3) possess many distinct properties such as antioxidant, cardioprotective, neuroprotective, anti-cancer, anti-inflammatory and anti-angiogenic, which are beneficial for the improvement of human health. However, there is limited data available on the safety assessment of T3 compared to tocopherols (T). A randomised, double-blinded, cross-over and placebo-controlled human clinical trial was conducted to determine the safety and tolerance of T3 supplementation in 31 subjects with metabolic syndrome. The subjects were supplemented with tocotrienol-rich fra tion (TRF) 200 mg or placebo capsules twice daily for two weeks followed by a post-intervention visit. Results showed that T3 supplementation had no significant adverse effect on the red blood cell (RBC), white blood cell (WBC) and platelet counts between TRF (5.10 ± 0.78 x 10"1"2 litre"-"1, 7.35 ± 1.59 x 10"9 litre"-"1, 279.45 ± 73.86 x 10"9 litre"-"1, respectively) and placebo interventions (5.13 ± 0.76 x 10"1"2 litre"-"1, 7.25 ± 1.95 x 10"9 litre"-"1, 267.45 ± 68.72 x 10"9 litre"-"1, respectively). Measures of serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT)) and albumin did not differ between TRF (25.68 ± 10.72 IU litre"-"1, 38.26 ± 24.74 IU litre"-"1, 43.61 ± 2.26 g litre"-"1, respectively) and placebo interventions (27.39 ± 16.44 IU litre"-"1, 42.23 ± 33.58 IU litre"-"1, 43.68 ± 2.15 g litre"-"1, respectively).This study indicated that supplementation with T3 at the dosage of 400 mg per day for 14 days did not induce haematoxicity and hepatotoxicity in subjects with metabolic syndrome. (author)

  12. [Influences of long-term application of organic and inorganic fertilizers on the composition and abundance of nirS-type denitrifiers in black soil].

    Science.gov (United States)

    Yin, Chang; Fan, Fen-Liang; Li, Zhao-Jun; Song, A-Lin; Zhu, Ping; Peng, Chang; Liang, Yong-Chao

    2012-11-01

    The objectives of this study were to explore the effects of long-term organic and inorganic fertilizations on the composition and abundance of nirS-type denitrifiers in black soil. Soil samples were collected from 4 treatments (i. e. no fertilizer treatment, CK; organic manure treatment, OM; chemical fertilizer treatment (NPK) and combination of organic and chemical fertilizers treatment (MNPK)) in Gongzhuling Long-term Fertilization Experiment Station. Composition and abundance of nirS-type denitrifiers were analyzed with terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR (Q-PCR), respectively. Denitrification enzyme activity (DEA) and soil properties were also measured. Application of organic fertilizers (OM and MNPK) significantly increased the DEAs of black soil, with the DEAs in OM and MNPK being 5.92 and 6.03 times higher than that in CK treatment, respectively, whereas there was no significant difference between NPK and CK. OM and MNPK treatments increased the abundances of nirS-type denitrifiers by 2.73 and 3.83 times relative to that of CK treatment, respectively. The abundance of nirS-type denitrifiers in NPK treatment was not significantly different from that of CK. The T-RFLP analysis of nirS genes showed significant differences in community composition between organic and inorganic treatments, with the emergence of a 79 bp T-RF, a significant decrease in relative abundance of the 84 bp T-RF and a loss of the 99 bp T-RF in all organic treatments. Phylogenetic analysis indicated that the airS-type denitrifiers in the black soil were mainly composed of alpha, beta and gamma-Proteobacteria. The 79 bp-type denitrifiers inhabiting exclusively in organic treatments (OM and MNPK) were affiliated to Pseudomonadaceae in gamma-Proteobacteria and Burkholderiales in beta-Proteobacteria. The 84 bp-types were related to Burkholderiales and Rhodocyclales. Correlation analysis indicated that pH, concentrations of total nitrogen

  13. Manejo comportamental em classe de crianças com transtorno de déficit de atenção e hiperatividade

    Directory of Open Access Journals (Sweden)

    Marcos Vinícius de Araújo

    2013-01-01

    Full Text Available Attention Deficit Hyperactivity Disorder (ADHD is characterized by a persistent pattern of inattention and / or hyperactivity. At school, it can contribute to academic and relationship difficulties. This work aimed to develop a guide for behavioral management of inattention and hyperactivity in the classroom and evaluate its use by a teacher in a class with student with ADHD. The instruments for pre-and post-intervention were: Scale Benczik of ADHD, Behaviors Inventory for Children form for parents (CBCL/6-18 and teachers (TRF/6-18 and a direct observation record of behaviors. The guide was conducted by a teacher for three months and accompanied by a psychologist every two weeks. The results showed a reduction in behaviors characteristic of ADHD that were directly observed and verified by behavioral inventories.

  14. Mechanisms of Disease Persistence in Chronic Myelogenous Leukemia (CML)

    Science.gov (United States)

    2006-10-01

    9.2 1.4 0.03 K-Ras* 6.7 1.3 0.04 RALA 5.4 1.1 0.13 Opioid receptor mu1 4.4 1.1 0.36 Jak2 4.3 1.2 0.14 TRF1 4.3 1.5 0.06 WT-1* n/a 1.3 0.008 c...2002 American Society of Hematology, Scientific Committee on Neoplasia 2000-2004 American Medical Informatics Association: Global Trial Bank...kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med 344:1038-1042

  15. Diferença de Classe no SUS é inconstitucional!

    Directory of Open Access Journals (Sweden)

    Alethele de Oliveira Santos

    2015-12-01

    Full Text Available A discussão sobre a ‘diferença de classe’ decorre do Recurso Extraordinário (RE 581488 interposto pelo Conselho Regional de Medicina do Rio Grande do Sul (Cremers em face da decisão do Tribunal Regional Federal da 4ªRegião (TRF –4ª, em favor do município de Canela (RS (1 junto ao Supremo Tribunal Federal (STF, com repercussão geral  reconhecida sob o número 579. Trata-se da possibilidade de que paciente do Sistema Único de Saúde (SUS pague valor financeiro por hotelaria diferenciada, ou ainda, médico de sua preferência.

  16. Gold nanoparticle immunochromatographic assay for quantitative detection of urinary RBP

    Directory of Open Access Journals (Sweden)

    XU Kuan

    2013-04-01

    Full Text Available A rapid quantitative detection of urinary RBP was established by using nano-gold immunochromatography (sandwich method and trisodium citrate reduction method and a rapid immunochromatographic test strip was developed. Theimmunochromatographic test strip can quantitatively detect RBP within 15 minutes. The detection limit was 150ng/mL and detection range was from 150 to 5000 ng/mL. There were no cross-reactions with others kidney disease markers,such as urinary albumin (ALB,transferrin protein (TRF,β2-microglobulin (β2-MG,urinary fiber connecting protein (FN,and lysozyme (LZM. The results indicate that it is a quick and simple method with strong specificity,high sensitivity,and wide detection range. The rapid detection method will have extensive clinical applications in the early diagnosis of proximal tubular damage,kidney disease,diabetic nephropathy,and process monitoring.

  17. Monitoring of Earth Rotation by VLBI

    Science.gov (United States)

    Ma., Chopo; Macmillan, D. S.

    2000-01-01

    Monitoring Earth rotation with Very Long Baseline Interferometry (VLBI) has unique potential because of direct access to the Celestial Reference System (CRF and Terrestrial Reference System (TRF) and the feasibility of re-analyzing the entire data set. While formal precision of better than 0.045 mas for pole and 0.002 ms for UT 1 has been seen in the best 24-hr data, the accuracy of the Earth Orientation Parameter (EOP) time series as a whole is subject to logistical, operational, analytical and conceptual constraints. The current issues related to the VLBI data set and the CORE program for greater time resolution such as analysis consistency, network jitter and reference frame stability will be discussed.

  18. Integration of Reference Frames Using VLBI

    Science.gov (United States)

    Ma, Chopo; Smith, David E. (Technical Monitor)

    2001-01-01

    Very Long Baseline Interferometry (VLBI) has the unique potential to integrate the terrestrial and celestial reference frames through simultaneous estimation of positions and velocities of approx. 40 active VLBI stations and a similar number of stations/sites with sufficient historical data, the position and position stability of approx. 150 well-observed extragalactic radio sources and another approx. 500 sources distributed fairly uniformly on the sky, and the time series of the five parameters that specify the relative orientation of the two frames. The full realization of this potential is limited by a number of factors including the temporal and spatial distribution of the stations, uneven distribution of observations over the sources and the sky, variations in source structure, modeling of the solid/fluid Earth and troposphere, logistical restrictions on the daily observing network size, and differing strategies for optimizing analysis for TRF, for CRF and for EOP. The current status of separately optimized and integrated VLBI analysis will be discussed.

  19. Acclimation of subsurface microbial communities to mercury

    DEFF Research Database (Denmark)

    de Lipthay, Julia R; Rasmussen, Lasse D; Øregaard, Gunnar

    2008-01-01

    of mercury tolerance and functional versatility of bacterial communities in contaminated soils initially were higher for surface soil, compared with the deeper soils. However, following new mercury exposure, no differences between bacterial communities were observed, which indicates a high adaptive potential......We studied the acclimation to mercury of bacterial communities of different depths from contaminated and noncontaminated floodplain soils. The level of mercury tolerance of the bacterial communities from the contaminated site was higher than those of the reference site. Furthermore, the level...... of the subsurface communities, possibly due to differences in the availability of mercury. IncP-1 trfA genes were detected in extracted community DNA from all soil depths of the contaminated site, and this finding was correlated to the isolation of four different mercury-resistance plasmids, all belonging...

  20. Telomere length of anterior crucial ligament after rupture

    DEFF Research Database (Denmark)

    Ponsot, Elodie; Langberg, Henning; Krogsgaard, Michael R

    2011-01-01

    The regeneration of ligaments following injury is a slow process compared to the healing of many other tissues and the underlying mechanisms remain unknown. The purpose of the study was to evaluate the proliferative potential of ligaments by assessing telomere length within three distinct parts...... of human anterior cruciate ligament (ACL) obtained during ACL reconstruction: the macroscopically injured proximal part and macroscopically noninjured mid- and distal portions in eight subjects (age 28 ± 8 years). The mean telomere length in ACL was within normal range of values usually reported for other...... tissues indicating that the endogenous machinery responsible for the proliferative potential of ligament is not implicated in its poor healing capacity. The three ACL parts showed similar mean TRF lengths (distal part: 11.5 ± 0.8 kbp, mid-portion: 11.8 ± 1.2 kbp, proximal part: 11.9 ± 1.6 kbp...

  1. Detritiation in Romania

    International Nuclear Information System (INIS)

    Stefan, Liviu; Stefanescu, Ioan

    2007-01-01

    Full text: Isotope separation in nuclear field started in Romania as a basic issue in the Romanian Nuclear Program, when the use of CANDU type nuclear power plant was adopted. Therefore, an objective was imposed namely to develop the Romanian technology for heavy water production at an industrial scale. In 1970 the heavy water program started by constructing the industrial plant - Uzina G - at Rm. Valcea. The activities at this plant resulted in obtaining the Romanian license for heavy water technology which was transposed later to industrial plant at ROMAG Drobeta Tr. Severin. Based on experience achieved in isotope separation for heavy water, since 1992 it was started a tritium program at ICSI Rm. Valcea (former Uzina G) with the objective of developing a Romanian technology for Tritium Removal Facility (TRF). (authors)

  2. Characterization of a "TRAMP-like" co-factor of the human RNA exosome

    DEFF Research Database (Denmark)

    Christensen, Marianne Skovgaard; Kristiansen, Maiken Søndergaard; Lubas, Michal Szymon

    Genome-wide studies in yeast, plants and humans have revealed numerous new transcripts in what was previously thought to be silent DNA or junk DNA. One class of non-coding transcript discovered recently is the PROMoter uPstream Transcripts (PROMPTs), which is only seen upon depletion of the RNA...... exosome, the major 3’-5’ exonuclease complex in human cells. PROMPTs have a lot in common with the yeast Cryptic Unstable Transcripts (CUTs), which are degraded by the concerted effort of the exosome, and its co-factor complex TRAMP (Trf4p/Air1p/Mtr4p). We have identified human proteins with functional...... similarities to components of the yeast TRAMP complex, and show that these are involved in the degradation of PROMPTs. While, these proteins form transient complexes with the exosome, our preliminary results also indicate that complex formation can occur directly with catalytic components of the exosome...

  3. Multi-informant reports of psychiatric symptoms among high-functioning adolescents with Asperger syndrome or autism.

    Science.gov (United States)

    Hurtig, Tuula; Kuusikko, Sanna; Mattila, Marja-Leena; Haapsamo, Helena; Ebeling, Hanna; Jussila, Katja; Joskitt, Leena; Pauls, David; Moilanen, Irma

    2009-11-01

    The aim of the study was to examine psychiatric symptoms in high-functioning adolescents with autism spectrum disorders reported by multiple informants. Forty-three 11- to 17-year-old adolescents with Asperger syndrome (AS) or high-functioning autism (HFA) and 217 typically developed adolescents completed the Youth Self-Report (YSR), while their parents completed the Child Behavior Checklist (CBCL). Teachers of adolescents with AS/HFA completed the Teacher Report Form (TRF). The informants reported significantly more psychiatric symptoms, especially withdrawn, anxious/depressed, social and attention problems, in adolescents with AS/HFA than in controls. In contrast to findings in the general population, the psychiatric problems of adolescents with AS/HFA are well acknowledged by multiple informants, including self-reports. However, anxiety and depressive symptoms were more commonly reported by adolescents with AS/HFA and their teachers than their parents, indicating that some emotional distress may be hidden from their parents.

  4. Implementation of Different Variants of Table-Based Frequency Synthesizers with Quadrature Output in VHDL

    Directory of Open Access Journals (Sweden)

    Daniel Kekrt

    2012-01-01

    Full Text Available This article describes the modelling and implementation of two different variants of direct frequency synthesizer, and evaluation of the performance of the finished design, in terms of memory and speed efficiency. The frequency synthesizer requirement comes from our complex radio transmission system design. The research activity has been focused on finding an optimal balance between simplicity, speed and memory consumption. The modelling was done in MATLAB environment in floating-point and fixed-point arithmetic, and the actual design was implemented and synthesized using the Xilinx ISE suite. The output has been connected to our customized radio front-end built on the Texas Instruments TRF2443 chip. The front-end output signal has been captured and compared with simulation results.

  5. Mice with Pulmonary Fibrosis Driven by Telomere Dysfunction

    Directory of Open Access Journals (Sweden)

    Juan M. Povedano

    2015-07-01

    Full Text Available Idiopathic pulmonary fibrosis (IPF is a degenerative disease of the lungs with an average survival post-diagnosis of 2–3 years. New therapeutic targets and treatments are necessary. Mutations in components of the telomere-maintenance enzyme telomerase or in proteins important for telomere protection are found in both familial and sporadic IPF cases. However, the lack of mouse models that faithfully recapitulate the human disease has hampered new advances. Here, we generate two independent mouse models that develop IPF owing to either critically short telomeres (telomerase-deficient mice or severe telomere dysfunction in the absence of telomere shortening (mice with Trf1 deletion in type II alveolar cells. We show that both mouse models develop pulmonary fibrosis through induction of telomere damage, thus providing proof of principle of the causal role of DNA damage stemming from dysfunctional telomeres in IPF development and identifying telomeres as promising targets for new treatments.

  6. Análisis molecular de la longitud telomérica en linfomas foliculares: Su participación en la progresión tumoral Molecular analysis of telomere length in follicular lymphomas. Its participation in tumor progression

    Directory of Open Access Journals (Sweden)

    Alejandra S. H. Cottliar

    2005-04-01

    Full Text Available Los telómeros son estructuras esenciales para el mantenimiento de la integridad cromosómica y la capacidad replicativa de la célula. La reducción de la longitud telomérica (LT aumenta la probabilidad de producir errores capaces de generar cambios genómicos importantes para el desarrollo neoplásico, determinando desbalances de material genético. En este trabajo se evaluó la LT mediante el análisis de fragmentos de restricción terminal (TRF en médula ósea y/o biopsia ganglionar de 36 pacientes (edad media: 54.2 años; rango 29-77 años; 21 varones: 29 con linfoma folicular (LF al diagnóstico y 7 con linfoma B difuso a células grandes secundario a LF (LBDCG-S. Se efectuó el análisis del rearreglo molecular del gen BCL-2 por PCR anidada y de larga distancia. Las medias de TRF en LF (4.18±0.18 Kb y LBDCG-S (3.31±0.25 Kb resultaron significativamente menores que en controles (8.50±0.50 Kb (pTelomeres are essential for maintaining chromosomal integrity and stability. We studied here telomere length (TL in bone marrow and/or lymph node from 36 patients: 29 with follicular lymphoma (FL at diagnosis and 7 with diffuse large B cell lymphoma secondary to FL (S-DLBCL. TL was evaluated using terminal restriction fragments (TRF assay. BCL-2 gene rearrangement was analyzed by nested and long distance PCR. Mean TRF values showed significant telomere shortening in FL (4.18±0.18 Kb and S-DLBCL (3.31±0.25 Kb respect to controls (8.50±0.50 Kb (p<0.001. Differences between both histological subtypes (p=0.036 were also detected. Moreover, the samples positive for BCL-2 rearrangements showed longer TL (4.25±0.19 Kb than the negative ones (3.39±0.30 Kb (p=0.023. A trend to telomere shortening was observed when Major Breakpoint Region (MBR-J H, minor cluster region (mcr-J H and BCL-2 negative patients were compared (4.35±0.21 Kb; 3.84±0.45 Kb and 3.39±0.30 Kb, respectively. Our results show a TL reduction in FL and S-DLBCL, with significant

  7. Use of fluorocarbons in the cooling of LHC experiments

    CERN Document Server

    Pimenta dos Santos, M

    2003-01-01

    Perfluorochemicals sold by 3M under the trade name 3M Fluorinert Electronic Liquids have been used for many years as heat transfer media in a variety of industries. The suitability of these liquids for the cooling of LHC experiment originates from their high dielectric strength as well as from their chemical stability under ionizing radiation. The Fluorinerts are clear, colorless, non-flammable with low toxicity and low corrosiveness. Additionally, they offer low global waming potential – GWP – and zero ozone-depletion potential – ODP. Some examples of fluorinert application in the cooling of LHC experiments will be presented : (a) the ATLAS Inner detector C3F8 evaporative cooling system (b) the ATLAS TRF C6F14 monophase cooling system and (c) the ALICE SPD “active heat pipe” C4F10 evaporative cooling system. A brief comparison of evaporative and monophase cooling systems will be outlined.

  8. Data and material of the Safe-Range-Inventory: An assistance tool helping to improve the charging infrastructure for electric vehicles.

    Science.gov (United States)

    Carbon, Claus-Christian; Gebauer, Fabian

    2017-10-01

    The Safe-Range-Inventory (SRI) was constructed in order to help public authorities to improve the charging infrastructures for electric vehicles [1; 10.1016/j.trf.2017.04.011]. Specifically, the impact of fast (vs slow) charging stations on people's range anxiety was examined. Ninety-seven electric vehicle users from Germany (81 male; M age =46.3 years, SD =12.1) were recruited to participate in the experimental design. Statistical analyses were conducted using ANOVA for repeated measures to test for interaction effects of available charging stations and remaining range with the dependent variable range anxiety . The full data set is publicly available via https://osf.io/bveyw/ (Carbon and Gebauer, 2017) [2].

  9. CrRNA-Protospacer Recognition during CRISPR- Directed DNA Interference Sulfolobus islandicus REY 15A and Structural Studies of CRISPR Binding Proteins (CBP) of Crenarchaeon Sulfolobus

    DEFF Research Database (Denmark)

    Mousaei, Marzieh

    The CRISPR-Cas (clustered regularly interspaced short palindromic repeats and associated proteins) is one of the important known immune mechanisms in archaea and bacteria. This adaptive immune system degrades invading genetic elements and protects the cell. Amongst 3 main types I, II and III...... of CRISPR system, two types (I and III) are found in archaea. However, in Sulfolobus species, subtypes IA, I-D, and III-B, III-D and rarely III-A are found. The model organism used for interference and structural studies is S. islandicus REY15A which carries subtypes I-A and III-B (α and β). Besides CRISPR...... ribonucleoprotein complex which is involved directly in defense, there are some less- known parts of the system including CPBs (CRISPR repeat-binding proteins) which are suggested to play a role in transcription. In the first part of my thesis, I provide a brief introduction to archaea and viruses that infect...

  10. Near-atomic model of microtubule-tau interactions.

    Science.gov (United States)

    Kellogg, Elizabeth H; Hejab, Nisreen M A; Poepsel, Simon; Downing, Kenneth H; DiMaio, Frank; Nogales, Eva

    2018-06-15

    Tau is a developmentally regulated axonal protein that stabilizes and bundles microtubules (MTs). Its hyperphosphorylation is thought to cause detachment from MTs and subsequent aggregation into fibrils implicated in Alzheimer's disease. It is unclear which tau residues are crucial for tau-MT interactions, where tau binds on MTs, and how it stabilizes them. We used cryo-electron microscopy to visualize different tau constructs on MTs and computational approaches to generate atomic models of tau-tubulin interactions. The conserved tubulin-binding repeats within tau adopt similar extended structures along the crest of the protofilament, stabilizing the interface between tubulin dimers. Our structures explain the effect of phosphorylation on MT affinity and lead to a model of tau repeats binding in tandem along protofilaments, tethering together tubulin dimers and stabilizing polymerization interfaces. Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

  11. Telomere erosion varies during in vitro aging of normal human fibroblasts from young and adult donors.

    Science.gov (United States)

    Figueroa, R; Lindenmaier, H; Hergenhahn, M; Nielsen, K V; Boukamp, P

    2000-06-01

    The life span of normal fibroblasts in vitro (Hayflick limit) depends on donor age, and telomere shortening has been proposed as a potential mechanism. By quantitative fluorescence in situ hybridization and Southern blot analysis, we show progressive telomere loss to about 5 kb mean telomere restriction fragment length in fibroblasts from two adult donors within 40 population doublings, whereas in fibroblasts from two infant donors, telomere erosion is reduced, leaving a mean telomere restriction fragment length of approximately 7 kb at senescence (after approximately 60 population doublings). Aging of fibroblasts from both infant and adult donors was not accompanied by chromosomal abnormalities but was correlated with increased telomere repeat-binding factor 2 expression at both the protein and transcriptional level.

  12. Space Geodetic Technique Co-location in Space: Simulation Results for the GRASP Mission

    Science.gov (United States)

    Kuzmicz-Cieslak, M.; Pavlis, E. C.

    2011-12-01

    The Global Geodetic Observing System-GGOS, places very stringent requirements in the accuracy and stability of future realizations of the International Terrestrial Reference Frame (ITRF): an origin definition at 1 mm or better at epoch and a temporal stability on the order of 0.1 mm/y, with similar numbers for the scale (0.1 ppb) and orientation components. These goals were derived from the requirements of Earth science problems that are currently the international community's highest priority. None of the geodetic positioning techniques can achieve this goal alone. This is due in part to the non-observability of certain attributes from a single technique. Another limitation is imposed from the extent and uniformity of the tracking network and the schedule of observational availability and number of suitable targets. The final limitation derives from the difficulty to "tie" the reference points of each technique at the same site, to an accuracy that will support the GGOS goals. The future GGOS network will address decisively the ground segment and to certain extent the space segment requirements. The JPL-proposed multi-technique mission GRASP (Geodetic Reference Antenna in Space) attempts to resolve the accurate tie between techniques, using their co-location in space, onboard a well-designed spacecraft equipped with GNSS receivers, a SLR retroreflector array, a VLBI beacon and a DORIS system. Using the anticipated system performance for all four techniques at the time the GGOS network is completed (ca 2020), we generated a number of simulated data sets for the development of a TRF. Our simulation studies examine the degree to which GRASP can improve the inter-technique "tie" issue compared to the classical approach, and the likely modus operandi for such a mission. The success of the examined scenarios is judged by the quality of the origin and scale definition of the resulting TRF.

  13. A homogeneous assay principle for universal substrate quantification via hydrogen peroxide producing enzymes

    International Nuclear Information System (INIS)

    Zscharnack, Kristin; Kreisig, Thomas; Prasse, Agneta A.; Zuchner, Thole

    2015-01-01

    Highlights: • Application of the TRF-based PATb system for universal oxidase substrate detection. • H 2 O 2 generated by choline or glucose oxidase quenches the TRF signal of PATb. • The assay time is only limited by the oxidase catalysis rate. • Glucose is precisely detected in human serum consistent to a commercial assay. • A reliable quantification of choline in infant formula is shown. - Abstract: H 2 O 2 is a widely occurring molecule which is also a byproduct of a number of enzymatic reactions. It can therefore be used to quantify the corresponding enzymatic substrates. In this study, the time-resolved fluorescence emission of a previously described complex consisting of phthalic acid and terbium (III) ions (PATb) is used for H 2 O 2 detection. In detail, glucose oxidase and choline oxidase convert glucose and choline, respectively, to generate H 2 O 2 which acts as a quencher for the PATb complex. The response time of the PATb complex toward H 2 O 2 is immediate and the assay time only depends on the conversion rate of the enzymes involved. The PATb assay quantifies glucose in a linear range of 0.02–10 mmol L −1 , and choline from 1.56 to 100 μmol L −1 with a detection limit of 20 μmol L −1 for glucose and 1.56 μmol L −1 for choline. Both biomolecules glucose and choline could be detected without pretreatment with good precision and reproducibility in human serum samples and infant formula, respectively. Furthermore, it is shown that the detected glucose concentrations by the PATb system agree with the results of a commercially available assay. In principle, the PATb system is a universal and versatile tool for the quantification of any substrate and enzyme reaction where H 2 O 2 is involved

  14. Functional characterization of replication and stability factors of an incompatibility group P-1 plasmid from Xylella fastidiosa.

    Science.gov (United States)

    Lee, Min Woo; Rogers, Elizabeth E; Stenger, Drake C

    2010-12-01

    Xylella fastidiosa strain riv11 harbors a 25-kbp plasmid (pXF-RIV11) belonging to the IncP-1 incompatibility group. Replication and stability factors of pXF-RIV11 were identified and used to construct plasmids able to replicate in X. fastidiosa and Escherichia coli. Replication in X. fastidiosa required a 1.4-kbp region from pXF-RIV11 containing a replication initiation gene (trfA) and the adjacent origin of DNA replication (oriV). Constructs containing trfA and oriV from pVEIS01, a related IncP-1 plasmid of the earthworm symbiont Verminephrobacter eiseniae, also were competent for replication in X. fastidiosa. Constructs derived from pXF-RIV11 but not pVEIS01 replicated in Agrobacterium tumefaciens, Xanthomonas campestris, and Pseudomonas syringae. Although plasmids bearing replication elements from pXF-RIV11 or pVEIS01 could be maintained in X. fastidiosa under antibiotic selection, removal of selection resulted in plasmid extinction after 3 weekly passages. Addition of a toxin-antitoxin addiction system (pemI/pemK) from pXF-RIV11 improved plasmid stability such that >80 to 90% of X. fastidiosa cells retained plasmid after 5 weekly passages in the absence of antibiotic selection. Expression of PemK in E. coli was toxic for cell growth, but toxicity was nullified by coexpression of PemI antitoxin. Deletion of N-terminal sequences of PemK containing the conserved motif RGD abolished toxicity. In vitro assays revealed a direct interaction of PemI with PemK, suggesting that antitoxin activity of PemI is mediated by toxin sequestration. IncP-1 plasmid replication and stability factors were added to an E. coli cloning vector to constitute a stable 6.0-kbp shuttle vector (pXF20-PEMIK) suitable for use in X. fastidiosa.

  15. Integration of replication-defective R68.45-like plasmids into the Pseudomonas aeruginosa chromosome.

    Science.gov (United States)

    Reimmann, C; Rella, M; Haas, D

    1988-06-01

    R68.45 and other similar broad-host-range (IncP) plasmids carrying a tandem repeat of the 2.1 kb insertion element IS21 mobilize the chromosome of many different Gram-negative bacteria. To analyse the structure of R68.45-chromosome cointegrates, whose involvement in the mobilization process had been postulated previously, we selected for the stable integration of R68.45-like plasmids into the Pseudomonas aeruginosa chromosome. Two plasmids were chosen: pME28, a transfer-deficient, mobilizable RP1 derivative with an inactive replication control (trfA) gene, and pME487, an R68.45 derivative with a trfA(ts) mutation causing temperature-sensitive replication. Chromosomally integrated pME28 and pME487 were found to be flanked by single IS21 elements. This structure is in agreement with a 'cut-and-paste' mode of R68.45 transposition. pME28 and pME487 showed a low specificity of insertion but rarely (less than 0.1%) induced auxotrophic mutations. Hfr (high-frequency-of-recombination) donors of P. aeruginosa could be obtained by chromosomal integration of pME487 or pME28; in the latter case, the transfer functions lacking from pME28 had to be provided in trans on an autonomous plasmid. Hfr donors gave higher conjugational linkage and transferred longer stretches of the P. aeruginosa chromosome than did R68.45 donors. This suggests that the integration of R68.45 into the donor chromosome is short-lived in P. aeruginosa.

  16. [Effect of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation: a randomized controlled trial].

    Science.gov (United States)

    Cai, Jun; Wang, Hua; Zhou, Sheng; Wu, Bin; Song, Hua-Rong; Xuan, Zheng-Rong

    2008-01-01

    To observe the effect of perioperative application of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation. In this prospective, single-blinded, controlled clinical trial, fifty-nine patients with gastric cancer were randomly divided into three groups: control group (n=20) and two study groups (group A, n=21; group B, n=18). Sjunzi Decoction (100 ml) was administered via nasogastric tube to the patients in the study group B from the second postoperation day to the 9th postoperation day. Patients in the two study groups were given an isocaloric and isonitrogonous enteral diet, which was started on the second day after operation, and continued for eight days. Patients in the control group were given an isocaloric and isonitrogonous parenteral diet for 9 days. All variables of nutritional status such as serum albumin (ALB), prealbumin (PA), transferrin (TRF) and T-cell subsets were measured one day before operation, and one day and 10 days after operation. All the nutritional variables and the levels of CD3(+), CD4(+), CD4(+)/CD8(+) were decreased significantly after operation. Ten days after operation, T-cell subsets and nutritional variables in the two study groups were increased as compare with the control group. The levels of ALB, TRF and T-cell subsets in the study group B were increased significantly as compared with the study group A (Pnutrition assisted with Sijunzi Decoction can positively improve and optimize cellular immune function and nutritional status in the patients with gastric cancer after operation.

  17. Filtering observations without the initial guess

    Science.gov (United States)

    Chin, T. M.; Abbondanza, C.; Gross, R. S.; Heflin, M. B.; Parker, J. W.; Soja, B.; Wu, X.

    2017-12-01

    Noisy geophysical observations sampled irregularly over space and time are often numerically "analyzed" or "filtered" before scientific usage. The standard analysis and filtering techniques based on the Bayesian principle requires "a priori" joint distribution of all the geophysical parameters of interest. However, such prior distributions are seldom known fully in practice, and best-guess mean values (e.g., "climatology" or "background" data if available) accompanied by some arbitrarily set covariance values are often used in lieu. It is therefore desirable to be able to exploit efficient (time sequential) Bayesian algorithms like the Kalman filter while not forced to provide a prior distribution (i.e., initial mean and covariance). An example of this is the estimation of the terrestrial reference frame (TRF) where requirement for numerical precision is such that any use of a priori constraints on the observation data needs to be minimized. We will present the Information Filter algorithm, a variant of the Kalman filter that does not require an initial distribution, and apply the algorithm (and an accompanying smoothing algorithm) to the TRF estimation problem. We show that the information filter allows temporal propagation of partial information on the distribution (marginal distribution of a transformed version of the state vector), instead of the full distribution (mean and covariance) required by the standard Kalman filter. The information filter appears to be a natural choice for the task of filtering observational data in general cases where prior assumption on the initial estimate is not available and/or desirable. For application to data assimilation problems, reduced-order approximations of both the information filter and square-root information filter (SRIF) have been published, and the former has previously been applied to a regional configuration of the HYCOM ocean general circulation model. Such approximation approaches are also briefed in the

  18. A two-level approach to VLBI terrestrial and celestial reference frames using both least-squares adjustment and Kalman filter algorithms

    Science.gov (United States)

    Soja, B.; Krasna, H.; Boehm, J.; Gross, R. S.; Abbondanza, C.; Chin, T. M.; Heflin, M. B.; Parker, J. W.; Wu, X.

    2017-12-01

    The most recent realizations of the ITRS include several innovations, two of which are especially relevant to this study. On the one hand, the IERS ITRS combination center at DGFI-TUM introduced a two-level approach with DTRF2014, consisting of a classical deterministic frame based on normal equations and an optional coordinate time series of non-tidal displacements calculated from geophysical loading models. On the other hand, the JTRF2014 by the combination center at JPL is a time series representation of the ITRF determined by Kalman filtering. Both the JTRF2014 and the second level of the DTRF2014 are thus able to take into account short-term variations in the station coordinates. In this study, based on VLBI data, we combine these two approaches, applying them to the determination of both terrestrial and celestial reference frames. Our product has two levels like DTRF2014, with the second level being a Kalman filter solution like JTRF2014. First, we compute a classical TRF and CRF in a global least-squares adjustment by stacking normal equations from 5446 VLBI sessions between 1979 and 2016 using the Vienna VLBI and Satellite Software VieVS (solution level 1). Next, we obtain coordinate residuals from the global adjustment by applying the level-1 TRF and CRF in the single-session analysis and estimating coordinate offsets. These residuals are fed into a Kalman filter and smoother, taking into account the stochastic properties of the individual stations and radio sources. The resulting coordinate time series (solution level 2) serve as an additional layer representing irregular variations not considered in the first level of our approach. Both levels of our solution are implemented in VieVS in order to test their individual and combined performance regarding the repeatabilities of estimated baseline lengths, EOP, and radio source coordinates.

  19. Assessing for suicidal behavior in youth using the Achenbach System of Empirically Based Assessment.

    Science.gov (United States)

    Van Meter, Anna R; Algorta, Guillermo Perez; Youngstrom, Eric A; Lechtman, Yana; Youngstrom, Jen K; Feeny, Norah C; Findling, Robert L

    2018-02-01

    This study investigated the clinical utility of the Achenbach System of Empirically Based Assessment (ASEBA) for identifying youth at risk for suicide. Specifically, we investigated how well the Total Problems scores and the sum of two suicide-related items (#18 "Deliberately harms self or attempts suicide" and #91 "Talks about killing self") were able to distinguish youth with a history of suicidal behavior. Youth (N = 1117) aged 5-18 were recruited for two studies of mental illness. History of suicidal behavior was assessed by semi-structured interviews (K-SADS) with youth and caregivers. Youth, caregivers, and a primary teacher each completed the appropriate form (YSR, CBCL, and TRF, respectively) of the ASEBA. Areas under the curve (AUCs) from ROC analyses and diagnostic likelihood ratios (DLRs) were used to measure the ability of both Total Problems T scores, as well as the summed score of two suicide-related items, to identify youth with a history of suicidal behavior. The Suicide Items from the CBCL and YSR performed well (AUCs = 0.85 and 0.70, respectively). The TRF Suicide Items did not perform better than chance, AUC = 0.45. The AUCs for the Total Problems scores were poor-to-fair (0.33-0.65). The CBCL Suicide Items outperformed all other scores (ps = 0.04 to youth's risk for suicidal behavior. The low burden of this approach could facilitate wide-spread screening for suicide in an increasingly at-risk population.

  20. [Effects of low-protein diet plus alpha-keto acid on micro-inflammation and the relationship between micro-inflammation and nutritional status in patients performing continuous ambulatory peritoneal dialysis: a randomized controlled trial].

    Science.gov (United States)

    Chen, Wei; Guo, Zhi-Yong; Wu, Hao; Sun, Li-Jing; Cai, Li-Li; Xu, Hai-Yan

    2008-05-01

    To investigate the effects of the combination of alpha-keto acid and low-protein diet on the levels of serum cytokines in patients performing continuous ambulatory peritoneal dialysis (CAPD) and to explore the relationship between inflammation and malnutrition in CAPD patients. Eighty-nine CAPD patients were randomized into three groups, and 78 cases completed a one-year follow-up and with complete data. There were 31 cases in low-protein diet plus alpha-keto acid group, 26 cases in low-protein diet group and 21 cases in routine-protein diet group. The levels of serum albumin (Alb), prealbumin (PA), retinol-binding protein (RBP), transferrin (TRF), cholesterol (TC), triglycerides (TG), leptin, and triceps skinfold thickness (TSF), mid-arm muscle circumference (MAMC), body mass index (BMI) were measured. The changes of serum interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and C-reactive protein (CRP) were also detected. Compared with low-protein diet group, serum levels of PA, RBP and TRF were significantly increased both in low-protein diet plus alpha-keto acid and routine-protein diet groups ( Pdiet plus alpha-keto acid group and routine-protein diet group. There was an increased tendency in the content of Alb, TC, TG, BMI, TSF and MAMC, but there were no significant differences. The plasma levels of IL-1alpha, IL-6 and TNF-alpha in low-protein diet plus alpha-keto acid group were decreased as compared with the routine-protein diet group, but there were no significant differences. The plasma level of CRP in low-protein diet plus alpha-keto acid group was lower than that in the routine-protein diet group ( Pketo acid and low-protein diet can ameliorate malnutrition and micro-inflammation in CAPD patients.

  1. Effects of different fertilizers on the abundance and community structure of ammonia oxidizers in a yellow clay soil.

    Science.gov (United States)

    Yao, Huaiying; Huang, Sha; Qiu, Qiongfen; Li, Yaying; Wu, Lianghuan; Mi, Wenhai; Dai, Feng

    2016-08-01

    Yellow clay paddy soil (Oxisols) is a typical soil with low productivity in southern China. Nitrification inhibitors and slow release fertilizers have been used to improve nitrogen fertilizer utilization and reduce environmental impaction of the paddy soil. However, their effects on ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in paddy soil have rarely been investigated. In the present work, we compared the influences of several slow release fertilizers and nitrification inhibitors on the community structure and activities of the ammonia oxidizers in yellow clay soil. The abundances and community compositions of AOA and AOB were determined with qPCR, terminal restriction fragment length polymorphism (T-RFLP), and clone library approaches. Our results indicated that the potential nitrification rate (PNR) of the soil was significantly related to the abundances of both AOA and AOB. Nitrogen fertilizer application stimulated the growth of AOA and AOB, and the combinations of nitrapyrin with urea (NPU) and urea-formaldehyde (UF) inhibited the growth of AOA and AOB, respectively. Compared with other treatments, the applications of NPU and UF also led to significant shifts in the community compositions of AOA and AOB, respectively. NPU showed an inhibitory effect on AOA T-RF 166 bp that belonged to Nitrosotalea. UF had a negative effect on AOB T-RF 62 bp that was assigned to Nitrosospira. These results suggested that NPU inhibited PNR and increased nitrogen use efficiency (NUE) by inhibiting the growth of AOA and altering AOA community. UF showed no effect on NUE but decreased AOB abundance and shifted AOB community.

  2. Effect of early nutrition support on postoperative rehabilitation in patients with oral cancer

    Directory of Open Access Journals (Sweden)

    Xing-An Zhang

    2016-08-01

    Full Text Available Objective: To explore the effect of early nutrition support on the postoperative nutrition metabolism and rehabilitation in patients with oral cancer. Methods: A total of 80 patients with oral cancer who were admitted in our hospital from January, 2013 to January, 2015 were included in the study, and divided into the observation group (early nutrition support and control group (routine treatments with 40 cases in each group. The postoperative basic nutrition requirement amount in the observation group was calculated according to HarrisNenedict formula. The appropriate pharmaceutics and nutritional pathway were selected. The patients in the control group were given routine diets after operation. The levels of ALB, PAB, TRF, TCL, IgM, IgG, and IgA before and after operation in the two groups were compared. The body mass and wound healing in the two groups were recorded and compared. Results: The body mass and serum TRF level after operation in the observation group were slightly reduced, but were not significantly different from those before operation (P>0.05. The serum ALB, PAB, and TLC levels after operation in the observation group were significantly reduced when compared with before operation (P0.05, but the above indicators were significantly elevated after operation (P<0.05. The peripheral blood IgM, IgG, and IgA levels 1 week after operation in the observation group were significantly higher than those in the control group (P<0.05. The stage I healing rate of the surgical wound in the observation group was significantly higher than that in the control group (P<0.05. Conclusions: The early nutrition support can effectively enhance the postoperative nutrition status and immunological function in patients with oral cancer, and is beneficial for the postoperative rehabilitation.

  3. Global diversification of a tropical plant growth form: environmental correlates and historical contingencies in climbing palms.

    Science.gov (United States)

    Couvreur, Thomas L P; Kissling, W Daniel; Condamine, Fabien L; Svenning, Jens-Christian; Rowe, Nick P; Baker, William J

    2014-01-01

    Tropical rain forests (TRF) are the most diverse terrestrial biome on Earth, but the diversification dynamics of their constituent growth forms remain largely unexplored. Climbing plants contribute significantly to species diversity and ecosystem processes in TRF. We investigate the broad-scale patterns and drivers of species richness as well as the diversification history of climbing and non-climbing palms (Arecaceae). We quantify to what extent macroecological diversity patterns are related to contemporary climate, forest canopy height, and paleoclimatic changes. We test whether diversification rates are higher for climbing than non-climbing palms and estimate the origin of the climbing habit. Climbers account for 22% of global palm species diversity, mostly concentrated in Southeast Asia. Global variation in climbing palm species richness can be partly explained by past and present-day climate and rain forest canopy height, but regional differences in residual species richness after accounting for current and past differences in environment suggest a strong role of historical contingencies in climbing palm diversification. Climbing palms show a higher net diversification rate than non-climbers. Diversification analyses of palms detected a diversification rate increase along the branches leading to the most species-rich clade of climbers. Ancestral character reconstructions revealed that the climbing habit originated between early Eocene and Miocene. These results imply that changes from non-climbing to climbing habits may have played an important role in palm diversification, resulting in the origin of one fifth of all palm species. We suggest that, in addition to current climate and paleoclimatic changes after the late Neogene, present-day diversity of climbing palms can be explained by morpho-anatomical innovations, the biogeographic history of Southeast Asia, and/or ecological opportunities due to the diversification of high-stature dipterocarps in Asian TRFs.

  4. Space geodesy validation of the global lithospheric flow

    Science.gov (United States)

    Crespi, M.; Cuffaro, M.; Doglioni, C.; Giannone, F.; Riguzzi, F.

    2007-02-01

    Space geodesy data are used to verify whether plates move chaotically or rather follow a sort of tectonic mainstream. While independent lines of geological evidence support the existence of a global ordered flow of plate motions that is westerly polarized, the Terrestrial Reference Frame (TRF) presents limitations in describing absolute plate motions relative to the mantle. For these reasons we jointly estimated a new plate motions model and three different solutions of net lithospheric rotation. Considering the six major plate boundaries and variable source depths of the main Pacific hotspots, we adapted the TRF plate kinematics by global space geodesy to absolute plate motions models with respect to the mantle. All three reconstructions confirm (i) the tectonic mainstream and (ii) the net rotation of the lithosphere. We still do not know the precise trend of this tectonic flow and the velocity of the differential rotation. However, our results show that assuming faster Pacific motions, as the asthenospheric source of the hotspots would allow, the best lithospheric net rotation estimate is 13.4 +/- 0.7 cm yr-1. This superfast solution seems in contradiction with present knowledge on the lithosphere decoupling, but it matches remarkably better with the geological constraints than those retrieved with slower Pacific motion and net rotation estimates. Assuming faster Pacific motion, it is shown that all plates move orderly `westward' along the tectonic mainstream at different velocities and the equator of the lithospheric net rotation lies inside the corresponding tectonic mainstream latitude band (~ +/-7°), defined by the 1σ confidence intervals.

  5. Associations between prenatal cigarette smoke exposure and externalized behaviors at school age among Inuit children exposed to environmental contaminants.

    Science.gov (United States)

    Desrosiers, Caroline; Boucher, Olivier; Forget-Dubois, Nadine; Dewailly, Eric; Ayotte, Pierre; Jacobson, Sandra W; Jacobson, Joseph L; Muckle, Gina

    2013-01-01

    Smoking during pregnancy is common among Inuit women from the Canadian Arctic. Yet prenatal cigarette smoke exposure (PCSE) is seen as a major risk factor for childhood behavior problems. Recent data also suggest that co-exposure to neurotoxic environmental contaminants can exacerbate the effects of PCSE on behavior. This study examined the association between PCSE and behavior at school age in a sample of Inuit children from Nunavik, Québec, where co-exposure to environmental contaminants is also an important issue. Interactions with lead (Pb) and mercury (Hg), two contaminants associated with behavioral problems, were also explored. Participants were 271 children (mean age=11.3years) involved in a prospective birth-cohort study. PCSE was assessed through maternal recall. Assessment of child behavior was obtained from the child's classroom teacher on the Teacher Report Form (TRF) and the Disruptive Behavior Disorders Rating Scale (DBD). Exposure to contaminants was assessed from umbilical cord and child blood samples. Other confounders were documented by maternal interview. After control for contaminants and confounders, PCSE was associated with increased externalizing behaviors and attention problems on the TRF and higher prevalence of attention deficit hyperactivity disorder (ADHD) assessed on the DBD. No interactions were found with contaminants. This study extends the existing empirical evidence linking PCSE to behavioral problems in school-aged children by reporting these effects in a population where tobacco use is normative rather than marginal. Co-exposure to Pb and Hg do not appear to exacerbate tobacco effects, suggesting that these substances act independently. © 2013. Published by Elsevier Inc. All rights reserved.

  6. Immunoassays for riboflavin and flavin mononucleotide using antibodies specific to d-ribitol and d-ribitol-5-phosphate.

    Science.gov (United States)

    Ravi, G; Venkatesh, Yeldur P

    2017-06-01

    Riboflavin (vitamin B 2 ), a water-soluble vitamin, plays a key role in maintaining human health. Though, numerous methods have been reported for the determination of total riboflavin (TRF) content in foods and biological samples, very few methods are reported for quantifying riboflavin and its coenzymes [flavin mononucleotide (FMN); flavin adenine dinucleotide (FAD)] individually. Recently, we have demonstrated that antibodies specific to d-ribitol and d-ribitol-5-phosphate also recognize riboflavin and FMN, respectively, and not vice-versa. In this study, we have evaluated these two antibodies for the analysis of riboflavin and FMN by indirect competitive ELISA (icELISA) in selected foods and pharmaceuticals. Under the optimal assay conditions, 50% inhibition concentration (IC 50 ) and limit of detection (LOD, IC 10 ) were 3.41ng/mL and 0.02ng/mL for riboflavin, and 7.84ng/mL and 0.24ng/mL for FMN, respectively, with detectable concentration range between 0.1 and 100ng of analytes and riboflavin and FMN) from the same food samples showed variation in their values compared to TRF, and were in good agreement with values obtained from HPLC and AOAC methods. Further, spiking and recovery analysis of food samples and pharmaceuticals showed no significant matrix effects. The immunoassays were validated in terms of accuracy and precision using inter- and intra-assays. The immunoassays developed in this study are sensitive and appears feasible for screening a large number of samples in the quantification of riboflavin and FMN in various biological samples, pharmaceuticals and natural/processed foods. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Diversity and ubiquity of thermophilic methanogenic archaea in temperate anoxic soils.

    Science.gov (United States)

    Wu, Xiao-Lei; Friedrich, Michael W; Conrad, Ralf

    2006-03-01

    Temperate rice field soil from Vercelli (Italy) contains moderately thermophilic methanogens of the yet uncultivated rice cluster I (RC-I), which become prevalent upon incubation at temperatures of 45-50 degrees C. We studied whether such thermophilic methanogens were ubiquitously present in anoxic soils. Incubation of different rice field soils (from Italy, China and the Philippines) and flooded riparian soils (from the Netherlands) at 45 degrees C resulted in vigorous CH(4) production after a lag phase of about 10 days. The archaeal community structure in the soils was analysed by terminal restriction fragment length polymorphism (T-RFLP) targeting the SSU rRNA genes retrieved from the soil, and by cloning and sequencing. Clones of RC-I methanogens mostly exhibited T-RF of 393 bp, but also terminal restriction fragment (T-RF) of 158 and 258 bp length, indicating a larger diversity than previously assumed. No RC-I methanogens were initially found in flooded riparian soils. However, these archaea became abundant upon incubation of the soil at 45 degrees C. Thermophilic RC-I methanogens were also found in the rice field soils from Pavia, Pila and Gapan. However, the archaeal communities in these soils also contained other methanogenic archaea at high temperature. Rice field soil from Buggalon, on the other hand, only contained thermophilic Methanomicrobiales rather than RC-I methanogens, and rice field soil from Jurong mostly Methanomicrobiales and only a few RC-I methanogens. The archaeal community of rice field soil from Zhenjiang almost exclusively consisted of Methanosarcinaceae when incubated at high temperature. Our results show that moderately thermophilic methanogens are common in temperate soils. However, RC-I methanogens are not always dominating or ubiquitous.

  8. AECB staff annual report of Darlington NGS for the year 1991

    International Nuclear Information System (INIS)

    1992-11-01

    Ontario Hydro operated Darlington in a safe manner in 1991. Ontario Hydro violated the Atomic Energy Control Regulations once and the physical security regulations three times in 1991. They failed to observe the Operating Licence conditions on ten occasions. The AECB did not find that the individual events had a significant impact on safety. There were no violations of the construction licence. None of the station staff received a radiation dose in excess of the regulatory limit. Radioactive emissions from the station were far below the regulatory limit. Special safety system performance was not fully satisfactory. Ontario Hydro failed to meet the unavailability targets for shutdown system one and the negative pressure containment system. Ontario Hydro reported seventeen incidents under conditions of the Operating and Construction licences. Units 1 and 2 remained shut down for most of 1991 because of unexplained fuel bundle damage in the reactor core. Ontario Hydro has decided to replace the main generator rotors because of cracks discovered on the rotor shaft. A fully modified rotor was installed on Unit 1. Ontario Hydro staff have a significant backlog of maintenance work. The Quality Improvement Program seemed to work well, resulting in some noticeable improvements. Three Shift Supervisors and four Control Operators were licensed this year. All planned emergency exercises and drills took place as scheduled. Ontario Hydro identified and are addressing several areas for improvement during the drills. Except for a power supply interruption to some IAEA equipment, Ontario Hydro achieved all its safeguards goals at Darlington in 1991. The Tritium Removal Facility (TRF) operated intermittently during 1991. Ontario Hydro is proceeding with the design and planning of an annex to the TRF to replace the present temporary facilities. (Author)

  9. JTRF2014, the JPL Kalman filter and smoother realization of the International Terrestrial Reference System

    Science.gov (United States)

    Abbondanza, Claudio; Chin, Toshio M.; Gross, Richard S.; Heflin, Michael B.; Parker, Jay W.; Soja, Benedikt S.; van Dam, Tonie; Wu, Xiaoping

    2017-10-01

    We present and discuss JTRF2014, the Terrestrial Reference Frame (TRF) the Jet Propulsion Laboratory constructed by combining space-geodetic inputs from very long baseline interferometry (VLBI), satellite laser ranging (SLR), Global Navigation Satellite Systems (GNSS), and Doppler orbitography and radiopositioning integrated by satellite submitted for the realization of ITRF2014. Determined through a Kalman filter and Rauch-Tung-Striebel smoother assimilating position observations, Earth orientation parameters, and local ties, JTRF2014 is a subsecular, time series-based TRF whose origin is at the quasi-instantaneous center of mass (CM) as sensed by SLR and whose scale is determined by the quasi-instantaneous VLBI and SLR scales. The dynamical evolution of the positions accounts for a secular motion term, annual, and semiannual periodic modes. Site-dependent variances based on the analysis of loading displacements induced by mass redistributions of terrestrial fluids have been used to control the extent of random walk adopted in the combination. With differences in the amplitude of the annual signal within the range 0.5-0.8 mm, JTRF2014-derived center of network-to-center of mass (CM-CN) is in remarkable agreement with the geocenter motion obtained via spectral inversion of GNSS, Gravity Recovery and Climate Experiment (GRACE) observations and modeled ocean bottom pressure from Estimating the Circulation and Climate of the Ocean (ECCO). Comparisons of JTRF2014 to ITRF2014 suggest high-level consistency with time derivatives of the Helmert transformation parameters connecting the two frames below 0.18 mm/yr and weighted root-mean-square differences of the polar motion (polar motion rate) in the order of 30 μas (17 μas/d).

  10. Behavioral and emotional problems on the Teacher's Report Form: a cross-national, cross-clinic comparative analysis of gender dysphoric children and adolescents.

    Science.gov (United States)

    Steensma, Thomas D; Zucker, Kenneth J; Kreukels, Baudewijntje P C; Vanderlaan, Doug P; Wood, Hayley; Fuentes, Amanda; Cohen-Kettenis, Peggy T

    2014-05-01

    For gender dysphoric children and adolescents, the school environment may be challenging due to peer social ostracism and rejection. To date, information on the psychological functioning and the quality of peer relations in gender dysphoric children and adolescents has been studied via parental report, peer sociometric methods, and social interactions in laboratory play groups. The present study was the first cross-national investigation that assessed behavior and emotional problems and the quality of peer relations, both measured by the Teacher's Report Form (TRF), in a sample of 728 gender dysphoric patients (554 children, 174 adolescents), who were referred to specialized gender identity clinics in the Netherlands and Canada. The gender dysphoric adolescents had significantly more teacher-reported emotional and behavioral problems than the gender dysphoric children. In both countries, gender dysphoric natal boys had poorer peer relations and more internalizing than externalizing problems compared to the gender dysphoric natal girls. Furthermore, there were significant between-clinic differences: both the children and the adolescents from Canada had more emotional and behavioral problems and a poorer quality of peer relations than the children and adolescents from the Netherlands. In conclusion, gender dysphoric children and adolescents showed the same pattern of emotional and behavioral problems in both countries. The extent of behavior and emotional problems was, however, higher in Canada than in the Netherlands, which appeared, in part, an effect of a poorer quality of peer relations. Per Bronfenbrenner's (American Psychologist, 32, 513-531, 1977) ecological model of human development and well-being, we consider various interpretations of the cross-national, cross-clinic differences on TRF behavior problems at the level of the family, the peer group, and the culture at large.

  11. DNA damage signaling and apoptosis in preinvasive tubal lesions of ovarian carcinoma.

    Science.gov (United States)

    Chene, Gautier; Ouellet, Veronique; Rahimi, Kurosh; Barres, Veronique; Caceres, Katia; Meunier, Liliane; Cyr, Louis; De Ladurantaye, Manon; Provencher, Diane; Mes Masson, Anne Marie

    2015-06-01

    High-grade serous ovarian cancer (HGSC) is the most life-threatening gynecological malignancy despite surgery and chemotherapy. A better understanding of the molecular basis of the preinvasive stages might be helpful in early detection and diagnosis. Genetic instability is 1 of the characteristics shared by most human cancers, and its level is variable through precancerous lesions to advanced cancer. Because DNA damage response (DDR) has been described as 1 of the first phases in genomic instability, we investigated the level of DDR activation and the apoptosis pathway in serous tubal intraepithelial carcinoma (STIC), the potential precursor of HGSC. A tissue microarray including 21 benign fallopian tubes, 21 STICs, 17 HGSCs from patients with STICs (associated ovarian cancer [AOC]) from the same individuals, and 30 HGSCs without STICs (non-AOC) was used in this study.Immunohistochemistry was performed to evaluate the level of DDR proteins (pATM, pChk2, γH2AX, 53BP1, and TRF2), apoptosis proteins (Bcl2, BAX, and BIM), and cyclin E. The expression of all DDR proteins increased from benign fallopian tubes to STICs. The level of expression of pATM, pChk2, γH2AX, and TRF2 was also increased in STICs in comparison with AOC. BAX, BIM, and cyclin E expressions were high in STICs, whereas Bcl2 expression was low. Immunohistochemical profiles of AOC and non-AOC were also different. These results suggest an activation of the DDR and apoptosis pathways in STICs, indicating that genomic instability may occur early in the precancerous lesions of HGSC.

  12. A Study of Rank Defect and Network Effect in Processing the CMONOC Network on Bernese

    Directory of Open Access Journals (Sweden)

    Weiwei Wu

    2018-02-01

    Full Text Available High-precision GPS data processing on Bernese has been employed to routinely resolve daily position solutions of GPS stations in the Crustal Movement Observation Network of China (CMONOC. The rank-deficient problems of the normal equation (NEQ system and the network effect on the frame alignment of NEQs in the processing of CMONOC data on Bernese still present difficulties. In this study, we diagnose the rank-deficient problems of the original NEQ, review the efficiency of the controlled datum removal (CDR method in filtering out the three frame-origin-related datum contents, investigate the reliabilities of the inherited frame orientation and scale information from the fixation of the GPS satellite orbits and the Earth rotation parameters in establishing the NEQ of the CMONOC network on Bernese, and analyze the impact of the network effect on the position time series of GPS stations. Our results confirm the nonsingularity of the original NEQ and the efficiency of the CDR filtering in resolving the rank-deficient problems; show that the frame origin parameters are weakly defined and should be stripped off, while the frame orientation and scale parameters should be retained due to their insufficient redefinition from the minimal constraint (MC implementation through inhomogeneous and asymmetrical fiducial networks; and reveal the superiority of a globally distributed fiducial network for frame alignment of the reconstructed NEQs via No-Net-Translation (NNT MC conditions. Finally, we attribute the two apparent discontinuities in the position time series to the terrestrial reference frame (TRF conversions of the GPS satellite orbits, and identify it as the orbit TRF effect.

  13. Differences in micronucleus frequency and acrylamide adduct levels with hemoglobin between vegetarians and non-vegetarians.

    Science.gov (United States)

    Kotova, Natalia; Frostne, Cecilia; Abramsson-Zetterberg, Lilianne; Tareke, Eden; Bergman, Rolf; Haghdoost, Siamak; Paulsson, Birgit; Törnqvist, Margareta; Segerbäck, Dan; Jenssen, Dag; Grawé, Jan

    2015-10-01

    Nutrients and food constituents can prevent or contribute to genotoxicity. In this study, the possible influence of a vegetarian/non-vegetarian diet on genotoxic effects was investigated in 58 non-smoking healthy vegetarians (V) and non-vegetarians (NV), age 21-37 years from the Stockholm area in Sweden. Physical activity and dietary habits were similar in both groups, with the exception of the intake of meat and fish. Using flow cytometry, we determined the formation of micronuclei (MN) in transferrin-positive immature peripheral blood reticulocytes (Trf-Ret) (Total: n = 53; V: n = 27; NV: n = 26). Dietary exposure to acrylamide was measured through hemoglobin (Hb) adducts in peripheral erythrocytes (Total: n = 53; V: n = 29; NV: n = 24). Hb adducts of both acrylamide and its genotoxic metabolite glycidamide were monitored as a measure of the corresponding in vivo doses. Our data demonstrated that compared with the non-vegetarians, the vegetarians exhibited lower frequencies of MN (fMN) in the Trf-Ret (p vegetarians and non-vegetarians. Furthermore, there were no significant relationships between the adduct levels and fMN in the individuals. The ratio of the Hb adduct levels from glycidamide and acrylamide, however, showed a significant difference (p vegetarian diet might be beneficial in lowering genomic instability in healthy individuals. The measured Hb adduct levels indicate that the total intake of acrylamide does not differ between the two studied groups and does not contribute to the observed difference in fMN, although an influence of the diet on the metabolic rates of acrylamide was indicated. In addition, the observed significant difference in the background fMN in the two groups demonstrated that the MN analysis method has a sensitivity applicable to the biomonitoring of human lifestyle factors.

  14. Neurodevelopmental retardation, as assessed clinically and with magnetoencephalography and electroencephalography, associated with perinatal dioxin exposure.

    Science.gov (United States)

    Ten Tusscher, G W; Leijs, M M; de Boer, L C C; Legler, J; Olie, K; Spekreijse, H; van Dijk, B W; Vulsma, T; Briët, J; Ilsen, A; Koppe, J G

    2014-09-01

    In 1980s Western Europe, human perinatal exposure to background levels of dioxins was rather high. We therefore evaluated the neurodevelopment of our cohort during the prepubertal period and in adolescence. At prepubertal age (7-12 years) 41 children were tested. Both neuromotor functioning and psychological testing were performed (Dutch version of the Wechsler Intelligence Scale for Children (WISC-R) and the Dutch version of the Child Behavior Checklist for ages 4-18 years (CBCL 4-18) and the Teacher Report Form (TRF)). Neurophysiological tests were performed using magnetoencephalography and electroencephalography. In adolescence (14-18 years) the behavior of 33 children was studied again (CBCL and TRF). And the levels of dioxins and dioxin-like PCBs (dl-PCBs) were measured in serum. At prepubertal age no association was found between perinatal dioxin exposure and verbal, performal and total IQ or with the Touwen's test for neuromotor development. There were behavioral problems associated with both prenatal and postnatal dioxin exposure. In adolescence there were problems associated with the current dioxin levels and dioxin-like-PCBs. Neurophysiological tests revealed clear negative dysfunction. An increase in latency time after a motion stimulus (N2b) of 13 ms (= a delay of 10%) is associated with the higher prenatal dioxin exposure. A similar delay was measured in testing cognitive ability by analyzing the odd ball measurements, N200 and P300, together with an amplitude decrease of 12 %. The delay is indicative of a defective myelinisation and the decrease in amplitude of a loss of neurons. We found effects on behavior in association with the perinatal dioxin exposure and in adolescence in association with the current dioxin levels. Neurophysiological testing is instrumental in the detection of effects of perinatal background levels of chemicals on brain development in normal, healthy children. The clinical, neurological and psychological tests commonly used are

  15. Nilai Karakter Anak Dalam Novel Karya Anak Usia 10 Tahun

    Directory of Open Access Journals (Sweden)

    Suyatno Suyatno

    2011-12-01

    Full Text Available Makalah ini mendeskripsikan nilai karakter anak dalam novel karya anak usia 10 tahun. Novel yang dibahas adalah The Rings of Friendship (TRF karya Vira (10 tahun, The Smart Girls Petualangan Seru Lima Sahabat (TSG karya Salma Syifa (10 tahun, dan The Special Day (TSD karya Yasyfa (10 tahun. Berdasarkan kajian deskriptif dengan teknik pembacaan berulang-ulang, ditemukan sejumlah informasi tentang nilai karakter anak berkaitan dengan nilai cerdas, nilai jujur, nilai peduli, dan nilai tangguh. Nilai karakter tersebut menyatu ke dalam narasi cerita tanpa menunjukkan secara eksplisit unsur nilai karakter melainkan berada pada alur cerita dari awal sampai akhir. Tema ketiga novel adalah petualangan dan persahabatan, alur maju, tokoh anak-anak, dan latarnya sekolah, rumah, serta alam lingkungan. Kemasan cerita bersifat sederhana, lugas, kalimat pendek, bergaya dialog, dan naratif. Abstract: This article describes the value of child character in novels written by ten year old children. The novels being discussed are The Rings f Friendship (TRF written by Vira (10 years old, The Smart Girls Petualangan Seru Lima Sahabat (TSG written by Salma Syifa (10 years old. Under the descriptive study by using continually reading technique, it was identified that there are several information about the value of child characters related to the value of intelligence, honesty, care , and strength. Those character values combine into the story narrative without explicitly exposing the character values component, but exist in the plot from the beginning to the end. The themes of the novels are adventure and friendship, flash-forward plot, child character. The settings are school, home, and the natural surrounding. The presentation of the story is simple, to the point, using short sentences, dialogic, and narrative. Key Words: character values, intelligence, honesty, care, strenght

  16. Solution structure of telomere binding domain of AtTRB2 derived from Arabidopsis thaliana

    International Nuclear Information System (INIS)

    Yun, Ji-Hye; Lee, Won Kyung; Kim, Heeyoun; Kim, Eunhee; Cheong, Chaejoon; Cho, Myeon Haeng; Lee, Weontae

    2014-01-01

    Highlights: • We have determined solution structure of Myb domain of AtTRB2. • The Myb domain of AtTRB2 is located in the N-terminal region. • The Myb domain of AtTRB2 binds to plant telomeric DNA without fourth helix. • Helix 2 and 3 of the Myb domain of AtTRB2 are involved in DNA recognition. • AtTRB2 is a novel protein distinguished from other known plant TBP. - Abstract: Telomere homeostasis is regulated by telomere-associated proteins, and the Myb domain is well conserved for telomere binding. AtTRB2 is a member of the SMH (Single-Myb-Histone)-like family in Arabidopsis thaliana, having an N-terminal Myb domain, which is responsible for DNA binding. The Myb domain of AtTRB2 contains three α-helices and loops for DNA binding, which is unusual given that other plant telomere-binding proteins have an additional fourth helix that is essential for DNA binding. To understand the structural role for telomeric DNA binding of AtTRB2, we determined the solution structure of the Myb domain of AtTRB2 (AtTRB2 1–64 ) using nuclear magnetic resonance (NMR) spectroscopy. In addition, the inter-molecular interaction between AtTRB2 1–64 and telomeric DNA has been characterized by the electrophoretic mobility shift assay (EMSA) and NMR titration analyses for both plant (TTTAGGG)n and human (TTAGGG)n telomere sequences. Data revealed that Trp28, Arg29, and Val47 residues located in Helix 2 and Helix 3 are crucial for DNA binding, which are well conserved among other plant telomere binding proteins. We concluded that although AtTRB2 is devoid of the additional fourth helix in the Myb-extension domain, it is able to bind to plant telomeric repeat sequences as well as human telomeric repeat sequences

  17. Solution structure of telomere binding domain of AtTRB2 derived from Arabidopsis thaliana

    Energy Technology Data Exchange (ETDEWEB)

    Yun, Ji-Hye [Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of); Lee, Won Kyung [Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of); Kim, Heeyoun [Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of); Kim, Eunhee; Cheong, Chaejoon [Magnetic Resonance Team, Korea Basic Science Institute (KBSI), Ochang, Chungbuk 363-883 (Korea, Republic of); Cho, Myeon Haeng [Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of); Lee, Weontae, E-mail: wlee@spin.yonsei.ac.kr [Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of)

    2014-09-26

    Highlights: • We have determined solution structure of Myb domain of AtTRB2. • The Myb domain of AtTRB2 is located in the N-terminal region. • The Myb domain of AtTRB2 binds to plant telomeric DNA without fourth helix. • Helix 2 and 3 of the Myb domain of AtTRB2 are involved in DNA recognition. • AtTRB2 is a novel protein distinguished from other known plant TBP. - Abstract: Telomere homeostasis is regulated by telomere-associated proteins, and the Myb domain is well conserved for telomere binding. AtTRB2 is a member of the SMH (Single-Myb-Histone)-like family in Arabidopsis thaliana, having an N-terminal Myb domain, which is responsible for DNA binding. The Myb domain of AtTRB2 contains three α-helices and loops for DNA binding, which is unusual given that other plant telomere-binding proteins have an additional fourth helix that is essential for DNA binding. To understand the structural role for telomeric DNA binding of AtTRB2, we determined the solution structure of the Myb domain of AtTRB2 (AtTRB2{sub 1–64}) using nuclear magnetic resonance (NMR) spectroscopy. In addition, the inter-molecular interaction between AtTRB2{sub 1–64} and telomeric DNA has been characterized by the electrophoretic mobility shift assay (EMSA) and NMR titration analyses for both plant (TTTAGGG)n and human (TTAGGG)n telomere sequences. Data revealed that Trp28, Arg29, and Val47 residues located in Helix 2 and Helix 3 are crucial for DNA binding, which are well conserved among other plant telomere binding proteins. We concluded that although AtTRB2 is devoid of the additional fourth helix in the Myb-extension domain, it is able to bind to plant telomeric repeat sequences as well as human telomeric repeat sequences.

  18. Karyological characterization of the endemic Iberian rock lizard, Iberolacerta monticola (Squamata, Lacertidae): insights into sex chromosome evolution.

    Science.gov (United States)

    Rojo, V; Giovannotti, M; Naveira, H; Nisi Cerioni, P; González-Tizón, A M; Caputo Barucchi, V; Galán, P; Olmo, E; Martínez-Lage, A

    2014-01-01

    Rock lizards of the genus Iberolacerta constitute a promising model to examine the process of sex chromosome evolution, as these closely related taxa exhibit remarkable diversity in the degree of sex chromosome differentiation with no clear phylogenetic segregation, ranging from cryptic to highly heteromorphic ZW chromosomes and even multiple chromosome systems (Z1Z1Z2Z2/Z1Z2W). To gain a deeper insight into the patterns of karyotype and sex chromosome evolution, we performed a cytogenetic analysis based on conventional staining, banding techniques and fluorescence in situ hybridization in the species I. monticola, for which previous cytogenetic investigations did not detect differentiated sex chromosomes. The karyotype is composed of 2n = 36 acrocentric chromosomes. NORs and the major ribosomal genes were located in the subtelomeric region of chromosome pair 6. Hybridization signals of the telomeric sequences (TTAGGG)n were visualized at the telomeres of all chromosomes and interstitially in 5 chromosome pairs. C-banding showed constitutive heterochromatin at the centromeres of all chromosomes, as well as clear pericentromeric and light telomeric C-bands in several chromosome pairs. These results highlight some chromosomal markers which can be useful to identify species-specific diagnostic characters, although they may not accurately reflect the phylogenetic relationships among the taxa. In addition, C-banding revealed the presence of a heteromorphic ZW sex chromosome pair, where W is smaller than Z and almost completely heterochromatic. This finding sheds light on sex chromosome evolution in the genus Iberolacerta and suggests that further comparative cytogenetic analyses are needed to understand the processes underlying the origin, differentiation and plasticity of sex chromosome systems in lacertid lizards. © 2013 S. Karger AG, Basel.

  19. Cytological maps of lampbrush chromosomes of European water frogs (Pelophylax esculentus complex) from the Eastern Ukraine

    Science.gov (United States)

    2013-01-01

    Background Hybridogenesis (hemiclonal inheritance) is a kind of clonal reproduction in which hybrids between parental species are reproduced by crossing with one of the parental species. European water frogs (Pelophylax esculentus complex) represent an appropriate model for studying interspecies hybridization, processes of hemiclonal inheritance and polyploidization. P. esculentus complex consists of two parental species, P. ridibundus (the lake frog) and P. lessonae (the pool frog), and their hybridogenetic hybrid – P. esculentus (the edible frog). Parental and hybrid frogs can reproduce syntopically and form hemiclonal population systems. For studying mechanisms underlying the maintenance of water frog population systems it is required to characterize the karyotypes transmitted in gametes of parental and different hybrid animals of both sexes. Results In order to obtain an instrument for characterization of oocyte karyotypes in hybrid female frogs, we constructed cytological maps of lampbrush chromosomes from oocytes of both parental species originating in Eastern Ukraine. We further identified certain molecular components of chromosomal marker structures and mapped coilin-rich spheres and granules, chromosome associated nucleoli and special loops accumulating splicing factors. We recorded the dissimilarities between P. ridibundus and P. lessonae lampbrush chromosomes in the length of orthologous chromosomes, number and location of marker structures and interstitial (TTAGGG)n-repeat sites as well as activity of nucleolus organizer. Satellite repeat RrS1 was mapped in centromere regions of lampbrush chromosomes of the both species. Additionally, we discovered transcripts of RrS1 repeat in oocytes of P. ridibundus and P. lessonae. Moreover, G-rich transcripts of telomere repeat were revealed in association with terminal regions of P. ridibundus and P. lessonae lampbrush chromosomes. Conclusions The constructed cytological maps of lampbrush chromosomes of P

  20. Mechanisms of telomere loss and their consequences for chromosome instability

    Energy Technology Data Exchange (ETDEWEB)

    Muraki, Keiko; Nyhan, Kristine; Han, Limei; Murnane, John P., E-mail: jmurnane@radonc.ucsf.edu [Department of Radiation Oncology, University of California at San Francisco, San Francisco, CA (United States)

    2012-10-04

    The ends of chromosomes in mammals, called telomeres, are composed of a 6-bp repeat sequence, TTAGGG, which is added on by the enzyme telomerase. In combination with a protein complex called shelterin, these telomeric repeat sequences form a cap that protects the ends of chromosomes. Due to insufficient telomerase expression, telomeres shorten gradually with each cell division in human somatic cells, which limits the number of times they can divide. The extensive cell division involved in cancer cell progression therefore requires that cancer cells must acquire the ability to maintain telomeres, either through expression of telomerase, or through an alternative mechanism involving recombination. It is commonly thought that the source of many chromosome rearrangements in cancer cells is a result of the extensive telomere shortening that occurs prior to the expression of telomerase. However, despite the expression of telomerase, tumor cells can continue to show chromosome instability due to telomere loss. Dysfunctional telomeres in cancer cells can result from oncogene-induced replication stress, which results in double-strand breaks (DSBs) at fragile sites, including telomeres. DSBs near telomeres are especially prone to chromosome rearrangements, because telomeric regions are deficient in DSB repair. The deficiency in DSB repair near telomeres is also an important mechanism for ionizing radiation-induced replicative senescence in normal human cells. In addition, DSBs near telomeres can result in chromosome instability in mouse embryonic stem cells, suggesting that telomere loss can contribute to heritable chromosome rearrangements. Consistent with this possibility, telomeric regions in humans are highly heterogeneous, and chromosome rearrangements near telomeres are commonly involved in human genetic disease. Understanding the mechanisms of telomere loss will therefore provide important insights into both human cancer and genetic disease.

  1. Co-located hAT transposable element and 5S rDNA in an interstitial telomeric sequence suggest the formation of Robertsonian fusion in armored catfish.

    Science.gov (United States)

    Glugoski, Larissa; Giuliano-Caetano, Lucia; Moreira-Filho, Orlando; Vicari, Marcelo R; Nogaroto, Viviane

    2018-04-15

    Co-located 5S rDNA genes and interstitial telomeric sites (ITS) revealed the involvement of multiple 5S rDNA clusters in chromosome rearrangements of Loricariidae. Interstitial (TTAGGG)n vestiges, in addition to telomeric sites, can coincide with locations of chromosomal rearrangements, and they are considered to be hotspots for chromosome breaks. This study aimed the molecular characterization of 5S rDNA in two Rineloricaria latirostris populations and examination of roles of 5S rDNA in breakpoint sites and its in situ localization. Rineloricaria latirostris from Brazil's Das Pedras river (2n = 46 chromosomes) presented five pairs identified using a 5S rDNA probe, in addition to a pair bearing a co-located ITS/5S rDNA. Rineloricaria latirostris from the Piumhi river (2n = 48 chromosomes) revealed two pairs containing 5S rDNA, without ITS. A 702-bp amplified sequence, using 5S rDNA primers, revealed an insertion of the hAT transposable element (TE), referred to as a degenerate 5S rDNA. Double-FISH (fluorescence in situ hybridization) demonstrated co-localization of 5S rDNA/degenerate 5S rDNA, 5S rDNA/hAT and ITS/5S rDNA from the Das Pedras river population. Piumhi river isolates possessed only 5S rDNA sites. We suggest that the degenerate 5S rDNA was generated by unequal crossing over, which was driven by invasion of hAT, establishing a breakpoint region susceptible to chromosome breakage, non-homologous recombination and Robertsonian (Rb) fusion. Furthermore, the presence of clusters of 5S rDNA at fusion points in other armored catfish species suggests its re-use and that these regions represent hotspots for evolutionary rearrangements within Loricariidae genomes. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. A calmodulin-like protein (LCALA) is a new Leishmania amazonensis candidate for telomere end-binding protein.

    Science.gov (United States)

    Morea, Edna G O; Viviescas, Maria Alejandra; Fernandes, Carlos A H; Matioli, Fabio F; Lira, Cristina B B; Fernandez, Maribel F; Moraes, Barbara S; da Silva, Marcelo S; Storti, Camila B; Fontes, Marcos R M; Cano, Maria Isabel N

    2017-11-01

    Leishmania spp. telomeres are composed of 5'-TTAGGG-3' repeats associated with proteins. We have previously identified LaRbp38 and LaRPA-1 as proteins that bind the G-rich telomeric strand. At that time, we had also partially characterized a protein: DNA complex, named LaGT1, but we could not identify its protein component. Using protein-DNA interaction and competition assays, we confirmed that LaGT1 is highly specific to the G-rich telomeric single-stranded DNA. Three protein bands, with LaGT1 activity, were isolated from affinity-purified protein extracts in-gel digested, and sequenced de novo using mass spectrometry analysis. In silico analysis of the digested peptide identified them as a putative calmodulin with sequences identical to the T. cruzi calmodulin. In the Leishmania genome, the calmodulin ortholog is present in three identical copies. We cloned and sequenced one of the gene copies, named it LCalA, and obtained the recombinant protein. Multiple sequence alignment and molecular modeling showed that LCalA shares homology to most eukaryotes calmodulin. In addition, we demonstrated that LCalA is nuclear, partially co-localizes with telomeres and binds in vivo the G-rich telomeric strand. Recombinant LCalA can bind specifically and with relative affinity to the G-rich telomeric single-strand and to a 3'G-overhang, and DNA binding is calcium dependent. We have described a novel candidate component of Leishmania telomeres, LCalA, a nuclear calmodulin that binds the G-rich telomeric strand with high specificity and relative affinity, in a calcium-dependent manner. LCalA is the first reported calmodulin that binds in vivo telomeric DNA. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. A Smart DNA Tweezer for Detection of Human Telomerase Activity.

    Science.gov (United States)

    Xu, Xiaowen; Wang, Lei; Li, Kan; Huang, Qihong; Jiang, Wei

    2018-03-06

    Reliable and accurate detection of telomerase activity is crucial to better understand its role in cancer cells and to further explore its function in cancer diagnosis and treatment. Here, we construct a smart DNA tweezer (DT) for detection of telomerase activity. The DT is assembled by three specially designed single-stranded oligonucleotides: a central strand dually labeled with donor/acceptor fluorophores and two arm strands containing overhangs complementary to telomerase reaction products (TRPs). It can get closed through hybridization with TRPs and get reopen through strand displacement reaction by TRPs' complementary sequences. First, under the action of telomerase, telomerase binding substrates (TS) are elongated to generate TRPs ended with telomeric repeats (TTAGGG) n . TRPs hybridize with the two arm overhangs cooperatively and strain DT to closed state, inducing an increased fluorescence resonance energy transfer (FRET) efficiency, which is utilized for telomerase activity detection. Second, upon introduction of a removal strand (RS) complementary to TRPs, the closed DT is relaxed to open state via the toehold-mediated strand displacement, inducing a decreased FRET efficiency, which is utilized for determination of TRP length distribution. The detection limit of telomerase activity is equivalent to 141 cells/μL for HeLa cells, and telomerase-active cellular extracts can be differentiated from telomerase-inactive cellular extracts. Furthermore, TRPs owning 1, 2, 3, 4, and ≥5 telomeric repeats are identified to account for 25.6%, 20.5%, 15.7%, 12.5%, and 25.7%, respectively. The proposed strategy will offer a new approach for reliable, accurate detection of telomerase activity and product length distribution for deeper studying its role and function in cancer.

  4. Augmented telomerase activity, reduced telomere length and the presence of alternative lengthening of telomere in renal cell carcinoma: plausible predictive and diagnostic markers.

    Science.gov (United States)

    Pal, Deeksha; Sharma, Ujjawal; Khajuria, Ragini; Singh, Shrawan Kumar; Kakkar, Nandita; Prasad, Rajendra

    2015-05-15

    In this study, we analyzed 100 cases of renal cell carcinoma (RCC) for telomerase activity, telomere length and alternative lengthening of telomeres (ALT) using the TRAP assay, TeloTTAGGG assay kit and immunohistochemical analysis of ALT associated promyelocytic leukemia (PML) bodies respectively. A significantly higher (P=0.000) telomerase activity was observed in 81 cases of RCC which was correlated with clinicopathological features of tumor for instance, stage (P=0.008) and grades (P=0.000) but not with the subtypes of RCC (P = 0.355). Notwithstanding, no correlation was found between telomerase activity and subtypes of RCC. Strikingly, the telomere length was found to be significantly shorter in RCC (P=0.000) to that of corresponding normal renal tissues and it is well correlated with grades (P=0.016) but not with stages (P=0.202) and subtypes (P=0.669) of RCC. In this study, telomere length was also negatively correlated with the age of patients (r(2)=0.528; P=0.000) which supports the notion that it could be used as a marker for biological aging. ALT associated PML bodies containing PML protein was found in telomerase negative cases of RCC. It suggests the presence of an ALT pathway mechanism to maintain the telomere length in telomerase negative RCC tissues which was associated with high stages of RCC, suggesting a prevalent mechanism for telomere maintenance in high stages. In conclusion, the telomerase activity and telomere length can be used as a diagnostic as well as a predictive marker in RCC. The prevalence of ALT mechanism in high stages of RCC is warranted for the development of anti-ALT inhibitors along with telomerase inhibitor against RCC as a therapeutic approach. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Identification of two new repetitive elements and chromosomal mapping of repetitive DNA sequences in the fish Gymnothorax unicolor (Anguilliformes: Muraenidae

    Directory of Open Access Journals (Sweden)

    E. Coluccia

    2011-05-01

    Full Text Available Muraenidae is a species-rich family, with relationships among genera and species and taxonomy that have not been completely clarified. Few cytogenetic studies have been conducted on this family, and all of them showed the same diploid chromosome number (2n=42 but with conspicuous karyotypic variation among species. The Mediterranean moray eel Gymnothorax unicolor was previously cytogenetically studied using classical techniques that allowed the characterization of its karyotype structure and the constitutive heterochromatin and argyrophilic nucleolar organizer regions (Ag-NORs distribution pattern. In the present study, we describe two new repetitive elements (called GuMboI and GuDdeI obtained from restricted genomic DNA of G. unicolor that were characterized by Southern blot and physically localized by in situ hybridization on metaphase chromosomes. As they are highly repetitive DNA sequences, they map in heterochromatic regions. However, while GuDdeI was localized in the centromeric regions, the GuMboI fraction was distributed on some centromeres and was co-localized with the nucleolus organizer region (NOR. Comparative analysis with other Mediterranean species such as Muraena helena pointed out that these DNA fractions are species-specific and could potentially be used for species discrimination. As a new contribution to the karyotype of this species, we found that the major ribosomal genes are localized on acrocentric chromosome 9 and that the telomeres of each chromosome are composed of a tandem repeat derived from a poly-TTAGGG DNA sequence, as it occurs in most vertebrate species. The results obtained add new information useful in comparative genomics at the chromosomal level and contribute to the cytogenetic knowledge regarding this fish family, which has not been extensively studied.

  6. Mechanisms of telomere loss and their consequences for chromosome instability

    International Nuclear Information System (INIS)

    Muraki, Keiko; Nyhan, Kristine; Han, Limei; Murnane, John P.

    2012-01-01

    The ends of chromosomes in mammals, called telomeres, are composed of a 6-bp repeat sequence, TTAGGG, which is added on by the enzyme telomerase. In combination with a protein complex called shelterin, these telomeric repeat sequences form a cap that protects the ends of chromosomes. Due to insufficient telomerase expression, telomeres shorten gradually with each cell division in human somatic cells, which limits the number of times they can divide. The extensive cell division involved in cancer cell progression therefore requires that cancer cells must acquire the ability to maintain telomeres, either through expression of telomerase, or through an alternative mechanism involving recombination. It is commonly thought that the source of many chromosome rearrangements in cancer cells is a result of the extensive telomere shortening that occurs prior to the expression of telomerase. However, despite the expression of telomerase, tumor cells can continue to show chromosome instability due to telomere loss. Dysfunctional telomeres in cancer cells can result from oncogene-induced replication stress, which results in double-strand breaks (DSBs) at fragile sites, including telomeres. DSBs near telomeres are especially prone to chromosome rearrangements, because telomeric regions are deficient in DSB repair. The deficiency in DSB repair near telomeres is also an important mechanism for ionizing radiation-induced replicative senescence in normal human cells. In addition, DSBs near telomeres can result in chromosome instability in mouse embryonic stem cells, suggesting that telomere loss can contribute to heritable chromosome rearrangements. Consistent with this possibility, telomeric regions in humans are highly heterogeneous, and chromosome rearrangements near telomeres are commonly involved in human genetic disease. Understanding the mechanisms of telomere loss will therefore provide important insights into both human cancer and genetic disease.

  7. Mechanisms of telomere loss and their consequences for chromosome instability

    Directory of Open Access Journals (Sweden)

    Keiko eMuraki

    2012-10-01

    Full Text Available The ends of chromosomes in mammals, called telomeres, are composed of a 6 base pair repeat sequence, TTAGGG, which is added on by the enzyme telomerase. In combination with a protein complex called shelterin, these telomeric repeat sequences form a cap that protects the ends of chromosomes. Due to insufficient telomerase expression, telomeres shorten gradually with each cell division in human somatic cells, which limits the number of times they can divide. The extensive cell division involved in cancer cell progression therefore requires that cancer cells must acquire the ability to maintain telomeres, either through expression of telomerase, or through an alternative mechanism involving recombination. It is commonly thought that the source of many chromosome rearrangements in cancer cells is a result of the extensive telomere shortening that occurs prior to the expression of telomerase. However, despite the expression of telomerase, tumor cells can continue to show chromosome instability due to telomere loss. Dysfunctional telomeres in cancer cells can result from oncogene-induced replication stress, which results in double-strand breaks (DSBs at fragile sites, including telomeres. DSBs near telomeres are especially prone to chromosome rearrangements, because telomeric regions are deficient in DSB repair. The deficiency in DSB repair near telomeres is also an important mechanism for ionizing radiation-induced replicative senescence in normal human cells. In addition, DSBs near telomeres can result in chromosome instability in mouse embryonic stem cells, suggesting that telomere loss can contribute to heritable chromosome rearrangements. Consistent with this possibility, telomeric regions in humans are highly heterogeneous, and chromosome rearrangements near telomeres are commonly involved in human genetic disease. Understanding the mechanisms of telomere loss will therefore provide important insights into both human cancer and genetic disease.

  8. Giardia telomeric sequence d(TAGGG)4 forms two intramolecular G-quadruplexes in K+ solution: effect of loop length and sequence on the folding topology.

    Science.gov (United States)

    Hu, Lanying; Lim, Kah Wai; Bouaziz, Serge; Phan, Anh Tuân

    2009-11-25

    Recently, it has been shown that in K(+) solution the human telomeric sequence d[TAGGG(TTAGGG)(3)] forms a (3 + 1) intramolecular G-quadruplex, while the Bombyx mori telomeric sequence d[TAGG(TTAGG)(3)], which differs from the human counterpart only by one G deletion in each repeat, forms a chair-type intramolecular G-quadruplex, indicating an effect of G-tract length on the folding topology of G-quadruplexes. To explore the effect of loop length and sequence on the folding topology of G-quadruplexes, here we examine the structure of the four-repeat Giardia telomeric sequence d[TAGGG(TAGGG)(3)], which differs from the human counterpart only by one T deletion within the non-G linker in each repeat. We show by NMR that this sequence forms two different intramolecular G-quadruplexes in K(+) solution. The first one is a novel basket-type antiparallel-stranded G-quadruplex containing two G-tetrads, a G x (A-G) triad, and two A x T base pairs; the three loops are consecutively edgewise-diagonal-edgewise. The second one is a propeller-type parallel-stranded G-quadruplex involving three G-tetrads; the three loops are all double-chain-reversal. Recurrence of several structural elements in the observed structures suggests a "cut and paste" principle for the design and prediction of G-quadruplex topologies, for which different elements could be extracted from one G-quadruplex and inserted into another.

  9. Intramolecular telomeric G-quadruplexes dramatically inhibit DNA synthesis by replicative and translesion polymerases, revealing their potential to lead to genetic change.

    Directory of Open Access Journals (Sweden)

    Deanna N Edwards

    Full Text Available Recent research indicates that hundreds of thousands of G-rich sequences within the human genome have the potential to form secondary structures known as G-quadruplexes. Telomeric regions, consisting of long arrays of TTAGGG/AATCCC repeats, are among the most likely areas in which these structures might form. Since G-quadruplexes assemble from certain G-rich single-stranded sequences, they might arise when duplex DNA is unwound such as during replication. Coincidentally, these bulky structures when present in the DNA template might also hinder the action of DNA polymerases. In this study, single-stranded telomeric templates with the potential to form G-quadruplexes were examined for their effects on a variety of replicative and translesion DNA polymerases from humans and lower organisms. Our results demonstrate that single-stranded templates containing four telomeric GGG runs fold into intramolecular G-quadruplex structures. These intramolecular G quadruplexes are somewhat dynamic in nature and stabilized by increasing KCl concentrations and decreasing temperatures. Furthermore, the presence of these intramolecular G-quadruplexes in the template dramatically inhibits DNA synthesis by various DNA polymerases, including the human polymerase δ employed during lagging strand replication of G-rich telomeric strands and several human translesion DNA polymerases potentially recruited to sites of replication blockage. Notably, misincorporation of nucleotides is observed when certain translesion polymerases are employed on substrates containing intramolecular G-quadruplexes, as is extension of the resulting mismatched base pairs upon dynamic unfolding of this secondary structure. These findings reveal the potential for blockage of DNA replication and genetic changes related to sequences capable of forming intramolecular G-quadruplexes.

  10. Structural Insights into the Quadruplex-Duplex 3' Interface Formed from a Telomeric Repeat: A Potential Molecular Target.

    Science.gov (United States)

    Russo Krauss, Irene; Ramaswamy, Sneha; Neidle, Stephen; Haider, Shozeb; Parkinson, Gary N

    2016-02-03

    We report here on an X-ray crystallographic and molecular modeling investigation into the complex 3' interface formed between putative parallel stranded G-quadruplexes and a duplex DNA sequence constructed from the human telomeric repeat sequence TTAGGG. Our crystallographic approach provides a detailed snapshot of a telomeric 3' quadruplex-duplex junction: a junction that appears to have the potential to form a unique molecular target for small molecule binding and interference with telomere-related functions. This unique target is particularly relevant as current high-affinity compounds that bind putative G-quadruplex forming sequences only rarely have a high degree of selectivity for a particular quadruplex. Here DNA junctions were assembled using different putative quadruplex-forming scaffolds linked at the 3' end to a telomeric duplex sequence and annealed to a complementary strand. We successfully generated a series of G-quadruplex-duplex containing crystals, both alone and in the presence of ligands. The structures demonstrate the formation of a parallel folded G-quadruplex and a B-form duplex DNA stacked coaxially. Most strikingly, structural data reveals the consistent formation of a TAT triad platform between the two motifs. This triad allows for a continuous stack of bases to link the quadruplex motif with the duplex region. For these crystal structures formed in the absence of ligands, the TAT triad interface occludes ligand binding at the 3' quadruplex-duplex interface, in agreement with in silico docking predictions. However, with the rearrangement of a single nucleotide, a stable pocket can be produced, thus providing an opportunity for the binding of selective molecules at the interface.

  11. Meningiomas, dicentric chromosomes, gliomas, and telomerase activity.

    Science.gov (United States)

    Carroll, T; Maltby, E; Brock, I; Royds, J; Timperley, W; Jellinek, D

    1999-08-01

    Lack of telomere maintenance during cell replication leads to telomere erosion and loss of function. This can result in telomere associations which probably cause the dicentric chromosomes seen in some tumour cells. One mechanism of telomere maintenance in dividing cells is the action of telomerase, a ribonucleoprotein enzyme that adds TTAGGG repeats onto telomeres and compensates for their shortening during cell division. Over 90 per cent of extracranial malignant neoplasms have been found to have telomerase activity. This study sought to determine if there was a relationship between absence of telomerase activity and presence of dicentric chromosomes in meningiomas and to what extent the other main group of central nervous system tumours, the gliomas, expressed telomerase activity. Telomerase activity was measured on 25 meningiomas and 29 gliomas. Four of the meningiomas were atypical variants and 11 were positive for dicentric chromosomes. Twenty-five of 29 gliomas were glioblastoma multiforme tumours. Measures were taken to ensure absence of false positives due to primer-dimer interaction and false negatives due to protein degradation or the presence of Taq polymerase inhibitors. All 25 meningiomas and the four low-grade gliomas (WHO grade II) were telomerase activity-negative. Seven (28 per cent) of the 25 glioblastoma multiforme tumours showed telomerase activity. The absence of telomerase activity in meningiomas and the high frequency of telomere associations support the hypothesis that these tumours are benign, transformed but pre-crisis. The relatively low frequency of telomerase activity in the malignant glioblastoma multiforme suggests that most of these tumours may have other mechanisms of telomere maintenance and that the potentially therapeutic telomerase inhibitors will not be of great value in the future management of the majority of patients suffering from these tumours. Copyright 1999 John Wiley & Sons, Ltd.

  12. Human telomeres are hypersensitive to UV-induced DNA Damage and refractory to repair.

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    Patrick J Rochette

    2010-04-01

    Full Text Available Telomeric repeats preserve genome integrity by stabilizing chromosomes, a function that appears to be important for both cancer and aging. In view of this critical role in genomic integrity, the telomere's own integrity should be of paramount importance to the cell. Ultraviolet light (UV, the preeminent risk factor in skin cancer development, induces mainly cyclobutane pyrimidine dimers (CPD which are both mutagenic and lethal. The human telomeric repeat unit (5'TTAGGG/CCCTAA3' is nearly optimal for acquiring UV-induced CPD, which form at dipyrimidine sites. We developed a ChIP-based technique, immunoprecipitation of DNA damage (IPoD, to simultaneously study DNA damage and repair in the telomere and in the coding regions of p53, 28S rDNA, and mitochondrial DNA. We find that human telomeres in vivo are 7-fold hypersensitive to UV-induced DNA damage. In double-stranded oligonucleotides, this hypersensitivity is a property of both telomeric and non-telomeric repeats; in a series of telomeric repeat oligonucleotides, a phase change conferring UV-sensitivity occurs above 4 repeats. Furthermore, CPD removal in the telomere is almost absent, matching the rate in mitochondria known to lack nucleotide excision repair. Cells containing persistent high levels of telomeric CPDs nevertheless proliferate, and chronic UV irradiation of cells does not accelerate telomere shortening. Telomeres are therefore unique in at least three respects: their biophysical UV sensitivity, their prevention of excision repair, and their tolerance of unrepaired lesions. Utilizing a lesion-tolerance strategy rather than repair would prevent double-strand breaks at closely-opposed excision repair sites on opposite strands of a damage-hypersensitive repeat.

  13. Re: Role of Telomeres and Telomerase in Cancer

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    Shay JW

    2016-03-01

    Full Text Available The most important difference between cancer and normall cells is the ability to continuous proliferation. This activation works due to telomeres and telomerase enzyme. Fifty years ago, Leonard Hayflick discovered that cultured normal humans cells have a limited capacity to divide. Today, this withdrawal from the cell cycle after a certain number of cellular divisions (replicative senescence is known to be triggered as a result of shortened telomeres. Studies on telomeres and telomerase have begun to provide additional information about aging and cancer development and have created new opportunities in the field of regenerative medicine for telomeropathies. Progressive telomere shortening from cell division (replicative aging provides a barrier for tumor progression. Continuous cell growth in malignancy correlates with the reactivation of telomerase. Telomerase is a cellular reverse transcriptase that adds new deoxyribonucleic acid (DNA onto the telomeres that are located at the ends of chromosomes. Telomeres consist of many kilobases of TTAGGG nucleotide repeats. The telomeric nucleotide repeats shorten with each cell division due to replication problems (DNA repair and oxidative damage. Quiescent/senescent state of the cell bypass can be accomplished by abrogating cell cycle checkpoint genes (such as TP53, p16INK4a, pRb. Telomerase is detected in approximately 90% of all malignant tumors. This telomerase activation has emerged as a target for cancer treatment. Telomerase therapeutics are classified as gene therapy (hTERT-telomerase catalytic protein component, hTR-telomerase functional, immunotherapy (Imetalstat-telomerase template antagonist, and small molecule inhibitors. In the near future, more specific researches on telomers and telomerase will contribute to aging/immortality studies (as stem cells and to discover new biomarkers for malignant tissue or anticancer therapeutics.

  14. Triagem cognitiva e comportamental de crianças com dificuldades de aprendizagem escolar: um estudo preliminar Selección cognitiva y conductual de niños con dificultades de aprendizaje escolar: un estudio preliminar Cognitive and behavioral screening of children with learning disabilities: a preliminary study

    Directory of Open Access Journals (Sweden)

    Lúcia Helena C. Santos

    2012-01-01

    Full Text Available OBJETIVO: Avaliar a aplicabilidade do Mini-Mental State Examination (MMSE e da lista de sintomas pediátricos (LSP como teste de triagem cognitiva e de problemas psicossociais em crianças com dificuldades de aprendizagem. MÉTODOS: Estudo descritivo e transversal envolvendo 103 crianças entre seis e nove anos de uma escola pública de Curitiba (PR com prováveis dificuldades de aprendizagem, realizado de 1º de março de 2002 a 30 de junho de 2009. Os dados foram obtidos simultaneamente por uma equipe multidisciplinar na avaliação inicial, com a aplicação do MMSE, da LSP e do Wechsler Intelligence Scale for Children (WISC III. Após 2007 o Child Behavior Checklist (CBCL e o Teacher's Report Form (TRF foram utilizados, sendo também comparados à LSP. Os coeficientes de correlação entre os testes foram calculados, sendo significante pOBJETIVO: Evaluar la aplicabilidad del mini-mental state examination (MMSE y de la lista de síntomas pediátricos (LSP como prueba de selección cognitiva y de problemas psicosociales en niños con dificultades de aprendizaje. MÉTODOS: Estudio descriptivo y transversal implicando a 103 niños entre seis y nueve años de una escuela pública de Curitiba (Paraná, Brasil con probables dificultades de aprendizaje, realizado desde el 1 de marzo de 2002 hasta el 30 de junio de 2009. Los datos se obtuvieron simultáneamente por un equipo multidisciplinario en la evaluación inicial, con la aplicación del MMSE, de la LSP y del Wechsler intelligence scale for children (WISC III. Después de 2007 el child behavior checklist (CBCL y el teacher's report form (TRF fueron utilizados, siendo también comparados a la LSP. Los coeficientes de correlación entre las pruebas fueron calculados, siendo significante pOBJECTIVE: To evaluate the applicability of the Mini-Mental State Examination (MMSE and the Pediatric Symptom Checklist (PSC as a screening test for cognitive and psychosocial problems in children with learning

  15. Characterization of primary biogenic aerosol particles in urban, rural, and high-alpine air by DNA sequence and restriction fragment analysis of ribosomal RNA genes

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    V. R. Després

    2007-12-01

    >
    Over 80% of the 53 bacterial sequences could be matched to one of the 19 T-RF peaks found in the PM2.5 samples, but only 40% of the T-RF peaks did correspond to one of the detected bacterial sequences. The results demonstrate that the T-RFLP analysis covered more of the bacterial diversity than the sequence analysis. Shannon-Weaver indices calculated from both sequence and T-RFLP data indicate that the bacterial diversity in the rural samples was higher than in the urban and alpine samples. Two of the bacterial sequences (Gammaproteobacteria and five of the T-RF peaks were found at all sampling locations.

  16. A homogeneous assay principle for universal substrate quantification via hydrogen peroxide producing enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Zscharnack, Kristin; Kreisig, Thomas; Prasse, Agneta A. [Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität Leipzig, Deutscher Platz 5, 04103 Leipzig (Germany); Zuchner, Thole, E-mail: Thole.Zuechner@octapharma.com [Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, Universität Leipzig, Deutscher Platz 5, 04103 Leipzig (Germany); Center for Biotechnology and Biomedicine, Universität Leipzig, Deutscher Platz 5, 04103 Leipzig (Germany)

    2015-01-07

    Highlights: • Application of the TRF-based PATb system for universal oxidase substrate detection. • H{sub 2}O{sub 2} generated by choline or glucose oxidase quenches the TRF signal of PATb. • The assay time is only limited by the oxidase catalysis rate. • Glucose is precisely detected in human serum consistent to a commercial assay. • A reliable quantification of choline in infant formula is shown. - Abstract: H{sub 2}O{sub 2} is a widely occurring molecule which is also a byproduct of a number of enzymatic reactions. It can therefore be used to quantify the corresponding enzymatic substrates. In this study, the time-resolved fluorescence emission of a previously described complex consisting of phthalic acid and terbium (III) ions (PATb) is used for H{sub 2}O{sub 2} detection. In detail, glucose oxidase and choline oxidase convert glucose and choline, respectively, to generate H{sub 2}O{sub 2} which acts as a quencher for the PATb complex. The response time of the PATb complex toward H{sub 2}O{sub 2} is immediate and the assay time only depends on the conversion rate of the enzymes involved. The PATb assay quantifies glucose in a linear range of 0.02–10 mmol L{sup −1}, and choline from 1.56 to 100 μmol L{sup −1} with a detection limit of 20 μmol L{sup −1} for glucose and 1.56 μmol L{sup −1} for choline. Both biomolecules glucose and choline could be detected without pretreatment with good precision and reproducibility in human serum samples and infant formula, respectively. Furthermore, it is shown that the detected glucose concentrations by the PATb system agree with the results of a commercially available assay. In principle, the PATb system is a universal and versatile tool for the quantification of any substrate and enzyme reaction where H{sub 2}O{sub 2} is involved.

  17. Postnatal arsenic exposure and attention impairment in school children.

    Science.gov (United States)

    Rodríguez-Barranco, Miguel; Gil, Fernando; Hernández, Antonio F; Alguacil, Juan; Lorca, Andres; Mendoza, Ramón; Gómez, Inmaculada; Molina-Villalba, Isabel; González-Alzaga, Beatriz; Aguilar-Garduño, Clemente; Rohlman, Diane S; Lacasaña, Marina

    2016-01-01

    Over the last few decades there has been an increased concern about the health risks from exposure to metallic trace elements, including arsenic, because of their potential neurotoxic effects on the developing brain. This study assessed whether urinary arsenic (UA) levels are associated with attention performance and Attention-Deficit/Hyperactivity Disorder (ADHD) in children living in an area with high industrial and mining activities in Southwestern Spain. A cross-sectional study was conducted on 261 children aged 6-9 years. Arsenic levels were determined in urine samples. Attention was measured by using 4 independent tools: a) tests from the Behavioral Assessment and Research System (BARS) designed to measure attention function: Simple Reaction Time Test (RTT), Continuous Performance Test (CPT) and Selective Attention Test (SAT); b) AULA Test, a virtual reality (VR)-based test that evaluates children's response to several stimuli in an environment simulating a classroom; c) Child Behavior Checklist (CBCL), administered to parents; and d) Teacher's Report Form (TRF), administered to teachers. Multivariate linear and logistic regression models, adjusted for potential confounders, were used to estimate the magnitude of the association between UA levels and attention performance scores. Higher UA levels were associated with an increased latency of response in RTT (β = 12.3; 95% confidence interval (CI): 3.5-21.1) and SAT (β = 3.6; 95% CI: .4-6.8) as well as with worse performance on selective and focalized attention in the AULA test (β for impulsivity = .6; 95% CI: .1-1.1; β for inattention = .5; 95% CI: .03-1.0). A dose-response relationship was observed between UA levels and inattention and impulsivity scores. In contrast, results from the CBCL and TRF tests failed to show a significant association with UA levels. In conclusion, UA levels were associated with impaired attention/cognitive function, even at levels considered safe. These results provide

  18. Burrow characteristics of the co-existing sibling species Mus booduga and Mus terricolor and the genetic basis of adaptation to hypoxic/hypercapnic stress

    Directory of Open Access Journals (Sweden)

    Narayan Gopeshwar

    2009-04-01

    Full Text Available Abstract Background The co-existing, sibling species Mus booduga and Mus terricolor show a difference in site-preference for burrows. The former build them in flat portion of the fields while the latter make burrows in earthen mounds raised for holding water in cultivated fields. In northern India which experiences great variation in climatic condition between summer and winter, M. booduga burrows have an average depth of 41 cm, as against 30 cm in southern India with less climatic fluctuation. M. terricolor burrows are about 20 cm deep everywhere. The three chromosomal species M. terricolor I, II and III have identical burrows, including location of the nest which is situated at the highest position. In contrast, in M. booduga burrows, the nest is at the lowest position. Results The nest chamber of M. booduga is located at greater depth than the nest chamber of M. terricolor. Also, in the burrows of M. booduga the exchange of air takes place only from one side (top surface in contrast to the burrows of M. terricolor where air exchange is through three sides. Hence, M. booduga lives in relatively more hypoxic and hypercapnic conditions than M. terricolor. We observed the fixation of alternative alleles in M. booduga and M. terricolor at Superoxide dismutase-1 (Sod-1, Transferrin (Trf and Hemoglobin beta chain (Hbb loci. All the three are directly or indirectly dependent on oxygen concentration for function. In addition to these, there are differences in burrow patterns and site-preference for burrows suggesting difference in probable adaptive strategy in these co-existing sibling species. Conclusion The burrow structure and depth of nest of the chromosomal species M. terricolor I, II and III are same everywhere probably due to the recency of their evolutionary divergence. Moreover, there is lack of competition for the well-adapted 'microhabitats' since they are non-overlapping in distribution. However, the co-existing sibling species M. booduga

  19. Effect of soil salinity and nutrient levels on the community structure of the root-associated bacteria of the facultative halophyte, Tamarix ramosissima, in southwestern United States.

    Science.gov (United States)

    Taniguchi, Takeshi; Imada, Shogo; Acharya, Kumud; Iwanaga, Fumiko; Yamanaka, Norikazu

    2015-01-01

    Tamarix ramosissima is a tree species that is highly resistant to salt and drought. The Tamarix species survives in a broad range of environmental salt levels, and invades major river systems in southwestern United States. It may affect root-associated bacteria (RB) by increasing soil salts and nutrients. The effects of RB on host plants may vary even under saline conditions, and the relationship may be important for T. ramosissima. However, to the best of our knowledge, there have been no reports relating to T. ramosissima RB and its association with salinity and nutrient levels. In this study, we have examined this association and the effect of arbuscular mycorrhizal colonization of T. ramosissima on RB because a previous study has reported that colonization of arbuscular mycorrhizal fungi affected the rhizobacterial community (Marschner et al., 2001). T. ramosissima roots were collected from five locations with varying soil salinity and nutrient levels. RB community structures were examined by terminal restriction fragment (T-RF) length polymorphism, cloning, and sequencing analyses. The results suggest that RB richness, or the diversity of T. ramosissima, have significant negative relationships with electrical conductivity (EC), sodium concentration (Na), and the colonization of arbuscular mycorrhizal fungi, but have a significant positive relationship with phosphorus in the soil. However, at each T-RF level, positive correlations between the emergence of some T-RFs and EC or Na were observed. These results indicate that high salinity decreased the total number of RB species, but some saline-tolerant RB species multiplied with increasing salinity levels. The ordination scores of nonmetric multidimensional scale analysis of RB community composition show significant relationships with water content, calcium concentration, available phosphorus, and total nitrogen. These results indicate that the RB diversity and community composition of T. ramosissima are affected

  20. Automatic registration of multi-modal microscopy images for integrative analysis of prostate tissue sections

    International Nuclear Information System (INIS)

    Lippolis, Giuseppe; Edsjö, Anders; Helczynski, Leszek; Bjartell, Anders; Overgaard, Niels Chr

    2013-01-01

    Prostate cancer is one of the leading causes of cancer related deaths. For diagnosis, predicting the outcome of the disease, and for assessing potential new biomarkers, pathologists and researchers routinely analyze histological samples. Morphological and molecular information may be integrated by aligning microscopic histological images in a multiplex fashion. This process is usually time-consuming and results in intra- and inter-user variability. The aim of this study is to investigate the feasibility of using modern image analysis methods for automated alignment of microscopic images from differently stained adjacent paraffin sections from prostatic tissue specimens. Tissue samples, obtained from biopsy or radical prostatectomy, were sectioned and stained with either hematoxylin & eosin (H&E), immunohistochemistry for p63 and AMACR or Time Resolved Fluorescence (TRF) for androgen receptor (AR). Image pairs were aligned allowing for translation, rotation and scaling. The registration was performed automatically by first detecting landmarks in both images, using the scale invariant image transform (SIFT), followed by the well-known RANSAC protocol for finding point correspondences and finally aligned by Procrustes fit. The Registration results were evaluated using both visual and quantitative criteria as defined in the text. Three experiments were carried out. First, images of consecutive tissue sections stained with H&E and p63/AMACR were successfully aligned in 85 of 88 cases (96.6%). The failures occurred in 3 out of 13 cores with highly aggressive cancer (Gleason score ≥ 8). Second, TRF and H&E image pairs were aligned correctly in 103 out of 106 cases (97%). The third experiment considered the alignment of image pairs with the same staining (H&E) coming from a stack of 4 sections. The success rate for alignment dropped from 93.8% in adjacent sections to 22% for sections furthest away. The proposed method is both reliable and fast and therefore well suited

  1. Plant management and biodiversity conservation in Náhuatl homegardens of the Tehuacán Valley, Mexico.

    Science.gov (United States)

    Larios, Carolina; Casas, Alejandro; Vallejo, Mariana; Moreno-Calles, Ana Isabel; Blancas, José

    2013-11-06

    The Tehuacán Valley is one of the areas of Mesoamerica with the oldest history of plant management. Homegardens are among the most ancient management systems that currently provide economic benefits to people and are reservoirs of native biodiversity. Previous studies estimated that 30% of the plant richness of homegardens of the region are native plant species from wild populations. We studied in Náhuatl communities the proportion of native plant species maintained in homegardens, hypothesizing to find a proportion similar to that estimated at regional level, mainly plant resources maintained for edible, medicinal and ornamental purposes. We analysed the composition of plant species of homegardens and their similarity with surrounding Cloud Forest (CF), Tropical Rainforest (TRF), Tropical Dry forest (TDF), and Thorn-Scrub Forest (TSF). We determined density, frequency and biomass of plant species composing homegardens and forests through vegetation sampling of a total of 30 homegardens and nine plots of forests, and documented ethnobotanical information on use, management, and economic benefits from plants maintained in homegardens. A total of 281 plant species was recorded with 12 use categories, 115 ornamental, 92 edible, and 50 medicinal plant species. We recorded 49.8 ± 23.2 (average ± S.D.) woody plant species (shrubs and trees) per homegarden. In total, 34% species are native to the Tehuacán Valley and nearly 16% are components of the surrounding forests. A total of 176 species were cultivated through seeds, vegetative propagules or transplanted entire individual plants, 71 tolerated, and 23 enhanced. The highest species richness and diversity were recorded in homegardens from the CF zone (199 species), followed by those from the TRF (157) and those from the TDF (141) zones. Homegardens provide a high diversity of resources for subsistence of local households and significantly contribute to conservation of native biodiversity. The highest diversity was

  2. High-resolution analysis of 16q22.1 in breast carcinoma using DNA amplifiable probes (multiplex amplifiable probe hybridization technique) and immunohistochemistry.

    Science.gov (United States)

    Rakha, Emad A; Armour, John A L; Pinder, Sarah E; Paish, Claire E; Ellis, Ian O

    2005-05-01

    Loss of the chromosomal material at 16q22.1 is one of the most frequent genetic aberrations found in both lobular and low-grade nonlobular invasive carcinoma of the breast, indicating the presence of a tumour suppressor gene (TSG) at this region in these tumours. However, the TSG (s) at the 16q22.1 in the more frequent nonlobular carcinomas is still unknown. Multiplex Amplifiable Probe Hybridisation (MAPH) is a simple, accurate and a high-resolution technique that provides an alternative approach to DNA copy-number measurement. The aim of our study was to examine the most likely candidate genes at 16q22.1 using MAPH assay combined with protein expression analysis by immunohistochemistry. We identified deletion at 16q22.1 that involves some or all of these genes. We also noticed that the smallest region of deletion at 16q22.1 could be delineated to a 3 Mb region centromeric to the P-cadherin gene. Apart from the correlation between E-cadherin protein expression and its gene copy number, no correlation was detected between the expression of E2F-4, CTCF, TRF2 or P-cadherin with their gene's copy number. In the malignant tissues, no significant loss or decrease of protein expression of any gene other than E-cadherin was seen in association with any specific tumour type. No expression of VE-cadherin or Ksp-cadherin was detected in the normal and/or malignant tissues of the breast in these cases. However, there was a correlation between increased nuclear expression of E2F-4 and tumours with higher histological grade (p = 0.04) and positive lymph node disease (p = 0.02), suggesting that it may have an oncogenic rather than a tumour suppressor role. The malignant breast tissues also showed abnormal cytoplasmic cellular localisation of CTCF, compared to its expression in the normal parenchymal cells. In conclusion, we have demonstrated that MAPH is a potential technique for assessment of genomic imbalances in malignant tissues. Although our results support E-cadherin as the

  3. Extracts from the edible seaweed, Ascophyllum nodosum, inhibit lipase activity in vitro: contributions of phenolic and polysaccharide components.

    Science.gov (United States)

    Austin, Ceri; Stewart, Derek; Allwood, J William; McDougall, Gordon J

    2018-01-24

    A polyphenol-rich extract (PRE) from the edible seaweed, Ascophyllum nodosum, inhibited pancreatic lipase activity in an oil-based turbidimetric assay with an IC 50 of 200 μg gallic acid equivalents (GAE) perassay) [∼230 μg DW] whereas the known inhibitor, Orlistat, gave an IC 50 at 0.4 μg per assay. A phlorotannin-enriched fraction (TRF) purified from the PRE was more potent with an IC 50 = 60 μg GAE per assay (∼65 μg DW). When the assay was started by the addition of lipase, both Orlistat and TRF were much less effective which suggests that pre-incubation of enzyme and inhibitor improved inhibition. Based on phenol content, water extracts from Ascophyllum were more potent lipase inhibitors than PRE (IC 50 ∼ 150 μg GAE per assay). However, this was equivalent to ∼580 μg DW and these extracts contained polysaccharides (e.g. alginate content = 110 μg mL -1 ) which may also contribute to inhibition. Indeed, a polysaccharide-enriched fraction obtained by ethanol precipitation gave an IC 50 of 1000 μg DW which was equivalent to 130 μg GAE and 420 μg alginate per assay. Therefore a >3 fold increase in alginate content did not markedly improve inhibition. Re-precipitation increased alginate content and reduced polyphenol content but lipase inhibition was markedly reduced (i.e. IC 50 at ∼1100 μg DW per assay, 700 μg alginate and 25 μg GAE). Purifying the polysaccharide fraction by ion exchange removed all phenolics but the IC 50 increased to >2500 μg DW, equivalent to >1970 μg alginate per assay. In conclusion, polysaccharides and phlorotannins may inhibit lipase in an additive fashion, with phlorotannins apparently more effective in vitro. However, interactions between these components may be important when food products containing this edible seaweed are consumed.

  4. The ILRS Reanalysis 1983 - 2009 Contributed To ITRF2008

    Science.gov (United States)

    Pavlis, E. C.; Luceri, V.; Sciarretta, C.; Kelm, R.

    2009-12-01

    For over two decades, Satellite Laser Ranging (SLR) data contribute to the definition of the Terrestrial Reference Frame (TRF). Until the development of ITRF2000, the contributions were submitted in the form of a set of normal equations or a covariance matrix of station coordinates and their linear rates at a standard epoch. The development of ITRF2005 ushered a new era with the use of weekly or session contributions, allowing greater flexibility in the relative weighting and the combination of information from various techniques. Moreover, the need of a unique, official, representative solution for each Technique Service, based on the rigorous combination of the various Analysis Centers’ contributions, gave the opportunity to all techniques to verify, as a first step, the intra-technique solution consistency and, immediately after, to engage in discussions and comparison of the internal procedures, leading to a harmonization and validation of these procedures and the adopted models in the inter-technique context. In many occasions, the time series approach joint with the intra- and inter-technique comparison steps also highlighted differences that previously went unnoticed, and corrected incompatibilities. During the past year we have been preparing the ILRS contribution to a second TRF developed in the same way, the ITRF2008. The ILRS approach is based strictly on the current IERS Conventions 2003 and our internal standards. The Unified Analysis Workshop in 2007 stressed a number of areas where each technique needed to focus more attention in future analyses. In the case of SLR, the primary areas of concern were tracking station biases, extending the data span used in the analysis, and target characteristics. The present re-analysis extends from 1983 to 2009, covering a 25-year period, the longest for any of the contributing techniques; although the network and data quality for the 1983-1993 period are significantly poorer than for the latter years, the overall

  5. The ILRS contribution to ITRF2008

    Science.gov (United States)

    Pavlis, E. C.; Luceri, V.; Sciarretta, C.; Kelm, R.

    2009-04-01

    Since over two decades, Satellite Laser Ranging (SLR) data contribute to the definition of the Terrestrial Reference Frame (TRF). Until the development of ITRF2000, the contributions were submitted in the form of a set of normal equations or a covariance matrix of station coordinates and their linear rates at a standard epoch. The development of ITRF2005 ushered a new era with the use of weekly or session contributions, allowing greater flexibility in the relative weighting and the combination of information from various techniques. Moreover, the need of a unique, official, representative solution for each Technique Service, based on the rigorous combination of the various Analysis Centers' contributions, gave the opportunity to all techniques to verify, as a first step, the intra-technique solution consistency and, immediately after, to engage in discussions and comparison of the internal procedures, leading to a harmonization and validation of these procedures and the adopted models in the inter-technique context. In many occasions, the time series approach joint with the intra- and inter-technique comparison steps also highlighted differences that previously went unnoticed, and corrected incompatibilities. During the past year we have been preparing the ILRS contribution to a second TRF developed in the same way, the ITRF2008. The ILRS approach is based strictly on the current IERS Conventions 2003 and our internal standards. The Unified Analysis Workshop in 2007 stressed a number of areas where each technique needed to focus more attention in future analyses. In the case of SLR, the primary areas of concern were tracking station biases, extending the data span used in the analysis, and target characteristics. The present re-analysis extends from 1983 to 2008, covering a 25-year period, the longest for any of the contributing techniques; although the network and data quality for the 1983-1993 period are significantly poorer than for the latter years, the overall

  6. Karyotypic diversity in four species of the genus Gymnotus Linnaeus, 1758 (Teleostei, Gymnotiformes, Gymnotidae): physical mapping of ribosomal genes and telomeric sequences

    Science.gov (United States)

    Scacchetti, Priscilla Cardim; Pansonato-Alves, José Carlos; Utsunomia, Ricardo; Oliveira, Claudio; Foresti, Fausto

    2011-01-01

    Abstract Conventional (Giemsa, C-Banding, Ag-NORs, CMA3) and molecular (5S rDNA, 18S rDNA, telomeric sequences) cytogenetic studies were carried out in specimens of ten distinct fish populations of the genus Gymnotus (Gymnotus sylvius Albert and Fernandes-Matioli, 1999, Gymnotus inaequilabiatus Valenciennes, 1839, Gymnotus pantherinus Steindachner, 1908, and G. cf. carapo Linnaeus, 1758) from different Brazilian hydrographic basins. Gymnotus sylvius presented a diploid number of 40 chromosomes (22m+12sm+6st), Gymnotus pantherinus presented 52 chromosomes (32m+18sm+2st), while Gymnotus inaequilabiatus (42m+10sm+2a)and Gymnotus cf. carapo (38m+12sm+4st) presented 54 chromosomes. The C-banding technique revealed centromeric marks in all chromosomes of all species. Besides that, conspicuous blocks of heterochromatin were found interstitially on the chromosomes of Gymnotus inaequilabiatus, Gymnotus cf. carapo,and Gymnotus pantherinus. All four species showed single nucleolus organizing regions confirmed by results obtained through Ag-NORs and FISH experiments using 18S rDNA probes, which showed the NORs localized on the first chromosome pair in Gymnotus inaequilabiatus, Gymnotus cf. carapo,and Gymnotus pantherinus, and on pair 2 in Gymnotus sylvius. CMA3 staining revealed additional unrelated NORs marks in Gymnotus sylvius and Gymnotus pantherinus. The 5S rDNA probes revealed signals on one pair in Gymnotus sylvius and two pairs in Gymnotus pantherinus; Gymnotus inaequilabiatus had about seventeen pairs marked, and Gymnotus cf. carapo had about fifteen pairs marked. It is considered that the high amount of heterochromatin identified in the chromosomes of Gymnotus inaequilabiatus and Gymnotus cf. carapo could have facilitated the dispersion of 5S rDNA in these species. Interstitial signals were detected on the first metacentric pair of Gymnotus sylvius by telomeric probes (TTAGGG)n indicating the possible occurrence of chromosomal fusions in this species. The present

  7. Three-dimensional telomere architecture of esophageal squamous cell carcinoma: comparison of tumor and normal epithelial cells.

    Science.gov (United States)

    Sunpaweravong, S; Sunpaweravong, P; Sathitruangsak, C; Mai, S

    2016-05-01

    Telomeres are repetitive nucleotide sequences (TTAGGG)n located at the ends of chromosomes that function to preserve chromosomal integrity and prevent terminal end-to-end fusions. Telomere loss or dysfunction results in breakage-bridge-fusion cycles, aneuploidy, gene amplification and chromosomal rearrangements, which can lead to genomic instability and promote carcinogenesis. Evaluating the hypothesis that changes in telomeres contribute to the development of esophageal squamous cell carcinoma (ESCC) and to determine whether there are differences between young and old patients, we compared the three-dimensional (3D) nuclear telomere architecture in ESCC tumor cells with that of normal epithelial cells obtained from the same patient. Patients were equally divided by age into two groups, one comprising those less than 45 years of age and the other consisting of those over 80 years of age. Tumor and normal epithelial cells located at least 10 cm from the border of the tumor were biopsied in ESCC patients. Hematoxylin and eosin staining was performed for each sample to confirm and identify the cancer and normal epithelial cells. This study was based on quantitative 3D fluorescence in situ hybridization (Q-FISH), 3D imaging and 3D analysis of paraffin-embedded slides. The 3D telomere architecture data were computer analyzed using 100 nuclei per slide. The following were the main parameters compared: the number of signals (number of telomeres), signal intensity (telomere length), number of telomere aggregates, and nuclear volume. Tumor and normal epithelial samples from 16 patients were compared. The normal epithelial cells had more telomere signals and higher intensities than the tumor cells, with P-values of P architecture and found no statistically significant differences in any parameter tested between the young and old patients in either the tumor or epithelial cells. The 3D nuclear telomeric signature was able to detect differences in telomere architecture

  8. Stabilization of Telomere G-Quadruplexes Interferes with Human Herpesvirus 6A Chromosomal Integration.

    Science.gov (United States)

    Gilbert-Girard, Shella; Gravel, Annie; Artusi, Sara; Richter, Sara N; Wallaschek, Nina; Kaufer, Benedikt B; Flamand, Louis

    2017-07-15

    Human herpesviruses 6A and 6B (HHV-6A/B) can integrate their genomes into the telomeres of human chromosomes using a mechanism that remains poorly understood. To achieve a better understanding of the HHV-6A/B integration mechanism, we made use of BRACO-19, a compound that stabilizes G-quadruplex secondary structures and prevents telomere elongation by the telomerase complex. First, we analyzed the folding of telomeric sequences into G-quadruplex structures and their binding to BRACO-19 using G-quadruplex-specific antibodies and surface plasmon resonance. Circular dichroism studies indicate that BRACO-19 modifies the conformation and greatly stabilizes the G-quadruplexes formed in G-rich telomeric DNA. Subsequently we assessed the effects of BRACO-19 on the HHV-6A initial phase of infection. Our results indicate that BRACO-19 does not affect entry of HHV-6A DNA into cells. We next investigated if stabilization of G-quadruplexes by BRACO-19 affected HHV-6A's ability to integrate its genome into host chromosomes. Incubation of telomerase-expressing cells with BRACO-19, such as HeLa and MCF-7, caused a significant reduction in the HHV-6A integration frequency ( P integration frequency in U2OS cells that lack telomerase activity and elongate their telomeres through alternative lengthening mechanisms. Our data suggest that the fluidity of telomeres is important for efficient chromosomal integration of HHV-6A and that interference with telomerase activity negatively affects the generation of cellular clones containing integrated HHV-6A. IMPORTANCE HHV-6A/B can integrate their genomes into the telomeres of infected cells. Telomeres consist of repeated hexanucleotides (TTAGGG) of various lengths (up to several kilobases) and end with a single-stranded 3' extension. To avoid recognition and induce a DNA damage response, the single-stranded overhang folds back on itself and forms a telomeric loop (T-loop) or adopts a tertiary structure, referred to as a G-quadruplex. In the

  9. Toward the design of new DNA G-quadruplex ligands through rational analysis of polymorphism and binding data.

    Science.gov (United States)

    Artese, Anna; Costa, Giosuè; Distinto, Simona; Moraca, Federica; Ortuso, Francesco; Parrotta, Lucia; Alcaro, Stefano

    2013-10-01

    Human telomeres play a key role in protecting chromosomal ends from fusion events; they are composed of d(TTAGGG) repeats, ranging in size from 3 to 15 kb. They form G-quadruplex DNA structures, stabilized by G-quartets in the presence of cations, and are involved in several biological processes. In particular, a telomere maintenance mechanism is provided by a specialized enzyme called telomerase, a reverse transcriptase able to add multiple copies of the 5'-GGTTAG-3' motif to the end of the G-strand of the telomere and which is over-expressed in the majority of cancer cells. The central cation has a crucial role in maintaining the stability of the structure. Based on its nature, it can be associated with different topological telomeric quadruplexes, which depend also on the orientation of the DNA strands and the syn/anti conformation of the guanines. Such a polymorphism, confirmed by the different structures deposited in the Protein Data Bank (PDB), prompted us to apply a computational protocol in order to investigate the conformational properties of a set of known G-quadruplex ligands and their molecular recognition against six different experimental models of the human telomeric sequence d[AG3(T2AG3)3]. The average AutoDock correlation between theoretical and experimental data yielded an r2 value equal to 0.882 among all the studied models. Such a result was always improved with respect to those of the single folds, with the exception of the parallel structure (r2 equal to 0.886), thus suggesting a key role of this G4 conformation in the stacking interaction network. Among the studied binders, a trisubstituted acridine and a dibenzophenanthroline derivative were well recognized by the parallel and the mixed G-quadruplex structures, allowing the identification of specific key contacts with DNA and the further design of more potent or target specific G-quadruplex ligands. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  10. THE EFFECT OF EMOTIONAL INTELLIGENCE TRAINING ON BEHAVIOR PROBLEMS OF BOYS WITH EXTERNALIZED BEHAVIOR DISORDER IN ELEMENTARY SCHOOLS

    Directory of Open Access Journals (Sweden)

    Kosar Moghaddam POUR

    2014-09-01

    Full Text Available The purpose of this study is to investigate the effect of emotional intelligence on the behavior problems of boys with Externalized behavior disorder in Primary Schools. Method: This quasi-experimental study was conducted along with a pre-test, post-test, with a control group and a follow-up test. For sampling, 40 students identified with Externalized behavioral problems through the Teacher Report Form (TRF and the Child Behavior Checklist (CBCL were chosen and randomly divided into two groups (20 in the experimental group and 20 in the control group. The experimental group received emotional intelligence training program in 17 sessions (2 sessions per week, 60 minutes per session and the control group received no training beyond their regular school program. After two months, in order to examine the stability (durability of training effect, the follow-up test was conducted. Finally, the data obtained were analyzed using the statistical method of generalized estimating equations. Results: The results showed that the intervention program had created a significant difference between the scores of the experimental and control group (p<0.001 and the rate of behavioral problems (aggression, rule breaking occurrence has dropped. This was true for the follow-up results too. Conclusions: It can be concluded that Emotional Intelligence Training decreases the behavior problems of boys with Externalized behavior disorder and helps to prevent high occurrence of these problems.

  11. Jason-1 and Jason-2 POD Using GPS

    Science.gov (United States)

    Melachroinos, Stavros; Lemoine, Frank G.; Zelensky, Nikita P.; Rowlands, David D.; Luthcke, Scott B.; Beckley, Brian D.

    2012-01-01

    The Jason-2 satellite, launched in June 2008, is the latest follow-on to the successful Jason-1 altimetry satellite mission launched in December 7, 2001. Both, Jason-2 and Jason-1 are equipped with a GPS dual-frequency receiver, a laser retroreflector array, and a DORIS receiver for precise orbit determination (POD). A series of dynamic and reduced-dynamic Jason-2 orbits computed at NASA GSFC, based on GPS-only data and the std0905 standards, have been completed till cy74through cycle 74 using the IGS05 framework. These orbits, now publicly available, have been shown to agree radially at 1 cm RMS with the GSFC std0905 SLR/DORIS orbits and in comparison with orbits produced by JPL, ESA and CNES. In this paper, we describe the implementation of the IGS08 and repro1 framework for the Jason-2 and Jason-1 GPS POD processing with the NASA GSFC GEODYN software. . In our updated GPS POD, ambiguity fixing and updated time variable and static gravity fields. We also evaluate the implementation of non-tidal and degree-1 loading displacement as forward modeling to the tracking stations. Reduced-dynamic versus dynamic orbit differences are used to characterize the remaining force model errors and TRF instability. In particular, we assess their consistency radially and the stability of the altimeter satellite reference frame in the North/South direction as a proxy to assess the consistency of the reference frame.

  12. THE VALIDATION OF THE RESULTS OF MICROARRAY STUDIES OF ASSOCIATION BETWEEN GENE POLYMORPHISMS AND THE FREQUENCY OF RADIATION EXPOSURE MARKERS

    Directory of Open Access Journals (Sweden)

    M. V. Khalyuzova

    2014-01-01

    Full Text Available The results from the selective validation research into the association between genetic polymorphisms and the frequency of cytogenetic abnormalities on a large independent sample are analyzed. These polymorphisms have been identified previously during own microarray studies. It has been shown an association with the frequency of dicentric and ring chromosomes induced by radiation exposure. The study was conducted among Siberian Group of Chemical Enterprises healthy employees (n = 573 exposed to professional irradiation in a dose range of 40–400 mSv. We have found that 5 SNP are confirmed to be associated with the frequency of dicentric and ring: INSR rs1051690 – insulin receptor gene; WRNrs2725349 – Werner syndrome gene, RecQ helicase-like; VCAM1 rs1041163 – vascular cell adhesion molecule 1 gene; PCTP rs2114443 – phosphatidylcholine transfer protein gene; TNKS rs7462102 – tankyrase gene; TRF1-interacting ankyrin-related ADP-ribose polymerase. IGF1 rs2373721 – insulin-like growth factor 1 gene has not confirmed to be associated with the frequency of dicentric and ring chromosomes.

  13. Detecting microsatellites within genomes: significant variation among algorithms

    Directory of Open Access Journals (Sweden)

    Rivals Eric

    2007-04-01

    Full Text Available Abstract Background Microsatellites are short, tandemly-repeated DNA sequences which are widely distributed among genomes. Their structure, role and evolution can be analyzed based on exhaustive extraction from sequenced genomes. Several dedicated algorithms have been developed for this purpose. Here, we compared the detection efficiency of five of them (TRF, Mreps, Sputnik, STAR, and RepeatMasker. Results Our analysis was first conducted on the human X chromosome, and microsatellite distributions were characterized by microsatellite number, length, and divergence from a pure motif. The algorithms work with user-defined parameters, and we demonstrate that the parameter values chosen can strongly influence microsatellite distributions. The five algorithms were then compared by fixing parameters settings, and the analysis was extended to three other genomes (Saccharomyces cerevisiae, Neurospora crassa and Drosophila melanogaster spanning a wide range of size and structure. Significant differences for all characteristics of microsatellites were observed among algorithms, but not among genomes, for both perfect and imperfect microsatellites. Striking differences were detected for short microsatellites (below 20 bp, regardless of motif. Conclusion Since the algorithm used strongly influences empirical distributions, studies analyzing microsatellite evolution based on a comparison between empirical and theoretical size distributions should therefore be considered with caution. We also discuss why a typological definition of microsatellites limits our capacity to capture their genomic distributions.

  14. Hydrogen isotope storage in zircaloy scrap

    Energy Technology Data Exchange (ETDEWEB)

    Lee, H. S.; Kuk, I. H.; Chung, H.; Paek, S. W.; Kang, H. S

    1999-08-01

    8 MCi of tritium a year will be produced after wolsong TRF is in operation. The metal hydride form is one of useful tritium storage. The metals in use for metal hydride are uranium, titanium, etc., however uranium is limited to use by regulation, and titanium is relatively costly. Both metals are not produced in country but whole amount is imported. On the other hand 2,000kg of zircaloy scrap is produced by CANDU nuclear fuel fabrication process, which is also useful for hydrogen storage. The purpose of this study is to evaluation of hydrogen absorption capacity for zircaloy scrap that is produced as waste by CANDU nuclear fuel fabrication process. The sample evacuated for an hour at 1000 deg C. The strip showed higher capacity : 0.7 at 25 deg C, 2.0 at 200 deg C, 2.0 at 200 deg C, 2.0 at 400 deg C, respectively. The H/M values for commercial zircaloy sponge were 2.0 at 25 deg C and 2.0 at 400 deg C.

  15. Telomerase gene therapy rescues telomere length, bone marrow aplasia, and survival in mice with aplastic anemia.

    Science.gov (United States)

    Bär, Christian; Povedano, Juan Manuel; Serrano, Rosa; Benitez-Buelga, Carlos; Popkes, Miriam; Formentini, Ivan; Bobadilla, Maria; Bosch, Fatima; Blasco, Maria A

    2016-04-07

    Aplastic anemia is a fatal bone marrow disorder characterized by peripheral pancytopenia and marrow hypoplasia. The disease can be hereditary or acquired and develops at any stage of life. A subgroup of the inherited form is caused by replicative impairment of hematopoietic stem and progenitor cells due to very short telomeres as a result of mutations in telomerase and other telomere components. Abnormal telomere shortening is also described in cases of acquired aplastic anemia, most likely secondary to increased turnover of bone marrow stem and progenitor cells. Here, we test the therapeutic efficacy of telomerase activation by using adeno-associated virus (AAV)9 gene therapy vectors carrying the telomerase Tert gene in 2 independent mouse models of aplastic anemia due to short telomeres (Trf1- and Tert-deficient mice). We find that a high dose of AAV9-Tert targets the bone marrow compartment, including hematopoietic stem cells. AAV9-Tert treatment after telomere attrition in bone marrow cells rescues aplastic anemia and mouse survival compared with mice treated with the empty vector. Improved survival is associated with a significant increase in telomere length in peripheral blood and bone marrow cells, as well as improved blood counts. These findings indicate that telomerase gene therapy represents a novel therapeutic strategy to treat aplastic anemia provoked or associated with short telomeres. © 2016 by The American Society of Hematology.

  16. The Tropospheric Products of the International VLBI Service for Geodesy and Astrometry

    Science.gov (United States)

    Heinkelmann, Robert; Schwatke, Christian

    2010-01-01

    The IVS runs two tropospheric products: The IVS tropospheric parameter rapid combination monitors the zenith wet delay (ZWD) and zenith total delay (ZTD) of the rapid turnaround sessions R1 and R4. Goal of the combination is the identification and the exclusion of outliers by comparison and the assessment of the precision of current VLBI solutions in terms of tropospheric parameters. The rapid combination is done on a weekly basis four weeks after the observation files are released on IVS Data Centers. Since tropospheric and geodetic parameters, such as vertical station components, can significantly correlate, the consistency of the ZTD can be a measure of the consistency of the corresponding TRF as well. The ZWD mainly rely on accurate atmospheric pressure data. Thus, besides estimation techniques, modeling and analyst s noise, ZWD reflects differences in the atmospheric pressure data applied to the VLBI analysis. The second product, called tropospheric parameter long-term combination, aims for an accurate determination of climatological signals, such as trends of the atmospheric water vapor observed by VLBI. Therefore, the long-term homogeneity of atmospheric pressure data plays a crucial role for this product. The paper reviews the methods applied and results achieved so far and describes the new maintenance through DGFI.

  17. Introduction to Wolsong Tritium Removal Facility (WTRF)

    International Nuclear Information System (INIS)

    Song, K. M.; Sohn, S. H.; Kang, D. W.; Chung, H. S.

    2005-01-01

    Four CANDU 6 reactors have been operated at Wolsong site. Tritium is primarily produced in heavywater-moderated-power reactors by neutron capture of deuterium nuclei in the heavy water moderator and coolant. The concentration of tritium in the reactor moderator and coolant systems increases with time of reactor operation. For CANDU 6 reactors, the estimated equilibrium values are ∼3 TBq/kg-D 2 O in the moderator and ∼74 GBq/kg-D 2 O in the coolant, where the production rate is balanced by tritium decay and water makeup and loss process. The tritium level in the moderator heavy water of Wolsong Unit-1 is getting higher for about 20-year operation and is over 2.22x10 12 Bq/kg at the end of 2003. It was known that the tritium levels in the moderators of the other units would be also steadily increased. In order to reduce the tritium activity, KHNP has committed to construct a Tritium Removal Facility (TRF) at the Wolsong site

  18. Hydrogen isotope storage in zircaloy scrap

    International Nuclear Information System (INIS)

    Lee, H. S.; Kuk, I. H.; Chung, H.; Paek, S. W.; Kang, H. S.

    1999-08-01

    8 MCi of tritium a year will be produced after wolsong TRF is in operation. The metal hydride form is one of useful tritium storage. The metals in use for metal hydride are uranium, titanium, etc., however uranium is limited to use by regulation, and titanium is relatively costly. Both metals are not produced in country but whole amount is imported. On the other hand 2,000kg of zircaloy scrap is produced by CANDU nuclear fuel fabrication process, which is also useful for hydrogen storage. The purpose of this study is to evaluation of hydrogen absorption capacity for zircaloy scrap that is produced as waste by CANDU nuclear fuel fabrication process. The sample evacuated for an hour at 1000 deg C. The strip showed higher capacity : 0.7 at 25 deg C, 2.0 at 200 deg C, 2.0 at 200 deg C, 2.0 at 400 deg C, respectively. The H/M values for commercial zircaloy sponge were 2.0 at 25 deg C and 2.0 at 400 deg C

  19. Effectiveness of Group Play Therapy on Symptoms of Oppositional Defiant Among Children

    Directory of Open Access Journals (Sweden)

    Narges Morshed

    2015-12-01

    Full Text Available Background and Objectives: With regard to the prevalence of Oppositional-Defiant Disorder in children and converting to the other disorders, if left untreated, this research aims to investigate the effectiveness of group play therapy on oppositional-defiant disorder symptoms among children. Materials and Methods: This study is interventional and quasi-experimental research. In this study based on cluster sampling method, 30 participants were selected and randomly assigned to the experimental and control groups. The tools discussed here included Child Behavior Checklist (CBCL, Raven's Progressive Matrixes, Teacher Report Form (TRF as well as a clinical interview with parents. Play therapy was provided weekly by group for the participants, in sixty-minute eight sessions. Participants were assessed in three stages of pre- interference post- interference and after two month intervals from completing sessions. SPSS18 and multi-variables covariance analysis method were used for analyzing data. Results: The results obtained by Mancova analysis showed that there was a significant decrease in oppositional defiant-disorder symptoms in comparison with control group reporting by parents and teacher (P < 0.001. In addition, the results indicated the same effect after two months. Conclusions: The results indicated the efficiency of group play therapy on decrease of oppositional defiant disorder symptoms among children. Accordingly using this treatment method on children was recommended to the therapists.

  20. An RFID-Based Closed-Loop Wireless Power Transmission System for Biomedical Applications.

    Science.gov (United States)

    Kiani, Mehdi; Ghovanloo, Maysam

    2010-04-01

    This brief presents a standalone closed-loop wireless power transmission system that is built around a commercial off-the-shelf (COTS) radio-frequency identification (RFID) reader (TRF7960) operating at 13.56 MHz. It can be used for inductively powering implantable biomedical devices in a closed loop. Any changes in the distance and misalignment between transmitter and receiver coils in near-field wireless power transmission can cause a significant change in the received power, which can cause either a malfunction or excessive heat dissipation. RFID circuits are often used in an open loop. However, their back telemetry capability can be utilized to stabilize the received voltage on the implant. Our measurements showed that the delivered power to the transponder was maintained at 11.2 mW over a range of 0.5 to 2 cm, while the transmitter power consumption changed from 78 mW to 1.1 W. The closed-loop system can also oppose voltage variations as a result of sudden changes in the load current.

  1. State-of-the-art and main options to improve fuel-energy complex of ferrous metallurgy

    Energy Technology Data Exchange (ETDEWEB)

    Rozenblit, G I; Pashkov, V D; Romanov, G M

    1981-01-01

    In 1980, the State Institute for the Design and Planning Metallury (Gripromez), elaborated ''The main options of Fuel and energy resources conservation (FERG) in ferrous metallurgy of the USSR program of works for the period 1981-1985 and up to 1990''. The Gipromez technical committee recommended: 1) elaborating feasibility studies and reports, developing branch schemes and starting complexes to separate out FERC measures; 2) inclusion of the FERC measures at the starting complexes as the first stages of main projects construction; 3) that the Ministy of Ferrous Metallurgy of the USSR, general designers and enterprises reconsider the starting complexes of the projects constructed during the present five-year period and incorporate in them the urgent FERC actions on heat-utilizing facilities. Changing the steel smelting process structure through more extensive use of the converter process and installation of continuous blank casting allows achievement of considerable fuel conservation, some 4 m trf per year as compared with its consumption in the scheme of open-hearth furnace - blooming mill (slabbing mill). During the 11th five-year-plan period introduction of metallurgy plant. An installation with discharge of the converter gas without its afterburning and successive utilization as a fuel in the converter shop of the Novolipetsk is planned.

  2. Effect of olanzapine combined with modified electroconvulsive therapy on cytokines, sTNFRs and neural electrophysiological characteristics in patients with schizophrenia

    Directory of Open Access Journals (Sweden)

    Wei Cheng

    2016-12-01

    Full Text Available Objective: To analyze the effect of olanzapine combined with modified electroconvulsive therapy on cytokines, sTNFRs and neural electrophysiological characteristics in patients with schizophrenia. Methods: Patients with schizophrenia treated in our hospital between March 2013 and March 2016 were selected and randomly divided into two groups, the observation group received olanzapine combined with modified electroconvulsive therapy, and the control group received olanzapine therapy. After 6 weeks of treatment, serum levels of soluble tumor necrosis factor receptor (sTNFR, acute phase reaction proteins and brain function indexes as well as the neural electrophysiological characteristics were compared between the two groups. Results: After 6 weeks of treatment, serum sTNFRs, CRP, CER and AAG content of observation group were lower than those of control group while TRF content was higher than that of control group; serum brain function indexes NGF and BDNF content were higher than those of control group while GFAP, S100B, NSE and Hcy content were lower than those of control group; nerve electrophysiology indexes P300, LPP and ERN amplitude were higher than those of control group while LPP amplitude was lower than that of control group. Conclusions: Olanzapine combined with modified electroconvulsive therapy can optimize the condition of schizophrenia, reduce the abnormal degree of nerve electrophysiology and help to improve treatment outcome.

  3. Thermal power terms in the Einstein-dilaton system

    International Nuclear Information System (INIS)

    Zuo, Fen

    2014-01-01

    We employ the gauge/string duality to study the thermal power terms of various thermodynamic quantities in gauge theories and the renormalized Polyakov loop above the deconfinement phase transition. We restrict ourselves to the five-dimensional Einstein gravity coupled to a single scalar, the dilaton. The asymptotic solutions of the system for a general dilaton potential are employed to study the power contributions of various quantities. If the dilaton is dual to the dimension-4 operator TrF μν 2 , no power corrections would be generated. Then the thermal quantities approach their asymptotic values much more quickly than those observed in lattice simulation. When the dimension of the dual operator is different from 4, various power terms are generated. The lowest power contributions to the thermal quantities are always quadratic in the dilaton, while that of the Polyakov loop is linear. As a result, the quadratic terms in inverse temperature for both the trace anomaly and the Polyakov loop, observed in lattice simulation, cannot be implemented consistently in the system. This is in accordance with the field theory expectation, where no gauge-invariant operator can accommodate such contributions. Two simple models, where the dilaton is dual to operators with different dimensions, are studied in detail to clarify the conclusion.

  4. Equivalent network for resistance and temperature coefficient of resistance versus temperature and composition of thick resistive films

    International Nuclear Information System (INIS)

    Kusy, A.

    1987-01-01

    Two types of elementary resistances in thick resistive films have been considered: (i) constriction resistance R/sub C/ determined by the bulk properties of conducting material and by the geometry of constriction, and (ii) barrier resistance R/sub B/ determined by the parameters of a thermally activated type of tunneling process and by the geometry of the metal-insulator-metal unit. On this basis a resistance network composed of a large number of the two types of resistances has been defined. The network has been considered as being equivalent to thick resistive film (TRF) from the point of view of the resistance and temperature coefficient of resistance (TCR). The parameters of this network have been evaluated by the computer-aided approximation of the experimental data found for RuO 2 -based TRFs. On the basis of the equations derived for the network as well as the results of the approximation process, it can be concluded that the small values of the network TCR result from the superposition of the TCR of the conducting component β/sub C/ and of the temperature coefficient of barrier resistance α/sub B/. In this superposition β/sub C/ is attenuated (by 1--2 orders of magnitude), while α/sub B/ is attenuated by only few percentages. The network has been found to be strongly barrier dominated

  5. Etiology of Stability and Growth of Internalizing and Externalizing Behavior Problems Across Childhood and Adolescence.

    Science.gov (United States)

    Hatoum, Alexander S; Rhee, Soo Hyun; Corley, Robin P; Hewitt, John K; Friedman, Naomi P

    2018-04-20

    Internalizing and externalizing behaviors are heritable, and show genetic stability during childhood and adolescence. Less work has explored how genes influence individual differences in developmental trajectories. We estimated ACE biometrical latent growth curve models for the Teacher Report Form (TRF) and parent Child Behavior Checklist (CBCL) internalizing and externalizing scales from ages 7 to 16 years in 408 twin pairs from the Colorado Longitudinal Twin Study. We found that Intercept factors were highly heritable for both internalizing and externalizing behaviors (a2 = .61-.92), with small and nonsignificant environmental influences for teacher-rated data but significant nonshared environmental influences for parent-rated data. There was some evidence of heritability of decline in internalizing behavior (Slopes for teacher and parent ratings), but the Slope genetic variance was almost entirely shared with that for the Intercept when different than zero. These results suggest that genetic effects on these developmental trajectories operate primarily on initial levels and stability, with no significant unique genetic influences for change. Finally, cross-rater analyses of the growth factor scores revealed moderate to large genetic and environmental associations between growth factors derived from parents' and teachers' ratings, particularly the Intercepts.

  6. Multiplex Brain Proteomic Analysis Revealed the Molecular Therapeutic Effects of Buyang Huanwu Decoction on Cerebral Ischemic Stroke Mice.

    Directory of Open Access Journals (Sweden)

    Hong-Jhang Chen

    Full Text Available Stroke is the second-leading cause of death worldwide, and tissue plasminogen activator (TPA is the only drug used for a limited group of stroke patients in the acute phase. Buyang Huanwu Decoction (BHD, a traditional Chinese medicine prescription, has long been used for improving neurological functional recovery in stroke. In this study, we characterized the therapeutic effect of TPA and BHD in a cerebral ischemia/reperfusion (CIR injury mouse model using multiplex proteomics approach. After the iTRAQ-based proteomics analysis, 1310 proteins were identified from the mouse brain with <1% false discovery rate. Among them, 877 quantitative proteins, 10.26% (90/877, 1.71% (15/877, and 2.62% (23/877 of the proteins was significantly changed in the CIR, BHD treatment, and TPA treatment, respectively. Functional categorization analysis showed that BHD treatment preserved the integrity of the blood-brain barrier (BBB (Alb, Fga, and Trf, suppressed excitotoxicity (Grm5, Gnai, and Gdi, and enhanced energy metabolism (Bdh, thereby revealing its multiple effects on ischemic stroke mice. Moreover, the neurogenesis marker doublecortin was upregulated, and the activity of glycogen synthase kinase 3 (GSK-3 and Tau was inhibited, which represented the neuroprotective effects. However, TPA treatment deteriorated BBB breakdown. This study highlights the potential of BHD in clinical applications for ischemic stroke.

  7. Effect of PDCA model on nutritional status in patients after laparoscopic repair of perforated peptic ulcer

    Directory of Open Access Journals (Sweden)

    Shu-Jing Hu

    2016-11-01

    Full Text Available Objective: To explore the effect of PDCA model on the nutritional status in patients after laparoscopic repair of perforated peptic ulcer. Methods: A total of 83 patients with gastric perforation who were admitted in our hospital from October, 2014 to December, 2015 for laparoscopic repair of perforated peptic ulcer were included in the study and randomized into the observation group (n=42 and the control group (n=41. The patients in the two groups were given routine treatments after operation. On this basis, the patients in the observation group were given additional PCDA model nursing. The gastrointestinal hormone levels and nutritional indicators after operation in the two groups were compared. Results: The difference of VIP, CCK, and GAS levels before operation between the two groups was not statistically significant (P>0.05. VIP, CCK, and GAS levels 3 d after operation were significantly reduced when compared with before operation (P0.05. WBC 1 d after operation in the observation group was significantly reduced, while TP, Hb, Alb, TRF, and BMI were significantly elevated (P0.05. Conclusions: PDCA nursing intervention can effectively improve the early nutritional status in patients after laparoscopic repair of perforated peptic ulcer and contribute to the postoperative rehabilitation.

  8. Isolation of Chromatin from Dysfunctional Telomeres Reveals an Important Role for Ring1b in NHEJ-Mediated Chromosome Fusions

    Directory of Open Access Journals (Sweden)

    Cristina Bartocci

    2014-05-01

    Full Text Available When telomeres become critically short, DNA damage response factors are recruited at chromosome ends, initiating a cellular response to DNA damage. We performed proteomic isolation of chromatin fragments (PICh in order to define changes in chromatin composition that occur upon onset of acute telomere dysfunction triggered by depletion of the telomere-associated factor TRF2. This unbiased purification of telomere-associated proteins in functional or dysfunctional conditions revealed the dynamic changes in chromatin composition that take place at telomeres upon DNA damage induction. On the basis of our results, we describe a critical role for the polycomb group protein Ring1b in nonhomologous end-joining (NHEJ-mediated end-to-end chromosome fusions. We show that cells with reduced levels of Ring1b have a reduced ability to repair uncapped telomeric chromatin. Our data represent an unbiased isolation of chromatin undergoing DNA damage and are a valuable resource to map the changes in chromatin composition in response to DNA damage activation.

  9. Solvent induced fluorescence enhancement of graphene oxide studied by ultrafast spectroscopy

    Science.gov (United States)

    Zhao, Litao; Chen, Jinquan; He, Xiaoxiao; Yu, Xiantong; Yan, Shujun; Zhang, Sanjun; Pan, Haifeng; Xu, Jianhua

    2018-05-01

    Femtosecond transient absorption (TA) spectroscopy combined with picosecond time resolved fluorescence (TRF) were used to reveal the fluorescence kinetics of graphene oxide (GO) in water, ethanol and water-ethanol mixtures. Size-independent fluorescence of GO were observed in water, and pH-dependent fluorescence spectra could be fitted well by a triple emission relaxation with peaks around 440 nm, 500 nm, and 590 nm respectively. The results indicate that polycyclic aromatic hydrocarbons (PAHs) linked by oxygen-containing functional groups dominate GO's fluorescence emission. GO's fluorescence quantum yield was measured to be 2.8% in ethanol but 1.2% in water. The three decay components fluorescence decay, as well as the transient absorption dynamics with an offset, confirmed this solvent induced fluorescence enhancement. GO's Raman spectral signals showed that GO in ethanol has a smaller average size of PAHs than that of GO in water. Therefore, besides other enhancement effects reported in literatures, we proposed that solvents could also change the size of PAHs, resulting in a photoluminescence enhancement. Our experimental data demonstrates that GO's quantum yield could be up to 2.8% in water and 8.4% in ethanol and this observation may help ones to improve GO's photoluminescence efficiency as well as its applications in solution.

  10. Effect of Shenkang injection combined with hemodialysis treatment on renal function, renal anemia and cytokine levels in patients with chronic renal failure

    Directory of Open Access Journals (Sweden)

    Rui Liu

    2016-10-01

    Full Text Available Objective: To study the effect of Shenkang injection combined with hemodialysis treatment on renal function, renal anemia and cytokine levels in patients with chronic renal failure. Methods: A total of 68 patients with chronic renal failure who received hemodialysis treatment in our hospital during between October 2013 and February 2016 were selected and randomly divided into two groups, the observation group received Shenkang injection treatment in the process of dialysis, and the control group only received conventional symptomatic and supportive treatment. 8 weeks after treatment, serum was collected to determine the levels of renal function indexes, nutritional status indexes, anemia indexes and cytokines, and urine was collected to determine renal function indexes. Results: β2-MG, UA, Cr, phosphorus, IL-17, IL-23, CTGF, TGF-β1, FGF-2 and FGF-23 levels in serum as well as NGAL, KIM-1 and RBP levels in urine of observation group were significantly lower than those of control group, and TP, Alb, PA, calcium, Hb, EPO, Fe, TRF and FER levels in serum were significantly higher than those of control group. Conclusion: Shenkang injection combined with hemodialysis treatment helps to improve renal function, nutritional status and renal anemia, and reduce the synthesis of inflammation and renal interstitial fibrosis-related cytokines in patients with chronic renal failure.

  11. Nitrification activity and community structure of nitrite-oxidizing bacteria in the bioreactors operated with addition of pharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Kraigher, Barbara, E-mail: barbara.kraigher@bf.uni-lj.si [University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Chair of Microbiology, Vecna pot 111, 1000 Ljubljana (Slovenia); Mandic-Mulec, Ines [University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Chair of Microbiology, Vecna pot 111, 1000 Ljubljana (Slovenia)

    2011-04-15

    Pharmaceuticals represent a group of the new emerging contaminants, which might influence microbial communities in the activated sludge. Nitrification activity and Nitrospira community structure in the small-scale reactors supplied with different concentrations (0, 50, 200, 500 {mu}g L{sup -1}) of the selected pharmaceuticals (ibuprofen, naproxen, ketoprofen, diclofenac and clofibric acid) were evaluated. Ammonia removal was not influenced by selected pharmaceuticals. However, in the two reactors operated with 50 {mu}g L{sup -1} of pharmaceuticals (R50 and R50P), the effluent concentration of N-(NO{sub 2}{sup -} + NO{sub 3}{sup -}) was significantly higher than in the other reactors. Nitrospira community structure was assessed by terminal restriction fragment length polymorphism (T-RFLP) and by cloning and sequencing of the partial genes for 16S rRNA. Nitrospira spp. were detected in all reactors. The two dominant T-RFs represented the sublineages I and II of the genus Nitrospira. Main shifts were observed in the reactors R50 and R50P, where the T-RF representing sublineage II was much higher as compared to the other reactors. Consistent with this, the Nitrospira sublineage II was detected only in the clone libraries from the reactors R50 and R50P. Our results suggest that the relative abundance of Nitrospira sublineage II could be related to the effluent N-(NO{sub 2}{sup -} + NO{sub 3}{sup -}) concentration.

  12. Parenting Styles in the Family of Origin in the Perception of Adolescents with Internalizing and Externalizing Disorders – Analysis of Dependencies

    Directory of Open Access Journals (Sweden)

    Barbara Ostafińska-Molik

    2015-06-01

    Full Text Available The authors have made an attempt at analyzing the mechanisms of internalizing, externalizing and mixed disorders, attitudes related to parenting (parenting styles of the parents: mother and father. The study was conducted among middle school pupils on the identified different types of disorders, and they related to the perception of the dominant parenting styles in the family of origin. The classification of parenting styles developed by Maria Ryś was adopted, who distinguished the following styles: democratic, autocratic, liberal loving and liberal unloving. The identification of the types of disorders was made based on the Teacher’s Report Form (TRF created by Thomas M. Achenbach. The analysis of dependencies between the types and dimensions (syndromes of adaptation disorders and parenting styles in the family of origin revealed that the attitude of parenting attitudes to a small extent determine the differences in the types of adaptation disorders, but specific trends are visible, which indicate a link between certain dimensions of parenting styles preferred by the mother and father: (1 internalizing disorders are significantly associated with the father’s parenting attitudes (autocratic and liberal loving attitude, causing the strengthening of problems in interpersonal functioning (social issues; (2 mixed disorders are associated significantly with the mother’s parenting attitudes (democratic, autocratic, liberal loving and unloving attitude, triggering disorders of withdrawal, anxiety-depressive and maladjusted behaviour; externalizing disorders are associated with parenting attitudes of both parents (democratic and liberal loving attitude, triggering mainly somatic complaints and anxiety-depressive disorders.

  13. Evaluation of effects of photooxidized Vespa orientalis venom on memory and learning in rats

    Directory of Open Access Journals (Sweden)

    H Mukund

    2011-01-01

    Full Text Available Wasp venom is mixture of complex proteins that have several physical and pharmacological properties. The photochemical detoxification of Vespa orientalis venom is expected to generate photooxidized venom sac extract (PVSE. Antigenically active PVSE is obtained by exposing the venom sac extract (VSE of Vespa orientalis to ultraviolet radiation in the presence of methylene blue. The aim of the present work was to evaluate the effect of PVSE on learning and memory of rats. Detoxification of PVSE was evident since treated mice had longer survival time than the group of mice treated with VSE. Photooxidized VSE of V. orientalis revealed enhancement on learning and memory by shortening the time to reach food (TRF in T-maze. In a 28-day study with rats, we observed that PVSE significantly decreased transfer latency (TL in elevated plus maze (EPM, significantly increased step down latency (SDL, diminished step down errors (SDE and time spent in shock zone (TSS in step down avoidance test. Thus, we concluded that although there is a possibility of employing PVSE in the treatment of Alzheimer, dementia or neurodegenerative illness as a non-herbal and non-synthetic alternative for patients who do not respond to available therapy, further investigation is still required.

  14. Structure and Composition of Leachfield Bacterial Communities: Role of Soil Texture, Depth and Septic Tank Effluent Inputs

    Directory of Open Access Journals (Sweden)

    Janet A. Atoyan

    2012-09-01

    Full Text Available Although groundwater quality depends on microbial processes in the soil treatment area (STA of onsite wastewater treatment systems (OWTS, our understanding of the development of these microbial communities is limited. We examined the bacterial communities of sand, sandy loam, and clay STAs at different depths in response to septic tank effluent (STE addition using mesocosms. Terminal restriction fragment length polymorphism (TRFLP analysis was used to compare the bacterial community structure and composition of STE, native soil prior to STE addition (UNX and soil exposed to STE (EXP. Principal component analysis separated communities with depth in sand but not in sandy loam or clay. Indices of richness, diversity, and evenness followed the order: sandy loam > sand > clay. Analysis of TRF peaks indicated that STE contributed least to the composition of STA bacterial communities (5%–16%, followed by UNX soil (18%–48%, with the highest proportion of the community made up of TRFs not detected previously in either UNX or STE (50%–82% for all three soils. Soil type and depth can have a marked effect on the structure and composition of STA bacterial communities, and on the relative contribution of native soil and STE to these communities.

  15. Telomere lengthening early in development.

    Science.gov (United States)

    Liu, Lin; Bailey, Susan M; Okuka, Maja; Muñoz, Purificación; Li, Chao; Zhou, Lingjun; Wu, Chao; Czerwiec, Eva; Sandler, Laurel; Seyfang, Andreas; Blasco, Maria A; Keefe, David L

    2007-12-01

    Stem cells and cancer cells maintain telomere length mostly through telomerase. Telomerase activity is high in male germ line and stem cells, but is low or absent in mature oocytes and cleavage stage embryos, and then high again in blastocysts. How early embryos reset telomere length remains poorly understood. Here, we show that oocytes actually have shorter telomeres than somatic cells, but their telomeres lengthen remarkably during early cleavage development. Moreover, parthenogenetically activated oocytes also lengthen their telomeres, thus the capacity to elongate telomeres must reside within oocytes themselves. Notably, telomeres also elongate in the early cleavage embryos of telomerase-null mice, demonstrating that telomerase is unlikely to be responsible for the abrupt lengthening of telomeres in these cells. Coincident with telomere lengthening, extensive telomere sister-chromatid exchange (T-SCE) and colocalization of the DNA recombination proteins Rad50 and TRF1 were observed in early cleavage embryos. Both T-SCE and DNA recombination proteins decrease in blastocyst stage embryos, whereas telomerase activity increases and telomeres elongate only slowly. We suggest that telomeres lengthen during the early cleavage cycles following fertilization through a recombination-based mechanism, and that from the blastocyst stage onwards, telomerase only maintains the telomere length established by this alternative mechanism.

  16. [Family-Based Trauma-Focused Cognitive Behavioral Therapy with Three Siblings of a Refugee Family].

    Science.gov (United States)

    Bohnacker, Isabelle; Goldbeck, Lutz

    2017-10-01

    Family-Based Trauma-Focused Cognitive Behavioral Therapy with Three Siblings of a Refugee Family The possibility and relevance of a joint trauma-therapy with siblings has yet received little attention in research and clinical practice. The following case study presents a joint family-based trauma-focused therapy process with a refugee family. All three siblings suffered from post-traumatic stress disorder (PTSD) before treatment. The treatment followed the manual of Trauma Focused Cognitive Behavioral Therapy (TF-CBT; Cohen, Mannarino, Deblinger, 2009). Measures were the short version of the Child and Adolescent Trauma Screen (CATS 7-17), as well as the Teacher's Report Form (TRF). After 18 treatment sessions together with the mother, all three children did no longer meet PTSD criteria. Benefits of the joint therapy were for all three siblings to be sharing and imitating each other's coping strategies. Furthermore, the protective factor of social support after experiencing a traumatic event became evident. The apprehension of the therapist not being sufficiently neutral towards all three siblings was not observed.

  17. Contribuição dos Indicadores de Problemas Emocionais e de Comportamento para o Rendimento Escolar

    Directory of Open Access Journals (Sweden)

    Bruna Mainardi Rosso Borba

    2017-07-01

    Full Text Available Este estudo investigou se há contribuição e em que medida os problemas e os indicadores comportamentais emocionais e de conduta explicam o rendimento escolar dos adolescentes. Setenta estudantes participaram, 70 famílias e 21 profes-sores. Os adolescentes responderam ao inventário de comportamentos da juventude (YSR, a família do inventário dos professores da criança e o comportamento dos adolescentes (CBCL e o inventário de conduta do Adolescente (TRF e crianças. O desempenho escolar foi avaliado a partir da média final nas disciplinas de português e matemática. Na ava-liação dos adolescentes, suas famílias e professores, os problemas de atenção explicam o mau desempenho escolar, bem como a terceirização dos problemas identificados pelos adolescentes e os sintomas de isolamento/depressão indicada pelos professores. É realçada a importância de atender tais indicadores, visto sua influência nos adolescentes.

  18. Transcriptome and proteome analyses and the role of atypical calpain protein and autophagy in the spliced leader silencing pathway in Trypanosoma brucei.

    Science.gov (United States)

    Hope, Ronen; Egarmina, Katarina; Voloshin, Konstantin; Waldman Ben-Asher, Hiba; Carmi, Shai; Eliaz, Dror; Drori, Yaron; Michaeli, Shulamit

    2016-10-01

    Under persistent ER stress, Trypanosoma brucei parasites induce the spliced leader silencing (SLS) pathway. In SLS, transcription of the SL RNA gene, the SL donor to all mRNAs, is extinguished, arresting trans-splicing and leading to programmed cell death (PCD). In this study, we investigated the transcriptome following silencing of SEC63, a factor essential for protein translocation across the ER membrane, and whose silencing induces SLS. The proteome of SEC63-silenced cells was analyzed with an emphasis on SLS-specific alterations in protein expression, and modifications that do not directly result from perturbations in trans-splicing. One such protein identified is an atypical calpain SKCRP7.1/7.2. Co-silencing of SKCRP7.1/7.2 and SEC63 eliminated SLS induction due its role in translocating the PK3 kinase. This kinase initiates SLS by migrating to the nucleus and phosphorylating TRF4 leading to shut-off of SL RNA transcription. Thus, SKCRP7.1 is involved in SLS signaling and the accompanying PCD. The role of autophagy in SLS was also investigated; eliminating autophagy through VPS34 or ATG7 silencing demonstrated that autophagy is not essential for SLS induction, but is associated with PCD. Thus, this study identified factors that are used by the parasite to cope with ER stress and to induce SLS and PCD. © 2016 John Wiley & Sons Ltd.

  19. TERRA Expression Levels Do Not Correlate With Telomere Length and Radiation Sensitivity in Human Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Alexandra eSmirnova

    2013-05-01

    Full Text Available Mammalian telomeres are transcribed into long non-coding telomeric RNA molecules (TERRA that seem to play a role in the maintenance of telomere stability. In human cells, CpG island promoters drive TERRA transcription and are regulated by methylation. It was suggested that the amount of TERRA may be related to telomere length. To test this hypothesis we measured telomere length and TERRA levels in single clones isolated from five human cell lines: HeLa (cervical carcinoma, BRC-230 (breast cancer, AKG and GK2 (gastric cancers and GM847 (SV40 immortalized skin fibroblasts. We observed great clonal heterogeneity both in TRF (Terminal Restriction Fragment length and in TERRA levels. However, these two parameters did not correlate with each other. Moreover, cell survival to γ-rays did not show a significant variation among the clones, suggesting that, in this cellular system, the intra-population variability in telomere length and TERRA levels does not influence sensitivity to ionizing radiation. This conclusion was supported by the observation that in a cell line in which telomeres were greatly elongated by the ectopic expression of telomerase, TERRA expression levels and radiation sensitivity were similar to the parental HeLa cell line.

  20. Nitrification activity and community structure of nitrite-oxidizing bacteria in the bioreactors operated with addition of pharmaceuticals

    International Nuclear Information System (INIS)

    Kraigher, Barbara; Mandic-Mulec, Ines

    2011-01-01

    Pharmaceuticals represent a group of the new emerging contaminants, which might influence microbial communities in the activated sludge. Nitrification activity and Nitrospira community structure in the small-scale reactors supplied with different concentrations (0, 50, 200, 500 μg L -1 ) of the selected pharmaceuticals (ibuprofen, naproxen, ketoprofen, diclofenac and clofibric acid) were evaluated. Ammonia removal was not influenced by selected pharmaceuticals. However, in the two reactors operated with 50 μg L -1 of pharmaceuticals (R50 and R50P), the effluent concentration of N-(NO 2 - + NO 3 - ) was significantly higher than in the other reactors. Nitrospira community structure was assessed by terminal restriction fragment length polymorphism (T-RFLP) and by cloning and sequencing of the partial genes for 16S rRNA. Nitrospira spp. were detected in all reactors. The two dominant T-RFs represented the sublineages I and II of the genus Nitrospira. Main shifts were observed in the reactors R50 and R50P, where the T-RF representing sublineage II was much higher as compared to the other reactors. Consistent with this, the Nitrospira sublineage II was detected only in the clone libraries from the reactors R50 and R50P. Our results suggest that the relative abundance of Nitrospira sublineage II could be related to the effluent N-(NO 2 - + NO 3 - ) concentration.

  1. Disappearance of the telomere dysfunction-induced stress response in fully senescent cells.

    Science.gov (United States)

    Bakkenist, Christopher J; Drissi, Rachid; Wu, Jing; Kastan, Michael B; Dome, Jeffrey S

    2004-06-01

    Replicative senescence is a natural barrier to cellular proliferation that is triggered by telomere erosion and dysfunction. Here, we demonstrate that ATM activation and H2AX-gamma nuclear focus formation are sensitive markers of telomere dysfunction in primary human fibroblasts. Whereas the activated form of ATM and H2AX-gamma foci were rarely observed in early-passage cells, they were readily detected in late-passage cells. The ectopic expression of telomerase in late-passage cells abrogated ATM activation and H2AX-gamma focus formation, suggesting that these stress responses were the consequence of telomere dysfunction. ATM activation was induced in quiescent fibroblasts by inhibition of TRF2 binding to telomeres, indicating that telomere uncapping is sufficient to initiate the telomere signaling response; breakage of chromosomes with telomeric associations is not required for this activation. Although ATM activation and H2AX-gamma foci were readily observed in late-passage cells, they disappeared once cells became fully senescent, indicating that constitutive signaling from dysfunctional telomeres is not required for the maintenance of senescence.

  2. Telomere dynamics, end-to-end fusions and telomerase activation during the human fibroblast immortalization process.

    Science.gov (United States)

    Ducray, C; Pommier, J P; Martins, L; Boussin, F D; Sabatier, L

    1999-07-22

    Loss of telomeric repeats during cell proliferation could play a role in senescence. It has been generally assumed that activation of telomerase prevents further telomere shortening and is essential for cell immortalization. In this study, we performed a detailed cytogenetic and molecular characterization of four SV40 transformed human fibroblastic cell lines by regularly monitoring the size distribution of terminal restriction fragments, telomerase activity and the associated chromosomal instability throughout immortalization. The mean TRF lengths progressively decreased in pre-crisis cells during the lifespan of the cultures. At crisis, telomeres reached a critical size, different among the cell lines, contributing to the peak of dicentric chromosomes, which resulted mostly from telomeric associations. We observed a direct correlation between short telomere length at crisis and chromosomal instability. In two immortal cell lines, although telomerase was detected, mean telomere length still continued to decrease whereas the number of dicentric chromosomes associated was stabilized. Thus telomerase could protect specifically telomeres which have reached a critical size against end-to-end dicentrics, while long telomeres continue to decrease, although at a slower rate as before crisis. This suggests a balance between elongation by telomerase and telomere shortening, towards a stabilized 'optimal' length.

  3. Getting in (and out of) the loop: regulating higher order telomere structures.

    Science.gov (United States)

    Luke-Glaser, Sarah; Poschke, Heiko; Luke, Brian

    2012-01-01

    The DNA at the ends of linear chromosomes (the telomere) folds back onto itself and forms an intramolecular lariat-like structure. Although the telomere loop has been implicated in the protection of chromosome ends from nuclease-mediated resection and unscheduled DNA repair activities, it potentially poses an obstacle to the DNA replication machinery during S-phase. Therefore, the coordinated regulation of telomere loop formation, maintenance, and resolution is required in order to establish a balance between protecting the chromosome ends and promoting their duplication prior to cell division. Until recently, the only factor known to influence telomere looping in human cells was TRF2, a component of the shelterin complex. Recent work in yeast and mouse cells has uncovered additional regulatory factors that affect the loop structure at telomeres. In the following "perspective" we outline what is known about telomere looping and highlight the latest results regarding the regulation of this chromosome end structure. We speculate about how the manipulation of the telomere loop may have therapeutic implications in terms of diseases associated with telomere dysfunction and uncontrolled proliferation.

  4. Getting in (and out of the loop: regulating higher order telomere structures

    Directory of Open Access Journals (Sweden)

    Sarah eLuke-Glaser

    2012-11-01

    Full Text Available The DNA at the ends of linear chromosomes (the telomere folds back onto itself and forms an intramolecular lariat-like structure. Although the telomere loop has been implicated in the protection of chromosome ends from nuclease-mediated resection and unscheduled DNA repair activities, it potentially poses an obstacle to the DNA replication machinery during S phase. Therefore, the coordinated regulation of telomere loop formation, maintenance and resolution is required in order to establish a balance between protecting the chromosome ends and promoting their duplication prior to cell division. Until recently, the only factor know to influence telomere looping in human cells was TRF2, a component of the shelterin complex. Recent work in yeast and mouse cells has uncovered additional regulatory factors that affect the loop structure at telomeres. In the following perspective we will outline what is known about telomere looping and highlight the latest results regarding the regulation of this chromosome end structure. We will speculate about how the manipulation of the telomere loop may have therapeutic implications in terms of diseases associated with telomere dysfunction and uncontrolled proliferation.

  5. Single-label kinase and phosphatase assays for tyrosine phosphorylation using nanosecond time-resolved fluorescence detection.

    Science.gov (United States)

    Sahoo, Harekrushna; Hennig, Andreas; Florea, Mara; Roth, Doris; Enderle, Thilo; Nau, Werner M

    2007-12-26

    The collision-induced fluorescence quenching of a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) by hydrogen atom abstraction from the tyrosine residue in peptide substrates was introduced as a single-labeling strategy to assay the activity of tyrosine kinases and phosphatases. The assays were tested for 12 different combinations of Dbo-labeled substrates and with the enzymes p60c-Src Src kinase, EGFR kinase, YOP protein tyrosine phosphatase, as well as acid and alkaline phosphatases, thereby demonstrating a broad application potential. The steady-state fluorescence changed by a factor of up to 7 in the course of the enzymatic reaction, which allowed for a sufficient sensitivity of continuous monitoring in steady-state experiments. The fluorescence lifetimes (and intensities) were found to be rather constant for the phosphotyrosine peptides (ca. 300 ns in aerated water), while those of the unphosphorylated peptides were as short as 40 ns (at pH 7) and 7 ns (at pH 13) as a result of intramolecular quenching. Owing to the exceptionally long fluorescence lifetime of Dbo, the assays were alternatively performed by using nanosecond time-resolved fluorescence (Nano-TRF) detection, which leads to an improved discrimination of background fluorescence and an increased sensitivity. The potential for inhibitor screening was demonstrated through the inhibition of acid and alkaline phosphatases by molybdate.

  6. [Evaluation of the Musical Concentration Training with Pepe (MusiKo mit Pepe) for children with attention deficits].

    Science.gov (United States)

    Rothmann, Kathrin; Hillmer, Jana-Mareike; Hosser, Daniela

    2014-09-01

    This study evaluates the Musical Concentration Training with Pepe ("MusiKo mit Pepe") for children aged 5 to 10 years with attention deficits. Using a pre-post-control design (N = 108), changes in attention capacity are measured by the Test of Attentional Performance for Children (KiTAP), whereas changes in the quality of life are assessed with the Children's Questionnaire (KINDL-R). Additionally, we utilized the Symptom Checklist for Attention Deficit Hyperactivity Disorders (FBB-ADHS) and for Conduct Disorder (FBB-SSV) of the Diagnostic System of Mental Disorders in Children and Adolescents II based on ICD-10 and DSM-IV as well as the Child Behavior Checklist and the Teacher's Report Form (CBCL, TRF). Significant pre-post effects were found in both attention and quality of life for children treated by the training compared to controls. Moreover, significant reductions were detected in ADHD symptomatology in parents' and teachers' ratings, and in internal problems in parents' ratings. The effectiveness of the intervention was not affected by age, sex, intelligence, or migration background. The music-based training "MusiKo mit Pepe" is an effective intervention for children with attention deficits, pending replication of these findings in future studies.

  7. The histone H4 lysine 20 monomethyl mark, set by PR-Set7 and stabilized by L(3mbt, is necessary for proper interphase chromatin organization.

    Directory of Open Access Journals (Sweden)

    Ayako Sakaguchi

    Full Text Available Drosophila PR-Set7 or SET8 is a histone methyltransferase that specifically monomethylates histone H4 lysine 20 (H4K20. L(3MBT has been identified as a reader of methylated H4K20. It contains several conserved domains including three MBT repeats binding mono- and dimethylated H4K20 peptides. We find that the depletion of PR-Set7 blocks de novo H4K20me1 resulting in the immediate activation of the DNA damage checkpoint, an increase in the size of interphase nuclei, and drastic reduction of cell viability. L(3mbt on the other hand stabilizes the monomethyl mark, as L(3mbt-depleted S2 cells show a reduction of more than 60% of bulk monomethylated H4K20 (H4K20me1 while viability is barely affected. Ploidy and basic chromatin structure show only small changes in PR-Set7-depleted cells, but higher order interphase chromatin organization is significantly affected presumably resulting in the activation of the DNA damage checkpoint. In the absence of any other known functions of PR-Set7, the setting of the de novo monomethyl mark appears essential for cell viability in the presence or absence of the DNA damage checkpoint, but once newly assembled chromatin is established the monomethyl mark, protected by L(3mbt, is dispensable.

  8. Plasma Membrane CRPK1-Mediated Phosphorylation of 14-3-3 Proteins Induces Their Nuclear Import to Fine-Tune CBF Signaling during Cold Response.

    Science.gov (United States)

    Liu, Ziyan; Jia, Yuxin; Ding, Yanglin; Shi, Yiting; Li, Zhen; Guo, Yan; Gong, Zhizhong; Yang, Shuhua

    2017-04-06

    In plant cells, changes in fluidity of the plasma membrane may serve as the primary sensor of cold stress; however, the precise mechanism and how the cell transduces and fine-tunes cold signals remain elusive. Here we show that the cold-activated plasma membrane protein cold-responsive protein kinase 1 (CRPK1) phosphorylates 14-3-3 proteins. The phosphorylated 14-3-3 proteins shuttle from the cytosol to the nucleus, where they interact with and destabilize the key cold-responsive C-repeat-binding factor (CBF) proteins. Consistent with this, the crpk1 and 14-3-3κλ mutants show enhanced freezing tolerance, and transgenic plants overexpressing 14-3-3λ show reduced freezing tolerance. Further study shows that CRPK1 is essential for the nuclear translocation of 14-3-3 proteins and for 14-3-3 function in freezing tolerance. Thus, our study reveals that the CRPK1-14-3-3 module transduces the cold signal from the plasma membrane to the nucleus to modulate CBF stability, which ensures a faithfully adjusted response to cold stress of plants. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Comparison of signaling interactions determining annual and perennial plant growth in response to low temperature

    Directory of Open Access Journals (Sweden)

    Astrid eWingler

    2015-01-01

    Full Text Available Low temperature inhibits plant growth despite the fact that considerable rates of photosynthetic activity can be maintained. Instead of lower rates of photosynthesis, active inhibition of cell division and expansion is primarily responsible for reduced growth. This results in sink limitation and enables plants to accumulate carbohydrates that act as compatible solutes or are stored throughout the winter to enable re-growth in spring. Regulation of growth in response to temperature therefore requires coordination with carbon metabolism, e.g. via the signaling metabolite trehalose-6-phosphate. The phytohormones gibberellins (GA and jasmonate (JA play an important role in regulating growth in response to temperature. Growth restriction at low temperature is mainly mediated by DELLA proteins, whose degradation is promoted by GA. For annual plants, it has been shown that the GA/DELLA pathway interacts with JA signaling and C-repeat binding factor (CBF dependent cold acclimation, but these interactions have not been explored in detail for perennials. Growth regulation in response to seasonal factors is, however, particularly important in perennials, especially at high latitudes. In autumn, growth cessation in trees is caused by shortening of the daylength in interaction with phytohormone signaling. In perennial grasses seasonal differences in the sensitivity to GA may enable enhanced growth in spring. This review provides an overview of the signaling interactions that determine plant growth at low temperature and highlights gaps in our knowledge, especially concerning the seasonality of signaling responses in perennial plants.

  10. Expression analysis and functional characterization of a novel cold-responsive gene CbCOR15a from Capsella bursa-pastoris.

    Science.gov (United States)

    Zhou, Mingqi; Wu, Lihua; Liang, Jing; Shen, Chen; Lin, Juan

    2012-05-01

    The cold-responsive (COR) genes involved in C-repeat binding factor signaling pathway function essentially in cold acclimation of higher plants. A novel COR gene CbCOR15a from shepherd's purse (Capsella bursa-pastoris) was predicted to be a homolog of COR15 in Arabidopsis. The analysis of tissue specific expression pattern as well as characterization of the CbCOR15a promoter revealed that the expression of CbCOR15a was induced by coldness not only in leaves and stem but also in roots. Sequence analysis showed that a 909 bp promoter region of CbCOR15a contained two CRT/DRE elements, two ABRE elements, one auxin-responsive TGA-element and one MeJA-responsive CGTCA-motif. In young seedlings the expression of CbCOR15a could be apparently increased by SA, ABA, MeJA and IAA, and transiently increased by GA(3) accompanied by obvious feedback suppression. According to the altered physiological index values in tobacco under cold treatments, the overexpression of CbCOR15a significantly increased the cold tolerance of transgenic tobacco plants. It can be suggested that CbCOR15a was involved in cold response of Capsella bursa-pastoris associated with SA, ABA, MeJA, IAA and GA(3) regulation and confers enhanced cold acclimation in transgenic plants.

  11. Coordinated Regulations of mRNA Synthesis and Decay during Cold Acclimation in Arabidopsis Cells.

    KAUST Repository

    Arae, Toshihiro

    2017-04-18

    Plants possess a cold acclimation system to acquire freezing tolerance through pre-exposure to non-freezing low temperatures. The transcriptional cascade of C-repeat binding factors (CBFs)/dehydration response element-binding factors (DREBs) is considered a major transcriptional regulatory pathway during cold acclimation. However, little is known regarding the functional significance of mRNA stability regulation in the response of gene expression to cold stress. The actual level of individual mRNAs is determined by a balance between mRNA synthesis and degradation. Therefore, it is important to assess the regulatory steps to increase our understanding of gene regulation. Here, we analyzed temporal changes in mRNA amounts and half-lives in response to cold stress in Arabidopsis cell cultures based on genome-wide analysis. In this mRNA decay array method, mRNA half-life measurements and microarray analyses were combined. In addition, temporal changes in the integrated value of transcription rates were estimated from the above two parameters using a mathematical approach. Our results showed that several cold-responsive genes, including Cold-regulated 15a, were relatively destabilized, whereas the mRNA amounts were increased during cold treatment by accelerating the transcription rate to overcome the destabilization. Considering the kinetics of mRNA synthesis and degradation, this apparently contradictory result supports that mRNA destabilization is advantageous for the swift increase in CBF-responsive genes in response to cold stress.

  12. Gene expression changes during short day induced terminal bud formation in Norway spruce.

    Science.gov (United States)

    Asante, Daniel K A; Yakovlev, Igor A; Fossdal, Carl Gunnar; Holefors, Anna; Opseth, Lars; Olsen, Jorunn E; Junttila, Olavi; Johnsen, Øystein

    2011-02-01

    The molecular basis for terminal bud formation in autumn is not well understood in conifers. By combining suppression subtractive hybridization and monitoring of gene expression by qRT-PCR analysis, we aimed to identify genes involved in photoperiodic control of growth cessation and bud set in Norway spruce. Close to 1400 ESTs were generated and their functional distribution differed between short day (SD-12 h photoperiod) and long day (LD-24 h photoperiod) libraries. Many genes with putative roles in protection against stress appeared differentially regulated under SD and LD, and also differed in transcript levels between 6 and 20 SDs. Of these, PaTFL1(TERMINAL FLOWER LIKE 1) showed strongly increased transcript levels at 6 SDs. PaCCCH(CCCH-TYPE ZINC FINGER) and PaCBF2&3(C-REPEAT BINDING FACTOR 2&3) showed a later response at 20 SDs, with increased and decreased transcript levels, respectively. For rhythmically expressed genes such as CBFs, such differences might represent a phase shift in peak expression, but might also suggest a putative role in response to SD. Multivariate analyses revealed strong differences in gene expression between LD, 6 SD and 20 SD. The robustness of the gene expression patterns was verified in 6 families differing in bud-set timing under natural light with gradually decreasing photoperiod. © 2010 Blackwell Publishing Ltd.

  13. Crosstalk of PmCBFs and PmDAMs Based on the Changes of Phytohormones under Seasonal Cold Stress in the Stem of Prunus mume

    Directory of Open Access Journals (Sweden)

    Kai Zhao

    2018-01-01

    Full Text Available Plants facing the seasonal variations always need a growth restraining mechanism when temperatures turn down. C-repeat binding factor (CBF genes work essentially in the cold perception. Despite lots of researches on CBFs, the multiple crosstalk is still interesting on their interaction with hormones and dormancy-associated MADS (DAM genes in the growth and dormancy control. Therefore, this study highlights roles of PmCBFs in cold-induced dormancy from different orgens. And a sense-response relationship between PmCBFs and PmDAMs is exhibited in this process, jointly regulated by six PmCBFs and PmDAM4–6. Meantime, GA3 and ABA showed negative and positive correlation with PmCBFs expression levels, respectively. We also find a high correlation between IAA and PmDAM1–3. Finally, we display the interaction mode of PmCBFs and PmDAMs, especially PmCBF1-PmDAM1. These results can disclose another view of molecular mechanism in plant growth between cold-response pathway and dormancy regulation together with genes and hormones.

  14. Comparative analyses reveal potential uses of Brachypodium distachyon as a model for cold stress responses in temperate grasses

    Directory of Open Access Journals (Sweden)

    Li Chuan

    2012-05-01

    Full Text Available Abstract Background Little is known about the potential of Brachypodium distachyon as a model for low temperature stress responses in Pooideae. The ice recrystallization inhibition protein (IRIP genes, fructosyltransferase (FST genes, and many C-repeat binding factor (CBF genes are Pooideae specific and important in low temperature responses. Here we used comparative analyses to study conservation and evolution of these gene families in B. distachyon to better understand its potential as a model species for agriculturally important temperate grasses. Results Brachypodium distachyon contains cold responsive IRIP genes which have evolved through Brachypodium specific gene family expansions. A large cold responsive CBF3 subfamily was identified in B. distachyon, while CBF4 homologs are absent from the genome. No B. distachyon FST gene homologs encode typical core Pooideae FST-motifs and low temperature induced fructan accumulation was dramatically different in B. distachyon compared to core Pooideae species. Conclusions We conclude that B. distachyon can serve as an interesting model for specific molecular mechanisms involved in low temperature responses in core Pooideae species. However, the evolutionary history of key genes involved in low temperature responses has been different in Brachypodium and core Pooideae species. These differences limit the use of B. distachyon as a model for holistic studies relevant for agricultural core Pooideae species.

  15. The Arabidopsis Mediator Complex Subunits MED16, MED14, and MED2 Regulate Mediator and RNA Polymerase II Recruitment to CBF-Responsive Cold-Regulated Genes[C][W][OPEN

    Science.gov (United States)

    Hemsley, Piers A.; Hurst, Charlotte H.; Kaliyadasa, Ewon; Lamb, Rebecca; Knight, Marc R.; De Cothi, Elizabeth A.; Steele, John F.; Knight, Heather

    2014-01-01

    The Mediator16 (MED16; formerly termed SENSITIVE TO FREEZING6 [SFR6]) subunit of the plant Mediator transcriptional coactivator complex regulates cold-responsive gene expression in Arabidopsis thaliana, acting downstream of the C-repeat binding factor (CBF) transcription factors to recruit the core Mediator complex to cold-regulated genes. Here, we use loss-of-function mutants to show that RNA polymerase II recruitment to CBF-responsive cold-regulated genes requires MED16, MED2, and MED14 subunits. Transcription of genes known to be regulated via CBFs binding to the C-repeat motif/drought-responsive element promoter motif requires all three Mediator subunits, as does cold acclimation–induced freezing tolerance. In addition, these three subunits are required for low temperature–induced expression of some other, but not all, cold-responsive genes, including genes that are not known targets of CBFs. Genes inducible by darkness also required MED16 but required a different combination of Mediator subunits for their expression than the genes induced by cold. Together, our data illustrate that plants control transcription of specific genes through the action of subsets of Mediator subunits; the specific combination defined by the nature of the stimulus but also by the identity of the gene induced. PMID:24415770

  16. The Arabidopsis mediator complex subunits MED16, MED14, and MED2 regulate mediator and RNA polymerase II recruitment to CBF-responsive cold-regulated genes.

    Science.gov (United States)

    Hemsley, Piers A; Hurst, Charlotte H; Kaliyadasa, Ewon; Lamb, Rebecca; Knight, Marc R; De Cothi, Elizabeth A; Steele, John F; Knight, Heather

    2014-01-01

    The Mediator16 (MED16; formerly termed SENSITIVE TO FREEZING6 [SFR6]) subunit of the plant Mediator transcriptional coactivator complex regulates cold-responsive gene expression in Arabidopsis thaliana, acting downstream of the C-repeat binding factor (CBF) transcription factors to recruit the core Mediator complex to cold-regulated genes. Here, we use loss-of-function mutants to show that RNA polymerase II recruitment to CBF-responsive cold-regulated genes requires MED16, MED2, and MED14 subunits. Transcription of genes known to be regulated via CBFs binding to the C-repeat motif/drought-responsive element promoter motif requires all three Mediator subunits, as does cold acclimation-induced freezing tolerance. In addition, these three subunits are required for low temperature-induced expression of some other, but not all, cold-responsive genes, including genes that are not known targets of CBFs. Genes inducible by darkness also required MED16 but required a different combination of Mediator subunits for their expression than the genes induced by cold. Together, our data illustrate that plants control transcription of specific genes through the action of subsets of Mediator subunits; the specific combination defined by the nature of the stimulus but also by the identity of the gene induced.

  17. C9orf72 nucleotide repeat structures initiate molecular cascades of disease.

    Science.gov (United States)

    Haeusler, Aaron R; Donnelly, Christopher J; Periz, Goran; Simko, Eric A J; Shaw, Patrick G; Kim, Min-Sik; Maragakis, Nicholas J; Troncoso, Juan C; Pandey, Akhilesh; Sattler, Rita; Rothstein, Jeffrey D; Wang, Jiou

    2014-03-13

    A hexanucleotide repeat expansion (HRE), (GGGGCC)n, in C9orf72 is the most common genetic cause of the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here we identify a molecular mechanism by which structural polymorphism of the HRE leads to ALS/FTD pathology and defects. The HRE forms DNA and RNA G-quadruplexes with distinct structures and promotes RNA•DNA hybrids (R-loops). The structural polymorphism causes a repeat-length-dependent accumulation of transcripts aborted in the HRE region. These transcribed repeats bind to ribonucleoproteins in a conformation-dependent manner. Specifically, nucleolin, an essential nucleolar protein, preferentially binds the HRE G-quadruplex, and patient cells show evidence of nucleolar stress. Our results demonstrate that distinct C9orf72 HRE structural polymorphism at both DNA and RNA levels initiates molecular cascades leading to ALS/FTD pathologies, and provide the basis for a mechanistic model for repeat-associated neurodegenerative diseases.

  18. Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS

    Energy Technology Data Exchange (ETDEWEB)

    Teplova, Marianna; Farazi, Thalia A.; Tuschl, Thomas; Patel, Dinshaw J.

    2015-09-08

    Abstract

    RNA-binding protein with multiple splicing (designated RBPMS) is a higher vertebrate mRNA-binding protein containing a single RNA recognition motif (RRM). RBPMS has been shown to be involved in mRNA transport, localization and stability, with key roles in axon guidance, smooth muscle plasticity, as well as regulation of cancer cell proliferation and migration. We report on structure-function studies of the RRM domain of RBPMS bound to a CAC-containing single-stranded RNA. These results provide insights into potential topologies of complexes formed by the RBPMS RRM domain and the tandem CAC repeat binding sites as detected by photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation. These studies establish that the RRM domain of RBPMS forms a symmetrical dimer in the free state, with each monomer binding sequence-specifically to all three nucleotides of a CAC segment in the RNA bound state. Structure-guided mutations within the dimerization and RNA-binding interfaces of RBPMS RRM on RNA complex formation resulted in both disruption of dimerization and a decrease in RNA-binding affinity as observed by size exclusion chromatography and isothermal titration calorimetry. As anticipated from biochemical binding studies, over-expression of dimerization or RNA-binding mutants of Flag-HA-tagged RBPMS were no longer able to track with stress granules in HEK293 cells, thereby documenting the deleterious effects of such mutationsin vivo.

  19. Redox Signaling and CBF-Responsive Pathway Are Involved in Salicylic Acid-Improved Photosynthesis and Growth under Chilling Stress in Watermelon

    Science.gov (United States)

    Cheng, Fei; Lu, Junyang; Gao, Min; Shi, Kai; Kong, Qiusheng; Huang, Yuan; Bie, Zhilong

    2016-01-01

    Salicylic acid (SA) plays an important role in plant response to abiotic stresses. This study investigated the potential role of SA in alleviating the adverse effects of chilling stress on photosynthesis and growth in watermelon (Citrullus lanatus). Chilling stress induced the simultaneous accumulation of free and conjugated SA in watermelon plants, and the chilling-induced SA production was attributed to the phenylalanine ammonia-lyase pathway. Applying SA at moderate concentrations induced chilling tolerance, whereas inhibition of SA biosynthesis by L-α-aminooxy-β-phenylpropionic acid (AOPP) increased the photooxidation of PS II under chilling stress in watermelon, resulting in reduced photosynthesis and growth. Chilling induced a transient increase in the ratios of reduced to oxidized glutathione and reduced ascorbate to dehydroascorbate. Then, the expression of antioxidant genes was upregulated, and the activities of antioxidant enzymes were enhanced. Furthermore, SA-induced chilling tolerance was associated with cellular glutathione and ascorbate homeostasis, which served as redox signals to regulate antioxidant metabolism under chilling stress. AOPP treatment stimulated the chilling-induced expression of cold-responsive genes, particularly via C-repeat binding factors CBF3 and CBF4. These results confirm the synergistic role of SA signaling and the CBF-dependent responsive pathway during chilling stress in watermelon. PMID:27777580

  20. Redox Signaling and CBF-Responsive Pathway are Involved in Salicylic Acid-Improved Photosynthesis and Growth under Chilling Stress in Watermelon

    Directory of Open Access Journals (Sweden)

    Fei Cheng

    2016-10-01

    Full Text Available Salicylic acid (SA plays an important role in plant response to abiotic stresses. This study investigated the potential role of SA in alleviating the adverse effects of chilling stress on photosynthesis and growth in watermelon (Citrullus lanatus. Chilling stress induced the simultaneous accumulation of free and conjugated SA in watermelon plants, and the chilling-induced SA production was attributed to the phenylalanine ammonia-lyase pathway. Applying SA at moderate concentrations induced chilling tolerance, whereas inhibition of SA biosynthesis by L-ɑ-aminooxy-β-phenylpropionic acid (AOPP increased the photooxidation of PS II under chilling stress in watermelon, resulting in reduced photosynthesis and growth. Chilling induced a transient increase in the ratios of reduced to oxidized glutathione and reduced ascorbate to dehydroascorbate. Then, the expression of antioxidant genes was upregulated, and the activities of antioxidant enzymes were enhanced. Furthermore, SA-induced chilling tolerance was associated with cellular glutathione and ascorbate homeostasis, which served as redox signals to regulate antioxidant metabolism under chilling stress. AOPP treatment stimulated the chilling-induced expression of cold-responsive genes, particularly via C-repeat binding factors CBF3 and CBF4. These results confirm the synergistic role of SA signaling and the CBF-dependent responsive pathway during chilling stress in watermelon.

  1. Melatonin-induced CBF/DREB1s are essential for diurnal change of disease resistance and CCA1 expression in Arabidopsis.

    Science.gov (United States)

    Shi, Haitao; Wei, Yunxie; He, Chaozu

    2016-03-01

    Melatonin (N-acetyl-5-methoxytryptamine) is an important regulator of circadian rhythms and immunity in animals. However, the diurnal changes of endogenous melatonin and melatonin-mediated diurnal change of downstream responses remain unclear in Arabidopsis. Using the publicly available microarray data, we found that the transcript levels of two melatonin synthesis genes (serotonin N-acetyltransferase (SNAT) and caffeate O-methyltransferase (COMT)) and endogenous melatonin level were regulated by diurnal cycles, with different magnitudes of change. Moreover, the transcripts of C-repeat-binding factors (CBFs)/Drought response element Binding 1 factors (DREB1s) were co-regulated by exogenous melatonin and diurnal changes, indicating the possible correlation among clock, endogenous melatonin level and AtCBFs expressions. Interestingly, diurnal change of plant immunity against Pst DC3000 and CIRCADIANCLOCK ASSOCIATED 1 (CCA1) expression were largely lost in AtCBFs knockdown line-amiR-1. Taken together, this study identifies the molecular pathway underlying the diurnal changes of immunity in Arabidopsis. Notably, the diurnal changes of endogenous melatonin may regulate corresponding changes of AtCBF/DREB1s expression and their underlying diurnal cycle of plant immunity and AtCCA1. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  2. Optimization of Photosynthetic Productivity in Contrasting Environments by Regulons Controlling Plant Form and Function

    Directory of Open Access Journals (Sweden)

    Barbara Demmig-Adams

    2018-03-01

    Full Text Available We review the role of a family of transcription factors and their regulons in maintaining high photosynthetic performance across a range of challenging environments with a focus on extreme temperatures and water availability. Specifically, these transcription factors include CBFs (C-repeat binding factors and DREBs (dehydration-responsive element-binding, with CBF/DREB1 primarily orchestrating cold adaptation and other DREBs serving in heat, drought, and salinity adaptation. The central role of these modulators in plant performance under challenging environments is based on (i interweaving of these regulators with other key signaling networks (plant hormones and redox signals as well as (ii their function in integrating responses across the whole plant, from light-harvesting and sugar-production in the leaf to foliar sugar export and water import and on to the plant’s sugar-consuming sinks (growth, storage, and reproduction. The example of Arabidopsis thaliana ecotypes from geographic origins with contrasting climates is used to describe the links between natural genetic variation in CBF transcription factors and the differential acclimation of plant anatomical and functional features needed to support superior photosynthetic performance in contrasting environments. Emphasis is placed on considering different temperature environments (hot versus cold and light environments (limiting versus high light, on trade-offs between adaptations to contrasting environments, and on plant lines minimizing such trade-offs.

  3. Género, Adversidad Familiar y Síntomas Emocionales en Preadolescentes Gender, Family Adversity and Emotional Symptoms in Preadolescents

    Directory of Open Access Journals (Sweden)

    Félix Cova

    2005-05-01

    Full Text Available Se evaluó la existencia de diferencias de género en preadolescentes en la presencia de problemas emocionales, así como la existencia de una vulnerabilidad diferencial de las niñas a presentar estos síntomas en condiciones de adversidad familiar. Se tomó una muestra de 133 niños y niñas de 11 a 13 años, elegidos al azar de 6 escuelas de alta vulnerabilidad social de la comuna de Concepción, Chile. Los problemas emocionales fueron valorados a través de cuestionarios aplicados a la madre (Child Behavior Checklist, CBCL y al profesor (Teacher Report Forms, TRF y a través de tres instrumentos de autoinforme (Youth Self Report, YRS; Cuestionario de Depresión Infantil, CDI; cuestionario de Ansiedad-Rasgo, STAIC/R. La presencia de indicadores de adversidad familiar fue evaluada a través de una entrevista semiestructurada desarrollada por la OMS, aplicada a la madre. Al considerar los autoinformes, se observó que las niñas informan experimentar más malestar emocional y sintomatología depresiva que los niños. Asimismo, se observó que las niñas expuestas a condiciones de adversidad familiar incrementan significativamente más que los niños expuestos a condiciones similares, su reporte de comportamientos internalizados, lo que indica la existencia de una mayor vulnerabilidad de las niñas a expresar sintomatología emocional en condiciones familiares negativas.This research evaluated both the existence of pre-teenager gender differences under emotional problems and the existence of a differential vulnerability in girls showing these symptoms in an adverse family environment. A sample of 133 boys and girls, aged 11 to 13, was randomly selected from high social risk schools from Concepción, Chile. The emotional disorders were evaluated by means of surveys to the mother (Child Behavior Checklist, CBCL and to the teacher (Teacher Report Forms, TRF and also taking into account three self-report instruments (Youth Self Report, YRS

  4. Consideration for a tritium removal facility at the Cernavoda Nuclear Power Station

    International Nuclear Information System (INIS)

    2006-01-01

    Full text: A pre-feasibility study considering process options for a Tritium Removal Facility at the Cernavoda Nuclear Power Station has been completed by ICIT and AECL. Three different process options were considered. These three options differ in the front-end process used to transfer tritium from heavy water to deuterium gas. All three options use cryogenic distillation (CD) as a back end process to extract tritium from the deuterium gas stream and concentrate it into a small volume stream of pure DT or T 2 that can be immobilized on a titanium sponge. The first option for the front-end process is Liquid Phase Catalytic Exchange (LPCE). The LPCE column is used to transfer the tritium from the heavy water to a recirculating stream of deuterium gas. The separation of hydrogen isotopes takes place in the cryogenic distillation column. Tritium-depleted deuterium gas from the CD system is fed back to the LPCE column. The cryogenic distillation system concentrates the tritium into a small volume of elemental tritium for storage. Tritiated heavy water that has been purified to remove catalyst poisons is fed to the top of the LPCE column. The heavy water leaving the column is depleted in deuterium. Both existing detritiation plants built to detritiate CANDU reactors (the Darlington TRF in Canada and the Wolsung TRF in Korea) use variations of the LPCE-CD process. The second option uses electrolysis to convert tritiated heavy water into oxygen and tritiated deuterium gas. The deuterium gas is sent to the Cryogenic Distillation system to extract and concentrate the tritium. The tritium depleted deuterium gas is recombined with the electrolytic oxygen to give a tritium-depleted heavy water product. The third option uses a Combined Electrolysis and Catalytic Exchange (CECE) front end. A CECE process concentrates the tritium in the water and, using water electrolysis, converts the concentrated tritium into deuterium gas. An overhead catalytic recombiner converts the

  5. Status of Precise Orbit Determination for Jason-2 Using GPS

    Science.gov (United States)

    Melachroinos, S.; Lemoine, F. G.; Zelensky, N. P.; Rowlands, D. D.; Pavlis, D. E.

    2011-01-01

    The JASON-2 satellite, launched in June 2008, is the latest follow-on to the successful TOPEX/Poseidon (T/P) and JASON-I altimetry missions. JASON-2 is equipped with a TRSR Blackjack GPS dual-frequency receiver, a laser retroreflector array, and a DORIS receiver for precise orbit determination (POD). The most recent time series of orbits computed at NASA GSFC, based on SLR/DORIS data have been completed using both ITRF2005 and ITRF2008. These orbits have been shown to agree radially at 1 cm RMS for dynamic vs SLRlDORIS reduced-dynamic orbits and in comparison with orbits produced by other analysis centers (Lemoine et al., 2010; Zelensky et al., 2010; Cerri et al., 2010). We have recently upgraded the GEODYN software to implement model improvements for GPS processing. We describe the implementation of IGS standards to the Jason2 GEODYN GPS processing, and other dynamical and measurement model improvements. Our GPS-only JASON-2 orbit accuracy is assessed using a number of tests including analysis of independent SLR and altimeter crossover residuals, orbit overlap differences, and direct comparison to orbits generated at GSFC using SLR and DORIS tracking, and to orbits generated externally at other centers. Tests based on SLR and the altimeter crossover residuals provide the best performance indicator for independent validation of the NASAlGSFC GPS-only reduced dynamic orbits. For the ITRF2005 and ITRF2008 implementation of our GPS-only obits we are using the IGS05 and IGS08 standards. Reduced dynamic versus dynamic orbit differences are used to characterize the remaining force model error and TRF instability. We evaluate the GPS vs SLR & DORIS orbits produced using the GEODYN software and assess in particular their consistency radially and the stability of the altimeter satellite reference frame in the Z direction for both ITRF2005 and ITRF2008 as a proxy to assess the consistency of the reference frame for altimeter satellite POD.

  6. Laminar Burning Velocities of Fuels for Advanced Combustion Engines (FACE) Gasoline and Gasoline Surrogates with and without Ethanol Blending Associated with Octane Rating

    KAUST Repository

    Mannaa, Ossama

    2016-05-04

    Laminar burning velocities of fuels for advanced combustion engines (FACE) C gasoline and of several blends of surrogate toluene reference fuels (TRFs) (n-heptane, iso-octane, and toluene mixtures) of the same research octane number are presented. Effects of ethanol addition on laminar flame speed of FACE-C and its surrogate are addressed. Measurements were conducted using a constant volume spherical combustion vessel in the constant pressure, stable flame regime at an initial temperature of 358 K and initial pressures up to 0.6 MPa with the equivalence ratios ranging from 0.8 to 1.6. Comparable values in the laminar burning velocities were measured for the FACE-C gasoline and the proposed surrogate fuel (17.60% n-heptane + 77.40% iso-octane + 5% toluene) over the range of experimental conditions. Sensitivity of flame propagation to total stretch rate effects and thermo-diffusive instability was quantified by determining Markstein length. Two percentages of an oxygenated fuel of ethanol as an additive, namely, 60 vol% and 85 vol% were investigated. The addition of ethanol to FACE-C and its surrogate TRF-1 (17.60% n-heptane + 77.40% iso-octane + 5% toluene) resulted in a relatively similar increase in the laminar burning velocities. The high-pressure measured values of Markstein length for the studied fuels blended with ethanol showed minimal influence of ethanol addition on the flame’s response to stretch rate and thermo-diffusive instability. © 2016 Taylor & Francis.

  7. Rapid screening of transferrin-binders in the flowers of Bauhinia blakeana Dunn by on-line high-performance liquid chromatography-diode-array detector-electrospray ionization-ion-trap-time-of-flight-mass spectrometry-transferrin-fluorescence detection system.

    Science.gov (United States)

    Liu, Meixian; Dong, Jing; Lin, Zongtao; Niu, Yanyan; Zhang, Xiaotian; Jiang, Haixiu; Guo, Ning; Li, Wei; Wang, Hong; Chen, Shizhong

    2016-06-10

    Transferrin (Transferrin, TRF, TF) has drawn increasing attention in cancer therapy due to its potential applications in drug delivery. TF receptor, highly expressed in tumor cells, recognizes and transports Fe(3+)-TF into cells to release iron into cytoplasm. Thus, discovering TF-binding compounds has become an active research area and is of great importance for target therapy. In this study, an on-line analysis method was established for screening TF-binding compounds from the flowers of Bauhinia blakeana Dunn using a high-performance liquid chromatography-diode-array detector-multi-stage mass spectrometry-transferrin-fluorescence detector (HPLC-DAD-MS(n)-TF-FLD) method. As a result, 33 of 80 identified or tentatively characterized compounds in the sample were TF-binding active. Twenty-five flavonol glycosides and eight phenolic acids were identified as TF-binders. Twelve of these active compounds together with six standard compounds were used to study the dose-response effects and structure-activity relationships of flavonoids and phenolic acids. The method was validated by vitexin with a good linearity in the range of concentrations used in the study. The limit of detection for vitexin was 0.1596 nmol. Our study indicated that the established method is simple, rapid and sensitive for screening TF-binding active compounds in the extract of Bauhinia blakeana Dunn, and therefore is important for discovering potential anti-cancer ingredients from complex samples for TF related drug delivery. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. First TSI observations of the new Compact Lightweight Absolute Radiometer (CLARA)

    Science.gov (United States)

    Walter, B.; Finsterle, W.; Koller, S.; Levesque, P. L.; Pfiffner, D.; Schmutz, W. K.

    2017-12-01

    Continuous and precise Total Solar Irradiance (TSI) measurements are indispensable to evaluate the influence of short- and long-term solar radiative emission variations on the Earth's energy budget. The existence of a potentially long-term trend in the suns activity and whether or not such a trend could be climate effective is still a matter of debate. The Compact Lightweight Absolute Radiometer (CLARA) is one of PMOD/WRC's future contributions to the almost seamless series of space borne TSI measurements since 1978. CLARA is one of three payloads of the Norwegian micro satellite NORSAT-1, along with Langmuir probes for space plasma research and an Automatic Identification System (AIS) receiver to monitor maritime traffic in Norwegian waters. NORSAT-1 was launched July 14th 2017 and the nominal operation of CLARA will start after the instrument commissioning beginning August 21st2017. We present the design, calibration and first TSI observations of CLARA, a new generation of active cavity Electrical Substitution Radiometers (ESR) comprising the latest radiometer developments of PMOD/WRC: i) A three-cavity design for degradation tracking and redundancy, ii) a digital control loop with feed forward system allowing for measurement cadences of 30s, iii) an aperture arrangement to reduce internal scattered light and iv) a new cavity and heatsink design to minimize non-equivalence, size and weight of the instrument. CLARA was end-to-end calibrated against the SI traceable cryogenic radiometer of the TSI Radiometer Facility (TRF) in Boulder (Colorado). The absolute measurement uncertainties for the three SI-traceable TSI detectors of CLARA are 567, 576 and 912 ppm (k = 1).

  9. [Reliability and Validity of the Behavioral Check List for Preschool Children to Measure Attention Deficit Hyperactivity Behaviors].

    Science.gov (United States)

    Tsuno, Kanami; Yoshimasu, Kouichi; Hayashi, Takashi; Tatsuta, Nozomi; Ito, Yuki; Kamijima, Michihiro; Nakai, Kunihiko

    2018-01-01

    Nowadays, attention deficit hyperactivity (ADH) problems are observed commonly among school-age children. However, questionnaires specific to ADH behaviors among preschool children are very few. The aim of this study was to investigate the reliability and validity of the 25-item Behavioral Check List (BCL), which was developed from interviews of parents with children who were diagnosed as having Attention-deficit/hyperactivity disorder (ADHD) and measures ADH behaviors in preschool age. We recruited 22 teachers from 10 nurseries/kindergartens in Miyagi Prefecture, Japan. A total of 138 preschool children were assessed using the BCL. To investigate inter-rater reliability, two teachers from each facility assess seven to twenty children in their class, and intraclass correlation coefficients (ICCs) were calculated. The teachers additionally answered questions in the 1/5-5 Caregiver-Teacher Report Form (C-TRF) to investigate the criterion validity of the BCL. To investigate structural validity, exploratory factor analysis with promax rotation and confirmatory factor analysis were performed. The internal consistency reliability of the BCL was good (α = 0.92) and correlation analyses also confirmed its excellent criterion validity. Although exploratory factor analysis for the BCL yielded a five-factor model that consisted of a factor structure different from that of the original one, the results were similar to the original six factors. The ICCs of the BCL were 0.38-0.99 and it was not high enough for inter-rater reliability in some facilities. However, there is a possibility to improve it by giving raters adequate explanations when using BCL. The present study showed acceptable levels of reliability and validity of the BCL among Japanese preschool children.

  10. Sequential sampling of visual objects during sustained attention.

    Directory of Open Access Journals (Sweden)

    Jianrong Jia

    2017-06-01

    Full Text Available In a crowded visual scene, attention must be distributed efficiently and flexibly over time and space to accommodate different contexts. It is well established that selective attention enhances the corresponding neural responses, presumably implying that attention would persistently dwell on the task-relevant item. Meanwhile, recent studies, mostly in divided attentional contexts, suggest that attention does not remain stationary but samples objects alternately over time, suggesting a rhythmic view of attention. However, it remains unknown whether the dynamic mechanism essentially mediates attentional processes at a general level. Importantly, there is also a complete lack of direct neural evidence reflecting whether and how the brain rhythmically samples multiple visual objects during stimulus processing. To address these issues, in this study, we employed electroencephalography (EEG and a temporal response function (TRF approach, which can dissociate responses that exclusively represent a single object from the overall neuronal activity, to examine the spatiotemporal characteristics of attention in various attentional contexts. First, attention, which is characterized by inhibitory alpha-band (approximately 10 Hz activity in TRFs, switches between attended and unattended objects every approximately 200 ms, suggesting a sequential sampling even when attention is required to mostly stay on the attended object. Second, the attentional spatiotemporal pattern is modulated by the task context, such that alpha-mediated switching becomes increasingly prominent as the task requires a more uniform distribution of attention. Finally, the switching pattern correlates with attentional behavioral performance. Our work provides direct neural evidence supporting a generally central role of temporal organization mechanism in attention, such that multiple objects are sequentially sorted according to their priority in attentional contexts. The results suggest

  11. Head and neck cancer. Usefulness of concurrent chemoradiation therapy with TPF

    International Nuclear Information System (INIS)

    Yoshida, Tomoyuki

    2010-01-01

    History of chemotherapy and radiotherapy for head and neck cancer (HNC) leading to the present regimen concurrent chemoradiation therapy (CCRT) with docetaxel, cisplatin, fluorouracil (5-FU), taxotere (docetaxel)+cisplatin+5-FU (TRF), is described together with authors' experience of its trials. History of the CCRT explains results of clinical trials of radio- and chemo-therapies conducted globally and in Japan with various regimens like neo-adjuvant, adjuvant, concurrent and alternating one. During the process, CCRT has been established as a standard therapy of nasopharyngeal cancer. Trials of CCRT further added with molecular targeting anticancer (cetuximab) are now in consideration and practice in HNC field. A phase I CCRT study conducted by authors with TPF at lower doses of the 3 agents than those reported abroad having resulted in outcomes with satisfactory tolerance, based on which phase II trials with 2 Gy/day for 5 days/week (total, 60 Gy) have been performed. The therapy is to be once discontinued for 2 weeks when the cumulative dose attains 40 Gy. Currently available results in 48 HNC patients (44 males, 4 females, middle-/hypo-pharynx and larynx cancers, age 48-72 y, stage III-N2b or more advanced) within 8-60 (mean 45.1) mo follow-up are: chemotherapy completion in 87.5%, total dose 60-70 Gy in all patients; adverse events grade 3-4, 14.9-54.2% (hematological), 50.0% (mucositis), 18.8% (nausea), and 6.3% (liver function); response rate of 87.5% including 77.1% complete response (CR); and 57 mo-survival and progression-free survival rates of 75.9 and 57.8%, respectively. The results suggest usefulness of CCRT with TPF for the disease. Similar clinical trials are now in progress globally. (T.T.)

  12. In Vivo toxicological assessment of biologically synthesized silver nanoparticles in adult Zebrafish (Danio rerio)

    Energy Technology Data Exchange (ETDEWEB)

    Krishnaraj, Chandran, E-mail: krishnarajbio@gmail.com [Department of Food Science & Technology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju 561-756 (Korea, Republic of); Harper, Stacey L. [Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR 97331 (United States); Yun, Soon-Il, E-mail: siyun@jbnu.ac.kr [Department of Food Science & Technology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju 561-756 (Korea, Republic of)

    2016-01-15

    Highlights: • Synthesis of AgNPs achieved using Malva crispa Linn., leaves extract. • 96 h LC{sub 50} concentration of AgNPs was observed at 142.2 μg/l in adult zebrafish. • Cytological changes and intrahepatic localization of AgNPs were demonstrated in tissues. • Presence of micronuclei and nuclear abnormalities were observed. • The mRNA expression of stress and immune response related genes were analyzed. - Abstract: The present study examines the deleterious effect of biologically synthesized silver nanoparticles in adult zebrafish. Silver nanoparticles (AgNPs) used in the study were synthesized by treating AgNO{sub 3} with aqueous leaves extract of Malva crispa Linn., a medicinal herb as source of reductants. LC{sub 50} concentration of AgNPs at 96 h was observed as 142.2 μg/l. In order to explore the underlying toxicity mechanisms of AgNPs, half of the LC{sub 50} concentration (71.1 μg/l) was exposed to adult zebrafish for 14 days. Cytological changes and intrahepatic localization of AgNPs were observed in gills and liver tissues respectively, and the results concluded a possible sign for oxidative stress. In addition to oxidative stress the genotoxic effect was observed in peripheral blood cells like presence of micronuclei, nuclear abnormalities and also loss in cell contact with irregular shape was observed in liver parenchyma cells. Hence to confirm the oxidative stress and genotoxic effects the mRNA expression of stress related (MTF-1, HSP70) and immune response related (TLR4, NFKB, IL1B, CEBP, TRF, TLR22) genes were analyzed in liver tissues and the results clearly concluded that the plant extract mediated synthesis of AgNPs leads to oxidative stress and immunotoxicity in adult zebrafish.

  13. The Use of a Combined Bioinformatics Approach to Locate Antibiotic Resistance Genes on Plasmids From Whole Genome Sequences of Salmonella enterica Serovars From Humans in Ghana

    Directory of Open Access Journals (Sweden)

    Egle Kudirkiene

    2018-05-01

    Full Text Available In the current study, we identified plasmids carrying antimicrobial resistance genes in draft whole genome sequences of 16 selected Salmonella enterica isolates representing six different serovars from humans in Ghana. The plasmids and the location of resistance genes in the genomes were predicted using a combination of PlasmidFinder, ResFinder, plasmidSPAdes and BLAST genomic analysis tools. Subsequently, S1-PFGE was employed for analysis of plasmid profiles. Whole genome sequencing confirmed the presence of antimicrobial resistance genes in Salmonella isolates showing multidrug resistance phenotypically. ESBL, either blaTEM52−B or blaCTX−M15 were present in two cephalosporin resistant isolates of S. Virchow and S. Poona, respectively. The systematic genome analysis revealed the presence of different plasmids in different serovars, with or without insertion of antimicrobial resistance genes. In S. Enteritidis, resistance genes were carried predominantly on plasmids of IncN type, in S. Typhimurium on plasmids of IncFII(S/IncFIB(S/IncQ1 type. In S. Virchow and in S. Poona, resistance genes were detected on plasmids of IncX1 and TrfA/IncHI2/IncHI2A type, respectively. The latter two plasmids were described for the first time in these serovars. The combination of genomic analytical tools allowed nearly full mapping of the resistance plasmids in all Salmonella strains analyzed. The results suggest that the improved analytical approach used in the current study may be used to identify plasmids that are specifically associated with resistance phenotypes in whole genome sequences. Such knowledge would allow the development of rapid multidrug resistance tracking tools in Salmonella populations using WGS.

  14. Differential effects of diet composition and timing of feeding behavior on rat brown adipose tissue and skeletal muscle peripheral clocks

    Directory of Open Access Journals (Sweden)

    Paul de Goede

    2018-01-01

    Full Text Available The effects of feeding behavior and diet composition, as well as their possible interactions, on daily (clock gene expression rhythms have mainly been studied in the liver, and to a lesser degree in white adipose tissue (WAT, but hardly in other metabolic tissues such as skeletal muscle (SM and brown adipose tissues (BAT. We therefore subjected male Wistar rats to a regular chow or free choice high-fat-high sugar (fcHFHS diet in combination with time restricted feeding (TRF to either the light or dark phase. In SM, all tested clock genes lost their rhythmic expression in the chow light fed group. In the fcHFHS light fed group rhythmic expression for some, but not all, clock genes was maintained, but shifted by several hours. In BAT the daily rhythmicity of clock genes was maintained for the light fed groups, but expression patterns were shifted as compared with ad libitum and dark fed groups, whilst the fcHFHS diet made the rhythmicity of clock genes become more pronounced. Most of the metabolic genes in BAT tissue tested did not show any rhythmic expression in either the chow or fcHFHS groups. In SM Pdk4 and Ucp3 were phase-shifted, but remained rhythmically expressed in the chow light fed groups. Rhythmic expression was lost for Ucp3 whilst on the fcHFHS diet during the light phase. In summary, both feeding at the wrong time of day and diet composition disturb the peripheral clocks in SM and BAT, but to different degrees and thereby result in a further desynchronization between metabolically active tissues such as SM, BAT, WAT and liver.

  15. Early enteral immune nutrition support after radical operation for gastric cancer on promoting the recovery of gastrointestinal function and immune function

    Directory of Open Access Journals (Sweden)

    Zhi-Gang Li

    2016-05-01

    Full Text Available Objective: To analyze the effect of early enteral immune nutrition support after radical operation for gastric cancer on the recovery of gastrointestinal function and immune function. Methods: A total of 106 cases of patients received radical operation for gastric cancer in our hospital were selected as research subjects, and according to different ways of postoperative nutrition intervention, all patients were divided into observation group (n=50 and control group (n=56. Control group received conventional enteral nutrition intervention, observation group received postoperative early enteral immune nutrition support, and then differences in postoperative intestinal mucosa barrier function, gastrointestinal hormone levels, immune function levels and nutrition-related indicator values were compared between two groups. Results: After observation group received enteral immune nutrition intervention, serum DAO, PS and D-lactate levels as well as urine L/M ratio were lower than those of control group; serum GAS, CCK, MTL and SP values of observation group after intervention were higher than those of control group, and GLU, VIP, GIP and SS values were lower than those of control group; CD4, IgG, NK cell, C3, C4, CH50 and S-IgA levels of observation group after intervention were higher than those of control group; serum ALB, PRE, TRF and RBP levels of observation group after intervention were higher than those of control group. Conclusion: Early enteral immune nutrition support after radical operation for gastric cancer is conducive to the recovery of gastrointestinal function and the promotion of immune state, eventually promotes patients’ postoperative overall recovery and has active clinical significance.

  16. T-oligo as an anticancer agent in colorectal cancer

    Energy Technology Data Exchange (ETDEWEB)

    Wojdyla, Luke; Stone, Amanda L. [Department of Biomedical Sciences, University of Illinois College of Medicine at Rockford, Rockford, IL (United States); Sethakorn, Nan [Department of Medicine, University of Chicago, Chicago, IL (United States); Uppada, Srijayaprakash B.; Devito, Joseph T. [Department of Biomedical Sciences, University of Illinois College of Medicine at Rockford, Rockford, IL (United States); Bissonnette, Marc [Department of Medicine, University of Chicago, Chicago, IL (United States); Puri, Neelu, E-mail: neelupur@uic.edu [Department of Biomedical Sciences, University of Illinois College of Medicine at Rockford, Rockford, IL (United States)

    2014-04-04

    Highlights: • T-oligo induces cell cycle arrest, senescence, apoptosis, and differentiation in CRC. • Treatment with T-oligo downregulates telomere-associated proteins. • T-oligo combined with an EGFR-TKI additively inhibits cellular proliferation. • T-oligo has potential as an effective therapeutic agent for CRC. - Abstract: In the United States, there will be an estimated 96,830 new cases of colorectal cancer (CRC) and 50,310 deaths in 2014. CRC is often detected at late stages of the disease, at which point there is no effective chemotherapy. Thus, there is an urgent need for effective novel therapies that have minimal effects on normal cells. T-oligo, an oligonucleotide homologous to the 3′-telomere overhang, induces potent DNA damage responses in multiple malignant cell types, however, its efficacy in CRC has not been studied. This is the first investigation demonstrating T-oligo-induced anticancer effects in two CRC cell lines, HT-29 and LoVo, which are highly resistant to conventional chemotherapies. In this investigation, we show that T-oligo may mediate its DNA damage responses through the p53/p73 pathway, thereby inhibiting cellular proliferation and inducing apoptosis or senescence. Additionally, upregulation of downstream DNA damage response proteins, including E2F1, p53 or p73, was observed. In LoVo cells, T-oligo induced senescence, decreased clonogenicity, and increased expression of senescence associated proteins p21, p27, and p53. In addition, downregulation of POT1 and TRF2, two components of the shelterin protein complex which protects telomeric ends, was observed. Moreover, we studied the antiproliferative effects of T-oligo in combination with an EGFR tyrosine kinase inhibitor, Gefitinib, which resulted in an additive inhibitory effect on cellular proliferation. Collectively, these data provide evidence that T-oligo alone, or in combination with other molecularly targeted therapies, has potential as an anti-cancer agent in CRC.

  17. SSU rDNA sequence diversity and seasonally differentiated distribution of nanoplanktonic ciliates in neritic Bohai and Yellow Seas as revealed by T-RFLP.

    Directory of Open Access Journals (Sweden)

    Jun Dong

    Full Text Available Nanociliates have been frequently found to be important players in the marine microbial loop, however, little is known about their diversity and distribution in coastal ecosystems. We investigated the molecular diversity and distribution patterns of nanoplanktonic oligotrich and choreotrich (OC ciliates in surface water of three neritic basins of northern China, the South Yellow Sea (SYS, North Yellow Sea (NYS, and Bohai Sea (BS in June and November 2011. SSU rRNA gene clone libraries generated from three summertime samples (sites B38, B4 and H8 were analyzed and revealed a large novel ribotype diversity, of which many were low-abundant phylotypes belonging to the subclass Oligotrichia, but divergent from described morphospecies. Based on the data of terminal-restriction fragment length polymorphism (T-RFLP analysis of all 35 samples, we found that the T-RF richness was generally higher in the SYS than in the BS, and negatively correlated with the molar ratio of P to Si. Overall, multidimensional scaling and permutational multivariate analysis of variance of the community turnover demonstrated a distinct seasonal pattern but no basin-to-basin differentiation across all samples. Nevertheless, significant community differences among basins were recognized in the winter dataset. Mantel tests showed that the environmental factors, P:Si ratio, water temperature and concentration of dissolved oxygen (DO, determined the community across all samples. However, both biogeographic distance and environment shaped the community in winter, with DO being the most important physicochemical factor. Our results indicate that the stoichiometric ratio of P:Si is a key factor, through which the phytoplankton community may be shaped, resulting in a cascade effect on the diversity and community composition of OC nanociliates in the N-rich, Si-limited coastal surface waters, and that the Yellow Sea Warm Current drives the nanociliate community, and possibly the

  18. SSU rDNA sequence diversity and seasonally differentiated distribution of nanoplanktonic ciliates in neritic Bohai and Yellow Seas as revealed by T-RFLP.

    Science.gov (United States)

    Dong, Jun; Shi, Fei; Li, Han; Zhang, Xiaoming; Hu, Xiaozhong; Gong, Jun

    2014-01-01

    Nanociliates have been frequently found to be important players in the marine microbial loop, however, little is known about their diversity and distribution in coastal ecosystems. We investigated the molecular diversity and distribution patterns of nanoplanktonic oligotrich and choreotrich (OC) ciliates in surface water of three neritic basins of northern China, the South Yellow Sea (SYS), North Yellow Sea (NYS), and Bohai Sea (BS) in June and November 2011. SSU rRNA gene clone libraries generated from three summertime samples (sites B38, B4 and H8) were analyzed and revealed a large novel ribotype diversity, of which many were low-abundant phylotypes belonging to the subclass Oligotrichia, but divergent from described morphospecies. Based on the data of terminal-restriction fragment length polymorphism (T-RFLP) analysis of all 35 samples, we found that the T-RF richness was generally higher in the SYS than in the BS, and negatively correlated with the molar ratio of P to Si. Overall, multidimensional scaling and permutational multivariate analysis of variance of the community turnover demonstrated a distinct seasonal pattern but no basin-to-basin differentiation across all samples. Nevertheless, significant community differences among basins were recognized in the winter dataset. Mantel tests showed that the environmental factors, P:Si ratio, water temperature and concentration of dissolved oxygen (DO), determined the community across all samples. However, both biogeographic distance and environment shaped the community in winter, with DO being the most important physicochemical factor. Our results indicate that the stoichiometric ratio of P:Si is a key factor, through which the phytoplankton community may be shaped, resulting in a cascade effect on the diversity and community composition of OC nanociliates in the N-rich, Si-limited coastal surface waters, and that the Yellow Sea Warm Current drives the nanociliate community, and possibly the microbial food webs

  19. Assessment of Current Global and Regional Mean Sea Level Estimates Based on the TOPEX/Poseidon Jason-1 and 2 Climate Data Record

    Science.gov (United States)

    Beckley, B. D.; Lemoine, F. G.; Zelensky, N. P.; Yang, X.; Holmes, S.; Ray, R. D.; Mitchum, G. T.; Desai, S.; Brown, S.; Haines, B.

    2011-01-01

    Recent developments in Precise Orbit Determinations (POD) due to in particular to revisions to the terrestrial reference frame realization and the time variable gravity (TVG) continues to provide improvements to the accuracy and stability of the PO directly affecting mean sea level (MSL) estimates. Long-term credible MSL estimates require the development and continued maintenance of a stable reference frame, along with vigilant monitoring of the performance of the independent tracking systems used to calculate the orbits for altimeter spacecrafts. The stringent MSL accuracy requirements of a few tenths of an mm/yr are particularly essential for mass budget closure analysis over the relative short time period of Jason-l &2, GRACE, and Argo coincident measurements. In an effort to adhere to cross mission consistency, we have generated a full time series of experimental orbits (GSFC stdlllO) for TOPEX/Poseidon (TP), Jason-I, and OSTM based on an improved terrestrial reference frame (TRF) realization (ITRF2008), revised static (GGM03s), and time variable gravity field (Eigen6s). In this presentation we assess the impact of the revised precision orbits on inter-mission bias estimates, and resultant global and regional MSL trends. Tide gauge verification results are shown to assess the current stability of the Jason-2 sea surface height time series that suggests a possible discontinuity initiated in early 2010. Although the Jason-2 time series is relatively short (approximately 3 years), a thorough review of the entire suite of geophysical and environmental range corrections is warranted and is underway to maintain the fidelity of the record.

  20. Fragile sites, dysfunctional telomere and chromosome fusions: What is 5S rDNA role?

    Science.gov (United States)

    Barros, Alain Victor; Wolski, Michele Andressa Vier; Nogaroto, Viviane; Almeida, Mara Cristina; Moreira-Filho, Orlando; Vicari, Marcelo Ricardo

    2017-04-15

    Repetitive DNA regions are known as fragile chromosomal sites which present a high flexibility and low stability. Our focus was characterize fragile sites in 5S rDNA regions. The Ancistrus sp. species shows a diploid number of 50 and an indicative Robertsonian fusion at chromosomal pair 1. Two sequences of 5S rDNA were identified: 5S.1 rDNA and 5S.2 rDNA. The first sequence gathers the necessary structures to gene expression and shows a functional secondary structure prediction. Otherwise, the 5S.2 rDNA sequence does not contain the upstream sequences that are required to expression, furthermore its structure prediction reveals a nonfunctional ribosomal RNA. The chromosomal mapping revealed several 5S.1 and 5S.2 rDNA clusters. In addition, the 5S.2 rDNA clusters were found in acrocentric and metacentric chromosomes proximal regions. The pair 1 5S.2 rDNA cluster is co-located with interstitial telomeric sites (ITS). Our results indicate that its clusters are hotspots to chromosomal breaks. During the meiotic prophase bouquet arrangement, double strand breaks (DSBs) at proximal 5S.2 rDNA of acrocentric chromosomes could lead to homologous and non-homologous repair mechanisms as Robertsonian fusions. Still, ITS sites provides chromosomal instability, resulting in telomeric recombination via TRF2 shelterin protein and a series of breakage-fusion-bridge cycles. Our proposal is that 5S rDNA derived sequences, act as chromosomal fragile sites in association with some chromosomal rearrangements of Loricariidae. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Estimating fuel octane numbers from homogeneous gas-phase ignition delay times

    KAUST Repository

    Naser, Nimal

    2017-11-05

    Fuel octane numbers are directly related to the autoignition properties of fuel/air mixtures in spark ignition (SI) engines. This work presents a methodology to estimate the research and the motor octane numbers (RON and MON) from homogeneous gas-phase ignition delay time (IDT) data calculated at various pressures and temperatures. The hypothesis under investigation is that at specific conditions of pressure and temperature (i.e., RON-like and MON-like conditions), fuels with IDT identical to that of a primary reference fuel (PRF) have the same octane rating. To test this hypothesis, IDTs with a detailed gasoline surrogate chemical kinetic model have been calculated at various temperatures and pressures. From this dataset, temperatures that best represent RON and MON have been correlated at a specified pressure. Correlations for pressures in the range of 10–50 bar were obtained. The proposed correlations were validated with toluene reference fuels (TRF), toluene primary reference fuels (TPRF), ethanol reference fuels (ERF), PRFs and TPRFs with ethanol, and multi-component gasoline surrogate mixtures. The predicted RON and MON showed satisfactory accuracy against measurements obtained by the standard ASTM methods and blending rules, demonstrating that the present methodology can be a viable tool for a first approximation. The correlations were also validated against an extensive set of experimental IDT data obtained from literature with a high degree of accuracy in RON/MON prediction. Conditions in homogeneous reactors such as shock tubes and rapid compression machines that are relevant to modern SI engines were also identified. Uncertainty analysis of the proposed correlations with linear error propagation theory is also presented.

  2. Multi-geodetic characterization of the seasonal signal at the CERGA geodetic reference, France

    Science.gov (United States)

    Memin, A.; Viswanathan, V.; Fienga, A.; Santamaría-Gómez, A.; Boy, J. P.

    2016-12-01

    Crustal deformations due to surface-mass loading account for a significant part of the variability in geodetic time series. A perfect understanding of the loading signal observed by geodetic techniques should help in improving terrestrial reference frame (TRF) realizations. Yet, discrepancies between crustal motion estimates from models of surface-mass loading and observations are still too large so that no model is currently recommended by the IERS for reducing the data. We investigate the discrepancy observed in the seasonal variations of the CERGA station, South of France.We characterize the seasonal motions of the reference geodetic station CERGA from GNSS, SLR and LLR. We compare the station motion observed with GNSS and SLR and we estimate changes in the station-to-the-moon distance using an improved processing strategy. We investigate the consistency between these geodetic techniques and compare the observed station motion with that estimated using models of surface-mass change. In that regard, we compute atmospheric loading effects using surface pressure fields from ECMWF, assuming an ocean response according to the classical inverted barometer (IB) assumption, considered to be valid for periods typically exceeding a week. We also used general circulation ocean models (ECCO and GLORYS) forced by wind, heat and fresh water fluxes. The continental water storage is described using GLDAS/Noah and MERRA-land models.Using the surface-mass models, we estimate the amplitude of the seasonal vertical motion of the CERGA station ranging between 5 and 10 mm with a maximum reached in August, mostly due to hydrology. The horizontal seasonal motion of the station may reach up to 3 mm. Such a station motion should induce a change in the distance to the moon reaching up to 10 mm, large enough to be detected in LLR time series and compared to GNSS- and SLR-derived motion.

  3. 伴有对立违抗障碍注意缺陷多动障碍患儿智力和行为特征及与血清 5- 羟色胺水平的关系%Intellectual and behavioral characteristics and their relations with serum 5 hydroxytryptamine level in children with attention deficit hyperactivity disorder complicated by oppositional defiant disorder

    Institute of Scientific and Technical Information of China (English)

    高雪屏; 苏林雁; 谢光荣; 黄春香; 李雪荣

    2005-01-01

    BACKGROUND:Oppositional defiant disorder(ODD) often occurs as a comorbid condition of attention deficit hyperactivity disorder(ADHD),characterized by defiant behaviors.ADHD children with ODD have more extensive impairments than those with ADHD alone. Some studies suggest that decreased serum 5 hydroxytryptamine(5 HT) level is related to aggressive behavior in ADHD, but no relevant report is available in China. OBJECTIVE:To investigate the clinical features of ADHD with ODD,and their relations with serum 5 HT. DESIGN:A randomized case controlled study taking the ADHD children with or without ODD as the subjects for study. SETTING:Mental Health Institute of the Second Xiangya Hospital. PARTICIPANTS:Sixty one ADHD children(53 boys and 8 girls) aged 7 to 14 years visiting the Children's Clinic of Mental Health Institute of Central South University from June 2002 to May 2003 were recruited and divided into two groups based on the symptomatic criteria of ODD recommended by the Diagnostic and Statistical Manual of Mental Disorder(DSM Ⅳ ):ADHD group(n=33,without ODD) and ADHD+ ODD group(n=28). INTERVENTIONS:The parents of the enrolled children(n=61) were asked to complete the Achenbach child behavior checklist (CBCL),and the teachers(n=31) completed the teacher's report form (TRF).Two milliliters of fasting venous blood was drawn from these children and the serum separated by centrifugation for quantification of 5 HT using external standard method,and whole blood 5 HT was analyzed by LD 10AD high performance liquid chromatography(HPLC). MAIN OUTCOME MEASURES:Scores of CBCL,TRF and Chinese Wechsler Intelligence Scale for Children(C WISC) and serum 5 HT levels. RESULTS:In the ADHD+ ODD group,the scores of CBCL and TRF for externalizing, aggressive behaviors and total scores for behavioral problems rated by the parents and teachers were significantly higher than those of the ADHD group(t=2.28 to 3.76,P< 0.05 to 0.01);the former group also had significantly higher scores of

  4. Analysis of molecular markers as predictive factors of lymph node involvement in breast carcinoma.

    Science.gov (United States)

    Paula, Luciana Marques; De Moraes, Luis Henrique Ferreira; Do Canto, Abaeté Leite; Dos Santos, Laurita; Martin, Airton Abrahão; Rogatto, Silvia Regina; De Azevedo Canevari, Renata

    2017-01-01

    Nodal status is the most significant independent prognostic factor in breast cancer. Identification of molecular markers would allow stratification of patients who require surgical assessment of lymph nodes from the large numbers of patients for whom this surgical procedure is unnecessary, thus leading to a more accurate prognosis. However, up to now, the reported studies are preliminary and controversial, and although hundreds of markers have been assessed, few of them have been used in clinical practice for treatment or prognosis in breast cancer. The purpose of the present study was to determine whether protein phosphatase Mg2+/Mn2+ dependent 1D, β-1,3-N-acetylglucosaminyltransferase, neural precursor cell expressed, developmentally down-regulated 9, prohibitin, phosphoinositide-3-kinase regulatory subunit 5 (PIK3R5), phosphatidylinositol-5-phosphate 4-kinase type IIα, TRF1-interacting ankyrin-related ADP-ribose polymerase 2, BCL2 associated agonist of cell death, G2 and S-phase expressed 1 and PAX interacting protein 1 genes, described as prognostic markers in breast cancer in a previous microarray study, are also predictors of lymph node involvement in breast carcinoma Reverse transcription-quantitative polymerase chain reaction analysis was performed on primary breast tumor tissues from women with negative lymph node involvement (n=27) compared with primary tumor tissues from women with positive lymph node involvement (n=23), and was also performed on primary tumors and paired lymph node metastases (n=11). For all genes analyzed, only the PIK3R5 gene exhibited differential expression in samples of primary tumors with positive lymph node involvement compared with primary tumors with negative lymph node involvement (P=0.0347). These results demonstrate that the PIK3R5 gene may be considered predictive of lymph node involvement in breast carcinoma. Although the other genes evaluated in the present study have been previously characterized to be involved with

  5. Intergenerational Transmission of Internalizing Behavior: The Role of Maternal Psychopathology, Child Responsiveness and Maternal Attachment Style Insecurity.

    Science.gov (United States)

    Reck, Corinna; Nonnenmacher, Nora; Zietlow, Anna-Lena

    Maternal depression and anxiety disorders are risk factors for the development of internalizing disorders in offspring. Maternal attachment has been discussed as one factor accounting for transmission. The aim of this study was to investigate child internalizing behavior at preschool age on a symptomatic and behavioral level and possible links to maternal mental health over time and maternal attachment style insecurity in a sample of postpartum depressed and anxious mothers. Child internalizing behavior at preschool age was rated by the Child Behavior Checklist (CBCL), the Caregiver-Teacher Report Form (C-TRF), and during a mother-child free-play situation. We focused on child responsiveness as it has been linked to child internalizing behavior. Maternal attachment style insecurity was tested to mediate the link between maternal mental health (assessed postpartum and at preschool age with the Structured Clinical Interview for DSM-IV Axis-I Disorders, SCID-I) and child internalizing behavior/child responsiveness. Of the overall sample (n = 58), 28 women were diagnosed with postpartum depression and/or anxiety disorders according to DSM-IV, and 30 were healthy controls. Data were collected 3-9 months after delivery and at preschool age (mean = 4.6 years). At preschool age, children of postpartum depressed and anxious mothers were rated significantly higher on child internalizing behavior by mothers, fathers, and additional caregivers compared to the control group. Child internalizing behavior rated by mothers was influenced by current psychiatric symptoms; maternal attachment style insecurity did not mediate this link. During interaction, children in the clinical group displayed significantly less child responsiveness compared to the control group. Maternal attachment style insecurity mediated the relationship between maternal mental health over time and child responsiveness. The results emphasize the need for interventions focusing on mother-child interaction and

  6. Genetic variation in GABRA2 moderates peer influence on externalizing behavior in adolescents.

    Science.gov (United States)

    Villafuerte, Sandra; Trucco, Elisa M; Heitzeg, Mary M; Burmeister, Margit; Zucker, Robert A

    2014-01-01

    Genetic predisposition and environmental influences are both important factors in the development of problematic behavior leading to substance use in adolescence. Involvement with delinquent peers also strongly predicts adolescent externalizing behavior. Several lines of evidence support a role of GABRA2 on externalizing behavior related to disinhibition. However, whether this genetic association is influenced by the environment such as peer behavior remains unknown. We examined the moderating role of GABRA2 genetic variation on the socialization model of delinquent peer affiliation (at ages 12-14 years) on externalizing behavior (at ages 15-17 years) in the Michigan Longitudinal Study (MLS) adolescent sample. The sample consisted of 244 adolescents (75 females and 152 with at least one parent with a DSM-IV lifetime alcohol dependence/abuse diagnosis). Peer delinquent activity reported by the participant and teacher-reported adolescent externalizing behavior (Teacher Report Form (TRF) were assessed. No main effect of the GABRA2 SNP rs279826, which tags a large haplotype, on externalizing behavior was observed. However, there was a statistically reliable GABRA2 × peer delinquency interaction. The effect of peer delinquent involvement on externalizing scores and the rule breaking subscale is significantly stronger for those with the GG genotype compared to A-carriers, whereas there was no effect of genotype on externalizing in the absence of peer delinquent involvement. No interaction was observed for the aggression subscale. Our results suggest that the genetic effect of GABRA2 on externalizing behavior, more specifically on rule breaking is, at least in part, due to its effect on susceptibility to environmental exposure (i.e., peer delinquency).

  7. SILENT MYOCARDIAL ISCHEMIA AND CARDIAC RHYTHM DISTURBANCES IN WOMEN WITH RHEUMATOID ARTHRITIS

    Directory of Open Access Journals (Sweden)

    D. S. Novikova

    2014-07-01

    Full Text Available Objective: to assess the rate of silent myocardial ischemia (SMI and the pattern of cardiac rhythm disturbances in women with rheumatoidarthritis (RA, their association with traditional risk factors (TRF for cardiovascular diseases (CVD, with subclinical structural and functionalchanges in the heart and vessels, with the activity and severity of rheumatoid inflammation.Subjects and methods. Two hundred and ninety-one female patients aged less than 60 years with a valid diagnosis of RA and no clinicalsigns were examined. A control group consisted of 125 women without rheumatic diseases. In addition to the clinical manifestations, activity,and severity of RA, the authors assessed major TRFs for CVD, performed Holter ECG monitoring, common carotid artery duplex scanning, transthoracic echocardiographic study, and determined the levels of serum inflammatory markers.Results. The women with RA differ from the control group in the higher incidence of SMI, supraventricular arrhythmias (SVA and highgradepremature ventricular contractions (PVC. The patients with RA and SMI are characterized in terms of age-adjustment by higher disease activity (DAS28, systemic manifestations, cumulative larger-dose glucocorticoids (GC and a higher percentage of patients receiving disease-modifying anti-rheumatic drugs as compared with those with RA and no SMI with adjustment for age. High disease activity(DAS28, level of inflammatory markers, IgM rheumatoid arthritis seropositivity, and GC therapy are SVA-associated factors in women with RA; larger left ventricular end-diastolic dimension and serositis are factors associated with high-grade PVC.Conclusion. The RA women without clinical manifestations of CVD are recorded to have high rates of SMI, SVA, and high-grade PVC, which is primarily due to the activity and severity of rheumatoid inflammation.

  8. Effect of traditional and integrative regimens on quality of life and early renal impairment in elderly patients with isolated systolic hypertension.

    Science.gov (United States)

    Li, Hao; Liu, Long-tao; Zhao, Wen-ming; Liu, Jian-gang; Yao, Ming-jiang; Han, Yong-xiang; Shen, Yan-peng; Liu, Xing-dong; Liu, Li; Wang, Xue-mei; Cai, Lin-lin; Guan, Jie

    2010-06-01

    To observe the effect of Chinese medical regimen and integrative medical regimen on quality of life and early renal impairment in elderly patients with isolated systolic hypertension (EISH). A multi-center, randomized, double-blinded controlled trail was adopted. A total of 270 cases of EISH were randomly divided into 3 groups: Chinese medicine group (CM), combination group and Western medicine group (WM). The course of treatment was 4 weeks. The clinical blood pressure, integral of quality of life (SF-36 scale), immunoglubin G (IgG), microalbumin (mALB), beta(2)-microglobulin (beta(2)-MG), transferrin (TRF) and N-acetyl-beta'-D-glucosa-minidase (NAG) in urine were determined before and after the treatment. After treatment, systolic blood pressure depressed significantly in each group (P<0.05), and the combination group was superior to CM or WM group in depressing SBP (P<0.05); in each group, integral of quality of life improved in different degree, and combination group was superior to WM group in all 8 dimensions (P<0.05). The level of mALB and beta(2)-MG in urine decreased in all groups (P<0.05), and the combination group was superior to CM group or WM group in decreasing mALB (P<0.05). Chinese medical regimen has affirmative effect in treating EISH patients, and could lower the systolic blood pressure, improve quality of life and early renal impairment of the patients, and integrative medical regimen has superiority on account of cooperation, and deserves further study.

  9. Molecular Detection, Phylogenetic Analysis, and Identification of Transcription Motifs in Feline Leukemia Virus from Naturally Infected Cats in Malaysia

    Directory of Open Access Journals (Sweden)

    Faruku Bande

    2014-01-01

    Full Text Available A nested PCR assay was used to determine the viral RNA and proviral DNA status of naturally infected cats. Selected samples that were FeLV-positive by PCR were subjected to sequencing, phylogenetic analysis, and motifs search. Of the 39 samples that were positive for FeLV p27 antigen, 87.2% (34/39 were confirmed positive with nested PCR. FeLV proviral DNA was detected in 38 (97.3% of p27-antigen negative samples. Malaysian FeLV isolates are found to be highly similar with a homology of 91% to 100%. Phylogenetic analysis revealed that Malaysian FeLV isolates divided into two clusters, with a majority (86.2% sharing similarity with FeLV-K01803 and fewer isolates (13.8% with FeLV-GM1 strain. Different enhancer motifs including NF-GMa, Krox-20/WT1I-del2, BAF1, AP-2, TBP, TFIIF-beta, TRF, and TFIID are found to occur either in single, duplicate, triplicate, or sets of 5 in different positions within the U3-LTR-gag region. The present result confirms the occurrence of FeLV viral RNA and provirus DNA in naturally infected cats. Malaysian FeLV isolates are highly similar, and a majority of them are closely related to a UK isolate. This study provides the first molecular based information on FeLV in Malaysia. Additionally, different enhancer motifs likely associated with FeLV related pathogenesis have been identified.

  10. [Cognitive, emotional and behavioral development of VLBW and ELBW preterm infants: diagnostic and therapeutic follow-up at preschool age].

    Science.gov (United States)

    Pomella, R; Baldino, R; Cravero, B

    2013-12-01

    Aims of the present study ware: to identify in preterm children of 4-6 years of age outcomes concerning cognitive, linguistic, emotional and behavioral development; to develop a therapeutic-rehabilitative project for those children in collaboration with the family and school. The study enrolled 20 children born prematurely at ≤32 weeks of gestational age and/or with a weight ≤1500 g, 12 VLBW (7 male e 5 female), 8 ELBW (4 male e 4 female), hospitalized at Novara Hospital "Maggiore della Carità" during the years 2003 and 2004, without severe outcomes. Psychodiagnostic evaluation was performed with standardized tests. On the final report results were discussed with parents, with specific indications for families and schools. Follow-up was at 6 months. Statistical elaboration of data was performed using Spss (Statistical Package for Social Sciences) version 16. Normal cognitive level resulted from the Griffiths Scale, without significant differences between VLBW and ELBW. The overall lowest score, in the "performance" subscale, especially for ELBW, was correlated with Vineland Scales (low scores in the subscales "everyday skills" and "motor ability"). The highest scores were detected in the "linguistic" subscale of the Griffiths Scales and in the "Communication" subscale of the Vineland Scales. The results at Bus Story Test (narrative language) were lower than average for that age. CBCL and TRF do not demonstrate clinical results in the emotional-behavioural area, but the teachers give a more critical assessment. Difficulties in emotional self-regulation interfere in the test, in the separation from the parents and in socializing. Monitoring development before starting primary school helps to discover potential problems and to activate supportive interventions. Early interventions allow to control and contain academic failure at school, which could have a negative impact on the child's image of himself and on the perception that the parents and school could have.

  11. Heterogeneity of pituitary and plasma prolactin in man: decreased affinity of big prolactin in a radioreceptor assay and evidence for its secretion

    International Nuclear Information System (INIS)

    Garnier, P.E.; Aubert, M.L.; Kaplan, S.L.; Grumbach, M.M.

    1978-01-01

    Molecular heterogeneity of immunoreactive human PRL (IR-hPRL) plasma was assessed by exclusion chromatography in blood from 4 normal adults, 3 newborn infants, 2 late gestational women, 3 patients with primary hypothyroidism and high PRL levels, 2 with functional hyperprolactinemia, 3 with acromegaly, and 10 with PRL-secreting tumors. Three forms of PRL were detected: big-big hPRL, big hPRL, and little hPRL. In normal subjects, the proportion of big-big, big, and little hPRL components was 5.1%, 9.1%, and 85.8%, respectively, without change in the distribution after TRF stimulation. In 8 of 10 patients with PRL-secreting tumors, we detected a significantly higher proportion of big PRL. In 2 additional patients with prolactinomas, the proportion of big PRL was much higher. In 3 of 10 patients, the molecular heterogeneity of the tumor PRL was similar to that in plasma. In 1 acromegalic, there was a very high proportion of big-big hPRL. The PRL fractions were tested in a radioreceptor assay (RRA) using membranes from rabbit mammary gland. Big PRL was much less active than little PRL in the RRA. The fractions were rechromatographed after storage. Big PRL partially distributed as little or big-big PRL, while little PRL remained unchanged. Big-big PRL from tumor extract partially converted into big and little PRL. The big PRL obtained by rechromatography had low activity in the RRA. These observations suggest at least part of the receptor activity of big PRL may arise from generation of or contamination by little PRL. The decreased binding affinity of big PRL in the RRA also indicates that big PRL has little, if any, biological activity. The evidence suggests big PRL is a native PRL dimer linked by intermolecular disulfide bonds which arises in the lactotrope as a postsynthetic product or derivative and is not a true precursor prohormone

  12. Detecting effects of the indicated prevention Programme for Externalizing Problem behaviour (PEP) on child symptoms, parenting, and parental quality of life in a randomized controlled trial.

    Science.gov (United States)

    Hanisch, Charlotte; Freund-Braier, Inez; Hautmann, Christopher; Jänen, Nicola; Plück, Julia; Brix, Gabriele; Eichelberger, Ilka; Döpfner, Manfred

    2010-01-01

    Behavioural parent training is effective in improving child disruptive behavioural problems in preschool children by increasing parenting competence. The indicated Prevention Programme for Externalizing Problem behaviour (PEP) is a group training programme for parents and kindergarten teachers of children aged 3-6 years with externalizing behavioural problems. To evaluate the effects of PEP on child problem behaviour, parenting practices, parent-child interactions, and parental quality of life. Parents and kindergarten teachers of 155 children were randomly assigned to an intervention group (n = 91) and a nontreated control group (n = 64). They rated children's problem behaviour before and after PEP training; parents also reported on their parenting practices and quality of life. Standardized play situations were video-taped and rated for parent-child interactions, e.g. parental warmth. In the intention to treat analysis, mothers of the intervention group described less disruptive child behaviour and better parenting strategies, and showed more parental warmth during a standardized parent-child interaction. Dosage analyses confirmed these results for parents who attended at least five training sessions. Children were also rated to show less behaviour problems by their kindergarten teachers. Training effects were especially positive for parents who attended at least half of the training sessions. CBCL: Child Behaviour Checklist; CII: Coder Impressions Inventory; DASS: Depression anxiety Stress Scale; HSQ: Home-situation Questionnaire; LSS: Life Satisfaction Scale; OBDT: observed behaviour during the test; PCL: Problem Checklist; PEP: prevention programme for externalizing problem behaviour; PPC: Parent Problem Checklist; PPS: Parent Practices Scale; PS: Parenting Scale; PSBC: Problem Setting and Behaviour checklist; QJPS: Questionnaire on Judging Parental Strains; SEFS: Self-Efficacy Scale; SSC: Social Support Scale; TRF: Caregiver-Teacher Report Form.

  13. p38 MAPK and JNK antagonistically control senescence and cytoplasmic p16INK4A expression in doxorubicin-treated endothelial progenitor cells.

    Directory of Open Access Journals (Sweden)

    Paolo Spallarossa

    Full Text Available Patients treated with low-dose anthracyclines often show late onset cardiotoxicity. Recent studies suggest that this form of cardiotoxicity is the result of a progenitor cell disease. In this study we demonstrate that Cord Blood Endothelial Progenitor Cells (EPCs exposed to low, sub-apoptotic doses of doxorubicin show a senescence phenotype characterized by increased SA-b-gal activity, decreased TRF2 and chromosomal abnormalities, enlarged cell shape, and disarrangement of F-actin stress fibers accompanied by impaired migratory ability. P16( INK4A localizes in the cytoplasm of doxorubicin-induced senescent EPCs and not in the nucleus as is the case in EPCs rendered senescent by different stimuli. This localization together with the presence of an arrest in G2, and not at the G1 phase boundary, which is what usually occurs in response to the cell cycle regulatory activity of p16(INK4A, suggests that doxorubicin-induced p16( INK4A does not regulate the cell cycle, even though its increase is closely associated with senescence. The effects of doxorubicin are the result of the activation of MAPKs p38 and JNK which act antagonistically. JNK attenuates the senescence, p16( INK4A expression and cytoskeleton remodeling that are induced by activated p38. We also found that conditioned medium from doxorubicin-induced senescent cardiomyocytes does not attract untreated EPCs, unlike conditioned medium from apoptotic cardiomyocytes which has a strong chemoattractant capacity. In conclusion, this study provides a better understanding of the senescence of doxorubicin-treated EPCs, which may be helpful in preventing and treating late onset cardiotoxicity.

  14. Comparison of intraoperative conditions and postoperative inflammatory response and immune response between patients with common bile duct stones undergoing ESBD and EST

    Directory of Open Access Journals (Sweden)

    Kun Zh

    2016-08-01

    Full Text Available Objective: To analyze the differences in intraoperative conditions and postoperative inflammatory response and immune response levels in patients with common bile duct stones undergoing ESBD and EST treatment. Methods: A total of 565 patients with common bile duct stones who underwent ERCP treatment in our hospital from January 2013 to December 2015 were included in the study, patients’ treatment and testing results were analyzed retrospectively, and then patients were divided into the observation group 300 cases that received ESBD treatment and control group 265 cases that received EST treatment. Differences in intraoperative stress levels as well as postoperative inflammation, nutrition-related index and immune response levels were compared between two groups. Results: Intraoperative N, Cor, C-P and NK cell levels of observation group were lower than those of control group, and FT3 level was higher than that of control group; postoperative inflammation-related factors such as IL-6, CRP, PCT, CA19-9 and CEA levels were lower than those of control group, protein nutrition indexes such as TP, Alb, PA, TRF and RBP values were higher than those of control group, and immune indexes such as CD4+, CD4+/CD8+, IgA, IgM and IgG levels were higher than those of control group while CD8+ level was lower than that of control group. Conclusion: ESBD for the treatment of patients with common bile duct stones is better than EST treatment in reducing surgical stress, optimizing postoperative physical status and other aspects.

  15. Telomere dynamics and homeostasis in a transmissible cancer.

    Science.gov (United States)

    Ujvari, Beata; Pearse, Anne-Maree; Taylor, Robyn; Pyecroft, Stephen; Flanagan, Cassandra; Gombert, Sara; Papenfuss, Anthony T; Madsen, Thomas; Belov, Katherine

    2012-01-01

    Devil Facial Tumour Disease (DFTD) is a unique clonal cancer that threatens the world's largest carnivorous marsupial, the Tasmanian devil (Sarcophilus harrisii) with extinction. This transmissible cancer is passed between individual devils by cell implantation during social interactions. The tumour arose in a Schwann cell of a single devil over 15 years ago and since then has expanded clonally, without showing signs of replicative senescence; in stark contrast to a somatic cell that displays a finite capacity for replication, known as the "Hayflick limit". In the present study we investigate the role of telomere length, measured as Telomere Copy Number (TCN), and telomerase and shelterin gene expression, as well as telomerase activity in maintaining hyperproliferation of Devil Facial Tumour (DFT) cells. Our results show that DFT cells have short telomeres. DFTD TCN does not differ between geographic regions or between strains. However, TCN has increased over time. Unlimited cell proliferation is likely to have been achieved through the observed up-regulation of the catalytic subunit of telomerase (TERT) and concomitant activation of telomerase. Up-regulation of the central component of shelterin, the TRF1-intercating nuclear factor 2 (TINF2) provides DFT a mechanism for telomere length homeostasis. The higher expression of both TERT and TINF2 may also protect DFT cells from genomic instability and enhance tumour proliferation. DFT cells appear to monitor and regulate the length of individual telomeres: i.e. shorter telomeres are elongated by up-regulation of telomerase-related genes; longer telomeres are protected from further elongation by members of the shelterin complex, which may explain the lack of spatial and strain variation in DFT telomere copy number. The observed longitudinal increase in gene expression in DFT tissue samples and telomerase activity in DFT cell lines might indicate a selection for more stable tumours with higher proliferative potential.

  16. Telomere dynamics and homeostasis in a transmissible cancer.

    Directory of Open Access Journals (Sweden)

    Beata Ujvari

    Full Text Available Devil Facial Tumour Disease (DFTD is a unique clonal cancer that threatens the world's largest carnivorous marsupial, the Tasmanian devil (Sarcophilus harrisii with extinction. This transmissible cancer is passed between individual devils by cell implantation during social interactions. The tumour arose in a Schwann cell of a single devil over 15 years ago and since then has expanded clonally, without showing signs of replicative senescence; in stark contrast to a somatic cell that displays a finite capacity for replication, known as the "Hayflick limit".In the present study we investigate the role of telomere length, measured as Telomere Copy Number (TCN, and telomerase and shelterin gene expression, as well as telomerase activity in maintaining hyperproliferation of Devil Facial Tumour (DFT cells. Our results show that DFT cells have short telomeres. DFTD TCN does not differ between geographic regions or between strains. However, TCN has increased over time. Unlimited cell proliferation is likely to have been achieved through the observed up-regulation of the catalytic subunit of telomerase (TERT and concomitant activation of telomerase. Up-regulation of the central component of shelterin, the TRF1-intercating nuclear factor 2 (TINF2 provides DFT a mechanism for telomere length homeostasis. The higher expression of both TERT and TINF2 may also protect DFT cells from genomic instability and enhance tumour proliferation.DFT cells appear to monitor and regulate the length of individual telomeres: i.e. shorter telomeres are elongated by up-regulation of telomerase-related genes; longer telomeres are protected from further elongation by members of the shelterin complex, which may explain the lack of spatial and strain variation in DFT telomere copy number. The observed longitudinal increase in gene expression in DFT tissue samples and telomerase activity in DFT cell lines might indicate a selection for more stable tumours with higher proliferative

  17. Applications of immunomagnetic capture and time-resolved fluorescence to detect outbreak Escherichia coli O157 and Salmonella in alfalfa sprouts

    Science.gov (United States)

    Tu, Shu-I.; Gordon, Marsha; Fett, William F.; Gehring, Andrew G.; Irwin, Peter L.

    2004-03-01

    Commercially available alfalfa seeds were inoculated with low levels (~ 4 CFU/g) of pathogenic bacteria. The inoculated seeds were then allowed to sprout in sterile tap water at 22°C. After 48 hours, the irrigation water and sprouts were separately transferred to bovine heart infusion (BHI) media. The microbes in the BHI samples were allowed to grow for 4 hours at 37°C and 160 rpm. Specific immunomagnetic beads (IMB) were then applied to capture the E.coli O157 and/or Salmonella in the growth media. Separation and concentration of IMB-captured pathogens were achieved using magnetic separators. The captured E. coli O157:H7 and Salmonella spp were further tagged with europium (Eu) labeled anti-E. coli O157 antibodies and samarium (Sm) labeled anti-Salmonella antibodies, respectively. After washing, the lanthanide labels were extracted out from the complexes by specific chelators to form strongly fluorescent chelates. The specific time-resolved fluorescence (TRF) associated with Eu or Sm was measured to estimate the extent of capture of the E. coli O157 and Salmonella, respectively. The results indicated that the approach could detect E. coli O157 and Salmonella enterica from the seeds inoculated with ~ 4 CFU/g of the pathogens. Non-targeted bacteria, e.g., Aeromonas and Citrobacter exhibited essentially no cross reactivity. Since the pathogen detection from the sprouts was achieved within 6 hours, the developed methodology could be use as a rapid, sensitive and specific screening process for E. coli O157 and Salmonella enterica in this popular salad food.

  18. Photophysical characterization and time-resolved spectroscopy of a anthradithiophene dimer: exploring the role of conformation in singlet fission

    KAUST Repository

    Dean, Jacob C.

    2017-08-18

    Quantitative singlet fission has been observed for a variety of acene derivatives such as tetracene and pentacene, and efforts to extend the library of singlet fission compounds is of current interest. Preliminary calculations suggest anthradithiophenes exhibit significant exothermicity between the first optically-allowed singlet state, S1, and 2 × T1 with an energy difference of >5000 cm−1. Given the fulfillment of this ingredient for singlet fission, here we investigate the singlet fission capability of a difluorinated anthradithiophene dimer (2ADT) covalently linked by a (dimethylsilyl)ethane bridge and derivatized by triisobutylsilylethynyl (TIBS) groups. Photophysical characterization of 2ADT and the single functionalized ADT monomer were carried out in toluene and acetone solution via absorption and fluorescence spectroscopy, and their photo-initiated dynamics were investigated with time-resolved fluorescence (TRF) and transient absorption (TA) spectroscopy. In accordance with computational predictions, two conformers of 2ADT were observed via fluorescence spectroscopy and were assigned to structures with the ADT cores trans or cis to one another about the covalent bridge. The two conformers exhibited markedly different excited state deactivation mechanisms, with the minor trans population being representative of the ADT monomer showing primarily radiative decay, while the dominant cis population underwent relaxation into an excimer geometry before internally converting to the ground state. The excimer formation kinetics were found to be solvent dependent, yielding time constants of ∼1.75 ns in toluene, and ∼600 ps in acetone. While the difference in rates elicits a role for the solvent in stabilizing the excimer structure, the rate is still decidedly long compared to most singlet fission rates of analogous dimers, suggesting that the excimer is neither a kinetic nor a thermodynamic trap, yet singlet fission was still not observed. The result

  19. ITRF2014 Evaluation with ILRS Data and Products

    Science.gov (United States)

    Pavlis, E. C.; Luceri, V.; Kuzmicz-Cieslak, M.; König, D.; Bianco, G.

    2015-12-01

    The development and release of the new realization of the International Terrestrial Reference Frame—ITRF2014 requires elaborate testing to ensure the quality of the final product. The evaluation effort ensures that the ITRF is of the indicated quality by its error estimates and the combination has not compromised the contributing techniques' input. The International Laser Ranging Service (ILRS) contributes unique information that only Satellite Laser Ranging—SLR is sensitive to: the definition of the origin, and in equal parts with VLBI, the scale of the model. The ILRS analysts adopted a revision of the internal standards and procedures in developing our contribution to ITRF2014 from our eight Analysis Centers. Anticipating the release of ITRF2014 we worked on designing and executing tests using data and products unique to ILRS. In addition to the data contributed to ITRF2014, ILRS has several other targets, in lower and higher orbits, the SLR tracking data of which are used as independent data for the evaluation process. Since SLR data are primarily sensitive to the origin and scale definition of the TRF model, these model attributes are the best to be validated using SLR data. LAGEOS and ETALON data collected outside the span of data used in ITRF2014 can also evaluate the quality of the estimated velocity vectors. The use of independent SLR data evaluates the model throughout the period that such data are available. SLR data from low altitude missions can validate the performance of the model from the late '70s all the way to present (using e.g. STARLETTE and LARES data). This presentation will give an overview of the new model's evaluation using exclusively ILRS tracking data and other ILRS products.

  20. Estimating fuel octane numbers from homogeneous gas-phase ignition delay times

    KAUST Repository

    Naser, Nimal; Sarathy, Mani; Chung, Suk-Ho

    2017-01-01

    Fuel octane numbers are directly related to the autoignition properties of fuel/air mixtures in spark ignition (SI) engines. This work presents a methodology to estimate the research and the motor octane numbers (RON and MON) from homogeneous gas-phase ignition delay time (IDT) data calculated at various pressures and temperatures. The hypothesis under investigation is that at specific conditions of pressure and temperature (i.e., RON-like and MON-like conditions), fuels with IDT identical to that of a primary reference fuel (PRF) have the same octane rating. To test this hypothesis, IDTs with a detailed gasoline surrogate chemical kinetic model have been calculated at various temperatures and pressures. From this dataset, temperatures that best represent RON and MON have been correlated at a specified pressure. Correlations for pressures in the range of 10–50 bar were obtained. The proposed correlations were validated with toluene reference fuels (TRF), toluene primary reference fuels (TPRF), ethanol reference fuels (ERF), PRFs and TPRFs with ethanol, and multi-component gasoline surrogate mixtures. The predicted RON and MON showed satisfactory accuracy against measurements obtained by the standard ASTM methods and blending rules, demonstrating that the present methodology can be a viable tool for a first approximation. The correlations were also validated against an extensive set of experimental IDT data obtained from literature with a high degree of accuracy in RON/MON prediction. Conditions in homogeneous reactors such as shock tubes and rapid compression machines that are relevant to modern SI engines were also identified. Uncertainty analysis of the proposed correlations with linear error propagation theory is also presented.

  1. An Improved Empirical Harmonic Model of the Celestial Intermediate Pole Offsets from a Global VLBI Solution

    Science.gov (United States)

    Belda, Santiago; Heinkelmann, Robert; Ferrándiz, José M.; Karbon, Maria; Nilsson, Tobias; Schuh, Harald

    2017-10-01

    Very Long Baseline Interferometry (VLBI) is the only space geodetic technique capable of measuring all the Earth orientation parameters (EOP) accurately and simultaneously. Modeling the Earth's rotational motion in space within the stringent consistency goals of the Global Geodetic Observing System (GGOS) makes VLBI observations essential for constraining the rotation theories. However, the inaccuracy of early VLBI data and the outdated products could cause non-compliance with these goals. In this paper, we perform a global VLBI analysis of sessions with different processing settings to determine a new set of empirical corrections to the precession offsets and rates, and to the amplitudes of a wide set of terms included in the IAU 2006/2000A precession-nutation theory. We discuss the results in terms of consistency, systematic errors, and physics of the Earth. We find that the largest improvements w.r.t. the values from IAU 2006/2000A precession-nutation theory are associated with the longest periods (e.g., 18.6-yr nutation). A statistical analysis of the residuals shows that the provided corrections attain an error reduction at the level of 15 μas. Additionally, including a Free Core Nutation (FCN) model into a priori Celestial Pole Offsets (CPOs) provides the lowest Weighted Root Mean Square (WRMS) of residuals. We show that the CPO estimates are quite insensitive to TRF choice, but slightly sensitive to the a priori EOP and the inclusion of different VLBI sessions. Finally, the remaining residuals reveal two apparent retrograde signals with periods of nearly 2069 and 1034 days.

  2. Molecular interaction of 2,4-diacetylphloroglucinol (DAPG) with human serum albumin (HSA): The spectroscopic, calorimetric and computational investigation

    Science.gov (United States)

    Pragna Lakshmi, T.; Mondal, Moumita; Ramadas, Krishna; Natarajan, Sakthivel

    2017-08-01

    Drug molecule interaction with human serum albumin (HSA) affects the distribution and elimination of the drug. The compound, 2,4-diacetylphloroglucinol (DAPG) has been known for its antimicrobial, antiviral, antihelminthic and anticancer properties. However, its interaction with HSA is not yet reported. In this study, the interaction between HSA and DAPG was investigated through steady-state fluorescence, time-resolved fluorescence (TRF), circular dichroism (CD), Fourier transform infrared (FT-IR) spectroscopy, isothermal titration calorimetry (ITC), molecular docking and molecular dynamics simulation (MDS). Fluorescence spectroscopy results showed the strong quenching of intrinsic fluorescence of HSA due to interaction with DAPG, through dynamic quenching mechanism. The compound bound to HSA with reversible and moderate affinity which explained its easy diffusion from circulatory system to target tissue. The thermodynamic parameters from fluorescence spectroscopic data clearly revealed the contribution of hydrophobic forces but, the role of hydrogen bonds was not negligible according to the ITC studies. The interaction was exothermic and spontaneous in nature. Binding with DAPG reduced the helical content of protein suggesting the unfolding of HSA. Site marker fluorescence experiments revealed the change in binding constant of DAPG in the presence of site I (warfarin) but not site II marker (ibuprofen) which confirmed that the DAPG bound to site I. ITC experiments also supported this as site I marker could not bind to HSA-DAPG complex while site II marker was accommodated in the complex. In silico studies further showed the lowest binding affinity and more stability of DAPG in site I than in site II. Thus the data presented in this study confirms the binding of DAPG to the site I of HSA which may help in further understanding of pharmacokinetic properties of DAPG.

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

    Directory of Open Access Journals (Sweden)

    So Yeon Kwon

    2016-04-01

    Full Text Available 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.

  4. P. berghei telomerase subunit TERT is essential for parasite survival.

    Directory of Open Access Journals (Sweden)

    Agnieszka A Religa

    Full Text Available Telomeres define the ends of chromosomes protecting eukaryotic cells from chromosome instability and eventual cell death. The complex regulation of telomeres involves various proteins including telomerase, which is a specialized ribonucleoprotein responsible for telomere maintenance. Telomeres of chromosomes of malaria parasites are kept at a constant length during blood stage proliferation. The 7-bp telomere repeat sequence is universal across different Plasmodium species (GGGTTT/CA, though the average telomere length varies. The catalytic subunit of telomerase, telomerase reverse transcriptase (TERT, is present in all sequenced Plasmodium species and is approximately three times larger than other eukaryotic TERTs. The Plasmodium RNA component of TERT has recently been identified in silico. A strategy to delete the gene encoding TERT via double cross-over (DXO homologous recombination was undertaken to study the telomerase function in P. berghei. Expression of both TERT and the RNA component (TR in P. berghei blood stages was analysed by Western blotting and Northern analysis. Average telomere length was measured in several Plasmodium species using Telomere Restriction Fragment (TRF analysis. TERT and TR were detected in blood stages and an average telomere length of ∼ 950 bp established. Deletion of the tert gene was performed using standard transfection methodologies and we show the presence of tert- mutants in the transfected parasite populations. Cloning of tert- mutants has been attempted multiple times without success. Thorough analysis of the transfected parasite populations and the parasite obtained from extensive parasite cloning from these populations provide evidence for a so called delayed death phenotype as observed in different organisms lacking TERT. The findings indicate that TERT is essential for P. berghei cell survival. The study extends our current knowledge on telomere biology in malaria parasites and validates further

  5. Laminar Burning Velocities of Fuels for Advanced Combustion Engines (FACE) Gasoline and Gasoline Surrogates with and without Ethanol Blending Associated with Octane Rating

    KAUST Repository

    Mannaa, Ossama; Mansour, Morkous S.; Roberts, William L.; Chung, Suk-Ho

    2016-01-01

    Laminar burning velocities of fuels for advanced combustion engines (FACE) C gasoline and of several blends of surrogate toluene reference fuels (TRFs) (n-heptane, iso-octane, and toluene mixtures) of the same research octane number are presented. Effects of ethanol addition on laminar flame speed of FACE-C and its surrogate are addressed. Measurements were conducted using a constant volume spherical combustion vessel in the constant pressure, stable flame regime at an initial temperature of 358 K and initial pressures up to 0.6 MPa with the equivalence ratios ranging from 0.8 to 1.6. Comparable values in the laminar burning velocities were measured for the FACE-C gasoline and the proposed surrogate fuel (17.60% n-heptane + 77.40% iso-octane + 5% toluene) over the range of experimental conditions. Sensitivity of flame propagation to total stretch rate effects and thermo-diffusive instability was quantified by determining Markstein length. Two percentages of an oxygenated fuel of ethanol as an additive, namely, 60 vol% and 85 vol% were investigated. The addition of ethanol to FACE-C and its surrogate TRF-1 (17.60% n-heptane + 77.40% iso-octane + 5% toluene) resulted in a relatively similar increase in the laminar burning velocities. The high-pressure measured values of Markstein length for the studied fuels blended with ethanol showed minimal influence of ethanol addition on the flame’s response to stretch rate and thermo-diffusive instability. © 2016 Taylor & Francis.

  6. A three-component gene expression system and its application for inducible flavonoid overproduction in transgenic Arabidopsis thaliana.

    Science.gov (United States)

    Feng, Yue; Cao, Cong-Mei; Vikram, Meenu; Park, Sunghun; Kim, Hye Jin; Hong, Jong Chan; Cisneros-Zevallos, Luis; Koiwa, Hisashi

    2011-03-08

    Inducible gene expression is a powerful tool to study and engineer genes whose overexpression could be detrimental for the host organisms. However, only limited systems have been adopted in plant biotechnology. We have developed an osmotically inducible system using three components of plant origin, RD29a (Responsive to Dehydration 29A) promoter, CBF3 (C-repeat Binding Factor 3) transcription factor and cpl1-2 (CTD phosphatase-like 1) mutation. The osmotic stress responsible RD29a promoter contains the CBF3 binding sites and thus RD29A-CBF3 feedforward cassette enhances induction of RD29a promoter under stress. The cpl1-2 mutation in a host repressor CPL1 promotes stress responsible RD29a promoter expression. The efficacy of this system was tested using PAP1 (Production of Anthocyanin Pigment 1) transgene, a model transcription factor that regulates the anthocyanin pathway in Arabidopsis. While transgenic plants with only one or two of three components did not reproducibly accumulate anthocyanin pigments above the control level, transgenic cpl1 plants containing homozygous RD29a-PAP1 and RD29a-CBF3 transgenes produced 30-fold higher level of total anthocyanins than control plants upon cold treatment. Growth retardation and phytochemical production of transgenic plants were minimum under normal conditions. The flavonoid profile in cold-induced transgenic plants was determined by LC/MS/MS, which resembled that of previously reported pap1-D plants but enriched for kaempferol derivatives. These results establish the functionality of the inducible three-component gene expression system in plant metabolic engineering. Furthermore, we show that PAP1 and environmental signals synergistically regulate the flavonoid pathway to produce a unique flavonoid blend that has not been produced by PAP1 overexpression or cold treatment alone.

  7. A three-component gene expression system and its application for inducible flavonoid overproduction in transgenic Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Yue Feng

    Full Text Available Inducible gene expression is a powerful tool to study and engineer genes whose overexpression could be detrimental for the host organisms. However, only limited systems have been adopted in plant biotechnology. We have developed an osmotically inducible system using three components of plant origin, RD29a (Responsive to Dehydration 29A promoter, CBF3 (C-repeat Binding Factor 3 transcription factor and cpl1-2 (CTD phosphatase-like 1 mutation. The osmotic stress responsible RD29a promoter contains the CBF3 binding sites and thus RD29A-CBF3 feedforward cassette enhances induction of RD29a promoter under stress. The cpl1-2 mutation in a host repressor CPL1 promotes stress responsible RD29a promoter expression. The efficacy of this system was tested using PAP1 (Production of Anthocyanin Pigment 1 transgene, a model transcription factor that regulates the anthocyanin pathway in Arabidopsis. While transgenic plants with only one or two of three components did not reproducibly accumulate anthocyanin pigments above the control level, transgenic cpl1 plants containing homozygous RD29a-PAP1 and RD29a-CBF3 transgenes produced 30-fold higher level of total anthocyanins than control plants upon cold treatment. Growth retardation and phytochemical production of transgenic plants were minimum under normal conditions. The flavonoid profile in cold-induced transgenic plants was determined by LC/MS/MS, which resembled that of previously reported pap1-D plants but enriched for kaempferol derivatives. These results establish the functionality of the inducible three-component gene expression system in plant metabolic engineering. Furthermore, we show that PAP1 and environmental signals synergistically regulate the flavonoid pathway to produce a unique flavonoid blend that has not been produced by PAP1 overexpression or cold treatment alone.

  8. Antioxidant therapy attenuates myocardial telomerase activity reduction in superoxide dismutase-deficient mice.

    Science.gov (United States)

    Makino, Naoki; Maeda, Toyoki; Oyama, Jun-ichi; Sasaki, Makoto; Higuchi, Yoshihiro; Mimori, Koji; Shimizu, Takahiko

    2011-04-01

    Oxidative stress plays a pathological role in the development of heart failure. This study examined telomere biology in heart/muscle-specific manganese superoxide dismutase-deficient mice (H/M-SOD2(-/-)), which develop progressive congestive heart failure and exhibit pathology typical of dilated cardiomyopathy. EUK-8 (25mg/kg/day), a superoxide dismutase and catalase mimetic, was administered to H/M-SOD2(-/-) mice for four weeks beginning at 8 weeks of age. Telomere length, telomerase activity, telomere-associated proteins, and cell death signals were assessed in hearts from control wild-type mice (H/M-Sod2 (lox/ lox)) and H/M-SOD2(-/-) mice either treated or untreated with EUK-8. While cardiac function was unchanged in these experimental mice, the end-diastolic dimension in H/M-SOD2(-/-) mice was notably dilated and could be significantly reduced by EUK-8 treatment. At the end of the study, no shortening of telomere length was observed in heart tissues from all mice tested, but telomerase activity was decreased in heart tissue from H/M-SOD2(-/-) mice compared to control mice. Protein expression for telomerase reverse transcriptase and telomere repeat binding factor 2 was also downregulated in H/M-SOD2(-/-) heart tissue as was expression of phospho-Akt, insulin-like growth factor, and endothelial nitric oxide synthase. Expression levels of Sirt1, a lifespan modulator, were enhanced while FoxO3a was depressed in H/M-SOD2(-/-) hearts. All of the changes seen in H/M-SOD2(-/-) heart tissue could be inhibited by EUK-8 treatment. Taken together, the results suggest that oxidant stress might affect myocardial telomerase activity and telomere-associated proteins. Telomerase may therefore play a pivotal role in antioxidant defense mechanisms, and may be useful as a novel therapeutic tool for treating human heart failure. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Targeted Genome Regulation and Editing in Plants

    KAUST Repository

    Piatek, Agnieszka

    2016-03-01

    The ability to precisely regulate gene expression patterns and to modify genome sequence in a site-specific manner holds much promise in determining gene function and linking genotype to phenotype. DNA-binding modules have been harnessed to generate customizable and programmable chimeric proteins capable of binding to site-specific DNA sequences and regulating the genome and epigenome. Modular DNA-binding domains from zinc fingers (ZFs) and transcriptional activator-like effectors (TALEs) are amenable to engineering to bind any DNA target sequence of interest. Deciphering the code of TALE repeat binding to DNA has helped to engineer customizable TALE proteins capable of binding to any sequence of interest. Therefore TALE repeats provide a rich resource for bioengineering applications. However, the TALE system is limited by the requirement to re-engineer one or two proteins for each new target sequence. Recently, the clustered regularly interspaced palindromic repeats (CRISPR)/ CRISPR associated 9 (Cas9) has been used as a versatile genome editing tool. This machinery has been also repurposed for targeted transcriptional regulation. Due to the facile engineering, simplicity and precision, the CRISPR/Cas9 system is poised to revolutionize the functional genomics studies across diverse eukaryotic species. In this dissertation I employed transcription activator-like effectors and CRISPR/Cas9 systems for targeted genome regulation and editing and my achievements include: 1) I deciphered and extended the DNA-binding code of Ralstonia TAL effectors providing new opportunities for bioengineering of customizable proteins; 2) I repurposed the CRISPR/Cas9 system for site-specific regulation of genes in plant genome; 3) I harnessed the power of CRISPR/Cas9 gene editing tool to study the function of the serine/arginine-rich (SR) proteins.

  10. A comparison of alternative 60-mer probe designs in an in-situ synthesized oligonucleotide microarray

    Directory of Open Access Journals (Sweden)

    Fairbanks Benjamin D

    2006-04-01

    Full Text Available Abstract Background DNA microarrays have proven powerful for functional genomics studies. Several technologies exist for the generation of whole-genome arrays. It is well documented that 25mer probes directed against different regions of the same gene produce variable signal intensity values. However, the extent to which this is true for probes of greater length (60mers is not well characterized. Moreover, this information has not previously been reported for whole-genome arrays designed against bacteria, whose genomes may differ substantially in characteristics directly affecting microarray performance. Results We report here an analysis of alternative 60mer probe designs for an in-situ synthesized oligonucleotide array for the GC rich, β-proteobacterium Burkholderia cenocepacia. Probes were designed using the ArrayOligoSel3.5 software package and whole-genome microarrays synthesized by Agilent, Inc. using their in-situ, ink-jet technology platform. We first validated the quality of the microarrays as demonstrated by an average signal to noise ratio of >1000. Next, we determined that the variance of replicate probes (1178 total probes examined of identical sequence was 3.8% whereas the variance of alternative probes (558 total alternative probes examined designs was 9.5%. We determined that depending upon the definition, about 2.4% of replicate and 7.8% of alternative probes produced outlier conclusions. Finally, we determined none of the probe design subscores (GC content, internal repeat, binding energy and self annealment produced by ArrayOligoSel3.5 were predictive or probes that produced outlier signals. Conclusion Our analysis demonstrated that the use of multiple probes per target sequence is not essential for in-situ synthesized 60mer oligonucleotide arrays designed against bacteria. Although probes producing outlier signals were identified, the use of ratios results in less than 10% of such outlier conclusions. We also determined that

  11. Activities and specificities of homodimeric TALENs in Saccharomyces cerevisiae

    KAUST Repository

    Aouida, Mustapha; Piatek, Marek J.; Bangarusamy, Dhinoth Kumar; Mahfouz, Magdy M.

    2013-01-01

    The development of highly efficient genome engineering reagents is of paramount importance to launch the next wave of biotechnology. TAL effectors have been developed as an adaptable DNA binding scaffold that can be engineered to bind to any user-defined sequence. Thus, TAL-based DNA binding modules have been used to generate chimeric proteins for a variety of targeted genome modifications across eukaryotic species. For example, TAL effectors fused to the catalytic domain of FokI endonuclease (TALENs) were used to generate site-specific double strand breaks (DSBs), the repair of which can be harnessed to dictate user-desired, genome-editing outcomes. To cleave DNA, FokI endonuclease must dimerize which can be achieved using a pair of TALENs that bind to the DNA targeted in a tail-to-tail orientation with proper spacing allowing the dimer formation. Because TALENs binding to DNA are dependent on their repeat sequences and nucleotides binding specificities, homodimers and heterodimers binding can be formed. In the present study, we used several TALEN monomers with increased repeats binding degeneracy to allow homodimer formation at increased number of genomic loci. We assessed their binding specificities and genome modification activities. Our results indicate that homodimeric TALENs could be used to modify the yeast genome in a site-specific manner and their binding to the promoter regions might modulate the expression of target genes. Taken together, our data indicate that homodimeric TALENs could be used to achieve different engineering possibilities of biotechnological applications and that their transcriptional modulations need to be considered when analyzing their phenotypic effects. © 2013 Springer-Verlag.

  12. Copy number and haplotype variation at the VRN-A1 and central FR-A2 loci are associated with frost tolerance in hexaploid wheat.

    Science.gov (United States)

    Zhu, Jie; Pearce, Stephen; Burke, Adrienne; See, Deven Robert; Skinner, Daniel Z; Dubcovsky, Jorge; Garland-Campbell, Kimberly

    2014-05-01

    The interaction between VRN - A1 and FR - A2 largely affect the frost tolerance of hexaploid wheat. Frost tolerance is critical for wheat survival during cold winters. Natural variation for this trait is mainly associated with allelic differences at the VERNALIZATION 1 (VRN1) and FROST RESISTANCE 2 (FR2) loci. VRN1 regulates the transition between vegetative and reproductive stages and FR2, a locus including several tandemly duplicated C-REPEAT BINDING FACTOR (CBF) transcription factors, regulates the expression of Cold-regulated genes. We identified sequence and copy number variation at these two loci among winter and spring wheat varieties and characterized their association with frost tolerance. We identified two FR-A2 haplotypes-'FR-A2-S' and 'FR-A2-T'-distinguished by two insertion/deletions and ten single nucleotide polymorphisms within the CBF-A12 and CBF-A15 genes. Increased copy number of CBF-A14 was frequently associated with the FR-A2-T haplotype and with higher CBF14 transcript levels in response to cold. Factorial ANOVAs revealed significant interactions between VRN1 and FR-A2 for frost tolerance in both winter and spring panels suggesting a crosstalk between vernalization and cold acclimation pathways. The model including these two loci and their interaction explained 32.0 and 20.7 % of the variation in frost tolerance in the winter and spring panels, respectively. The interaction was validated in a winter wheat F 4:5 population segregating for both genes. Increased VRN-A1 copy number was associated with improved frost tolerance among varieties carrying the FR-A2-T allele but not among those carrying the FR-A2-S allele. These results suggest that selection of varieties carrying the FR-A2-T allele and three copies of the recessive vrn-A1 allele would be a good strategy to improve frost tolerance in wheat.

  13. TMPyP4 porphyrin distorts RNA G-quadruplex structures of the disease-associated r(GGGGCC)n repeat of the C9orf72 gene and blocks interaction of RNA-binding proteins.

    Science.gov (United States)

    Zamiri, Bita; Reddy, Kaalak; Macgregor, Robert B; Pearson, Christopher E

    2014-02-21

    Certain DNA and RNA sequences can form G-quadruplexes, which can affect genetic instability, promoter activity, RNA splicing, RNA stability, and neurite mRNA localization. Amyotrophic lateral sclerosis and frontotemporal dementia can be caused by expansion of a (GGGGCC)n repeat in the C9orf72 gene. Mutant r(GGGGCC)n- and r(GGCCCC)n-containing transcripts aggregate in nuclear foci, possibly sequestering repeat-binding proteins such as ASF/SF2 and hnRNPA1, suggesting a toxic RNA pathogenesis, as occurs in myotonic dystrophy. Furthermore, the C9orf72 repeat RNA was recently demonstrated to undergo the noncanonical repeat-associated non-AUG translation (RAN translation) into pathologic dipeptide repeats in patient brains, a process that is thought to depend upon RNA structure. We previously demonstrated that the r(GGGGCC)n RNA forms repeat tract length-dependent G-quadruplex structures that bind the ASF/SF2 protein. Here we show that the cationic porphyrin (5,10,15,20-tetra(N-methyl-4-pyridyl) porphyrin (TMPyP4)), which can bind some G-quadruplex-forming sequences, can bind and distort the G-quadruplex formed by r(GGGGCC)8, and this ablates the interaction of either hnRNPA1 or ASF/SF2 with the repeat. These findings provide proof of concept that nucleic acid binding small molecules, such as TMPyP4, can distort the secondary structure of the C9orf72 repeat, which may beneficially disrupt protein interactions, which may ablate either protein sequestration and/or RAN translation into potentially toxic dipeptides. Disruption of secondary structure formation of the C9orf72 RNA repeats may be a viable therapeutic avenue, as well as a means to test the role of RNA structure upon RAN translation.

  14. Constitutive expression of DaCBF7, an Antarctic vascular plant Deschampsia antarctica CBF homolog, resulted in improved cold tolerance in transgenic rice plants.

    Science.gov (United States)

    Byun, Mi Young; Lee, Jungeun; Cui, Li Hua; Kang, Yoonjee; Oh, Tae Kyung; Park, Hyun; Lee, Hyoungseok; Kim, Woo Taek

    2015-07-01

    Deschampsia antarctica is an Antarctic hairgrass that grows on the west coast of the Antarctic peninsula. In this report, we have identified and characterized a transcription factor, D. antarctica C-repeat binding factor 7 (DaCBF7), that is a member of the monocot group V CBF homologs. The protein contains a single AP2 domain, a putative nuclear localization signal, and the typical CBF signature. DaCBF7, like other monocot group V homologs, contains a distinct polypeptide stretch composed of 43 amino acids in front of the AP2 motif. DaCBF7 was predominantly localized to nuclei and interacted with the C-repeat/dehydration responsive element (CRT/DRE) core sequence (ACCGAC) in vitro. DaCBF7 was induced by abiotic stresses, including drought, cold, and salinity. To investigate its possible cellular role in cold tolerance, a transgenic rice system was employed. DaCBF7-overexpressing transgenic rice plants (Ubi:DaCBF7) exhibited markedly increased tolerance to cold stress compared to wild-type plants without growth defects; however, overexpression of DaCBF7 exerted little effect on tolerance to drought or salt stress. Transcriptome analysis of a Ubi:DaCBF7 transgenic line revealed 13 genes that were up-regulated in DaCBF7-overexpressing plants compared to wild-type plants in the absence of cold stress and in short- or long-term cold stress. Five of these genes, dehydrin, remorin, Os03g63870, Os11g34790, and Os10g22630, contained putative CRT/DRE or low-temperature responsive elements in their promoter regions. These results suggest that overexpression of DaCBF7 directly and indirectly induces diverse genes in transgenic rice plants and confers enhanced tolerance to cold stress. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  15. Supra-optimal expression of the cold-regulated OsMyb4 transcription factor in transgenic rice changes the complexity of transcriptional network with major effects on stress tolerance and panicle development

    KAUST Repository

    Park, Myoungryoul

    2010-09-28

    The R2R3-type OsMyb4 transcription factor of rice has been shown to play a role in the regulation of osmotic adjustment in heterologous overexpression studies. However, the exact composition and organization of its underlying transcriptional network has not been established to be a robust tool for stress tolerance enhancement by regulon engineering. OsMyb4 network was dissected based on commonalities between the global chilling stress transcriptome and the transcriptome configured by OsMyb4 overexpression. OsMyb4 controls a hierarchical network comprised of several regulatory sub-clusters associated with cellular defense and rescue, metabolism and development. It regulates target genes either directly or indirectly through intermediary MYB, ERF, bZIP, NAC, ARF and CCAAT-HAP transcription factors. Regulatory sub-clusters have different combinations of MYB-like, GCC-box-like, ERD1-box-like, ABRE-like, G-box-like, as1/ocs/TGA-like, AuxRE-like, gibberellic acid response element (GARE)-like and JAre-like cis-elements. Cold-dependent network activity enhanced cellular antioxidant capacity through radical scavenging mechanisms and increased activities of phenylpropanoid and isoprenoid metabolic processes involving various abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), ethylene and reactive oxygen species (ROS) responsive genes. OsMyb4 network is independent of drought response element binding protein/C-repeat binding factor (DREB/CBF) and its sub-regulons operate with possible co-regulators including nuclear factor-Y. Because of its upstream position in the network hierarchy, OsMyb4 functions quantitatively and pleiotrophically. Supra-optimal expression causes misexpression of alternative targets with costly trade-offs to panicle development. © 2010 Blackwell Publishing Ltd.

  16. Multi-dimensional virtual system introduced to enhance canonical sampling

    Science.gov (United States)

    Higo, Junichi; Kasahara, Kota; Nakamura, Haruki

    2017-10-01

    When an important process of a molecular system occurs via a combination of two or more rare events, which occur almost independently to one another, computational sampling for the important process is difficult. Here, to sample such a process effectively, we developed a new method, named the "multi-dimensional Virtual-system coupled Monte Carlo (multi-dimensional-VcMC)" method, where the system interacts with a virtual system expressed by two or more virtual coordinates. Each virtual coordinate controls sampling along a reaction coordinate. By setting multiple reaction coordinates to be related to the corresponding rare events, sampling of the important process can be enhanced. An advantage of multi-dimensional-VcMC is its simplicity: Namely, the conformation moves widely in the multi-dimensional reaction coordinate space without knowledge of canonical distribution functions of the system. To examine the effectiveness of the algorithm, we introduced a toy model where two molecules (receptor and its ligand) bind and unbind to each other. The receptor has a deep binding pocket, to which the ligand enters for binding. Furthermore, a gate is set at the entrance of the pocket, and the gate is usually closed. Thus, the molecular binding takes place via the two events: ligand approach to the pocket and gate opening. In two-dimensional (2D)-VcMC, the two molecules exhibited repeated binding and unbinding, and an equilibrated distribution was obtained as expected. A conventional canonical simulation, which was 200 times longer than 2D-VcMC, failed in sampling the binding/unbinding effectively. The current method is applicable to various biological systems.

  17. Activities and specificities of homodimeric TALENs in Saccharomyces cerevisiae

    KAUST Repository

    Aouida, Mustapha

    2013-10-01

    The development of highly efficient genome engineering reagents is of paramount importance to launch the next wave of biotechnology. TAL effectors have been developed as an adaptable DNA binding scaffold that can be engineered to bind to any user-defined sequence. Thus, TAL-based DNA binding modules have been used to generate chimeric proteins for a variety of targeted genome modifications across eukaryotic species. For example, TAL effectors fused to the catalytic domain of FokI endonuclease (TALENs) were used to generate site-specific double strand breaks (DSBs), the repair of which can be harnessed to dictate user-desired, genome-editing outcomes. To cleave DNA, FokI endonuclease must dimerize which can be achieved using a pair of TALENs that bind to the DNA targeted in a tail-to-tail orientation with proper spacing allowing the dimer formation. Because TALENs binding to DNA are dependent on their repeat sequences and nucleotides binding specificities, homodimers and heterodimers binding can be formed. In the present study, we used several TALEN monomers with increased repeats binding degeneracy to allow homodimer formation at increased number of genomic loci. We assessed their binding specificities and genome modification activities. Our results indicate that homodimeric TALENs could be used to modify the yeast genome in a site-specific manner and their binding to the promoter regions might modulate the expression of target genes. Taken together, our data indicate that homodimeric TALENs could be used to achieve different engineering possibilities of biotechnological applications and that their transcriptional modulations need to be considered when analyzing their phenotypic effects. © 2013 Springer-Verlag.

  18. A novel Zea mays ssp. mexicana L. MYC-type ICE-like transcription factor gene ZmmICE1, enhances freezing tolerance in transgenic Arabidopsis thaliana.

    Science.gov (United States)

    Lu, Xiang; Yang, Lei; Yu, Mengyuan; Lai, Jianbin; Wang, Chao; McNeil, David; Zhou, Meixue; Yang, Chengwei

    2017-04-01

    The annual Zea mays ssp. mexicana L., a member of the teosinte group, is a close wild relative of maize and thus can be effectively used in maize improvement. In this study, an ICE-like gene, ZmmICE1, was isolated from a cDNA library of RNA-Seq from cold-treated seedling tissues of Zea mays ssp. mexicana L. The deduced protein of ZmmICE1 contains a highly conserved basic helix-loop-helix (bHLH) domain and C-terminal region of ICE-like proteins. The ZmmICE1 protein localizes to the nucleus and shows sumoylation when expressed in an Escherichia coli reconstitution system. In addition, yeast one hybrid assays indicated that ZmmICE1 has transactivation activities. Moreover, ectopic expression of ZmmICE1 in the Arabidopsis ice1-2 mutant increased freezing tolerance. The ZmmICE1 overexpressed plants showed lower electrolyte leakage (EL), reduced contents of malondialdehyde (MDA). The expression of downstream cold related genes of Arabidopsis C-repeat-binding factors (AtCBF1, AtCBF2 and AtCBF3), cold-responsive genes (AtCOR15A and AtCOR47), kinesin-1 member gene (AtKIN1) and responsive to desiccation gene (AtRD29A) was significantly induced when compared with wild type under low temperature treatment. Taken together, these results indicated that ZmmICE1 is the homolog of Arabidopsis inducer of CBF expression genes (AtICE1/2) and plays an important role in the regulation of freezing stress response. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  19. Association of Candidate Genes With Submergence Response in Perennial Ryegrass

    Directory of Open Access Journals (Sweden)

    Xicheng Wang

    2017-05-01

    Full Text Available Perennial ryegrass is a popular cool-season grass species due to its high quality for forage and turf. The objective of this study was to identify associations of candidate genes with growth and physiological traits to submergence stress and recovery after de-submergence in a global collection of 94 perennial ryegrass accessions. Accessions varied largely in leaf color, plant height (HT, leaf fresh weight (LFW, leaf dry weight (LDW, and chlorophyll fluorescence (Fv/Fm at 7 days of submergence and in HT, LFW and LDW at 7 days of recovery in two experiments. Among 26 candidate genes tested by various models, single nucleotide polymorphisms (SNPs in 10 genes showed significant associations with traits including 16 associations for control, 10 for submergence, and 8 for recovery. Under submergence, Lp1-SST encoding sucrose:sucrose 1-fructosyltransferase and LpGA20ox encoding gibberellin 20-oxidase were associated with LFW and LDW, and LpACO1 encoding 1-aminocyclopropane-1-carboxylic acid oxidase was associated with LFW. Associations between Lp1-SST and HT, Lp6G-FFT encoding fructan:fructan 6G-fructosyltransferase and Fv/Fm, LpCAT encoding catalase and HT were also detected under submergence stress. Upon de-submergence, Lp1-SST, Lp6G-FFT, and LpPIP1 encoding plasma membrane intrinsic protein type 1 were associated with LFW or LDW, while LpCBF1b encoding C-repeat binding factor were associated with HT. Nine significant SNPs in Lp1-SST, Lp6G-FFT, LpCAT, and LpACO1 resulted in amino acid changes with five substitutions found in Lp1-SST under submergence or recovery. The results indicated that allelic diversity in genes involved in carbohydrate and antioxidant metabolism, ethylene and gibberellin biosynthesis, and transcript factor could contribute to growth variations in perennial ryegrass under submergence stress and recovery after de-submergence.

  20. Cross-talk between abscisic acid-dependent and abscisic acid-independent pathways during abiotic stress.

    Science.gov (United States)

    Roychoudhury, Aryadeep; Paul, Saikat; Basu, Supratim

    2013-07-01

    Salinity, drought and low temperature are the common forms of abiotic stress encountered by land plants. To cope with these adverse environmental factors, plants execute several physiological and metabolic responses. Both osmotic stress (elicited by water deficit or high salt) and cold stress increase the endogenous level of the phytohormone abscisic acid (ABA). ABA-dependent stomatal closure to reduce water loss is associated with small signaling molecules like nitric oxide, reactive oxygen species and cytosolic free calcium, and mediated by rapidly altering ion fluxes in guard cells. ABA also triggers the expression of osmotic stress-responsive (OR) genes, which usually contain single/multiple copies of cis-acting sequence called abscisic acid-responsive element (ABRE) in their upstream regions, mostly recognized by the basic leucine zipper-transcription factors (TFs), namely, ABA-responsive element-binding protein/ABA-binding factor. Another conserved sequence called the dehydration-responsive element (DRE)/C-repeat, responding to cold or osmotic stress, but not to ABA, occurs in some OR promoters, to which the DRE-binding protein/C-repeat-binding factor binds. In contrast, there are genes or TFs containing both DRE/CRT and ABRE, which can integrate input stimuli from salinity, drought, cold and ABA signaling pathways, thereby enabling cross-tolerance to multiple stresses. A strong candidate that mediates such cross-talk is calcium, which serves as a common second messenger for abiotic stress conditions and ABA. The present review highlights the involvement of both ABA-dependent and ABA-independent signaling components and their interaction or convergence in activating the stress genes. We restrict our discussion to salinity, drought and cold stress.

  1. Supra-optimal expression of the cold-regulated OsMyb4 transcription factor in transgenic rice changes the complexity of transcriptional network with major effects on stress tolerance and panicle development.

    Science.gov (United States)

    Park, Myoung-Ryoul; Yun, Kil-Young; Mohanty, Bijayalaxmi; Herath, Venura; Xu, Fuyu; Wijaya, Edward; Bajic, Vladimir B; Yun, Song-Joong; De Los Reyes, Benildo G

    2010-12-01

    The R2R3-type OsMyb4 transcription factor of rice has been shown to play a role in the regulation of osmotic adjustment in heterologous overexpression studies. However, the exact composition and organization of its underlying transcriptional network has not been established to be a robust tool for stress tolerance enhancement by regulon engineering. OsMyb4 network was dissected based on commonalities between the global chilling stress transcriptome and the transcriptome configured by OsMyb4 overexpression. OsMyb4 controls a hierarchical network comprised of several regulatory sub-clusters associated with cellular defense and rescue, metabolism and development. It regulates target genes either directly or indirectly through intermediary MYB, ERF, bZIP, NAC, ARF and CCAAT-HAP transcription factors. Regulatory sub-clusters have different combinations of MYB-like, GCC-box-like, ERD1-box-like, ABRE-like, G-box-like, as1/ocs/TGA-like, AuxRE-like, gibberellic acid response element (GARE)-like and JAre-like cis-elements. Cold-dependent network activity enhanced cellular antioxidant capacity through radical scavenging mechanisms and increased activities of phenylpropanoid and isoprenoid metabolic processes involving various abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), ethylene and reactive oxygen species (ROS) responsive genes. OsMyb4 network is independent of drought response element binding protein/C-repeat binding factor (DREB/CBF) and its sub-regulons operate with possible co-regulators including nuclear factor-Y. Because of its upstream position in the network hierarchy, OsMyb4 functions quantitatively and pleiotrophically. Supra-optimal expression causes misexpression of alternative targets with costly trade-offs to panicle development. © 2010 Blackwell Publishing Ltd.

  2. Diversity of the marine picocyanobacteria Prochlorococcus and Synechococcus assessed by terminal restriction fragment length polymorphisms of 16S-23S rRNA internal transcribed spacer sequences Diversidad de las picocianobacterias marinas Prochlorococcus y Synechococcus por medio de polimorfismos de longitud de fragmentos de restricción terminal en secuencias del espaciador transcrito interno del ARNr 16S - 23S

    Directory of Open Access Journals (Sweden)

    PARIS LAVIN

    2008-12-01

    Full Text Available In order to assess the appropriateness of the use of internal transcribed spacer (ITS sequences for the study of population genetics of marine cyanobacteria, we amplified and cloned the 16S rRNA gene plus the 16S-23S ITS regions of six strains of Prochlorococcus and Synechococcus. We analyzed them by denaturing gradient gel electrophoresis (DGGE and terminal restriction fragment length polymorphisms (T-RFLP. When using the standard application of these techniques, we obtained more than one band or terminal restriction fragment (T-RF per strain or cloned sequence. Reports in literature have suggested that these anomalies can result from the formation of secondary structures. Secondary structures of the ITS sequences of Prochlorococcus and Synechococcus strains were computationally modelled at the different temperatures that were used during the polymerase chain reaction (PCR. Modelling results predicted the existence of hairpin loops that would still be present at the extensión temperature; it is likely that these loops produced incomplete and single stranded PCR products. We modified the standard T-RFLP procedure by adding the labelled ITS primer in the last two cycles of the PCR reaction; this resulted, in most cases, in only one T-RF per ribotype. Application of this technique to a natural picoplankton community in marine waters off northern Chile, showed that it was possible to identify the presence, and determine the relative abundance, of several phylogenetic lineages within the genera Prochlorococcus and Synechococcus inhabiting the euphotic zone. Phylogenetic analysis of ITS sequences obtained by cloning and sequencing DNA from the same sample confirmed the presence of the different genotypes. With the proposed modification, T-RFLP profiles should therefore be suitable for studying the diversity of natural populations of cyanobacteria, and should become an important tool to study the factors influencing the genetic structure and

  3. Expression of human telomerase reverse transcriptase protein in oral epithelial dysplasia and oral squamous cell carcinoma: An immunohistochemical study

    Science.gov (United States)

    Raghunandan, Bangalore Nagarajachar; Sanjai, Karpagaselvi; Kumaraswamy, Jayalakshmi; Papaiah, Lokesh; Pandey, Bhavna; Jyothi, Bellur MadhavaRao

    2016-01-01

    Background: Telomerase is an RNA-dependent DNA polymerase that synthesizes TTAGGG telomeric DNA sequences and almost universally provides the molecular basis for unlimited proliferative potential. The telomeres become shorter with each cycle of replication and reach a critical limit; most cells die or enter stage of replicative senescence. Telomere length maintenance by telomerase is required for all the cells that exhibit limitless replicative potential. It has been postulated that reactivation of telomerase expression is necessary for the continuous proliferation of neoplastic cells to attain immortality. Use of immunohistochemistry (IHC) is a useful, reliable method of localizing the human telomerase reverse transcriptase (hTERT) protein in tissue sections which permits cellular localization. Although there exists a lot of information on telomerase in oral cancer, little is known about their expression in oral epithelial dysplasia and their progression to oral squamous cell carcinoma (OSCC) compared to normal oral mucosa. This study addresses this lacuna. Aims: To compare the expression of hTERT protein in oral epithelial dysplasia and OSCC with normal oral mucosa by Immunohistochemical method. Subjects and Methods: In this preliminary study, IHC was used to detect the expression of hTERT protein in OSCC (n = 20), oral epithelial dysplasia (n = 21) and normal oral mucosa (n = 10). The tissue localization of immunostain, cellular localization of immunostain, nature of stain, intensity of stain, percentage of cells stained with hTERT protein were studied. A total number of 100 cells were counted in each slide. Statistical Analysis: All the data were analyzed using SPSS software version 16.0. The tissue localization, cellular localization of cytoplasmic/nuclear/both of hTERT stain, staining intensity was compared across the groups using Pearson's Chi-square test. The mean percentage of cells stained for oral epithelial dysplasia, OSCC and normal oral mucosa were

  4. Combination of precise GNSS orbit and clock solutions in a multi-constellation, multi-frequency environment

    Science.gov (United States)

    Ortiz Geist, Estefania

    2015-04-01

    Precise GNSS orbit and clock solutions are essential for the generation of the Terrestrial Reference Frame (TRF) and required for a broad variety of applications. Over the last decades the combination products of the International GNSS Service (IGS) have become the standard for all kinds of GNSS applications requiring highest accuracy. The emerging new GNSS constellations Galileo, BeiDou and the QZSS as well as the modernization of the already established GPS and GLONASS constellations will stimulate a new development in the GNSS data processing in order to gain be best benefit from the new signals and systems for geodetic and geodynamic applications. This introduces the question regarding the influence of this development on the orbit and clock products. What are the consequences for the consistency of the contributions from the Analysis Centres (ACs) of the IGS and how does the combination procedure need to react on his development? Another set of questions is related to the expected scenario in which not all IGS ACs will likely include all GNSS. The algorithm for the orbit and clock combination needs to be adapted for a multi-system combination to keep on one hand the internal consistency between the GNSS during the combination procedure but also consider the differences in the expected orbit qualities between the satellite systems (e.g., due to the number of satellites or network coverage). To investigate these questions ESOC and AIUB have agreed on a joint research fellowship for three years. The objective of this research is to analyse the capabilities and challenges when combining hybrid multi-GNSS solutions and to develop a concept, which compares and combines orbit and clock contributions to come up with a consistent, reliable, truly combined multi-GNSS combination product. Well-defined test scenarios shall be constructed and analysed based on the GNSS data processing software packages in the two institutions, namely "NAPEOS" and "Bernese GNSS Software

  5. A conjugative 38 kB plasmid is present in multiple subspecies of Xylella fastidiosa.

    Science.gov (United States)

    Rogers, Elizabeth E; Stenger, Drake C

    2012-01-01

    A ≈ 38kB plasmid (pXF-RIV5) was present in the Riv5 strain of Xylella fastidiosa subsp. multiplex isolated from ornamental plum in southern California. The complete nucleotide sequence of pXF-RIV5 is almost identical to that of pXFAS01 from X. fastidiosa subsp. fastidiosa strain M23; the two plasmids vary at only 6 nucleotide positions. BLAST searches and phylogenetic analyses indicate pXF-RIV5 and pXFAS01 share some similarity to chromosomal and plasmid (pXF51) sequences of X. fastidiosa subsp. pauca strain 9a5c and more distant similarity to plasmids from a wide variety of bacteria. Both pXF-RIV5 and pXFAS01 encode homologues of a complete Type IV secretion system involved in conjugation and DNA transfer among bacteria. Mating pair formation proteins (Trb) from Yersinia pseudotuberculosis IP31758 are the mostly closely related non-X. fastidiosa proteins to most of the Trb proteins encoded by pXF-RIV5 and pXFAS01. Unlike many bacterial conjugative plasmids, pXF-RIV5 and pXFAS01 do not carry homologues of known accessory modules that confer selective advantage on host bacteria. However, both plasmids encode seven hypothetical proteins of unknown function and possess a small transposon-associated region encoding a putative transposase and associated factor. Vegetative replication of pXF-RIV5 and pXFAS01 appears to be under control of RepA protein and both plasmids have an origin of DNA replication (oriV) similar to that of pRP4 and pR751 from Escherichia coli. In contrast, conjugative plasmids commonly encode TrfA and have an oriV similar to those found in IncP-1 incompatibility group plasmids. The presence of nearly identical plasmids in single strains from two distinct subspecies of X. fastidiosa is indicative of recent horizontal transfer, probably subsequent to the introduction of subspecies fastidiosa to the United States in the late 19(th) century.

  6. Two New Loci for Body-Weight Regulation Identified in a Joint Analysis of Genome-Wide Association Studies for Early-Onset Extreme Obesity in French and German Study Groups

    Science.gov (United States)

    Scherag, André; Dina, Christian; Hinney, Anke; Vatin, Vincent; Scherag, Susann; Vogel, Carla I. G.; Müller, Timo D.; Grallert, Harald; Wichmann, H.-Erich; Balkau, Beverley; Heude, Barbara; Jarvelin, Marjo-Riitta; Hartikainen, Anna-Liisa; Levy-Marchal, Claire; Weill, Jacques; Delplanque, Jérôme; Körner, Antje; Kiess, Wieland; Kovacs, Peter; Rayner, Nigel W.; Prokopenko, Inga; McCarthy, Mark I.; Schäfer, Helmut; Jarick, Ivonne; Boeing, Heiner; Fisher, Eva; Reinehr, Thomas; Heinrich, Joachim; Rzehak, Peter; Berdel, Dietrich; Borte, Michael; Biebermann, Heike; Krude, Heiko; Rosskopf, Dieter; Rimmbach, Christian; Rief, Winfried; Fromme, Tobias; Klingenspor, Martin; Schürmann, Annette; Schulz, Nadja; Nöthen, Markus M.; Mühleisen, Thomas W.; Erbel, Raimund; Jöckel, Karl-Heinz; Moebus, Susanne; Boes, Tanja; Illig, Thomas; Froguel, Philippe; Hebebrand, Johannes; Meyre, David

    2010-01-01

    Meta-analyses of population-based genome-wide association studies (GWAS) in adults have recently led to the detection of new genetic loci for obesity. Here we aimed to discover additional obesity loci in extremely obese children and adolescents. We also investigated if these results generalize by estimating the effects of these obesity loci in adults and in population-based samples including both children and adults. We jointly analysed two GWAS of 2,258 individuals and followed-up the best, according to lowest p-values, 44 single nucleotide polymorphisms (SNP) from 21 genomic regions in 3,141 individuals. After this DISCOVERY step, we explored if the findings derived from the extremely obese children and adolescents (10 SNPs from 5 genomic regions) generalized to (i) the population level and (ii) to adults by genotyping another 31,182 individuals (GENERALIZATION step). Apart from previously identified FTO, MC4R, and TMEM18, we detected two new loci for obesity: one in SDCCAG8 (serologically defined colon cancer antigen 8 gene; p = 1.85×10−8 in the DISCOVERY step) and one between TNKS (tankyrase, TRF1-interacting ankyrin-related ADP-ribose polymerase gene) and MSRA (methionine sulfoxide reductase A gene; p = 4.84×10−7), the latter finding being limited to children and adolescents as demonstrated in the GENERALIZATION step. The odds ratios for early-onset obesity were estimated at ∼1.10 per risk allele for both loci. Interestingly, the TNKS/MSRA locus has recently been found to be associated with adult waist circumference. In summary, we have completed a meta-analysis of two GWAS which both focus on extremely obese children and adolescents and replicated our findings in a large followed-up data set. We observed that genetic variants in or near FTO, MC4R, TMEM18, SDCCAG8, and TNKS/MSRA were robustly associated with early-onset obesity. We conclude that the currently known major common variants related to obesity overlap to a substantial degree between

  7. Impact of intermittent fasting on health and disease processes.

    Science.gov (United States)

    Mattson, Mark P; Longo, Valter D; Harvie, Michelle

    2017-10-01

    Humans in modern societies typically consume food at least three times daily, while laboratory animals are fed ad libitum. Overconsumption of food with such eating patterns often leads to metabolic morbidities (insulin resistance, excessive accumulation of visceral fat, etc.), particularly when associated with a sedentary lifestyle. Because animals, including humans, evolved in environments where food was relatively scarce, they developed numerous adaptations that enabled them to function at a high level, both physically and cognitively, when in a food-deprived/fasted state. Intermittent fasting (IF) encompasses eating patterns in which individuals go extended time periods (e.g., 16-48h) with little or no energy intake, with intervening periods of normal food intake, on a recurring basis. We use the term periodic fasting (PF) to refer to IF with periods of fasting or fasting mimicking diets lasting from 2 to as many as 21 or more days. In laboratory rats and mice IF and PF have profound beneficial effects on many different indices of health and, importantly, can counteract disease processes and improve functional outcome in experimental models of a wide range of age-related disorders including diabetes, cardiovascular disease, cancers and neurological disorders such as Alzheimer's disease Parkinson's disease and stroke. Studies of IF (e.g., 60% energy restriction on 2days per week or every other day), PF (e.g., a 5day diet providing 750-1100kcal) and time-restricted feeding (TRF; limiting the daily period of food intake to 8h or less) in normal and overweight human subjects have demonstrated efficacy for weight loss and improvements in multiple health indicators including insulin resistance and reductions in risk factors for cardiovascular disease. The cellular and molecular mechanisms by which IF improves health and counteracts disease processes involve activation of adaptive cellular stress response signaling pathways that enhance mitochondrial health, DNA repair

  8. CLINICAL STUDY OF THE EFFECTS OF LOW-PROTEIN DIET AND SUPPLEMENTED WITH α-KETOACIDS THERAPY ON NUTRITION STATUS AND RESIDUAL RENAL FUNCTION IN CONTINUOUS AMBULATORY PERITONEAL DIALYSIS(CAPD PATIENTS

    Directory of Open Access Journals (Sweden)

    Juan Huang

    2012-06-01

    Full Text Available It is critical to preserve residual renal function (RRF in CAPD, as RRF is associated with lower morbidity and mortality. low- protein diet supplemented with α-keto acids was reported to have an important roles in delaying in follow-up period progression of renal failure and relieving malnutritional status in non-dialysis patients with chronic renal failure. We evaluate the effects on the nutritional status and RRF of a low-protein diet supplemented with α-keto acids on CAPD patients prospectively. Seventy eight CAPD patients who were randomly assigned to a low-protein diet supplemented with α-keto acid group(keto acid group,31 patients,low-protein diet group(LPD group,26 patientsand routine protein diet group(RPD group,21 patientswere investigated and followed up for one year. The nutritional parameters were measured, and examinations of residual renal function (RRF, Kt/v, clearance of creatinine (Ccr and levels of serum amino acids, mean arterial pressure (MAP, peritoneal ultrafiltration(UF,residual urine volume(RUV and the status of water-sodium retentio were performed. Compared to LPD group, serum levels of prealbumin(PA, transferrin (TRF retinal binding protein (RBP had a significant increment both in keto acid group and RPD group(P〈0.01,but there was no significance between these two groups(p〉0.05□Compared to RPD group, There was an incremental tendency in albumin(Alb, total cholesterol(TC, triglyceride (TG, Triceps skinfoles (TSF,midarm riuscle circumference(MAMC,body mass index(BMI in keto acid group, but no significance(p〉0.05□The serum concentrations of Valine(Val,Leucine(Leu, Isoleucine (Ile were significantly increased in keto acid group(p〈0.01,compared to the other groups. Levels of RRF, Kt/V, Ccr, RUV were significantly higher both in keto acid group and LPD group than in RPD group(p〈0.01□There were no significant differences in MAP, UF and peritoneal dialysate albumin loss between these groups, However, the

  9. Moldagem por compressão a frio do polietileno de ultra alto peso molecular. Parte 1: influência do tamanho, distribuição e morfologia da partícula na densidade a verde Cold compression molding of ultra high molecular weight polyethylene. Part 1: influence of the size, distribution and morphology of particles on the green density

    Directory of Open Access Journals (Sweden)

    Bernadete A. Bittencourt

    2009-01-01

    Full Text Available Neste trabalho foram investigadas as características de amostras de pó de polietileno de ultra alto peso molecular (PEUAPM, tais como porosidade, morfologia, tamanho médio e distribuição de partícula, que são importantes na moldagem por compressão a frio. Também foi avaliada a influência dessas características na densidade a verde de pré-formas. As amostras dos pós foram caracterizadas por calorimetria diferencial de varredura (DSC, análise granulométrica, absorção de óleo, área superficial, porosimetria de mercúrio, fluidez do pó, densidade de compactação, densidade aparente e microscopia eletrônica de varredura (MEV. Através das técnicas de caracterização estudadas ficou evidenciado que as características da partícula citadas anteriormente, assim como o parâmetro de densificação (DP, que é função direta da porosidade interparticular, favorecem a densidade a verde relativa (DVR e consequentemente a tensão de resistência à flexão (TRF.In this paper an investigation was made of the characteristics of Ultra High Molecular Weight Polyethylene (UHMWPE powder samples, including porosity, particles average size, size distribution and morphology, which are important in cold compression molding. The influence of these characteristics on the green density of molded pre-shapes was also investigated. The UHMWPE powder samples were characterized by Differential Scanning Calorimetry (DSC, granulometric analysis, oil absorption, surface area, mercury porosity, density compaction, apparent density and Scanning Electron Microscopy (SEM. The characterization techniques used demonstrate that the UHMWPE particles characteristics cited above as well as the densification parameter (DP, which is a direct function of the interparticles porosity, affect the relative green density (RGD and hence, the flexural tensile strength (FTS.

  10. Complete nucleotide sequence and analysis of two conjugative broad host range plasmids from a marine microbial biofilm.

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    Peter Norberg

    Full Text Available The complete nucleotide sequence of plasmids pMCBF1 and pMCBF6 was determined and analyzed. pMCBF1 and pMCBF6 form a novel clade within the IncP-1 plasmid family designated IncP-1 ς. The plasmids were exogenously isolated earlier from a marine biofilm. pMCBF1 (62 689 base pairs; bp and pMCBF6 (66 729 bp have identical backbones, but differ in their mercury resistance transposons. pMCBF1 carries Tn5053 and pMCBF6 carries Tn5058. Both are flanked by 5 bp direct repeats, typical of replicative transposition. Both insertions are in the vicinity of a resolvase gene in the backbone, supporting the idea that both transposons are "res-site hunters" that preferably insert close to and use external resolvase functions. The similarity of the backbones indicates recent insertion of the two transposons and the ongoing dynamics of plasmid evolution in marine biofilms. Both plasmids also carry the insertion sequence ISPst1, albeit without flanking repeats. ISPs1is located in an unusual site within the control region of the plasmid. In contrast to most known IncP-1 plasmids the pMCBF1/pMCBF6 backbone has no insert between the replication initiation gene (trfA and the vegetative replication origin (oriV. One pMCBF1/pMCBF6 block of about 2.5 kilo bases (kb has no similarity with known sequences in the databases. Furthermore, insertion of three genes with similarity to the multidrug efflux pump operon mexEF and a gene from the NodT family of the tripartite multi-drug resistance-nodulation-division (RND system in Pseudomonas aeruginosa was found. They do not seem to confer antibiotic resistance to the hosts of pMCBF1/pMCBF6, but the presence of RND on promiscuous plasmids may have serious implications for the spread of antibiotic multi-resistance.

  11. Dyslipidemia and chronic inflammation markers are correlated with telomere length shortening in Cushing's syndrome.

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    Anna Aulinas

    Full Text Available Cushing's syndrome (CS increases cardiovascular risk (CVR and adipocytokine imbalance, associated with an increased inflammatory state. Telomere length (TL shortening is a novel CVR marker, associated with inflammation biomarkers. We hypothesized that inflammatory state and higher CVR in CS might be related to TL shortening, as observed in premature aging.To evaluate relationships between TL, CVR and inflammation markers in CS.In a cross-sectional study, 77 patients with CS (14 males, 59 pituitary-, 17 adrenal- and 1 ectopic-origin; 21 active disease and 77 age-, gender-, smoking-matched controls were included. Total white blood cell TL was measured by TRF-Southern technique. Clinical data and blood samples were collected (lipids, adrenal function, glucose. Adiponectin, interleukin-6 (IL6 and C-reactive protein (CRP were available in a subgroup of patients (n=32. Correlations between TL and clinical features were examined and multiple linear regression analysis was performed to investigate potential predictors of TL.Dyslipidemic CS had shorter TL than non-dyslipidemic subjects (7328±1274 vs 7957±1137 bp, p<0.05. After adjustment for age and body mass index, cured and active CS dyslipidemic patients had shorter TL than non-dyslipidemic CS (cured: 7187±1309 vs 7868±1104; active: 7203±1262 vs 8615±1056, respectively, p<0.05. Total cholesterol and triglycerides negatively correlated with TL (r-0.279 and -0.259, respectively, p<0.05, as well as CRP and IL6 (r-0.412 and -0.441, respectively, p<0.05. No difference in TL according the presence of other individual CVR factors (hypertension, diabetes mellitus, obesity were observed in CS or in the control group. Additional TL shortening was observed in dyslipidemic obese patients who were also hypertensive, compared to those with two or less CVR factors (6956±1280 vs 7860±1180, respectively, p<0.001. Age and dyslipidemia were independent negative predictors of TL.TL is shortened in dyslipidemic CS

  12. The effect of meteorological data on atmospheric pressure loading corrections in VLBI data analysis

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    Balidakis, Kyriakos; Glaser, Susanne; Karbon, Maria; Soja, Benedikt; Nilsson, Tobias; Lu, Cuixian; Anderson, James; Liu, Li; Andres Mora-Diaz, Julian; Raposo-Pulido, Virginia; Xu, Minghui; Heinkelmann, Robert; Schuh, Harald

    2015-04-01

    well as the inconsistencies between them, and the interpolation scheme which yields the elastic deformations. Differences of the resulting TRF (Terrestrial Reference Frame) determinations and other products derived from VLBI analysis between the approach followed here and the one employing NWM's data for obtaining the input pressure fields, are illustrated. The providers of the atmospheric pressure loading models employed for our comparisons are GSFC/NASA, the University of Luxembourg, the University of Strasbourg, the Technical University of Vienna and GeoForschungsZentrum of Potsdam.

  13. A conjugative 38 kB plasmid is present in multiple subspecies of Xylella fastidiosa.

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    Elizabeth E Rogers

    Full Text Available A ≈ 38kB plasmid (pXF-RIV5 was present in the Riv5 strain of Xylella fastidiosa subsp. multiplex isolated from ornamental plum in southern California. The complete nucleotide sequence of pXF-RIV5 is almost identical to that of pXFAS01 from X. fastidiosa subsp. fastidiosa strain M23; the two plasmids vary at only 6 nucleotide positions. BLAST searches and phylogenetic analyses indicate pXF-RIV5 and pXFAS01 share some similarity to chromosomal and plasmid (pXF51 sequences of X. fastidiosa subsp. pauca strain 9a5c and more distant similarity to plasmids from a wide variety of bacteria. Both pXF-RIV5 and pXFAS01 encode homologues of a complete Type IV secretion system involved in conjugation and DNA transfer among bacteria. Mating pair formation proteins (Trb from Yersinia pseudotuberculosis IP31758 are the mostly closely related non-X. fastidiosa proteins to most of the Trb proteins encoded by pXF-RIV5 and pXFAS01. Unlike many bacterial conjugative plasmids, pXF-RIV5 and pXFAS01 do not carry homologues of known accessory modules that confer selective advantage on host bacteria. However, both plasmids encode seven hypothetical proteins of unknown function and possess a small transposon-associated region encoding a putative transposase and associated factor. Vegetative replication of pXF-RIV5 and pXFAS01 appears to be under control of RepA protein and both plasmids have an origin of DNA replication (oriV similar to that of pRP4 and pR751 from Escherichia coli. In contrast, conjugative plasmids commonly encode TrfA and have an oriV similar to those found in IncP-1 incompatibility group plasmids. The presence of nearly identical plasmids in single strains from two distinct subspecies of X. fastidiosa is indicative of recent horizontal transfer, probably subsequent to the introduction of subspecies fastidiosa to the United States in the late 19(th century.

  14. Telomerase Repeated Amplification Protocol (TRAP).

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    Mender, Ilgen; Shay, Jerry W

    2015-11-20

    Telomeres are found at the end of eukaryotic linear chromosomes, and proteins that bind to telomeres protect DNA from being recognized as double-strand breaks thus preventing end-to-end fusions (Griffith et al. , 1999). However, due to the end replication problem and other factors such as oxidative damage, the limited life span of cultured cells (Hayflick limit) results in progressive shortening of these protective structures (Hayflick and Moorhead, 1961; Olovnikov, 1973). The ribonucleoprotein enzyme complex telomerase-consisting of a protein catalytic component hTERT and a functional RNA component hTR or hTERC - counteracts telomere shortening by adding telomeric repeats to the end of chromosomes in ~90% of primary human tumors and in some transiently proliferating stem-like cells (Shay and Wright, 1996; Shay and Wright, 2001). This results in continuous proliferation of cells which is a hallmark of cancer. Therefore, telomere biology has a central role in aging, cancer progression/metastasis as well as targeted cancer therapies. There are commonly used methods in telomere biology such as Telomere Restriction Fragment (TRF) (Mender and Shay, 2015b), Telomere Repeat Amplification Protocol (TRAP) and Telomere dysfunction Induced Foci (TIF) analysis (Mender and Shay, 2015a). In this detailed protocol we describe Telomere Repeat Amplification Protocol (TRAP). The TRAP assay is a popular method to determine telomerase activity in mammalian cells and tissue samples (Kim et al. , 1994). The TRAP assay includes three steps: extension, amplification, and detection of telomerase products. In the extension step, telomeric repeats are added to the telomerase substrate (which is actually a non telomeric oligonucleotide, TS) by telomerase. In the amplification step, the extension products are amplified by the polymerase chain reaction (PCR) using specific primers (TS upstream primer and ACX downstream primer) and in the detection step, the presence or absence of telomerase is

  15. Two new Loci for body-weight regulation identified in a joint analysis of genome-wide association studies for early-onset extreme obesity in French and german study groups.

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    André Scherag

    2010-04-01

    Full Text Available Meta-analyses of population-based genome-wide association studies (GWAS in adults have recently led to the detection of new genetic loci for obesity. Here we aimed to discover additional obesity loci in extremely obese children and adolescents. We also investigated if these results generalize by estimating the effects of these obesity loci in adults and in population-based samples including both children and adults. We jointly analysed two GWAS of 2,258 individuals and followed-up the best, according to lowest p-values, 44 single nucleotide polymorphisms (SNP from 21 genomic regions in 3,141 individuals. After this DISCOVERY step, we explored if the findings derived from the extremely obese children and adolescents (10 SNPs from 5 genomic regions generalized to (i the population level and (ii to adults by genotyping another 31,182 individuals (GENERALIZATION step. Apart from previously identified FTO, MC4R, and TMEM18, we detected two new loci for obesity: one in SDCCAG8 (serologically defined colon cancer antigen 8 gene; p = 1.85x10(-8 in the DISCOVERY step and one between TNKS (tankyrase, TRF1-interacting ankyrin-related ADP-ribose polymerase gene and MSRA (methionine sulfoxide reductase A gene; p = 4.84x10(-7, the latter finding being limited to children and adolescents as demonstrated in the GENERALIZATION step. The odds ratios for early-onset obesity were estimated at approximately 1.10 per risk allele for both loci. Interestingly, the TNKS/MSRA locus has recently been found to be associated with adult waist circumference. In summary, we have completed a meta-analysis of two GWAS which both focus on extremely obese children and adolescents and replicated our findings in a large followed-up data set. We observed that genetic variants in or near FTO, MC4R, TMEM18, SDCCAG8, and TNKS/MSRA were robustly associated with early-onset obesity. We conclude that the currently known major common variants related to obesity overlap to a substantial

  16. Preventing Behavioral Disorders via Supporting Social and Emotional Competence at Preschool Age.

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    Schell, Annika; Albers, Lucia; von Kries, Rüdiger; Hillenbrand, Clemens; Hennemann, Thomas

    2015-09-25

    13-18% of all preschool children have severe behavioral problems at least transiently, sometimes with long-term adverse consequences. In this study, the social training program "Lubo aus dem All! - Vorschulalter" (Lubo from Outer Space, Preschool Version) was evaluated in a kindergarten setting. 15 kindergartens were randomly assigned to either an intervention group or a control group, in a 2:1 ratio. The intervention was designed to strengthen emotional knowledge and regulation, the ability to take another person's point of view, communication skills, and social problem solving. The control group continued with conventional kindergarten activities. The primary endpoint was improvement in social-cognitive problem solving strategies, as assessed with the Wally Social Skills and Problem Solving Game (Wally). Secondary endpoints were improvement in prosocial behavior and reduction in problematic behavior, as assessed with the Preschool Social Behavior Questionnaire (PSBQ) and the Caregiver-Teacher Report Form (C-TRF). Data were collected before and after the intervention and also 5 months later. Mixed models were calculated with random effects to take account of the cluster design and for adjustment for confounding variables. 221 children in kindergarten, aged 5-6 years, were included in the study. Randomization was unsuccessful: the children in the intervention group performed markedly worse on the tests carried out before the intervention. Five months after the end of the intervention, the social-cognitive problem solving strategies of the children in the intervention group had improved more than those of the children in the control group: the intergroup difference in improvement was 0.79 standard deviations of the Wally test (95% confidence interval [CI] 0.13-1.46). This effect was just as marked 5 months later (0.63, 95% CI 0.03-1.23). Prosocial behavior, as measured by the PSBQ, also improved more in the intervention group, with an intergroup difference of 0

  17. Identification of active dehalorespiring microbial populations in anoxic river sediment by RNA-based stable isotope probing

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    Kittelmann, S.; Friedrich, M. W.

    2005-12-01

    Tetrachloroethene (perchloroethylene, PCE), a persistent contaminant in aquifers, soils and sediments, can be reductively dechlorinated by anaerobic microorganisms in a process referred to as dehalorespiration. However, the biodiversity of dehalorespiring microorganisms and their distribution especially in pristine environments is largely unexplored. Therefore, the aim of this study was to detect potentially novel PCE-dehalorespiring microorganisms by using stable isotope probing (SIP), a technique that allows to directly identify the function of uncultivated microbial populations. We simulated a PCE contamination by incubating pristine river sediment in the presence of PCE at a steady, low aqueous concentration (20 μM). Dehalogenation activity in microcosms (20 nmol cis-dichloroethene per ml slurry per day formed) was detected already after 4 weeks at 20°C with sediment indigenous electron donors. The microbial community in sediment incubations was probed with 13C-labelled acetate (0.5 mM) as electron donor and carbon source at 15°C for 3 days. After RNA extraction, "heavy" 13C-rRNA and light 12C-rRNA were separated by isopycnic centrifugation, and Bacteria-related populations in gradient fractions were characterised by terminal restriction fragment length polymorphism analysis and cloning. In heavy gradient fractions from the microcosm with PCE, we detected a prominent 506-bp terminal restriction fragment (T-RF) and a few minor T-RFs only. In contrast, in the control without PCE, Bacteria-specific rRNA was restricted to light gradient fractions, and the prominent T-RFs found in the PCE-dechlorinating microcosm were of minor importance. Apparently, 13C-acetate was incorporated into bacterial rRNA more effectively in PCE-respiring microcosms. Thus, rRNA-SIP provides strong evidence for the presence of PCE-dehalorespiring, 13C-acetate-utilising populations in river sediment microcosms. Cloning/sequencing analysis identified the prominent members of the heavy

  18. The ICRF-3: Status, Plans, and Multi-wavelength Progress on the next generation Celestial Reference Frame.

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    Jacobs, Christopher

    2015-08-01

    ICRF-3 seeks to improve upon the highly successful ICRF-2. Our goals are to improve the precision, spatial and frequency coverage relative to the ICRF-2 by 2018. This date is driven by the desire to create radio frames that are ready for comparison with the Gaia optical frame.Several specific actions are underway. A collaboration to improve at S/X-band precision of the Very Long Baseline Array (VLBA) Calibrator Survey's ~2200 sources, which are typically 5 times less precise than the rest of the ICRF-2, is bearing fruit and is projected to yield a factor of 3 improvement in precision. S/X-band southern hemisphere precision improvements are underway with observations using southern antennas such as the AuScope, Warkworth, and HartRAO, South Africa.We also seek to improve radio frequency coverage with X/Ka-band and K-band work. An X/Ka frame of 660 sources now has full sky coverage from the addition of a 2nd southern station in Argentina which is strengthening the southern hemisphere in general. The X/Ka-band frame's precision is now comparable to the ICRF-2 for the 530 sources in common. A K-band collaboration has formed with similar coverage and southern precision goals. By the time of this meeting, we expect K-band to complete full sky coverage with south polar cap observations and to improve spatial density north of -30 deg declination with VLBA observations.On the analysis front, special attention is being given to combination techniques both of Very Long Baseline Interferometry (VLBI) frames and of multiple data types. Consistency of the Celestial Reference Frame (CRF) with the Terrestrial Reference Frame (TRF) and Earth Oreintation Parameters (EOP) is another area of concern. Comparison of celestial frame solutions from various groups is underway in order to identify and correct systematic errors. We will discuss evidence emerging for 100 µas zonal errors in the ICRF2 in the declination range from 0 to -30 deg.Finally, work is underway to identify and

  19. Feature genes in metastatic breast cancer identified by MetaDE and SVM classifier methods.

    Science.gov (United States)

    Tuo, Youlin; An, Ning; Zhang, Ming

    2018-03-01

    The aim of the present study was to investigate the feature genes in metastatic breast cancer samples. A total of 5 expression profiles of metastatic breast cancer samples were downloaded from the Gene Expression Omnibus database, which were then analyzed using the MetaQC and MetaDE packages in R language. The feature genes between metastasis and non‑metastasis samples were screened under the threshold of PSVM) classifier training and verification. The accuracy of the SVM classifier was then evaluated using another independent dataset from The Cancer Genome Atlas database. Finally, function and pathway enrichment analyses for genes in the SVM classifier were performed. A total of 541 feature genes were identified between metastatic and non‑metastatic samples. The top 10 genes with the highest betweenness centrality values in the PPI network of feature genes were Nuclear RNA Export Factor 1, cyclin‑dependent kinase 2 (CDK2), myelocytomatosis proto‑oncogene protein (MYC), Cullin 5, SHC Adaptor Protein 1, Clathrin heavy chain, Nucleolin, WD repeat domain 1, proteasome 26S subunit non‑ATPase 2 and telomeric repeat binding factor 2. The cyclin‑dependent kinase inhibitor 1A (CDKN1A), E2F transcription factor 1 (E2F1), and MYC interacted with CDK2. The SVM classifier constructed by the top 30 feature genes was able to distinguish metastatic samples from non‑metastatic samples [correct rate, specificity, positive predictive value and negative predictive value >0.89; sensitivity >0.84; area under the receiver operating characteristic curve (AUROC) >0.96]. The verification of the SVM classifier in an independent dataset (35 metastatic samples and 143 non‑metastatic samples) revealed an accuracy of 94.38% and AUROC of 0.958. Cell cycle associated functions and pathways were the most significant terms of the 30 feature genes. A SVM classifier was constructed to assess the possibility of breast cancer metastasis, which presented high accuracy in several

  20. Targeted Genetic Screen in Amyotrophic Lateral Sclerosis Reveals Novel Genetic Variants with Synergistic Effect on Clinical Phenotype

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    Johnathan Cooper-Knock

    2017-11-01

    Full Text Available Amyotrophic lateral sclerosis (ALS is underpinned by an oligogenic rare variant architecture. Identified genetic variants of ALS include RNA-binding proteins containing prion-like domains (PrLDs. We hypothesized that screening genes encoding additional similar proteins will yield novel genetic causes of ALS. The most common genetic variant of ALS patients is a G4C2-repeat expansion within C9ORF72. We have shown that G4C2-repeat RNA sequesters RNA-binding proteins. A logical consequence of this is that loss-of-function mutations in G4C2-binding partners might contribute to ALS pathogenesis independently of and/or synergistically with C9ORF72 expansions. Targeted sequencing of genomic DNA encoding either RNA-binding proteins or known ALS genes (n = 274 genes was performed in ALS patients to identify rare deleterious genetic variants and explore genotype-phenotype relationships. Genomic DNA was extracted from 103 ALS patients including 42 familial ALS patients and 61 young-onset (average age of onset 41 years sporadic ALS patients; patients were chosen to maximize the probability of identifying genetic causes of ALS. Thirteen patients carried a G4C2-repeat expansion of C9ORF72. We identified 42 patients with rare deleterious variants; 6 patients carried more than one variant. Twelve mutations were discovered in known ALS genes which served as a validation of our strategy. Rare deleterious variants in RNA-binding proteins were significantly enriched in ALS patients compared to control frequencies (p = 5.31E-18. Nineteen patients featured at least one variant in a RNA-binding protein containing a PrLD. The number of variants per patient correlated with rate of disease progression (t-test, p = 0.033. We identified eighteen patients with a single variant in a G4C2-repeat binding protein. Patients with a G4C2-binding protein variant in combination with a C9ORF72 expansion had a significantly faster disease course (t-test, p = 0.025. Our data are

  1. Multiple activities of LigB potentiate virulence of Leptospira interrogans: inhibition of alternative and classical pathways of complement.

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    Henry A Choy

    Full Text Available Microbial pathogens acquire the immediate imperative to avoid or counteract the formidable defense of innate immunity as soon as they overcome the initial physical barriers of the host. Many have adopted the strategy of directly disrupting the complement system through the capture of its components, using proteins on the pathogen's surface. In leptospirosis, pathogenic Leptospira spp. are resistant to complement-mediated killing, in contrast to the highly vulnerable non-pathogenic strains. Pathogenic L. interrogans uses LenA/LfhA and LcpA to respectively sequester and commandeer the function of two regulators, factor H and C4BP, which in turn bind C3b or C4b to interrupt the alternative or classical pathways of complement activation. LigB, another surface-proximal protein originally characterized as an adhesin binding multiple host proteins, has other activities suggesting its importance early in infection, including binding extracellular matrix, plasma, and cutaneous repair proteins and inhibiting hemostasis. In this study, we used a recent model of ectopic expression of LigB in the saprophyte, L. biflexa, to test the hypothesis that LigB also interacts with complement proteins C3b and C4b to promote the virulence of L. interrogans. The surface expression of LigB partially rescued the non-pathogen from killing by 5% normal human serum, showing 1.3- to 48-fold greater survival 4 to 6 d following exposure to complement than cultures of the non-expressing parental strain. Recombinant LigB7'-12 comprising the LigB-specific immunoglobulin repeats binds directly to human complement proteins, C3b and C4b, with respective K(ds of 43±26 nM and 69±18 nM. Repeats 9 to 11, previously shown to contain the binding domain for fibronectin and fibrinogen, are also important in LigB-complement interactions, which interfere with the alternative and classical pathways measured by complement-mediated hemolysis of erythrocytes. Thus, LigB is an adaptable interface

  2. Long term change in atmospheric dust absorption, dust scattering and black carbon aerosols scattering coefficient parameters over western Indian locations

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    Satoliya, Anil Kumar; Vyas, B. M.; Shekhawat, M. S.

    2018-05-01

    The first time satellite space based measurement of atmospheric black carbon (BC) aerosols scattering coefficient at 550nm (BC SC at 550nm), dust aerosols scattering and dust aerosols extinction coefficient (DSC at 550nm and DEC at 550nm) parameters have been used to understand their long term trend of natural and anthropogenic aerosols behavior with its close association with ground based measured precipitation parameters such as Total Rain Fall (TRF), and Total Number of Rainy Days (TNRD) for the same period over western Indian regions concerned to the primary aerosols sources of natural activities. The basic objective of this study is an attempt to investigate the inter-correlation between dust and black carbon aerosols loading characteristics with a variation of rainfall pattern parameters as indirect aerosols induced effect i.e., aerosols-cloud interaction. The black carbon aerosols generated by diverse anthropogenic or human made activities are studied by choosing of measured atmospheric BC SC at 550nm parameter, whereas desert dust mineral aerosols primarily produced by varieties of natural activities pre-dominated of dust mineral desert aerosols mainly over Thar de